Tag: agriculture

Although we acknowledge this system as complex, it makes some sense to try and simplify it a little to perhaps gain some deeper insight into its operation. A glance at figure 12 suggests that there are two generalized groups of ant predators on the berry borer: the big ants and the small ants . So we can think of it as a two-predator, one-prey system. But there is an obvious indirect effect because the bigger ants negatively affect the ability of the smaller ants to be predators, as was discussed above. Depending on nest densities, it would seem that because the smaller ants prey on all three life stages of the borer and throughout the year for those that forage in infected berries on the ground, they might, in the end, be more efficient predators than the larger ants. Furthermore, the larger ants have an indirect trait-mediated effect on the smaller ants, reducing their effectiveness. One might argue that it is the existence of the smaller ants that potentially regulates the borers over the long run. Unpublished evidence even supports the idea that the berry borer actually seeks areas that are under protection from the ants, supporting the speculation of Gonthier and colleagues that the berry borer gains protection from smaller predators by preferring to attack berries under the protection of larger ants. This basic speculation was put to an unintended test in 2012. Because of a major outbreak of the coffee rust disease , the coffee landscape where we work was heavily sprayed with a combination of calcium carbonate and copper sulfate, hydroponic bucket a permitted activity for organic agriculture. In figure 13, we show the distribution of P. synanthropica as a heat map based on how many tuna fish baits placed in coffee bushes had swarms of P. synanthropica after about 30 minutes.

It is clear that in a single year, a population of thousands of nests of P. synanthropica simply disappeared. Surveys in subsequent years indicated that the smaller ants in the system began to recuperate from the reduced state they had been in, apparently because of the indirect effects of P. synanthropica . Those small ants that attack the borer within the seed, both on the tree and on the ground, especially increased over the next few years. In figure 14, we show the distribution of several of the species in a 50 × 50 meter subplot within the 45-hectare plot. Note how, during the years 2009–2012, the distribution of P. synanthropica remained relatively constant, perhaps slowly increasing in its area of dominance, at the expense of P. protensa on the ground and S. picea arboreally. Then, after the collapse of P. synanthropica between 2012 and 2013, both of those smaller species began to move into the area previously dominated by P. synanthropica. If the above speculations about how the ant community affects the borer are true, we might expect that the elimination of one of the borer’s predators would result in better overall control of the berry borer. In surveys of the berry borer in 2005 and then repeated in approximately the same area in 2018, the attack rate of the borer went from an average of about 15% of berries infected with borers to less than 1%. Insect populations are notoriously variable and respond to many cues in the environment by increasing and decreasing population numbers, frequently in unpredictable ways. Therefore, although this dramatic decline in borer numbers cannot be directly linked to the change in the ant community structure, it is nevertheless worth noting that the underlying narrative of how that community functions as a system of biological control concords perfectly with the changes observed.In the early 1980s, a specter haunted the coffee growing regions of Central America.

The infamous coffee rust disease had arrived in Brazil, and its eventual spread all the way to Mexico was expected, causing extreme worry among farmers and technical advisors. This worry was certainly justified on the basis of the history of the coffee rust disease in Asia . Great Britain’s expansion in what was then called Ceylon was qualitatively distinct from many of its previous imperial adventures. Planting what was effectively a monoculture of coffee, along with a great deal of infrastructure for the time, it was a remarkable centrally planned agricultural development plan. However, the plan effectively created ideal conditions for any disease that could get a foothold, with its virtually shadeless monoculture and networks of roads and railroads that could help distribute the fungal spores widely. When the disease arrived, it took hold and spread throughout the entire island, eventually causing a complete loss of coffee production . However, the rust scare of the 1980s Mesoamerica turned out to be a bit of a false alarm, at least until 2012. Before that year, the rust was always an irksome constraint on production, but the complete devastation that had been feared when it was discovered in the early 1980s never came to pass; it was a problem, to be sure, but not one to get overly agitated about. But then, without much warning, there was an explosion of coffee rust in the 2012–2013 cycle. Countries in the zone declared emergencies as one of their main sources of income seemed to be threatened with severe disruption. Local governments throughout the affected area provided emergency support to coffee producers and both the United Kingdom and the United States came up with significant international aid, specifically for what rapidly came to be called the most devastating emergency in the history of coffee production throughout the region . There are two ecological questions associated with this episode. First, why did the disease not become rampant for approximately 30 years after its introduction, and, second, what caused the very sudden explosion?

Although the answers to either of these questions remain enigmatic, using tools from complexity science provides us with some ideas. The relevant biology of the rust disease is well known. A windblown spore adheres to the under surface of a leaf and encounters a small amount of moisture, causing germination directly into a stoma. The mycelia grow intercellularly and produce haustoria, which penetrate into the plant cell and absorb nutrients, effectively killing the cell. As the fungus grows within the leaf tissue it eventually forms uridia that contain new spores, exiting the leaf from other stomata, causing the characteristic yellow spots on the under surface of the leaves . The transmission dynamics of the disease are dual , stackable planters with some close plant-to-plant dispersion of spores , especially when plants are close enough to touch one another, along with propagule rain from the general spore load that exists in the atmosphere, especially in areas of high concentration of coffee production, when that coffee is attacked by the rust. From the perspective of an individual coffee plant, there are two sources of rust spores: its local neighbors and the general accumulation of spores in its region—that is, from the overall spore load in the atmosphere. But it is also the case that this coffee plant and all others over a very large region contribute to the spores in the overall spore rain from the atmosphere. Given this narrative, it is easy to imagine a situation in which a generally traditional shade coffee landscape would receive a particular rate of spore rain each year and would contribute a bit to the general pool, but because the shade trees act as windbreaks, much of the spore load is never delivered to the coffee plants. One could imagine an equilibrium in which the rust disease would be endemic but not severe, partly because the wind-borne spores have limited access to the coffee trees, meaning that the increment of spore load in the general atmosphere would be limited. Focusing on the large landscape level, if the abundance of spores in the atmosphere is low, it is likely that the incidence of the disease will also be low. But each epidemic will increase the spore density in the atmosphere. The probability that a given farm will become epidemic is a function of both the spore density in the atmosphere and the dispersion rate from the atmosphere to the farm. Changing focus to the local level, the rate of spread of spores from coffee bush to coffee bush on an average individual farm will partially determine whether the rust within that farm will become epidemic. From the point of view of an individual coffee bush, the danger of being infected by a spore comes from two sources: the atmosphere and neighboring infected plants—a regional source and a local source. Imagine that a forested ecosystem is gradually deforested of both shade trees in the coffee farms and the trees in the natural forest around them, and ask what proportion of the farms could be susceptible to an epidemic of coffee rust? According to a simple model that incorporates both regional and local dispersal , the initial deforestation will generate an increase in the number of farms experiencing an epidemic. That increase is likely to be slow and steady at first, but there will be a specific point at which a critical transition will occur and a large number of farms will suddenly become highly infected. This will happen in the complete absence of any other environmental driver, such as climate change or a new more virulent strain of the disease. Indeed, one study in Costa Rica showed that the incidence of rust disease was correlated with the amount of sun coffee and pasture in the surrounding landscape. It could very well be that the sudden outbreak of coffee rust in 2012 is an example of the inevitability of surprise arising from the formality of a critical transition that we have come to associate with highly nonlinear complex systems .

A cartoon version of this theoretical process is presented in figure 16. An important component of the rust disease system, not yet completely understood, is the existence of several natural enemies of the rust . Providing an example of the sorts of ecological complexity of popular literature, the fungal disease of the first pest we discussed, the green coffee scale , is caused by the same species of fungus that, when given the chance, attacks the coffee rust fungus. That same white halo fungus that attacks the green coffee scale, now acts as a mycoparasite . Because this natural enemy is also a natural enemy of the green coffee scale, the connection to the Azteca ant became obvious early on ; Azteca creates conditions under which the scale insect becomes highly concentrated locally, which attracts the infestation of the white halo fungus and creates local hot spots of spores that disperse locally and attack the rust. Correlative evidence for this hypothesis, prior to the 2012 epidemic of rust, comes from multiple sources . Indeed, there has been considerable discussion at international conferences on the potential of L. lecanii as a spray for the rust disease. Our work suggests that partial control of the rust may naturally occur through this and other agents , although the epidemic throughout Mesoamerica in the 2012–2013 growing season shows the potential for the disease to escape such control, if, in fact, it did exist before that. It is quite a remarkable qualitative impression one gets when examining the rust and its control comparatively. It is endemic but rarely epidemic in Puerto Rico but has maintained a relatively severe status in much of Mesoamerica since 2012 . Examining coffee leaves in Mexico easily reveals the presence of L. lecanii but only after considerable searching effort, whereas in Puerto Rico, it is almost inevitable that, if one encounters the rust on a leaf, it is almost certain that one encounters L. lecanii also. What seems epidemic in Puerto Rico is the L. lecanii that seems to keep the rust under control. In addition to the white halo fungus, the larval form of a small fly, Mycodiplosis hamaelae, preys on the spores directly on the coffee leaf but probably also acts as a dispersal agent, at least locally . The coffee leaf rust continues to plague Latin American coffee farmers.

The emergence of the #MeToo movement also appears to have brought the parties together in an effort to change the prevailing culture in the legislature. The most publicized controversy of the legislative session was the emotionally charged debate surrounding the expulsion of a Democratic legislator following multiple sexual harassment allegations. Representative Steve Lebsock was the subject of multiple allegations of sexual harassment. Three formal complaints were made against him, including one from fellow Representative Faith Winter . Following an independent investigation, which found credible evidence that he had engaged in harassment and retaliation, and a full day of emotional speeches on the House floor, the chamber voted 52 to 9 to expel Lebsock from the legislature. The outcome was far from certain prior to the historic vote, as the expulsion resolution required a two-thirds majority vote in order to remove the incumbent representative. Many who supported removing Lebsock cited a need to change the culture in the state legislature. As a result of the vote, Lebsock became the first state representative to be expelled in more than 100 years. In a strange twist, Lebsock submitted the paperwork to change his party affiliation from Democrat to Republican minutes before the expulsion vote. According to state law, this handed the House vacancy to the state’s Republican Party to fill instead of the Democrats who previously held the seat. This act was clearly intended to spite fellow Democrats who abandoned their support of Lebsock once the extent of his harassing behavior became apparent. A spokesperson for the Colorado GOP initially floated the idea of not filling the seat within 30 days, hydroponic nft system which would allow the Democratic governor the opportunity to appoint a successor.

However, the party ultimately decided to appoint a Republican to temporarily hold the seat, which many expected to be regained by the Democrats in the next election cycle. This did not alter the majority party in the chamber since Democrats hold an eight-seat majority after Lebsock’s expulsion. Lebsock was not the only state legislator rebuked for inappropriate behavior, as an independent investigation found credible evidence that Senator Randy Baumgardner sexually harassed a legislative aide in 2016. Senator Baumgardner resigned a key committee chair position while maintaining his innocence in advance of an expulsion vote in the Senate. While the vote to expel Lebsock was supported by representatives of both parties, the parties appeared more divided in the Senate with regard to the allegations against Baumgardner. Republican leadership ordered Baumgardner to undergo sensitivity training, while Democrats have called for more severe sanctions including expulsion. In early April, Baumgardner survived an expulsion vote with 17 voting in favor and 17 voting opposed. While this fell short of the two thirds majority necessary to remove a lawmaker, potential expulsions from the Senate threatened larger reverberations on policy and the budget as the Republicans held a narrow one-seat majority until Senator Cheri Jahn changed her party affiliation from Democrat to unaffiliated in December 2017. On the legislative front, the second session of the 71st General Assembly convened on January 10, 2018 after a special session called by Governor John Hickenlooper in October 2017 failed to produce any accomplishments. Hickenlooper previously called a special session in 2012 to debate a civil unions bill, which ultimately failed to pass. The 2017 special session similarly adjourned amid partisan gridlock. The governor called this special session to address what was described as a bill drafting error contained in a major reform that passed at the end of the General Assembly’s regular session in 2017. Senate Bill 17-267 imposed many substantial reforms .

Among the most notable of these was the repeal of the existing hospital provider fee structure and the creation of the Colorado Healthcare Affordability and Sustainability Enterprise as a government enterprise fund to collect healthcare sustainability and affordability fees. Importantly, this change made it so hospital provider fees do not count as state revenue for the purposes of the Taxpayer’s Bill of Rights . The bill drafting error concerned a modification to the taxes levied on recreational marijuana sales. When Colorado voters approved Amendment 64 to legalize recreational marijuana use in 2012, the amendment included the following language governing the General Assembly’s authority to levy taxes on retail marijuana sales, “The General Assembly shall enact an excise tax to be levied upon marijuana sold or otherwise transferred by a marijuana cultivation facility to a marijuana product manufacturing facility or to a retail marijuana store at a rate not to exceed fifteen percent prior to January 1, 2017 and at a rate to be determined by the General Assembly thereafter, and shall direct the department to establish procedures for the collection of all taxes levied.” Senate Bill 17-267 collapsed the existing 2.9 percent sales tax on retail marijuana sales and the 10 percent special tax into a single 15 percent state tax. The drafting error occurred because legislators intended to only remove the statewide sales tax, while an exemption allowed municipal governments to impose their own sales taxes on marijuana sales where permitted. The bill, however, failed to include a similar exemption for special districts such as the Denver metro area’s Regional Transportation District and the Scientific and Cultural Facilities District , among several others. This omission meant that these districts could no longer continue collecting tax revenue on marijuana sales as they had before Senate Bill 17-267 went into effect. While the SCFD supports scores of organizations and programs including the Denver Zoo, Denver Art Museum, and Denver Museum of Nature & Science, the funding losses incurred by RTD were arguably more likely to affect Colorado residents.

The RTD special district encompasses 2,342 square miles across eight counties. The district includes nearly three million residents in its service area. Ridership statistics from the past three years show that annual ridership on RTD commuter rail, light rail, and bus services exceeded 100 million trips . Analysts projected that the bill drafting error would cost RTD nearly $600,000 per month. SCFD monthly losses were estimated at about $56,000. The special session to correct the bill drafting error was opposed by many Republicans. Some argued that modifying the existing tax structure would require a vote of state residents consistent with TABOR’s requirements. The session ended after two measures to correct the error were rejected in the Senate after passage in the House with mostly Democratic support. Following an unsuccessful special session, Governor John Hickenlooper enters the final year of his second term as governor. Term limits prevent him from running again, although there are several indications that he is preparing to launch a bid for the presidency in 2020. A November 1 letter to the General Assembly’s Joint Budget Committee provided details on the key aspects of the governor’s budget request. The budget proposal for the 2018–2019 fiscal year proposed a total funds budget of $30.5 billion, which is an increase of 3.7 percent from the prior year. The General Fund spending budget of $11.5 billion represents a 2.6 percent increase. After legislators considered nearly 200 amendments to the long bill, the state budget totals enacted into law were close to the numbers proposed by the governor. The state’s final General Fund budget was $11.42 billion with $30.63 billion in total fund spending . The letter to the JBC stated that the governor’s budget request “reflects the dynamic factors that are occurring in Colorado’s economy, nft channel constitutional and statutory requirements, and demand for services from State government” . It further noted that the “passage of S.B. 17-267 has materially and positively changed the State’s financial outlook compared with one year ago, when the request had to close a $500 million funding gap in the General Fund. We believe the request reflects the priorities that Colorado citizens expect and accordingly the request addresses important needs in K-12 and higher education as well as certain essential criminal justice system needs” . In percentage terms, spending on K-12 education represents the largest share of the general fund request at 36 percent, while the spending allotment for higher education budget is 9 percent. Nearly every state department was slated to receive increased funding relative to the prior fiscal year with just two exceptions. The budget approved by the General Assembly ultimately increased funding for every state department except for Labor and Employment. The three sections that follow discuss the state’s demographics, revenue, and spending. Colorado’s population growth continues to present both opportunities and challenges for policymakers. While a growing population provides a larger tax base, affordable housing and transportation problems have also accompanied this rapid population growth.

The 2017 update places the number of Colorado residents at slightly greater than 5.6 million . This figure represents an increase of 11.5 percent since 2010 and a 1.2 percent increase since 2016. Of the 223,260 individuals who moved to Colorado in 2016, the Census estimates that 14 percent immigrated from abroad. The size of this group is larger than the number of new residents who moved from any state in the union. Among current U.S. residents who moved to Colorado, the largest shares arrived from California , Texas , Florida , Arizona , Illinois , New York , Virginia , Missouri , Georgia , and North Carolina . The most popular destinations for Coloradoans moving out of state were California, Texas, Washington, Arizona, and Florida. Because most of these population gains have occurred along the Front Range and in the Denver metro area, the housing market in the state’s largest city remains one of the least affordable in the nation. Among U.S. metropolitan areas, the Denver housing market is the most expensive of any city not located in a coastal state. Median home prices in Denver exceeded the national average by more than $100,000 in 2017, and the average price of a single-family homes sold in Denver exceeded $500,000 for the first time in February 2018 . Many residents priced out of the home-buying market have also encountered affordability challenges in the rental market. According to one real estate research firm, apartment rental rates in Denver have increased by nearly 50 percent since 2010. This is the largest increase in rent for any city outside of the San Francisco Bay area. Increased demand for housing units has spurred development in the Denver metro area as evidenced by the construction of 23,000 new apartments between 2016 and 2017 . A growth in the number of available housing units has increased the vacancy rate in the metro area to 6.4 percent in 2018. This figure is the highest seen in Denver since the 2009 recession. It has also resulted in a modest reversal in the long-term trend of rent increases, as the median monthly rental price in Denver decreased to $1,353 from $1,370 at the end of the third quarter in 2017. Table 1 reports selected data from the Census Bureau and Bureau of Labor Statistics  for Denver County, Colorado, and the United States. As seen in Table 1, per capita and household income in Denver and Colorado remain above the national average. The OSBP projects personal income in Colorado to outpace the national average of 3.1 percent in 2017 with a 5.4 percent growth rate . The 2017 population estimate for Colorado is slightly greater than 5.6 million people. Colorado remains one of the fastest growing states in the nation, although population growth has slowed during the past year. After reaching nearly 2 percent in 2015, population growth fell to 1.4 percent in 2017. This figure remains twice the national growth rate of 0.7 percent. The state projects the number of residents to increase to 5.8 million by the 2020 Census. For most racial minority groups, the state is less diverse than the nation as a whole. Native Americans are one exception to this, as the share of those with American Indian heritage is slightly greater in Denver County and Colorado compared to their percentage nationally . The proportion of residents who identify as Hispanic or Latino is also greater in Colorado and Denver County than in the entire country . The Census estimates that 13.2 percent of current U.S. residents were born abroad. Among all Coloradoans, the percentage of foreign-born residents is about 3 percentage points lower, while the percent foreign born residing in Denver County is nearly 3 percentage points greater.

The antioxidant properties of CGA have been associated with preventing various chronic diseases.To better understand the biosynthesis of antioxidants in blueberry fruit, we identified homologs of previously characterized genes in other species involved in ascorbate, flavonols, chlorogenic acid, and anthocyanin biosynthesis. The key biosynthetic genes for these compounds exhibited a distinct developmental-specific pattern of expression . For example, genes involved in the conversion of leucoanthocyanidins to proanthocyanidins are highly expressed in the earliest and middle developmental fruit stages but not in ripening fruit . Conversely, genes involved in the conversion of leucoanthocyanidins to anthocyanins were highly expressed in mature and ripe fruit but not during early fruit developmental stages . Additionally, paralogs encoding the same anthocyanin pathway enzymes and genes involved in vacuolar localization of proanthcyanidins exhibited similar developmental stage-specific expression patterns. For example, the transcription factor complex MYB-bHLH-WD regulates expression of anthocyanin biosynthetic genes in eudicots. Using the Plant Transcription Factor Database v.4.0 , we identified homologs of transcription factors belonging to 55 gene families, and members of some of these gene families were predicted to be involved in the developmental regulation of flavonoid biosynthesis during blueberry fruit growth , including R2- R3-MYBs, R3-MYBs, bHLHs, and WDRs . These transcription factors also exhibit fruit development-specific expression patterns. In addition, we performed a gene co-expression network analysis to identify meta modules of genes that appear coregulated during fruit development, specifically genes that are associated with phytonutrient biosynthesis. Our analysis identified 1,988 meta modules of co-expressed genes, nft hydroponic of which 428 metamodules contained at least one of the 57 Pfam domains that have been previously categorized as associated with specialized metabolic pathways in plants.

Our analysis revealed that 142 of 428 meta modules were more highly expressed in developing fruit compared to other plant Thissues. Some meta modules showed clear trends of being highly expressed during either early or late fruit development. For example, METAMOD00377 is expressed early in fruit development and contains homologs to known anthocyanin genes OMT, HCT, PAL, and HQT as well as 31 homologs to known transcription factors. In contrast, METAMOD01221 is expressed late in fruit development and contains homologs of HCT, TT19, UFGT, and OMT and contains 10 homologs to known transcription factors. Moreover, we also examined meta modules for genes associated with other biosynthetic pathways that impart unique blueberry fruit characteristics. We identified two meta modules where genes appear to be co-regulated. Metamodule METAMOD00377, which contains Pfam domains associated with terpene, saccharide, and alkaloid specialized metabolism, and METAMOD01221, which contains terpene and saccharide metabolism. These meta modules contained genes that are differentially expressed during fruit development. Overall, the developmental-specific expression patterns of key bio-synthetic genes and their putative transcriptional regulators emphasize the tight regulation of production, conversion, and transport of precursor compounds that lead to the accumulation of antioxidant-related metabolites in blue Berry.The coregulation of genes involved in the biosynthesis of terpenes and saccharides during early and late fruit development described above reflects a coordinated interplay between these metabolites during fruit growth. Both terpenes and sugars contribute to the characteristic flavor of ripened fruit . In blueberry, two components play a central role in flavor perception: taste, which is a balance of sweetness and acidity, and aroma. Blueberry aroma is a complex blend of volatiles that include aldehydes, esters, terpenes, ketones, and alcohols. Previous reports in blueberry showed that the aroma profile varies greatly across different blueberry ecotypes and cultivars . For example, the aroma of high bush blueberry is primarily driven by terpene hydrocarbons and aldehydes -2-hexenal, -2-hexenol, -3-hexenol. Both linalool and geraniol are associated with sweet floral flavor. However, linalool was reported to largely impart the characteristic blueberry flavor when combined with certain aldehydes . Here, we also identified and examined the expression of genes involved in the biosynthesis of linalool. Four of the linalool synthase homologs in tetraploid blueberry are highly expressed during late fruit development .

This pattern of expression coincides with previous reports of linalool accumulation in ripened blueberry fruit . On the other hand, one homolog of linalool synthase, although it was expressed during fruit growth, did not show a clear fruit development-specific pattern. Investigating the underlying factors regulating these enzymes will facilitate genetic manipulations that may lead to further improving blueberry flavor in the future.Superior fruit quality is also associated with sugar levels. During fruit ripening, sugar levels of the endocarp increase by importing hexose symplastically and/or apoplastically. Sugar transporters , sucrose transporter, and tonoplast sugar transporter have been demonstrated to regulate intercellular sugar transport in phloem and fruit. In A. thaliana, all clade III SWEET play a role in sucrose transport, with AtSWEET9 primarily functioning in nectary secretion, while AtSWEET15 is required for seed filling by acting with SWEET11 and SWEET12. In blueberry, the clade III SWEET transporters 9 and 10 were highly expressed during early fruit growth, while clade III SWEET transporter 15 was mainly expressed in ripe fruit . interestingly, one of the blueberry SWEET15 homologs showed a distinct pattern of expression compared to the other three homologs. To the best of our knowledge, we are the first to report on the potential role of these genes during blueberry fruit development. In addition, homologs of A. thaliana TST1 and watermelon ClTST1 and ClTST3 were expressed during fruit ripening in blueberry. Elevated expression of a ClTST1 homolog was observed throughout fruit development, but the ClTST3 homolog showed very low expression. Another gene that is highly expressed during fruit maturation is vacuolar invertase. As described in other systems, its upregulation during fruit ripening coincided with the breakdown of starch to sucrose or a mixture of glucose and fructose, suggesting that it may be involved in the regulation of sugar accumulation in blueberry fruit. It was previously reported that vacuolar invertase modulates the hexose to sucrose ratio in ripening frui. In addition, there are also two sugar transport protein homologs that exhibited developmental specific expression. However, their function remains largely unknown, thus, their potential role in sugar accumulation in the developing berry requires further investigation.

Tandemly duplicated genes arise as a result of unequal crossing over or template slippage during DNA repair, exhibit high birth-death rates, and typically are in co-regulated clusters in the genome. Smallerscale duplications , which include tandem duplicates, are highly biased toward certain gene families including those involved in specialized metabolism. Furthermore, tandem duplications often results in the increased dosage of gene products and may improve the metabolic flux of rate limiting steps in certain bio-synthetic pathways. Most genes associated with the biosynthesis of antioxidants have at least one tandem duplicate present in the high bush blueberry genome, with tandem array sizes ranging from 2 to 10 gene copies . The largest tandem arrays were found for HQT and HCT genes, which are co-regulated and involved in the CGA pathway . Differences in tandem array sizes were also observed between homoeologous chromosomes for various genes. For example, the C3H gene, which is involved in CGA biosynthesis , was present on all four homoeologous chromosomes but with varying tandem array sizes. One of the homoeologous chromosomes had two copies of C3H, while the other three homoeologous chromosomes had four copies. This suggests that copy number differences of C3H among sub genomes may be due to either selection for gene duplication or loss or, in the case of allopolyploidy, may be due to preexisting gene content differences among the diploid progenitor species. Genes in the anthocyanin pathway with other unique duplication patterns include CHS, CHI, OMT, and UFGT. The gene CHS, involved in the conversion of 4-coumaryl-CoA to naringenin chalcone, has two copies, and both have tandem duplicates in at least three of the homoeologous chromosomes. interestingly, the gene CHI has a single preserved tandem gene duplicate on only one of the homoeologous chromosomes. However, nft system additional copies of CHI were also identified more distantly away from the syntenic ortholog on another homoeologous chromosome, likely involving a transposition event following tandem duplication. The OMT and UFGT genes all have tandem duplicates on all of the homoeologous chromosomes, although with varying array sizes, while the ANR gene involved in the conversion of anthocyanidin to proanthocyanidin is single copy on all homoeologous chromosomes. DFR gene, which is involved inthe conversion of dihydroquercetin/dihyromyricetin to leucoanthocyanidin, has a single tandem duplicate on only one of the homoeologous chromosomes. These findings suggest that there may have been greater selective pressure to retain tandem duplicates for genes encoding enzymes involved in anthocyanin production than conversion to proanthocyanidins. The vast majority of tandem duplicates are eventually lost ; however, in rare instances, some may undergo functional diversification. Gene expression analysis revealed that 83.4% of the tandem duplicates were expressed in at least one transcriptome library with 73.5% expressed in at least one of the fruit developmental stages. This suggests that a subset of these duplicate genes have non-functionalized, subfunctionalized, or neofunctionalized. Future studies are needed to more thoroughly investigate the functions of these genes with more diverse libraries and additional transcriptome analyses.Despite the economic importance of blueberry, molecular breeding approaches to produce superior cultivars have been greatly hampered by inadequate genomic resources and a limited understanding of the underlying genetics encoding important traits. This has resulted in breeders having to solely rely on traditional approaches to generate new cultivars, each with widely varying fruit quality characteristics. For example, our analysis of a diversity panel consisting of 84 cultivars and wild species revealed that ”Draper” has antioxidant levels that are up to 19x higher than other cultivars. Thus, the genome of ”Draper” should serve as a powerful resource to the blueberry community for guiding future breeding efforts aimed at improving antioxidant levels among other important fruit quality traits.

Furthermore, to our knowledge, this is not only the first genome assembly of the cultivated high bush blueberry but is also the first chromosome-scale and haplotype-phased genome for any species in the order Ericales. Ericales includes several other high-value crops and wild species with unique lift history traits . Thus, we anticipate that this reference genome, plus associated datasets, will be useful for a wide variety of evolutionary studies. Here, we also leveraged the genome to identify candidate genes and pathways that encode superior fruit quality in blueberry, including those associated with pigmentation, sugar, and antioxidant levels. Furthermore, we found that genes encoding key bio-synthetic steps in various antioxidant pathways are enriched with tandem gene duplicates. For example, tandem gene duplications have expanded gene families that are involved in the biosynthesis of anthocyanins. This suggests that, in addition to a recent whole genome duplication, tandem duplications may have greatly contributed to the metabolic diversity observed in blueberry . These tandem duplicates may have evolved new functions , possibly involved in the biosynthesis of novel compounds, and/or were selected to improve the metabolic flux of specific bio-synthetic steps that alter the dosage of certain endpoint metabolites. Future studies are needed to further investigate the possible role of tandem duplications in having modified metabolite levels and composition in wild and cultivated blueberry. Our analyses also revealed that high bush blueberry, a tetraploid, likely arose from the hybridization of two distinct parents, possibly allopolyploidy, based on the sequence divergence, unique transposable element insertions, and sub genome expression patterns. Our analyses revealed that the sub genomes in high bush blueberry may be controlling a distinct set of genetic programs . The dominantly expressed sub genome in most surveyed Thissues becomes the lowest expressed during fruit development. This observation is similar to findings in allopolyploid wheat where developmental and adaptive traits were shown to be controlled by different sub genomes. For example, cell type- and stage dependent sub genome expression dominance was observed in the developing wheat grain. We argue that both high bush blueberry and hexaploid wheat, each now with high-quality reference genomes, make excellent systems to further investigate these underlying mechanisms of sub genome dominance. sub genome dominance has far-reaching implications to numerous research areas including breeding efforts. For example, marker-assisted breeding needs to target the correct set of dominant homoeologs given the trait in polyploids that exhibit sub genome dominance. Thus, we anticipate that this genome, combined with improved insights into sub genome dominance, will greatly accelerate molecular breeding efforts in the cultivated high bush blueberry.High-molecular-weight genomic DNA was isolated from young leaf Thissue, following a 72-hour dark treatment, using a modified nuclei preparation method.

Likewise, Nicolás et al. found a better photosynthetic performance after inoculating Crimson grapevines grown in a commercial vineyard. Indeed, a recent meta-analysis demonstrated that AMF exert a positive influence on photosynthetic rates, stomatal conductance, and water use efficiency on both C3 and C4 plants subjected to salt stress . Merlot grapevines did not show changes on their berry primary metabolites as affected by the treatments applied . Similarly, a recent study evaluating the effect of different sustained deficit irrigation and RDI showed no differences in must pH and TSS in Merlot berries in a 4-year field experiment conducted in a hot climate . This lack of effect of the irrigation systems on berry primary metabolism might be due to grapevines were not subjected to a severe water stress . On the other hand, previous studies showed that inoculation with AMF of grapevines vineyards did not affect TSS or TA under field conditions or under controlled conditions and our results corroborated these findings. Regarding secondary metabolism, neither irrigation systems nor AMF inoculation modified flavonol and anthocyanin total content at harvest . Similarly, a 2-year field study conducted in Central valley in Califtornia with Merlot did not report differences on flavonol or anthocyanin skin content due to different irrigation amounts . A previous study conducted on Cabernet Sauvignon subjected to water deficit reported that although flavonol synthesis related genes were up-regulated after the onset of fruit ripening, this did not affect berry flavonol concentration at harvest . Similarly, hydroponic channel previous studies with Tempranillo grown under controlled conditions did not observed differences due to AMF inoculation on the total content of flavonol and anthocyanins in berry skins .

Flavonol composition was affected by treatments. Thus, HII grapevines increased quercetin and decreased syringetin contents in berry skins at harvest in accordance to a previous study . Indeed, it is known that AMF inoculation up-regulated phenyl-propanoid biosynthesis key genes in grapevines in response to pathogens . On the other hand, HI led to decreased contents of quercetins, laricitrins, kaempferols, syringetins, and isorhamnetins. Likewise, Martínez-Lüscher et al. found that in spite of the increase in O-methyl-transferase transcript level, methylated flavonols did not increase under water deficit. These authors suggested that given the higher affinity of OMT for quercetins, the lower concentration of quercetins under water deficit could act as a limiting factor for the synthesis of methylated forms, and our findings corroborated this hypothesis. Regarding anthocyanin composition, berry skins from HI grapevines showed lower contents of di-substituted anthocyanins than the ones of FI grapevines. It is well known that water deficit regulates the expression of key genes of the flavonoid pathway such as the flavonoid 30 -hydroxylase, flavonoid 30 ,50 -hydroxylase, and O-methyltransferase in red cultivars . Therefore, these decreased contents of disubstituted anthocyanins were likely explained by a different regulation of these genes when grapevines are subjected to water deficit. The role of AMF for enhancing phenolic compounds was reported in several studies with potted grapevines. Thus, AMF grapevines showed increased content of resveratrol, viniferins, and pterostilbene , total phenols and quercetin content , and total flavonoids in leaves of different grapevine varieties facing different biotic and/or abiotic stresses. Moreover, increased anthocyanin contents were reported in berries from grapevines grown under water deficit and warming conditions . Similarly, we found a strong relationship between the percentage of mycorrhizal colonization and some flavonoids .

The economic analysis data indicated that AMF inoculation and water management did not affect the cost of labor operations, in spite of irrigating with half amount may lead to decreases in yield. However, this came with reductions of the water footprint that have to be taken into account. It is noteworthy that extreme weather recorded in 2020 could modulate the effects described in this work. Moreover, the mycorrhizal extraradical mycelium coexists with soil microbial communities and the synergistic activity between the AMF, the bacterial communities, and the grapevine modulates the benefits of symbiosis on nitrogen fixation, P solubilization, and production of phytohormones, siderophores, and antibiotics . On the other hand, previous studies demonstrated that the microbiome of vineyards is shaped by cropping management , and little is known about whether these communities stimulate or suppress the extraradicular mycelium activity . Therefore, given the effect of AMF inoculation and different irrigation amounts had on grapevine physiology and berry composition, further studies should consider the potential effects of these management practices on vineyard soil living microbiota. Current research aimed to study how Merlot grapevines responded to AMF inoculation and different water amounts in their first productive year in situ. Our results highlighted the role of AMF inoculation for improving vegetative growth, photosynthetic activity, and water status of grapevines, especially when facing mild water deficits in field grown grapevines. Additionally, a strong relationship between the mycorrhizal colonization of roots and some flavonoids was found, corroboration the effect of AMF for regulating anthocyanin and flavonol metabolisms. Finally, although some berry quality traits and grapevine performance were improved by AMF inoculation under water deficit, AMF inoculation was not sufficient to avoid the yield losses due to water deficit in the first productive year of Merlot when facing a hyper-arid growing season.

It is noteworthy that these results may be affected by edaphoclimatic characteristics and living microbiota in vineyard soils, which should be taken into account before making the decision of inoculating the vineyard. Therefore, this study offer a starting point to assess the effect of AMF inoculation on young vines under real field conditions. However, benefits distinct from simple nutrition, such as phytochemicals have recently become clear. Diets rich in a plethora of phytochemicals can promote a healthy and diverse gut microbiota, reduce intestinal and systemic inflammation, and decrease the risk of colorectal cancer and type 2 diabetes melliThus. Some of these benefits can be observed around the world. Many parts of India have historically low colon cancer incidence rates. The Indian subcontinent has been continuously settled for millennia. Ancient cities in the Indus valley have been dated to the third and fourth millennia BC and some sites are even older. Archaeological evidence of grain cultivation, including several varieties of barley and wheat, has been found in excavations dated to the sixth millennium BC. Wheat is still a staple crop in northern India, and many other grains, including barley, were commonly cultivated and eaten until the 1950s, when wheat and white rice became dominant. Although the country has many diverse cultures, some customs remain common and conventional throughout the nation. One such tradition is the form of main meals where a large round platter, the thali, holds rice or bread and several smaller bowls, or katori, which hold a separate condiment or curry to be eaten with the rice or bread at the diner’s preference. Typical dishes include, but are not limited to, dal , yogurt , and assorted spices and vegetables. The development of agriculture early in its history has allowed India to develop rich traditions around food. These traditions have been deeply influenced by Ayurveda, the ancient Indian system of medicine. In Ayurvedic practice, food is a source of nourishment and medicine, used to both prevent and treat illness. Maintaining a proper balance of Ayurvedic elements through diet is considered an effective way to live a healthy liftestyle. According to the Ayurvedic principles, each meal should contain a balance of the six major flavors. This calls for the many small portions of a thali meal which also easily incorporate variety. A variety of flavors in a meal often indicates the presence of many classes of bio-active compounds . Although these substances may not be macronutrients, vitamins, or minerals, they still impact human health. Polyphenols are perhaps the largest class of bio-active compounds, containing subclasses such as flavonoids, isoflavones, stilbenes, lignans, and tannins. As a flavonoid subgroup, anthocyanins are included in this class. Anthocyanins are of interest in the food industry as nontoxic and water-soluble pigments, as most are colored red, purple, or blue, and many display antioxidant and anti-inflammatory activity. A class of phytochemicals called polyphenols is also found in virtually all plant foods, though their quantity may be reduced by preparation methods. Rich sources of anthocyanins include deeply colored fruits and vegetables, such as blueberries, eggplants, hydroponic dutch buckets and certain carrot and potato cultivars. Given that many phytochemicals exert anti-inflammatory activity via promoting gut bacterial diversity, there is a growing interest in a food-based approach to countering the growing epidemic of inflammation-promoted chronic diseases such as colon cancer.We have learned that no discussion of diet is complete without consideration of the intestinal microbiota. Trillions of bacteria, distributed throughout the gastrointestinal tract from mouth to anus, facilitate digestion and intestinal homeostasis. Structural factors greatly impact the overall makeup of each community. For example, low pH prohibits many pathogenic bacteria from colonizing the stomach and the upper small intestine. The depths of the large intestine, on the other hand, is an ideal habitat for many anaerobes. The gut microbiota is a dynamic community, composed of living organisms that can alter in response to diet, disease, and other environmental pressures. Changes in the intestinal microbiota were first correlated with illness in 1681 when Anton van Leeuwenhoek recorded that the microbial composition of his diarrhea differed from normal fecal samples. Since then, the intestinal microbiome has been closely studied to show how it can be implicated in a variety of conditions ranging from obesity to colon cancer. A great deal of investigation into microbiota has been accomplished in the last decade. Many of these observed changes result in an overall loss of bacterial diversity in the microbiota, indicating that species diversity is associated with health. However, the opposite may be true for cause-consequence relations, but not enough research has been brought to light. High-throughput technologies have driven advances in identifying the trillions of microbes and the metabolic functions that live in the colon.

This led to a critical insight that gut plays as dynamic of a role in metabolism as the liver. The proximity of these microbes to the intestinal mucosa and gut lymphoid Thissue explains the critical role they play in health and disease. Indeed, dysbiosis plays a significant role in the development of inflammatory bowel disease, obesity, and colon cancer. Emerging evidence suggests that diet can directly influence the content and composition of gut microbiota. Thus, understanding the complex interactions between diet, gut microbiota, and the host are crucial in prevention and treatment of chronic diseases that plague our society. Studies in murine models have shown rapid changes in the gut bacteria of mice being switched quickly from a standard diet to a high-calorie diet back to a standard diet. In humans, surveys show that diets high in fiber correlate with higher microbial diversity and reduced populations of Enterobacteriaceae, including Escherichia and Shigella species. Marked differences are also seen during consumption of animal- vs. plant-based diets. While nutrients in the diet will affect intestinal microbes, other substances present in food may also have an effect. For example, most anthocyanins are not absorbed into the bloodstream in the small intestine, and so they stay in the gastrointestinal tract until they reach the colon. There, they can affect the colonic microbiota in multiple ways. Firstly, anthocyanins have antioxidant activity that can reduce inflammation-induced oxidative stress on the gut bacteria. Secondly, anthocyanins are a potential carbon source, which bacteria can metabolize, resulting in increased growth of certain microbes. Lastly, bacterial metabolism of anthocyanins produces a variety of metabolite byproducts, some of which have antimicrobial effects on enteric pathogen species including Escherichia coli.Chronic intestinal inflammation is a hallmark of certain bowel disorders, such as ulcerative coliThis and Crohn’s disease, which are two major forms of inflammatory bowel diseases , and IBD is also considered a risk factor for colorectal cancer. In the latter, inflammation is generally low-grade but persists over a long period of time. Diet composition can promote or suppress chronic inflammation. Low-fiber high-calorie diets, which are typical in Western countries, may directly promote inflammation, or as already discussed, indirectly promote this through dysbiosis. Indeed, some dietary patterns associated with chronic inflammation are also linked to the reduction of total microbial diversity and imbalances in intestinal microbial groups. Furthermore, some bacteria, including E. coli, can flourish during low-grade inflammation, where thinning of the intestinal mucus layer occurs and allows for more direct interaction between the host’s cells and the intestinal bacteria.

Commercial standards of epicatechin, malvidin-3-O-glucoside, and quercetin-3-O-glucoside were used for the quantification of flavan-3-ols, anthocyanins, and flavonols, respectively.Cost estimates on labor operations and gross income per hectare were calculated based on yield and net returns per hectare . Water footprint was calculated as described by Zotou and Tsihrintzis . Briefly, for the green component of the WF , precipitation data during the growing season was obtained from the CIMIS Station and estimated as m3 /ha to obtain the total green consumed water volume . Then the value was divided by the yield expressed as ton/ha. The blue component of the WF was calculated with the total irrigation water amount that grapevines received per hectare, and this blue consumed water volume value was divided by the yield . The gray component of the WF was not calculated given that our experimental conditions avoided the use of fertilizers. Then, the total WF was estimated as the sum of green WF and blue WF. statistical analyses were performed in R-Studio version 3.6.1 for Windows. All the monitored parameters were fit in linear mixed-effect models by using the lmer function from lme4 package with AMF inoculation , irrigation treatment , and their combination as fixed factors, and replicate as random factor . Then, pairwise contrasts were conducted with function lsmeans from lsmeans package using the Kenward–Roger method and Tukey adjustment for p-values. Previously, for gas exchange parameters, stem water potential, mycorrhizal colonization, and flavonoid contents a mixed-effect model including sampling date as fixed factor was run . However, as the treatment effect seemed to be independent in the sampling date , sampling date was removed from the analysis to assess the effect of treatments for each sampling date. Finally, blueberry packaging box correlations between the percentage of mycorrhizal inoculation and flavonoid contents were calculated with the Pearson’s test using the same software.

The comparison between the growing season of the experiment and the reference data for the same period within the last 20 years showed that 2019–2020 was warmer and drier . Thus, average daily temperature was 0.5◦C higher, especially in August, which reached 1.8◦C more, and precipitation of 530 mm less compared to the average, hence, the 2020 growing season was an extreme year regarding temperature and rainfall. Native mycorrhizal colonization was determined before treatment application and no differences between them were observed . The mycorrhizal colonization intensity was analyzed after 3 months of treatment application to ensure the establishment of the mycorrhizal symbiosis, which frequently take place after 2–4 months of inoculation. Similar patterns in AMF colonization intensity were observed in both, 3 months after inoculation and at harvest, where roots from inoculated grapevines showed percentages of colonization values threefold higher than non-inoculated ones . In addition, we observed increased AMF colonization rates along the growing season as shows the significant effect of the sampling date and its interaction with the AMF inoculation . Relative mycorrhizal dependency index allows assessing the dependency of a crop on the mycorrhizal symbiosis to achieve its maximum growth at a given environmental condition. Under FI conditions, RMD values were lower than 100% indicating that the mycorrhizal association impairs the vegetative growth of grapevines; however, RMD values for HI conditions highlighted the role of the mycorrhizal symbiosis for improving grapevine growth under water deficit conditions . Grapevine vegetative growth was also monitored during the 2020 growing season by measuring the green pruning weight, trunk diameter, and leaf area . Measurements before treatment showed no differences between the different plants concerning trunk diameter , corroborating the effect of treatments modulating vegetative growth of vines. Irrigation amount was the main factor affecting both vegetative growth and yield, with grapevines subjected to HI decreasing them . However, as RMD reported AMF inoculation impair the grapevine growth estimated as trunk diameter and as green pruning weight when vines were FI, whereas under conditions, inoculated vines improved their growth . Finally, the leaf area to fruit ratio was not affected by treatments applied. The contents of minerals measured in leaf blades were not affected by AMF inoculation or applied water amount in our experiment . Plant water status was determined by monitoring the SWP each 2 weeks at noon during the growing season.

The SWP values ranged between −0.8 and −1.3 MPa at harvest suggesting that the amount of applied water was successful in reaching the SWP target during the growing season. Irrigation amount was the main factor affecting the water status of vines especially at the end of the growing season. However, before veraison AMF inoculation could increase the grapevine water status under HI conditions . The calculation of the seasonal integral of SWP showed the same pattern; hence, siSWP was mainly affected by irrigation system with HI plants being the most stressed vines . Gas exchange parameters monitored during the season are shown in Figure 3. Carbon assimilation rates increased through the growing season, and were affected by the interaction between AMF inoculation and irrigation amounts . Thus, FII plants showed the highest values of AN at fruit set and harvest, while FINI grapevines increased AN after veraison. Leaf evapotranspiration was slightly modified by treatments at the beginning on the season but no effect was observed later in the season. On the other hand, although no differences in instantaneous water use efficiency were recorded at harvest, AMF inoculated plants showed a better WUE during berry development and ripening . Finally, stomatal conductance was highly affected by the interaction between AMF inoculation and irrigation system during the whole season . Thereby, AMF inoculation of HI plants mitigated the reduction of gs . Primary metabolites and berry fresh weight are presented in Table 4. Must pH, TA, and TSS were not affected by treatments. However, BFW was modified by treatments; hence, AMF inoculation increased BFW of FI plants and decreased in HI . Flavonols and anthocyanins were monitored through berry ripening. The effect of AMF inoculation and irrigation systems on berry skin flavonol content and composition was modulated during the growing season as indicated by the significant interaction of treatments with the sampling dates . At mid ripening, the berry skin flavonol content increased in HINI grapevines . Similarly, quercetin-3-O-glucoside and laricitrin- 3-O-glucoside decreased with AMF inoculation under HIconditions . At harvest, myricetin and quercetin derivatives were the most abundant flavonols found in Merlot berry skins, accounting for more than 40% of the total flavonols. Irrigation treatment was the main factor affecting flavonol content and composition as indicated by the decrease in quercetin, laricitrin, kaempferol, isorhanmetin, and syringetin derivative contents in HI grapevines . It is noteworthy to highlight the increased content of quercetin-3-O-galactoside in HII grapevines . At mid ripening the main anthocyanin was cyanidin-3- O-glucoside, which accounted for ca. 20% .

The total anthocyanin content of Merlot berry skins was not affected by treatments but HI treatment decreased the contents of some anthocyanin derivatives . At harvest, the total anthocyanin content in Merlot berry skins was not affected by different treatments . Malvidin was the most abundant anthocyanin detected in Merlot berry skins , with contents ranged between 23.1% for HINI plants and 28.7% from FII but none of the malvidin derivatives were affected by treatments . The main changes in anthocyanin composition were due to irrigation treatments, thus, HI ledto decreased contents of cyanidin and peonidin derivatives . Finally, an analysis of the relationship between the percentage of AMF colonization and the main flavonoid contents was conducted . The intensity of the AMF colonization had a significant positive relationship with total cyanidins , total peonidins , and total quercetins . In the last decades, warming trends in viticulture areas have been described worldwide . Likewise, weather data recorded during 2020 growing season in Oakville, CA, United States , suggested more stressful conditions for grapevines comparing to the average of last 20 years, blueberry packaging containers challenging their production and quality. Indeed, a recent study based on climate indices suggested a reduction of 8,700 km2 for the Califtornia land suitable for grapevine cultivation by mid-21st century . Within this scenario, smart-farming techniques are mandatory for adaptation and mitigation to guarantee the future of the wine making industry and for reducing potential water conservation issues. Colonization analysis of Merlot grapevine roots indicated that AMF inoculated integrated with the native communities colonizing grapevine roots . Thus, we found that the percentage of mycorrhizal colonization was two to three fold higher in mycorrhizal inoculated treatments compared to non-inoculated ones. However, no differences in mycorrhizal colonization due to water amount received by plants were evident in accordance with a previous study conducted on fruit bearing cuttings . In contrast, a study conducted on own-rooted Cabernet Sauvignon field grapevines reported increased frequency of arbuscules and reduced fine root production when an additional water deficit was applied to the regulated deficit irrigation plot, suggesting that plants could compensate the lower density of fine roots in vines facing water deficit by increasing AMF colonization . These discrepancies between studies may be explained by the fact that grapevines responded to the degree of water deficit from the previous growing season. Thus, Schreiner et al. observed increased arbuscular colonization at bloom, before the onset of differences between the treatments they applied whereas under our experimental conditions, water amounts received by Merlot grapevines the previous season did not differ. AMF colonization data also confirmed the seasonality effect on mycorrhizal colonization and the reinforcement that AMF inoculation exerts on native mycorrhizal colonization . Without imposed water stress, AMF inoculation impaired vegetative growth as indicated in the RMD index. However, when grapevines were subjected to HI treatment, AMF-inoculated vines grew better as indicated by the RMD, green pruning, and trunk diameter. Nevertheless, leaf area was not enhanced after AMF inoculation according to previous studies , which would explain that AMF inoculation was not sufficient to avoid the yield loss due to HI treatment. It is well established that AMF inoculation enhances mineral nutrition of grapevines presumably by a greater exploration of soil by the external hyphal network of the AMF resulting in more efficient roots for obtaining nutrients from soils . Moreover, it was recently reported that the inoculation of grapevines with AMF under controlled conditions led to the upregulation of nutrient transport genes . In spite of the consensus about AMF enhancing grapevine nutrient uptake, contradictory results are reported about increased mineral nutrient content due to the symbiosis . Leaf or petiole mineral nutrient content might be useful for the diagnostic of soil mineral deficiencies allowing growers to manage them. However, concentration of mineral nutrients does not provide accurate information on nutrient uptake or allocation of nutrient in various organs . Therefore, although no differences on the mineral nutrient content in leaf blades were observed, mineral uptake was presumably enhanced by AMF inoculation given the growth promotion recorded in mycorrhizal plants under HI conditions. Furthermore, Balestrini et al. recently reported that although mineral nutrient uptake genes were upregulated after inoculation with different inoculants , the degree of upregulation differed between them, suggesting a specific response to a specific inoculum. Similarly, Nogales et al. did not find accumulation of minerals in grapevine leaves after AMF inoculation with the exception of P, which was enhanced and decreased after F. mosseae and R. irregulare inoculations, respectively. Grapevine water status monitored during the growing season showed that irrigation amounts were the main factor affecting the plant water status. Thus, according to previous work FI plants were maintained under well-watered conditions with values of midday SWP higher than −0.9 MPa and/or gs higher than 200 mmol m−2 s −1 . On the other hand, grapevines subjected to HI were not exposed to a severe water stress as they never reached values of SWP and gs lower than −1.5 MPa and 50 mmol m−2 s −1 , respectively, considered detrimental for grapevine development . We did not measure any SWP differences due to the AMF inoculation when plants were subjected to FI. However, within HI plants, AMF inoculation tended to result in higher SWP values in accordance with previous studies . Therefore, a higher AMF occurrence in the root zone has been related to improve water status of vines by increasing water uptake presumably by increasing the mycorrhizal structures, mainly arbuscules . Accordingly, we observed that photosynthetic performance of AMF inoculated Merlot grapevines was improved .

Others on the farm believe that the courses are open to all workers except pickers. This unofficial, yet effective exclusion of pickers from the English classes inadvertently shores up segregation on the farm.Mateo is 29 years old, a Mixteco father of two young children. He has worked on the farm for 12 years and has taken English classes for 5 years. His family had enough money to allow him to finish high school in Oaxaca before emigrating. He is fluent in his native language, Mixteco Alto, and Spanish, and is the only Oaxacan person on the farm who speaks English. He is also the only Oaxacan with a job other than picker. He oversees pickers in the strawberry and blueberry harvests. He hopes to continue studying English and be promoted on the farm until he can ‘‘work with his mind instead of his body’’ . Mateo worries about the pregnant women in his crew picking long days in pesticide-covered plants. He explains that many give birth prematurely due to the difficulty of their work. He also worries about the low pay of the pickers. The pay for strawberries has gone up only a few cents per pound in a decade and the pay for blueberries has gone down in the past several years. Barbara and Mateo manifest the common desire to treat workers well even though the structures within which they all work are ‘‘unfair’’ . Mateo’s position as the only Oaxacan crew boss shows the importance of the ability to study English in order to be promoted and helps illuminate the contours of the structures leading to vulnerability.

During my second summer on the farm, a white, wholesale grow bags female college student came up to me and said, ‘‘So, I hear you’re writing a book.’’ Laura grew up in the area and worked assigning pickers to rows and checking ID badges. She is studying Spanish in college in Seattle and enjoys talking with and learning about the pickers. She has been frustrated with the way one supervisor, Shelly, sees the Mexican pickers. She explained, ‘‘One day we were walking back to the cars, one girl was talking to one of the pickers, practicing her Spanish. I don’t know if they were even talking to each other but Shelly said something to her, she didn’t want her to talk to pickers. It’s like she doesn’t trust them. She gets frazzled a lot. I was surprised, like, ‘why didn’t she want you to talk to them?’ ’’ Although the higher farm management sees the employment of white teenager checkers to be developing positive values toward agriculture and diversity in the valley, checkers also learn that they deserve to have power over Mexicans, even those old enough to be their parents or grandparents. The teenagers are paid minimum wage while being allowed to talk and sit most of the time, while the pickers have to kneel constantly and work as fast as possible in order to keep their jobs. The checkers are given power over the number of pounds marked for pickers. They are allowed to treat the pickers as people who do not deserve equal respect. This experience serves to develop the lenses through which symbolic violence, the naturalization of inequality, is affected . In addition, Laura points out that the farm management sometimes works directly to keep labor positions and ethnicities segregated.Several small groups of field workers are paid per hour.

All live in labor camps and work seven days a week from approximately five in the morning until the early evening. Approximately a dozen men, mostly mestizo Mexicans along with a few Mixteco Oaxacans, drive tractors between the fields and the processing plant. The tractors carry stacks of berry containers several feet high, and the drivers are exposed to direct sun or rain all day. In addition, small groups of mostly mestizo Mexican men and women, and a handful of Mixtecos, work in other capacities, from tying off the new raspberry growth to covering blueberry bushes with plastic, from spraying chemical or concentrated vinegar pesticides to using hoes between rows of plants. Thirty-some raspberry pickers work 12 to 18 hours a day, 7 days a week for approximately 1 month. Two or three people work on each raspberry harvester, which is approximately one-story tall, bright yellow, and shaped like an upside down ‘‘U’’ tall enough for the row of raspberry bushes to pass beneath its middle. The machine shakes the bushes such that the ripe berries fall onto a conveyor belt and then onto a crate. One worker drives the machine; the others move the full berry crates and remove bad berries and leaves. They are all seated and have minimal shade from umbrellas attached to the machine. All the raspberry pickers are US Citizen Latinos from Texas; most are relatives of the raspberry crop manager.Pickers are the only group not paid by the hour. Instead, they are considered ‘‘contract workers’’ and are paid a certain amount per pound of fruit harvested. Most live in the camp furthest from farm headquarters and some live in the next furthest camp. Each day, they are told a minimum amount of fruit to pick. If they pick less, they are fired and kicked out of the camp. The first contract picker I met, a Triqui man named Abelino, explained, ‘‘The hourly jobs, the salaried jobs are better because you can count on how much you will make. But, they don’t give those jobs to us.’’ Approximately 25 people, mostly mestizo Mexican with a few Mixteco and Triqui people, pick apples.

The field boss, Abby, explained to me that picking apples is the hardest job on the farm. Apple pickers work 5 to 10 hours a day, 7 days a week, carrying a heavy bag of apples over their shoulders. They repeatedly climb up and down ladders to reach the apples. This job is sought after because it is known to be the highest paid picking position. However, the majority—350 to 400—of pickers, often called simply ‘‘farm workers,’’ work in the strawberry fields for one month, followed by three months in the blueberry fields. Other than a few Mixtecos, they are almost all Triqui men, women, and children; agricultural workers can legallybe 14 years or older. Most pickers come with other family members. The official contract for strawberry pickers is 14 cents per pound of strawberries. This means that pickers must bring in 50 pounds of de-leafed strawberries every hour because the farm is required to pay Washington State minimum wage . In order to meet this minimum, pickers take few or no breaks from 5 a.m. until the afternoon when that field is completed. Nonetheless, they are often reprimanded and called perros , burros, Oaxacos, or indios estupidos. Many do not eat or drink before work so they do not have to take time to use the bathroom. They work as hard and fast as they can, arms flying in the air as they kneel in the dirt, picking and running with their buckets of berries to the checkers. Although they are referred to as ‘‘contract workers,’’ this is misleading. The pay per unit may be changed by the crop managers without warning or opportunity for negotiation. Strawberry pickers work simultaneously with both hands in order to make the minimum. They pop off the green stem and leaves from each strawberry and avoid the green and the rotten berries. During my fieldwork, I picked once or twice a week and experienced gastriThis, headaches, and knee, back, grow bags for gardening and hip pain for days afterward. I wrote in a field note after picking, ‘‘It honestly felt like pure torture.’’ Triqui pickers work seven days a week, rain or shine, without a day off until the last strawberry is processed. Occupying the bottom of the ethnic-labor hierarchy, Triqui pickers bear an unequal share of health problems, from idiopathic musculoskeletal pains to slipped vertebral disks, from type 2 diabetes to premature births and developmental malformations . Most Triqui workers on this farm are from one village, San Miguel, located in the mountains of Oaxaca, Mexico. Next, I highlight the economic and physical hardships of the pickers on the farm and on the US-Mexico border, touching on the importance of language, ethnicity, and education in the organization of the farm labor hierarchy. I also indicate the importance of immigration and border policies in determining the structural vulnerability of farm workers. Marcelina is a 28-year-old Triqui mother of two. A local non-profit organized a seminar on farm labor for which I invited Marcelina to speak about her experiences migrating and picking. Shyly, she approached the translator, holding her one year-old daughter, speaking in Spanish, her second language.My first day picking, the only people who picked as slowly as I did were two Latina US citizen girls from Califtornia and one Latino US citizen manwho commuted from Seattle. After the first week, the two Latina girls began picking into the same bucket in order to make the minimum and keep one paycheck.

The second week, I no longer saw the man from Seattle. I asked a supervisor where he had gone, assuming he had decided the work was too difficult and given up. She told me the farm made a deal with him that if he could make it through a week picking, they would give him a job paid hourly in the processing plant. He has been ‘‘one of the hardest workers’’ in the plant since then. I inquired as to why indigenous Mexicans could not get processing plant jobs. The supervisor replied, ‘‘People who live in migrant camps cannot have those jobs, they can only pick.’’ She considered it farm policy without any need for explanation. Thus, marginalization begets marginalization. Structural vulnerability increases along the labor hierarchy and is reinforced by official and unofficial policies, practices, and prejudices . The indigenous Mexicans live in the migrant camps because they do not have the resources to rent apartments in town. Because they live in the camps, they are given only the worst jobs on the farm. Unofficial farm policies subtly reinforce labor and ethnic hierarchies. These profiles show that the position of the Triqui workers at the bottom of the hierarchy is multiply determined by poverty, education level, language, citizenship status, and ethnicity. In addition, these factors produce each other. For example, a family’s poverty cuts short an individual’s education, which limits one’s ability to learn Spanish , which limits one’s ability to leave the bottom rung of labor and housing. Poverty, at the same time, is determined in large part by the institutional racism at work against Triqui people in the first place. Segregation on the farm is the result of a complex system of feedback and feed forward loops organized around these multiple nodes. Late in my second summer on the farm, the pickers walked out of the field just after the pay per weight was lowered. The pickers listed over 20 grievances about the working conditions, from low pay to racist statements from supervisors, lack of lunch breaks to unfair promotions of mestizo and Latino workers over indigenous pickers. Over the next week, several executives and a dozen pickers held meetings to discuss the grievances. The executives were visibly surprised and upset at the explicit racist treatment and differential promotions on the farm. They promptly instructed the crop managers to pass along the message to treat all workers respectfully. Lunch breaks and higher pay were instituted, but were silently rescinded the following summer. The pickers called the document a ‘‘contract’’ and requested signatures from the executives. The farm president filed it as a ‘‘memo.’’ This strike, the temporary nature of its results, and the conversion of the contract into a memo highlight the differential demands and pressures at all levels of the farm hierarchy. The executives demand that all workers are treated with respect at the same time that their real anxieties over farm survival prohibit them from effectively addressing the primary, economic concerns of the pickers. Although everyone on the farm works for and is paid by the same business, they do not share vulnerability evenly. The pay and working conditions of the pickers function as variables semicontrollable by the farm executives as partial buffers between market changes and the viability of the rest of the farm.

Since tuition increases were capped at 6 percent in 2015 by virtue of a budgetary agreement, larger increases next year could make it more challenging for families to afford college. In addition to higher education, the corrections and revenue departments will also likely experience substantial funding reductions. The proposed reduction in funding the Department of Corrections is less than 1 percent of its previous year’s budget. This follows several years of in- creased funding for correctional facilities and programs. Over $6 million in savings from the correctional budget came as a result of the state’s total prison population for the year being significantly less than projected. This substantial reduction was partially offset by proposed inflationary and other rate increases. In absolute terms, the proposed increase in health care spending is second only to K-12. The Department of Health Care Policy and Planning oversees the state’s Medicaid and children’s health insurance programs in addition to numerous other public health programs and initiatives. The proposed $135.5 million in new health care spending would be an increase of greater than 5 percent from the previous year. As usual, greater spending in this area is driven in part by greater caseload and program participation. The state’s Medicaid and children’s health insurance caseloads are each projected to grow by nearly 5 percent. Such an increase would bring the Medicaid caseload to include more than 1.3 million Coloradans. This number corresponds to about one out of every four state residents, more than half of whom are either children or senior citizens. Colorado is now several years into the legalized marijuana experiment. According to the Department of Revenue’s Marijuana Enforcement Division, plastic nursery plant pot the state has approved recreational marijuana licenses for 424 dispensaries as of March 2016 .

In addition to these retail outlets, officials have approved licenses for over 500 recreational marijuana cultivation operations and nearly 175 marijuana product-manufacturing facilities. Licenses awarded for medical marijuana sales , cultivation , and manufacturing exceed these recreational numbers, although the disparity between the two continues to shrink . The increase in the number of marijuana business licenses has correlated positively with sales. By the end of 2014, recreational and medicinal marijuana sales totaled nearly $700 million. In total, state marijuana sales in 2015 approached $1 billion. The year’s official cumulative sales totaled $996,184,788. Though tax revenues have been considerable, the actual amounts generated have fallen short of most projections. Figure 1 provides monthly sales data for recreational and medical marijuana in 2014 and 2015. Figure 2 reports monthly tax, licensing, and fee revenue data for the two markets. All data are from the Colorado Department of Revenue. As seen in Figure 1, recreational sales lagged behind medical sales for the first seven months of 2014. This is partially because retail dispensary licenses were initially limited to those with a preexisting medical marijuana business license. Over time, the number of retail outlets has grown considerably. Sales have increased as well. Recreational sales have surpassed medical sales every month since November 2014. Medical sales exhibited more stability over the course of 2014 through the first half of 2015. Sales increased over the summer months and surpassed $40 million for the first time in August before falling again through the winter. In November, the number of residents on the medical marijuana registry dipped below 110,000 for the first time in 28 months. Figure 2 reports the tax, fee, and licensing revenue from the recreational and medical marijuana markets during 2014 and 2015. As mandated by the constitutional amendment ratified by voters, the first $40 million in marijuana excise tax revenue is mandated to go toward school construction. The legislature has more control over revenues beyond that amount. The original estimate from the governor’s office was that marijuana tax revenue in the first fiscal year would exceed $130 million.

Though the projection was lowered on several subsequent occasions, combined tax revenues totaled $76 million—$63.4 million from taxes and $12.6 million from licensing and fees . Revenue from the more modest medical marijuana taxes has been relatively stable over this two-year span, ranging from $1.41 million to $1.99 million. Though it has demonstrated more variability in recent months, the growth in recreational marijuana tax revenue has been fairly linear. Over these two years, recreational tax revenue has grown by an average of $400,000 every month. Tax revenue surpassed $10 million in four of the last six months in 2015. The $11.28 million collected in December 2015 is the highest amount to date. One would expect the growth in sales and taxes to taper off at some point. The decline in sales during the fall of 2015 was perhaps a sign that the markets were reaching a plateau; however, the trend reversed with strong sales reports in the final months of the year. In sum, the state continues to receive a reliable amount of medical tax revenue coupled with increasingly larger sums from recreational taxes and fees. If sales continue to climb, the tax revenues in the current fiscal year may approach what was originally forecast for the 2014 fiscal year as the market becomes more developed. Many expected the uncertainty surrounding Colorado’s budget for the next fiscal year to come into better focus upon the resolution of the hospital provider fee controversy. House Speaker Dickey Lee Hulling horst sponsored legislation to reclassify the fee into an enterprise just one day after Attorney General Coffman issued the formal opinion supporting the proposal’s constitutionality. Though supporters of the reform had some reasons to be more optimistic about the prospects for success in 2016, chief among them the supportive opinion from the state’s Republican attorney general, the proposal to reclassify the hospital provider fee died in the Senate once again as it had the year before. In May, the Senate Finance Committee voted along party lines to defeat the measure . Substantial parThisan disagreement on the issue is emblematic of broader conflicts in the legislature on matters of policy and spending. In many respects the governor’s budget for the upcoming fiscal year is cautious, partially out of necessity given the uncertainty that exists regarding the hospital provider fee, potential income tax rebates triggered by TABOR, and slowed economic growth. Legislators are scheduled to begin work on the budget “long bill” beginning on March 28. To further complicate matters, the Legislative Council’s chief economist recently informed JBC that General Fund revenue for the next fiscal year will likely be $90 million less than previously expected. This will almost certainly result in further funding cuts to different parts of the budget, some of which have already been targeted for reductions. State officials continue to study and refine the legal and regulatory framework surrounding the medical and recreational marijuana industries in the state. In addition to those designated for school construction, legislators have used marijuana taxes to oversee the state’s marijuana industry. Tax revenue has also been allocated to youth drug prevention, public safety, and public education campaigns. Last year, legislators adopted stricter packaging rules for edible marijuana products. In March 2016, the state announced the largest recall to date of marijuana products when laboratory testing reported the presence of a non-approved pesticide. Officials have not released data on the scope of the recall in terms of number of plants or products affected. During the same week that the recall was announced, the Supreme Court announced that it would not hear a case filed by officials in Oklahoma and Nebraska challenging the legalization of recreational marijuana in Colorado. As the market becomes more developed, plastic planters officials in other states considering legalization will have guidance in the form of Colorado and Washington.Coffee is native to Ethiopia and was introduced to the Americas in 1723 . It was first found in Brazil in 1922 , spread to Central America and Mexico between 1971 and 1978 , and was first found in Colombia in 1988 . Like other Scolytinae, CBB females form galleries for the development of progeny . In coffee, a female CBB bores into the calyx end of a single berry and forms galleries in the seed. At maturity, new CBB adults leave the host berry to infest other berries.

Considerable effort has occurred to develop strategies for control of CBB, including classical and augmentative biological control using introduced parasitic wasps , nematodesand entomo-pathogens. Here we focus on the biological control of CBB using parasitic wasps. Three parasitoids species were released in the Americas:two betilids Cephalonomia stephanoderis and Prorops nasuta , and the eulophid Phymastichus coffea . In Colombia, C. stephanoderis, P. nasuta and P. coffea were released , but only P. nasuta established . In Brazil C. stephanoderis and P. nasuta were established , and P. coffea was introduced in 1999 but its establishment is unknown. A third betilid, Cephalonomia hyalinipennis was reviewed as a possible control agent, but research suggests it could have a detrimental effect on control of CBB because of its negative interaction with the other betilids , and hence has not been released in the Americas. The braconid Heterospilus coffeicola is reported from CBB in Africa , but it has not been reared successfully for field releases in South America , and it is not considered in this study.Gutierrez et al. developed a tritrophic coffee-CBB-three parasitoid demographic system model based on the distributed maturation time demographicmodel proposed byManetsch and Vansickle, . Rodríguez et al. updated the models for coffee and CBB with new laboratory data, and tested them against field data. Here, we update the parasitoid models, and add the parasitoid C. hyalinipennis to the system. All the models use the physiologically based demographic modeling approach developed by Gutierrez and Baumgärtner . We first review the biology of the parasitoids.Adult females of C. stephanoderis and P. nasuta enter one CBB gallery during their lifte time where they host-feed on CBB adults and immature stages, and oviposit in mature larvae and pupae . The sex ratio of C. stephanoderis decreases with temperature being 25 females: one male at 17 ◦C and 7.3:1 at 32 ◦C. The sex ratio of P. nasuta increases from 4.5:1 to 5.8:1 over the same range of temperature . The reproductive strategy of C. stephanoderis and P. nasuta is characterized by sibling-mating. At maturity, new adult parasitoid females leave host berries in search of CBB infested berries. SuperparasiThism by both parasitoids has been observed under laboratory conditions, but not in the field , and intraguild predation between C. stephanoderis and P. nasuta occurs . C. hyalinipennis is a facultative predator and hyperparasite of C. stephanoderis and P. nasuta immature stages, and in the laboratory super parasitizes CBB larvae and hyperparasitizes conspecifics. A female invades only one infested berry, sibling mating is the norm, and it has not been observed attacking CBB adults . Dyadic contest was defined by Pérez-Lachaud et al. as direct competition by females of different betilid species for the resources of a CBB infested berry. Triadic contest has not been observed in the field, and it is not considered in this study.Table 1 provides coordinates and climatic details of the five locations simulated in this study, but the simulation results reported in the figures are for Buenavista and Londrina . The studies were conducted using weather data for the period 1990–1995 for Colombia and 1982–1986 for Londrina, PR, Brazil. Daily maximum and minimum temperatures , relative humidity , precipitation , and hours of sunshine for Colombia were obtained from data published in the Anuario Meterológico Cafetero . Daily solar radiation was estimated from recorded hours of sunshine using the relationship developed by Prescott . Weather data for Londrina, Brazil were obtained from the Instituto Agronômico do Paraná. To assess the effects of weather on system dynamics,the six years of weather data were shuffled across the four Colombian localities to increase the number of weather scenarios.Competition between individual parasitoids of two different species occurs in coffee berries when a betilid female enters a berry previously colonized by a female of another species . This behavior is more likely to occur when CBB infested berries are scarce. In the model, dyadic contest occurs at a rate dependent on the shortfall of CBB infested berries attacked /D) > 0) .

Black pepper is the most important one found in Berenike. Black peppers are either sun-dried or dried in special smoke rooms. Black pepper is very vulnerable to damage from mold and should be dried quickly and thoroughly. If peppercorns are stored in a dry environment, they may be kept for many years without losing their quality. Maybe it is for that reason that traders of Berenike chose to leave behind a large supply of black pepper safely stored in a buried dolium on the assumption that they would return within a few years.The group of fruits and nuts is well represented in the archaeobotanical record of Berenike, both qualitatively and quantitatively. The classification of raw food is mostly based on its use in food preparation, but its categorization is not unequivocal, as can be seen for food products classified as fruits and nuts. Some fruits, such as okra and tomatoes are even classified as vegetables. The matter is even more confused by the inclusion of seeds such as stone pine , pigeon pea , and chickpea within the group classified as nuts . From a biological point of view, nuts are a special group of fruits. A classification of fruits, nuts, and a single seed based on their anatomy and moisture content makes more sense with respect to their storage life and will serve here as a guideline. A group that is easily preserved concerns nut like seeds and nuts: stone pine , hazel , and sacred lotus . The first two were imported from the Mediterranean and the last one from the Nile Valley. Long-distance transport of these food products would not have been problematic as they can be stored for considerably long periods without spoilage. It is striking that very often seed coats of stone pine were found together with cone scales, drain trays for plants implying that they were transported as whole cones on a regular basis.

A reason for this might be that in this way all seeds could be transported, whereas after threshing some seeds would become damaged and unsuitable for transportation. It should be realized that import and consumption of seed contents only cannot be ruled out, but their possible share cannot be established as no traces are left. The baobab and the bentree have dry fruits that contain many seeds. Trading whole fruits of the baobab makes sense as both seeds and pulp are edible, and the indehiscent fruit does not easily shed its seeds. The fruit of the bentree is a three-valved capsule that bursts open when ripe. To prevent seed loss, it is believed that fruits were threshed to release the seeds. This kind of processing is still practiced by the Ma’aza bedouins living in the northern part of the Eastern Desert . No information on storage life is available, but it is assumed that both fruits can be stored for considerable periods. Most fruits found at Berenike are classified as drupes. This fruit type is characterized by one or more seeds enclosed by a fruit wall that consists of three different layers. A dichotomy between dry and fleshy drupes is based on the moisture content of the middle layer of the fruit: the mesocarp. The coconut , the almond , the walnut , and the fruit of the balsam tree are classified as dry fruits. Only the first three are edible, and it is the seed that is consumed. All three species are evidenced by their hard endocarps, indicating that these fruits were, at least partly, traded destoned. The presence of the hard endocarp is advantageous as it protects the seeds against decay and insects. The fibrous mesocarp of the coconut is highly valued as crude material for mats and ropes and explains the import of whole fruits to Berenike. The coconut was traded over long distances as whole fruits and maybe also as dried copra, although the presence of the latter could not be demonstrated. With or without the fibrous husks, coconuts can be kept until the liquid endosperm has completely transformed into solid “meat,” as is mentioned for example by Cosmas. For long-distance transport, fruits must therefore have been harvested in an unripe condition.

The drying process of coconuts depends on the temperature to which they are exposed, but will take at least several months. Although coconuts should be harvested at intervals of two months from a particular tree, in practice a yield of coconuts can be obtained from a coconut plantation every month. The logistics of transporting coconuts from India to Rome were, therefore, not problematic with respect to the short, fixed period during which Roman ships visited the Indian ports but may have been critical with respect to the duration of the voyages from Muziris to Rome, which would have taken some three months without any unforeseen delays. The fruits of the sugar date and doam palm are semisoft and take up a middle position in terms of preservation. The mesocarp of both species is sugary and edible. The archaeobotanical remains of both species indicate that they were transported as whole fruits. Although the seeds of both species also have an economic value, there is clear evidence for the latter species only that the seeds were extracted from the fruits. Both species are adapted to desert conditions and could be obtained from reasonable short distances. Hence their preservation might not have been problematic. It should be noticed, however, that fruits of the sugar date were partly infected by an insect that feeds on the seeds. Such infected fruits are still among the fruits offered for sale at markets today, which implies that the consumer does not discriminate in this case. Fruits having a soft mesocarp make up the largest part of those categorized as a drupe. The only fruit that could have been gathered from short distances, even from locally cultivated specimens, is nabq . Three other species originate from the Gebel Elba area or beyond: Cordia nevillii/sinensis, Grewia, and Cocculus pendulus. All others must have been brought from more-remote areas. Transported from the Mediterranean were the cherry plum , the domestic or bullace plum , the olive , the apricot , the peach , and the Egyptian plum . The last four species might also have been obtained from orchards in Egypt, for example in the Fayum. Finally, the soft fruits of the emblic had their origin in India. Although the high vitamin C content of emblic fruits has an antiscorbutic property and may have been transported in syrup, it is more likely that the dried fruits were traded.

Very perishable fruits are the apricot and the peach, ones that are only sweet and juicy if harvested in a ripe condition. Galen states that the juice and flesh of apricots rot easily and that these fruits are bad in general . He even advises against the consumption of these perishable fruits after other foods, as they will initiate rotting. A similar warning is given by Pliny , cited earlier. From his remark it can be concluded that fresh, sweet peaches were offered for sale, and people must have appreciated their sweet taste. The only way to transport such a fruit in an unspoiled condition is to add preservatives. Today, pickled peaches still consist of whole fruits that are peeled but not destoned. When apricots and peaches are dried, the stones can be removed. Dried apricots that are sold in Egypt today are available with or without stones. Those containing the stone are strongly dehydrated and have lost their characteristic color and are cheaper as a result. This kind of preservation also occurs quite easily under natural conditions after windfalls. Many well-preserved fruits can be collected under trees without any extra effort. The presence of a stone fragment in Berenike from a peach could be indicative of pickled fruit which must have been a rare item judging by the single fragment found so far. Considering the same fragment as a contamination in a load of dried peaches, it would leave open the possibility that they were available on a more regular scale. The apricots could originate from either pickled or dried specimens. The few remains indicate that the fruit probably was not commonly available. Plum stones, especially those of the cherry plum, are better represented in the archaeobotanical record of Berenike than peach and apricot and indicate that plums most probably were traded with their stones. Whole plums are easily dried but were also preserved in honey, as is mentioned by Apicius . The storage life of dried plums depends on the moisture content. Well-dried plums, having lost two-thirds of their weight, can be stored for about three years. Members of Cordia would have been preserved in a similar way. Some whole specimens were found, in addition to calyxes and huge numbers of stones, 4 x 8 grow tray and support the assumption that they were transported as dried fruits. Dried fruits were among the products offered for sale in Iran , but are probably no longer part of the assortment in Egyptian bazaars. According to Theophrastus , the inhabitants of Thebes made dried destoned fruits of the Egyptian plum into cakes. Fruits of the emblic can be preserved by drying or by pickling in a sugary syrup. Pickled fruits concern whole specimens, whereas recent samples of dried fruits that have been analyzed consist for the most part of fragments of the mesocarp. Segments of the stony endocarp were only present as a contamination. Thus, the few inedible stone fragments found in Berenike indicate a substantial import of the emblic if we assume that they were traded in a dried condition. Berries are fleshy, mostly indehiscent fruits and are represented by the date , the pomegranate , the grape , the persea , the caper bush and members of the Cucurbitaceae. The date contains a single seed; with all other berries the seed count ranges from a few to many. The less-juicy varieties of the date are easily sun-dried and have a suitable sugar-towater proportion of two to one. When they are kept for a long period, they become candied and lose some quality.

Dry varieties are known from the Sudan and are easily transported and stored. Mostly, dried dates still have their seed inside, which explains the large quantities of seeds in the trash dumps of Berenike. Seeds may also be present in a paste made from compressed, soft dates, which is called agwah . The storage life of fresh grapes is limited, but raisins and currants, being a Greek variety from Corinth, can easily be transported and stored. Whole fresh bunches stored in clay jars with chaff were also transported in classical times and they could keep for a long time. However, both jars and such fruits were vulnerable, which is why we may consider the grape more of a luxury item. Due to their sturdy leathery skin, pomegranates can be stored for several weeks despite their juicy content. It is important that carefully picked, unbruised fruits are selected for long-distance transport. Special treatments, such as dipping the fruit in tar or seawater before drying, will benefit the longevity of the fruit . Saltwater also prevents discoloration of the fruit. Fruits that are kept too long will become completely dehydrated and inedible. Transport of pomegranates by caravan trade was possible over quite long distances, as is demonstrated by the trade between Kabul and Peshawar in Afghanistan, a distance of about 250 km . No particular information is available on the preservation of the persea fruits. Judging by their less-juicy consistency and their availability within relatively short distances, it is assumed that preserving the persea would not have caused many problems. Galen mentions that fruits were transported from Persia to Egypt, and were ripe and became edible when they reached the land of destination . Although it is the unripe flower buds of caper bush that enter the commercial trade today, ripe fruits are also edible. Gathering ripe fruits should be done before they burst open. Once the pulp is exposed, the fruits are eaten by animals, most probably by birds that are attracted to the red-colored pulp. Caper fruits are juicy and easily rot when they are ripe. Galen mentions that fruits subjected to transport are sprinkled with salt as a means of preservation .

Biological nitrogen fixation as a bacterial process in symbiosis with plants accounts for a major fraction of nitrogen input in terrestrial ecosystems . The spectrum ranges from obligate endosymbioses, such as arbuscular mycorrhizal fungi , to generally facultative nitrogen-fixing symbioses; the latter, including the rhizobia-legume nodules , Frankia-actinorhizal nodules , and cyanobacteriumGunnera associations . Nitrogen-fixing root nodule symbiosis began with one evolutionary event that occurred in the common ancestor of a clade of angiosperms known as the nitrogen- fixing clade approximately 100 million years ago . The Frankia-actinorhizal symbioses are of particular interest in the interpretation of the evolutionary underpinnings of RNS because these symbioses developed, probably independently, within several distinct lineages across the NFC . Genetically distinct subgroups within the genus Frankia form compatible associations with different lineages of plants within the NFC , while cluster IV includes atypical non-symbiotic and/or non-nitrogen-fixing strains . Of these subgroups, cluster II Frankia strains have the broadest host range among nitrogen-fixing plants: species in four different families in two orders form RNS with cluster II Frankia strains. Representatives of these families are particularly abundant in western North America, where the current study has been carried out. Comparisons of representative sequenced genomes from these subgroups suggested a general correlation of Frankia genomic features with the corresponding host bio-geographic distribution . That being said, since RNS is first established between the host-plant root and the bacterial symbiont by the two partners coming into physical contact with each other in the rhizosphere, procona valencia it is to be expected that all of the symbiotic Frankia subgroups must have at least some capability to persist in a free living state in soil, in addition to living in symbiosis.

Clusters I, III, and IV are known to have a free-living phase in soil: infective units of clusters I and III are ubiquitously distributed in soils . Moreover, infective units of cluster I Frankia can be as abundant, or more abundant, in nearby non-host rhizosphere soil than in hosts . Similarly, strains of cluster III Frankia are well known to occur in soil independent of host-plant presence . Cluster II Frankia strains, however, have been characterized as highly host dependent, with only one report of nodule formation from soil devoid of a host, 18 months after inoculation . Previous DNA-based surveys of cluster II Frankia strain distribution have been conducted using field-collected nodules . Thus, there is only limited knowledge about the soil ecology of cluster II Frankia strains in relation to their host plants, particularly concerning the degree of association with the host presence. Since the cluster II Frankia strains have been shown to be phylogenetically early divergent within the genus , a better understanding of host association in this subgroup may shed light on the process of evolutionary acquisition of RNS. The paucity of evidence for cluster II Frankia strains living independently of host plant influences compared with the other subgroups led us to hypothesize that cluster II Frankia strains may be unusually dependent on the unique environment provided by the host plant for their long-term persistence . To test this, we conducted a comparative survey to determine the soil distribution pattern of free-living cluster II Frankia strains in three geographical locations in northern Califtornia; in different soil types, including serpentine soils, a depauperate soil type that harbors several actinorhizal hosts; and under different climatic conditions. The distribution of cluster II Frankia strains was assessed through detection of DNA sequences unique to cluster II Frankia strains in host and non-host rhizosphere soils, sampled in sites with species of host and non-host plants. The microbiome of a subset of these soil samples was also analyzed via high-throughput sequencing.

Because of the seasonally arid climate and soil environment, which has been shown to limit nitrogen fixation activity in Ceanothus , nitrogen-stable isotope analysis was conducted to confirm an active state of nitrogen-fixing symbiosis in each host plant sampled. In addition, stable isotope analysis enabled us to assess the degree of site-level influence of nitrogen-fixing host plants on cluster II Frankia strain distribution.Our study has demonstrated, based both on presence/absence of glnA sequences and relative abundance of OThus in rhizosphere soil, a significant correlation between the presence of cluster II Frankia strains and the presence of host plants in a particular site, that is consistent across all three experimental locations. There was complete correlation between cluster II strains and host plant presence in two of the three locations . Even though a high proportion of host-plant-absent site samples in MNR were positive for cluster II Frankiastrains , this was nevertheless statistically significantly different from the host-plant-present site samples . This confirms and expands upon previous observations of increased abundance of cluster II Frankia strains in soil of one host-plant-present site relative to the soil of an adjacent host-plant-absent site, using a bait plant bio-assay . While the effect of host-plant presence was consistent, the following ecological factors did not correlate with cluster II Frankia strain presence/ absence across the three locations: host species present , soil type , elevation, geographic location, summer or annual precipitation, average monthly temperature, or soil pH. The consistency of the host influence is evidence that a host-plant-derived factor present in the soil of host-plant-present sites is the major determinant of cluster II Frankia distribution in these ecosystems. We found that pH was not a significant ecological factor determining presence/ absence of cluster II Frankia strains. The pH differed significantly among the three locations, but within each location, the pattern was consistent between sites, regardless of host presence or absence. Earlier studies found pH to have an influence on Frankia nodulation units in cluster I strains , but, on the other hand, both cluster I Frankia and cluster II Frankia strains have been reported to nodulate successfully in a wide range of pH between 3.5 to 8.0 and 5.0 to 9.0 , respectively. The range of soil pH in our study was between 5.0 and 7.2. Although no studies to date have examined the effect of pH directly on cluster II Frankia strain populations in soil, in culture, the optimal growing condition seems to be neutral to basic.

The cultured cluster II Frankia strain BMG5.1 was shown to grow optimally at pH between 9.0 and 10.0 , and in another study, Coriaria myrtifolia had optimal growth and nodulation at pH between 7.0 and 9.0 . It seems, therefore, that in general Frankia spp. can tolerate a range of pH while having a more specific optimum. It is possible that at a more basic pH, where cluster II Frankia spp. are known to grow optimally, there will be a difference in cluster II Frankia abundance; however, Frankia presence/absence is not affected.ANOVA of the rhizosphere microbiomes showed that while host-plant presence/absence at a given site significantly correlated with the relative abundance of cluster II Frankia spp., this correlation did not differ significantly between the rhizospheres of C. jepsonii and H. arbutifolia , when H. arbutifolia was growing in the host-plant-present site. This suggests that the effect of the host plant is not limited to its rhizosphere but can extend to the site level. A site-level influence was also observed for nitrogen fixation, determined by the ANOVA of 15N. While host plants all showed significantly more negative 15N values than the co-occurring non-host plants, flower bucket indicating active engagement in nitrogen fixation via RNS , the non-host species in host-plant-present sites also showed significantly more negative 15N values in host-plant-present sites than in the same species in host-plant-absent sites. This pattern of 14N stable isotope enrichment at the site level is similar to the facilitative effect of nitrogen-fixing plants on primary succession, reported in other nitrogen-fixing ecosystems . It was shown previously that Thissue extracts from both roots and shoots of a host plant Casuarina cunninghamiana enhanced growth of a range of Frankia spp. grown in culture but particularly enhanced the in vitro growth of the Casuarina-compatible strains . This suggests that some factor derived from the host plant, either as leaf litter or root exudate, can explain the site-level effect observed in our experiments.Our findings suggest that the abundance of cluster II Frankia spp. in a given site is associated with the strength of the influence that a host plant exerts. In the case of MNR, where the host-plant-absent site had a relatively close proximity to the nearest host plant growing in a similar vegetation type , cluster II Frankia spp. were detectable; however, no cluster II Frankia spp. were detectable in the CC site in ALCP where no host plant was proximate , despite similarities in vegetation types. At the SEF site, cluster II Frankia spp. were not detected in the wetland site despite the high proximity to the nearest host plants ; however, the wetland site had dramatically different soil conditions and vegetation type than the adjacent montane chaparral. Cluster II Frankia itself can survive in moist wetland soil: DaThisca glomerata, another host for cluster II Frankia spp. found in Califtornia, Nevada, and Baja Califtornia, grows and nodulates in seasonally flooded streambeds or washes throughout its range . Rather, it is likely that cluster II Frankia spp. were absent in the wetland site at SEF because the more xeric-adapted host plants could not grow in wetland soil, thus eliminating a host effect. The strength of the influence that the host plants may have on a given site might be affected by factors other than proximity or vegetation type. Since the cluster II Frankia enhancement factor may be able to persist in the soil over time, a legacy effect from host plants that were previously present might play a role. For example, cluster II Frankia spp. have been found in sites devoid of host plants for more than 100 years ; Elaeagnus-compatible Frankia spp. were present at a site that had not had Elaeagnus for many years , and Comptonia peregrina seed was viable after being in a pine-dominated forest seed bank for more than 70 years . Alternatively, since Ceanothus and Cercocarpus support mycorrhizal symbiosis , the mycorrhizal mycelia may be a means of conveying a chemical signal.There was low glnA sequence variability throughout this study with all samples within 0.01 substitution/site from each other; in fact, of the 74 sequences generated in this study, 56 had at least one other sequence that was identical .

Low genetic diversity of cluster II Frankia spp. based on DNA from field nodule collections, rather than from rhizosphere soil DNA, has been well documented in Oregon with repetitive sequence PCR or PCR-restriction fragment length polymorphism and in Califtornia and Nevada using single-sequence analysis . interestingly, a glnA sequence identical to one found in the current study has been detected in DNA extracted from a nodule of Dryas drummondii collected in Alaska . On the other hand, DNA from nodules collected in the southeastern United States showed a substantial divergence, based on RFLP, from that of nodules from western North America . A study of nodules collected in a circumscribed area in southern coastal Califtornia found diversity of cluster II Frankia at the species level, using rep-PCR . The sampling area in the southern coastal Califtornia study corresponds to a zone of biodiversity for the genus Ceanothus as documented previously . Taken together, these findings suggest that a suite of closely related strains of cluster II Frankia is dominant in major portions of western North America, with some localized diversity. Various factors have been suggested to account for the observed strain diversity within cluster II, however low it may be. Previously, a weak geographic pattern of strain diversity by Ceanothus collection sites was found . In reference 15, diversity was attributed to two ecological groupings of the host plants , but the sequence groupings were separated by 1%. Both of these findings reflect the generally low cluster II Frankia strain sequence diversity in western North America. Whole-genome sequencing has been carried out on strains of cluster II Frankia from Pakistan and Japan ; additional whole-genome sequencing from a greater number of sources will be valuable for assessing the degree of global geographic and genetic diversity within this clade.It has been postulated that cluster II Frankia is an obligate symbiont, based on the difficulty of isolating any strain in this group and the difficulty of detecting it in soils devoid of host plants .

The fruits are produced in the axils of the leaves and on the upper portion of the stem and branches where they ripen in an upward direction. Owing to the long period of fruit ripening and the fact that the capsules remain open during ripening, seed losses can be sizeable during harvesting. Strictly speaking, varieties that still have open capsules are not fully domesticated. To prevent seed losses, the plants can be uprooted before the first fruits start to scatter their seeds. The uprooted plants are shocked and left to dry, a process in which the water and minerals present in the stem allow further ripening. Drying and threshing of sesame is done at suitable spots that may be found at quite some distance from the fields before harvesting. This is still practiced in Egypt today. The color of the seeds varies from white to black. Fried seeds can be eaten in soups and, after removal of the seed coat, they are also frequently used to garnish bread and the like. From the dried ground white seeds, a paste called tehinah or halva is made. The oil, traded as oil of benne, can be kept for a long period owing to its resistance to oxidation and rancidity, subject to temperatures being rather stable. Also the high vitamin E content, high grade protein, and good taste attribute to its quality . Sesame oil is used as a salad and cooking oil. Furthermore, both seeds and oil are also used in medicine. Pliny , for example, recommends them for a variety of disorders, including the bite of a gecko and inflammation of the ears. Strabo mentions that it was a custom in Mesopotamia to anoint the body with sesame oil. The uprooted sesame stems, which are leftover after threshing, have little stock-feeding value and are used as fuel, 30 litre pots bulk as is still practiced, for example, in Egypt and Syria. In India, sesame seeds have a long tradition in religious ceremonies. Depending on the associated food products with which it is offered, it gratifies ancestral souls for one month, with for example rice and barley , and for infinity if combined with rhinoceros flesh .

Literature dealing with the origin of sesame is incomplete in its compilation of archaeobotanical records, that of Egypt being no exception. All Egyptian finds are dated from the New Kingdom onward with the exception of two records from Naqada, located some 25 km north of Luxor. One record concerns unspecified remains and is dated to the First Intermediate Period or even to predynastic times . The other record concerns pollen from either S. indicum or S. alatum Thonning and is dated to the predynastic period . S. alatum is a wild species from Sudan, but is also locally cultivated for its edible seeds, which are sometimes used as an adulterant of sesame . If both records from Naqada concern Predynastic S. indicum indeed, they predate the one from Harappa. Historical sources indicate that the Greek and the Romans also were very well acquainted with sesame. During the Greco-Roman period, sesame was especially cultivated for the consumption of the whole seeds rather than for oil production . According to the Periplus Maris Erythraei, only sesame oil could be obtained from the so-called farside ports along the Somalian coast, this oil originating from Skythia, the alluvial plain of the Indus in northwest India . Obviously, the import of sesame oil to Berenike would have been on a modest scale. The seeds found at Berenike were probably imported from the Nile Valley and used as a garnish, among other things. Of particular interest is a piece of what might be a sesame cake, found in trench 33. It consists of an indefinable matrix in which many sesame seeds are incorporated. According to Galen , such a cake was made of raw honey and sesame seeds .Sorghum is a typical domesticated grain in the African savanna belt, south of the Sahara where the wild sorghum is still widely distributed . The plant has a C4-metabolic pathway, making its photosynthesis very efficient in an environment characterized by high temperatures and intense light. In comparison with other cereals, such as barley, wheat, and rice, it has a considerable yield.

Two subspecies are distinguished: wild sorghum and domesticated sorghum . Additionally, four races of wild sorghum and five races of domesticated sorghum have been described. The start of sorghum domestication is still disputed. Some are of the opinion that sorghum is a latecomer and opt for a domestication not earlier than the late first millennium BC . Others, such as Harlan and Fuller , do not rule out the possibility of a much earlier African domestication and accept the validity of some of the records from Pakistan and northern India, dating to the first half of the second millennium BC. This would imply that soon after its domestication, sorghum reached this area through coastal trade. The oldest record of Egyptian sorghum comes from the early Neolithic site Nabta Playa and is considered as wild sorghum . Several records of sorghum are dated to the New Kingdom , without further indication on its wild or cultivated status. From several of these finds, the dates are doubted. It should be realized, however, that the tough and glossy husks of sorghum may retain a surprisingly fresh appearance, comparable with that of well-preserved rachis fragments from barley and wheat, as is the case with many specimens from Berenike. The abundant remains of cultivated sorghum that have been excavated from Berenike indicate that this cereal played a substantial role in the food supply in the city from the fifth century AD onward. There are several possibilities with regard to the origin of the sorghum found at Berenike: import from the East African coast south of the Sahara, import from the Nile Valley, and from local cultivation. Import from Ethiopia and Eritrea might have been possible via Adulis, which according to the Periplus was the nearest port south of Berenike. It was the only port of the Axsumite kingdom. Recent excavations at Axum, the capital of the kingdom and located some 150 km southwest of Adulis, have so far revealed only one single grain of Sorghum cf. bicolor . As far as the Nile Valley is concerned as an area from which sorghum was obtained, it is possible that sorghum originated from Nubia. Archaeobotanical evidence from this area is relatively rich and indicates that sorghum was cultivated here at the start of the Christian era.

Clapham and Rowley-Conwy found cultivated sorghum, race bicolor, in Qasr Ibrim from the Roman period onward. Trade contacts between Berenike and this important urban site is also evidenced from black pepper . Furthermore, most of the amphora vessels unearthed from trench BE95/96/97-5 are very similar to those recorded from Qasr Ibrim . Direct or indirect import of sorghum from India seems unlikely, as we are dealing with a cereal that predominates in late Roman Berenike and is, therefore, probably related to the presence of native populations. Moreover, the crop was available from neighboring areas and was probably also cultivated locally on a small scale. Both humans and animals may have consumed sorghum. Because the seeds do not contain gluten, it cannot be used for making bread. Mostly, the fl our is mixed with water or fat and made into a porridge. Schweinfurth mentions that camels thrived if fed on sorghum . The sorghum that is occasionally cultivated near Arab Saleh, wholesale plant containers some 10 km northwest of Berenike is also used as camel fodder. An old necklace obtained from an Ababda nomad, made of woven leather strands and decorated with glass beads and braided tassels, proved to be filled with sorghum seeds . Other specimens were stuffed with fibers or fenugreek could be the tamarind is only based on a description of the leaves and is not convincing as this description also applies to other members of the legume family . Also the archaeobotanical record is scanty. The specimens unearthed at Berenike are the first ones from the African continent. Additionally, only two archaeobotanical finds are recorded from India. They originate from Nevasa and Kolhapur, both located in the northwest coastal area of India in the vicinity of Bombay and are dated to the Satavahana period . The 66 tamarind seeds from Berenike were found in 11 different loci spread over 3 trenches. Assuming that the fruit part was consumed, the presence of its seeds indicates that the fruits of tamarind were at least partly traded without the seeds removed. Ripe fruits can be kept for a long time and are easily traded if compressed together in cakes or balls. Such tamarind balls are still offered for sale in Egyptian bazaars. According to Von Maydell , it is recommended that besides the inedible fruit wall and the fibers, the seeds also should be removed from fruits meant for storage over a long period of time, as the seeds may contain seed feeding insects that can cause heavy losses. Because the seeds themselves also have an economic value, and the production of pulp without seeds would have increased the price, it seems likely that whole fruits were brought to Berenike. The fruits might have been imported from either northeast Africa or India. Deducing from the information available, it seems most likely that they were obtained from the southern savanna of Sudan and Ethiopia, from where they might have been shipped to Berenike via Adulis.Ajowan occurs in south Europe, northeast Africa, and west and central Asia, but it is only recorded as a cultivated plant or as one that has strayed from cultivation. Archaeobotanical remains could be helpful in identifying its area of origin, however, records are scarce, probably due to the small size of the otherwise characteristic fruits. The only sub-fossil remains of ajowan that are known so far originate from Egypt. Besides the fruit from Berenike, it has been recorded from Mons Claudianus in the Eastern Desert , Amarna in the lower Nile Valley , and there is a sample of unknown origin and date stored at the Florence Museum .

The find from Mons Claudianus is contemporary with the one from Berenike, whereas the fruits from Amarna are dated to the Eighteenth Dynasty . Although the find of ajowan at Amarna suggests that the plant probably already had a long tradition in Egyptian horticulture, it remains unclear whether the plant once belonged to the native flora. The many synonyms that have been used for this plant indicate its close resemblance with several other umbelliferous plant species, such as khella , carrot , caraway , black caraway , and stone parsley . Most of these plants, including ajowan, are still used for flavoring food. Among others, Dalby assumes that ajowan is already mentioned by classical writers such as Pliny, Galen, and Dioscorides. But the descriptions of the concerning plant mainly deal with its uses, which are quite universal, and do not mention diagnostic features specific to ajowan. Nevertheless, we cannot know for sure if ajowan was meant indeed. Ajowan is mainly cultivated for its fruits, which are used both as a flavoring spice for food and as a medicine, for example, in the treatment of kidney calculi and dysentery. Its medical properties can be attributed to the presence of thymol, a powerful antiseptic that can be used both internally and externally . Several plant species are exploited as a source of thymol, but ajowan is the most favored one as it yields the highest percentage of this valued compound. Today, however, thymol can be prepared synthetically, which decreasing the demand for natural sources. Ajowan could have been imported from the Nile Valley, but it is also possible that it was grown in local kitchen gardens at Berenike. Although it takes 5 to 6 months before the fruits are ripe, they can be harvested earlier to prevent loss due to seed dispersal, without a reduction in quantity of the fragrant oil .The wild relatives of fenugreek grow in the Mediterranean area and in the Near East, and it is assumed that this pulse crop had been cultivated somewhere within this area .