Several of the cache pits were made entirely of flat slabs of quartz diorite that evidently had fallen from the ceiling of the shelter. In each case, these tabular rocks were carefully overlapped to make as tight a construction as possible .Other slab-lined cache pits differed in that the rocks were placed in a mosaic fashion rather than overlapping. No small chinking rocks were noted in any of these features. One of these was constructed globular, as opposed to tabular, form. In some cases, smaller rocks were used to chink spaces between the larger rocks . One cache pit , constructed of irregularly shaped rocks, lacked any small rocks or predominantly of tabular rocks and was placed such that two massive boulders were part of the overall construction. Incorporated into its construction were two metate fragments, both of granitic material. The fill of the feature consisted of miscellaneous rocks and soil, a mano and two mano fragments, and one complete unifacial block metate. Some ofthe milling implements or other rocks from the fill may have been used in its construction. The majority of the cache pits excavated at Indian Hill Rock shelter were constructed of medium- to large-sized rocks of overall chinking, and there were large spaces between the rocks. Some rocks may have been taken from this feature and reused in the construction of the overlapping cache pit or for another purpose. One of the rocks used in the feature is a bifacial block metate; also present was a rock with a patch of red hematite paint. Feature 19 was a well-constructed rock-Hned basin. It was fairly shallow and apparently represented the floor of a cache pit. Construction material consisted of about 40 rocks,hydroponic vertical garden including one core-hammerstone, nine block metate fragments of granitic material , and one unifacial mano.
This feature was unique among the cache pits in that it contained so many large metate fragments. The situation at Indian Hill Rock shelter, then, is one in which rock-lined cache pits were built in sandy matrix using either flat slabs or other rocks, including metate fragments. Care was taken in some of the cache pits to overlap the slabs used in their construction, and in others to chink interstices between rocks. This suggests attempts to make the structures rodent-proof. We therefore conclude that bags or baskets of foodstuffs, rather than equipment, most likely were stored in these cache pits. We are inclined to classify these cache pits as group specific, permanent furnishings at Indian Hill Rock shelter. Each cache pit may have been used many times, each time carefully roofed over, perhaps with bunch grass to keep the sand out, and then rocks, and concealed beneath the sand that formed the floor of the shelter. Concealment may have been considered important in the event other groups used the shelter. We thus interpret the cache pits as more or less “permanent” site furnishings of whatever group used Indian HiU Rock shelter, but not publicly available site furniture. In all cases the contents of the cache pits had been removed in antiquity. In most cases, only the rock-paved floors of the cache pits remained intact, and most of the rocks that formed the walls were lacking. In some cases large rocks all but filled the cache pits, apparently having been discarded there upon abandonment of the cache. The roofing rocks and wall rocks, which may have extended to near the former ground surface, must have been pulled up out of the site matrix and used elsewhere for the same or another purpose. The ethnographic description of “pulling up” the site furniture , or “site furnishings,” in this case using it elsewhere at the site for the same or for another purpose, appears to describe the situation at Indian Hill Rock shelter. The levels from which cache pits are believed to have been dug have few or no ceramic sherds, but ceramics are common in overlying levels.
This distribution leads to the conclusion that ceramic ollas may have replaced rock-lined cache pits after the inception of ceramic technology in the region. Ollas may have been buried in the same sheltered sites that formerly contained rock lined cache pits. Rock-lined cache pits are reported from two sites, Chapman Rock shelter No. 1 and Resurrection Shelter . Four rock lined cache pits were excavated at Chapman Rock shelter No. 1. These cache pits were lined with basalt slabs, and metates were used in the construction of two of them. They ranged from ca. 75 to 95 cm. in diameter and from 45 to 60 cm. in depth. Two of these cache pits contained historic artifacts ; the third was built from a comparable level below the surface . Linings in these cache pits included such material as bunch grass; buckwheat plant parts, Joshua tree fiber, tule matting, and twined basketry. The fourth cache pit possibly may be older, as it was built far below the others, at about 1.2 m. below the present surface. This cache pit was not lined, but contained debitage, a small biface, a slate pendant fragment, a basalt mano, a bone bead, pinyon hulls, and twined basketry. A large, rock-lined cache pit was partially exposed during test excavations at Resurrection Shelter on the eastern slope of the Coso Range near Darwin Wash. The maximum diameter of this feature is estimated to be about 2 m., the depth about 40 cm. It was constructed of large, blocky rocks. None of these appeared to be metates, nor was any chinking with smaller rocks noted. The pit was lined with alternate layers of bunch grass plants and Joshua treebark. A covering of rocks was not discernible, but a large pile of rocks just inside the entrance of the shelter and next to this cache pit once may have covered it. Artifacts in and around the cache pit include basket fragments, a remnant of a fiber brush, fiber cordage, pinyon nut shells, fiber cordage wrapped with strips of skin, and painted reed fragments . The shelter is very small, and it may have been used only for storage. Some or all of the artifacts could have found their way into the cache when opened. Since pinyon trees grow several kilometers away at higher elevations, the nut shells apparently did not get into the cache pit accidentally. The available evidence suggests the cache served for storage of pine nuts rather than equipment. A detailed report by the investigators is in progress, and radiocarbon analyses are planned for selected materials from this cache pit .
Insufficient information is available to make a determination of the organizational strategy of the people that built the cache pits in the Coso Range. Ethnographically, the region was occupied by Panamint Shoshoni, who shifted from a dispersed forager strategy during the spring and summer to a clustered collector strategy during the fall and winter. Crown gall of grapevine reduces the grape yield by 20% to 40% and trunk diameter approximately by 9%, compared to uninfected healthy grapevines . The causal agent of the grapevine crown gall, Agrobacterium vitis, is a different biovar from other Agrobacterium spp. because it can induce a grapevine specific crown gall . A. vitis has host specificity-related genes, such as pehA, which encodes polygalacturonase that is hypothesized to facilitate A. vitis attachment and systemic colonization on susceptible grapevine tissues . Another host specificity-related gene function is tartrate utilization, and the host specificity is explained by the use of tartaric acid from specific organic acids in grapevines . Crown gall disease is one of the difficult diseases to eradicate by chemicals in vineyards. As a strategy to prevent crown gall disease without chemical sprays, vertical vegetable tower the use of resistance plant cultivars and antagonistic microbial species have been developed. Some grapevine species, mostly Vitis vinifera, are susceptible to crown gall disease, and thus, root stocks from resistant species or crossed seedlings are used by many vineyards to prevent crown gall disease on the scions of the susceptible grapevine cultivars . Another prevention strategy is the use of a non-tumorigenic A. vitis strain as host-specific biological control of grapevine crown gall, in which its pretreatment inhibits transformation by tumorigenic A. vitis strains on the grapevine, leading to the inhibition of crown gall induction . Recently, various platforms have been used to attempt to identify disease resistance-related metabolites in grapevine, including nuclear magnetic resonance spectroscopy and liquid chromatography mass spectrometry . However, when using gas chromatography mass spectrometry , extracted metabolite-profiling data can annotate information from the metabolite libraries . Derivatization techniques are required to process GC-MS metabolite profiling . Silylation is the most suitable derivatization method for non-volatile metabolites, such as hydroxyl and amino compounds . One silylation agent, Nmethyl-N–trifluoro-acetamide , has been used to reveal the whole range of metabolites in plants . However, the highest advantages of GCMS analysis in metabolomics are noted as well-organized stable protocols from sampling to data analysis, covering relatively a broad range of compound classes .
Principal component analysis is an unsupervised method that attempts to create a model of the data without a priori information, which is able to provide an overview of the whole metabolite profile and find the differential metabolite profiling within the group using a qualitative analysis . Projections of latent structures analysis is a supervised analytical method that enhances the separation between groups, combining variable importance for projection scores to obtain a proper cutoff value and increase its performance . Orthogonal projections of latent structures analysis add a single component to the PLS analysis as a predictor of the model, and the other components describe the variation orthogonal to the first predictive component . OPLS discriminant analysis provides useful information about putative biomarkers using an S-plot that combines the covariation and correlation from the model in a scatter plot . The VIP score in PLS-DA and correlation value in OPLS-DA provide cutoff values for finding significant metabolites. This experimental method using GC-MS and multivariate data analysis can identify new aspects of pathogenicity-related metabolites. Genetic, physiological and morphological changes of the host plant to the infection of Agrobacterium species and in the crown gall development have been well documented in relation to plant responses to the tumorigenic bacterium before T-DNA transfer and during gall development . However, relatively little has been studied on those changes occurring in the non-tumorigenic plant tissues that have been influenced by the disease development , which may broaden the holistic understandings of plant responses to the crown gall pathogen infection and disease development. This may enhance their practical applicability in the disease control; e.g., use of crown gall-resistant root stocks. Therefore, in the present study, we examined metabolite profiling in the intermodal tissues of grapevine species in relation to plant responses to the crown gall development in grapevines induced by the infection of A. vitis using GC-MS and statistical multivariate data analysis. Grapevines used in our study were obtained from the National Clonal Germplasm Repository . For Vitis spp., 11 to 22 green shoot cuttings were collected from NCGR fields in June 2015 and placed in a perlite bed for rooting at 25o C with mist for one month. The green shoot cuttings with root systems were transferred to a pot with bed soil for pathogen inoculation test. The pathogen, kindly provided by Dr. Burr, Cornell University, was A. vitis. K305. The bacterium was cultured on potato-dextrose agar at 25o C for three days. The bacterial cultures grown on PDA were collected by the agar surface scratching in sterile distilled water , and the population density was examined by dilution plate method , which was diluted with SDW to make a bacterial suspension with a concentration of 109 colony-forming units ml–1. For inoculation, a wound was made in the internode of the green shoot using a 2-mm-day drill and 10 µl of the bacterial suspension was injected into the wound using a micropipette. After inoculation, the wound site was immediately wrapped with sealing film and the green shoot cuttings were grown at 25o C in a greenhouse. Symptom development was examined every other day after inoculation throughout the experimental period. The GC-MS system consisting of a gas chromatograph and mass detector was used for GC-MS analysis of the metabolites extracted from the tissue samples.