Specialized relationship between specialized pollinator and host-plant flowers is commonly mediated by specific volatiles

The Hazara University, Mansehra, Pakistan is situated 14 km to the north of Mansehra town on the right side of Karakurum highway in a serene, peaceful, rural and pollution-free environment. The vast plain of Pakhal with its agricultural wealth irrigated by the river Siran and tea gardens surrounds the University. The ladybird beetles  are bright in colors. Female is larger than males. The haemolymph is repellent by having a repulsive smell with containing various alkaloid toxins. The bright red on black or black on red color of some adults is aposematic. Both are defensive mechanisms against predators. They are associated with good fortune in many myths and legends. Scientists increasingly prefer the names of ladybird beetles or lady beetles, as they have great economic importance as predators to many pests and environmental or bio-indicator. They may be entomophagous or phytophagous and expose both potential and real effects on the life of their hosts. Cannibalism of eggs, larvae and pupae are common, especially when prey is scarce. They have several generations in a year and reproduction is slowed in winter, when adults may hibernate. They require 190.3 ± 10.2 days to complete a generation under laboratory conditions. The adonis ladybird, Hippodamia variegate  completes 4 generations per season on the alfalfa crop, Medicago sativa L. . There have typical differences in their behavior between 2 trophic groups. First one is feeding on aphids; development, age and movement will be faster, and is also typically larger and lays their eggs in clusters. Second one is feeding on scale insects which develops more slowly, lives longer, moves more slowly, and is typically smaller and lays their eggs singly .During the present survey, the study area was divided into 3 quadrated, i.e., residential area, administration area and main campus, and visited 2 h in each morning and evening. In total 300 individuals, 7 species from 6 genera belonging to subfamily Coccinellinea and tribe Coccinellini were collected.

Different species of the ladybird beetles were reported by Gilani from Faisal Abad, Shah from Peshawar, Irshad and Rehman et al.from Pakistan. The present study reported them from Hazara University, Pakistan. Trehan and Malhotra had reported from India and Zhang and Han from China. Therefore, these studies showed that their species are cosmopolitan. However, no new species was collected. At the present, the distributions of these insects were different in three quadrants of the study sites, however, ecological conditions of these areas were more or less the same, and moreover, human activities were the highest in administration area and the lowest in residential area than main campus. When comparing the vegetation, mobile vertical grow tables we found that in residential area, the ladybird beetles were the densest, but in administration area, they were sparsely grown compared with main campus. There might be possibility that residential area was the most frequently visited and administration area was the most rarely visited. During the present study, the most encountered species was C. septempunctata whose in which 220 individuals were collected: 150 were from residential area and 70 were from main campus. Rahatullah et al. reported total 50 specimens of C. septempunctata collected during a survey was conducted in district Dir Lower over a period of 2 years. This species was previously reported by Rehman et al., Gilani, Shah and Irshad from Pakistan. Due to their broad habitat range and voracity, there has been concern that C7  may pose a threat to native Aphidophagous coccinellids. Sometimes C. septempunctata has been used as a model for polyphagous predators. Habitat choice and foraging behaviour have attracted considerable interest in studies of the ecology and life history of C. septempunctata. Since it is a polyphagous species, Vet and Dicke hypothesized it, as a capacity for learning. Therefore, the present studies provide us awareness, education and learning. At the present, the second most encountered species was the H. variegate, in which 32 individuals were collected: eight were from residential and 24 were from administration area of Hazara University. Gilani, Shah and Irshad also reported it from Pakistan. Therefore, this species is common in Pakistan. An extensive survey of predatory Coccinellid beetles  was conducted in the Chitral district, Pakistan, over a period of 7 months .

Total 2600 individuals of Coccinellids were collected from 12 different localities having altitudes from 1219.40 – 2651.63 m. Twelve different species belonging to 9 genera of 3 tribes and 2 sub-families were recorded. Two sub-families, viz., Coccinellinae and Chilocorinae were identified. However, the present survey was conducted in terms of short duration , limited study area  and no finding, moreover, a total 300 specimens, 7 species from 6 genera belonging to subfamily Coccinellinea and tribe Coccinellini were collected. Further, only 4 more species were found by Khan et al. If an extensive survey will be done in the present study area, it will provide more information about the ladybird beetles. Sathe and Bhosale reported 21 species of ladybird beetles feeding on aphids and several soft-bodied homopterous pests of agricultural and forest plants from Maharashtra. Kandibane et al. reported 7 species of predatory ladybird beetles in an irrigated rice ecosystem at Madurai, Tamil Nadu. Fish et al. stated globally invasive hemipteran pests have adversely affected the production of many food, fiber and ornamental crops. The globalization and the concomitant increase in international travel and commerce are directly-correlated to the resultant enhancement in introduction and spread of exotic invasive species, which cause loss of biodiversity, modify the habitat and cause extensive environmental and economic harm. While predatory species are often used as biological control agents, introduced species of ladybirds such as Coccinella septempunctata in North America was outcompeted, displaced native coccinellids, and became pests in their own right. The present results suggested that the ladybird beetles community structure in 3 study sites with different vegetation differs greatly as described above. The number of species found in the administration area was minimum compared to main campus and residential area because of more construction with greater number of people there. The diversity of ladybird beetles seen at 3 sites in the current study was in accord with Omkar and Bind that reported only 6 – 7 species of the ladybird beetles from agricultural and horticultural plants of Utra Pradesh. Most species of ladybirds are considered beneficial because they are predators of Homoptera, many of which are considered to be pests. These predatory ladybirds contribute to the regulation of populations of their prey, and in some situations contribute a high level of regulation.

Many insect groups are important mutualistic partners of plants, mostly due to their roles as pollinators. Volatiles from plant flowers attract nectar-seeking and pollen-collecting pollinators to visit and pollinate by supplying rewards. Meanwhile, volatiles from floral branches prevent flower-robbers from foraging and pollinating . Components from floral scents and herbivore induced flower-volatiles include alcohols, aldehydes, ketones, organic acids, ester, terpenes, nitrogen compounds, hydrocarbons, as well as other chemicals varying with plant species and volatile collection types. Floral scents from 4 cultivated varieties Prunus persica were analysed , but the effect of these volatiles on pollinator attraction is unknown. The evolution of floral traits in the context of attracting mutualist or antagonist pollinators may place conflicting selection pressures on plant traits, and the effect of these volatiles on pollinator attraction is indeterminate. Many flowers produce volatiles at times when pollinators are most active; VOCs such as a kind of muurolene are characteristics of the syconium pollen receptive diurnal phase, and may attract the diurnally-active pollinating wasps. In some specialized pollination context, only one or a few specific floral volatiles mediate communication with specific pollinator species, and pollinators are attracted by a blend of widespread volatiles.Pollinator visitation is influenced by composition and quantity of flower volatile in long distance, dependent on the pollinator rewards, i.e., pollen and nectar in short distance, and volatile emission by flowers changed upon feeding by herbivores on the leaves when Pieris brassicae caterpillars were allowed to feed only on the leaves of Brassica nigra plants. The volatile composition of floral scents is not static, spatiotemporal variability in the identity and complexity of scent bouquets could provide critical information for the mediation of plant-insect communication because floral visitors can use subtle difference involatiles to make foraging choices. Floral scents have the advantage in that they can supply pollinators a species-specific identity to flowers with their potentially infinite composition and diversity. That Prunus persica fruits need flower-visiting insects, and floral scents play a key role in pollinating. We investigated the floral scent volatile bouquets of Prunus persica, and tested VOCs composition, similarity of different trees, dominant floral volatile dynamic, floral scents correlation with flower-visiting insect community richness, mobile vertical farm abundance and diversity before, on or after volatiles sampled day with dividing sampled insects visiting floral branches by odd or even method. We report the use of living flowers captured by closely and circularly head space way in adsorbent tubes and ATD-GC/MS analysed 20 VOCs from Prunus persica flowers, and floral scents correlation with flower-visiting insect richness, abundance and diversity, which are special and dominant floral scents and what the similarity from Prunus persica in different spatial and temporal branches.

Are the dominant floral odors temporal dynamic varied differently, and floral odors with difference from temporal trees. We expected that get the special and dominant floral odors from different sampled branches, which led us to hypothesize the potential attractive floral odors that insects locating on trees. Does VOCs correlate with floral insect richness, abundance and diversity, we want to know that the insects visiting odd or even flower branches correlated with floral odor closely, so that definite the branches kept when pruning Prunus persica, and promote harvest to get economic benefit.Sample sites that the 1280.0 m altitude of sunny and shady slope habitats in Wan Bailin Ecological Garden, Shanxi Province, China , with temperature 9˚C – 31˚C, humidity 27% – 48%, precipitation probability 5% – 90%, sunshine duration 2 – 12 h, 2 – 3 level wind, UV intensity weak-strong, atmospheric pressure 1002 – 1016 Pa, 05:00-5:52 sunrise, 19:06-19:53 sundown. 186 plants Prunus persica are surrounded by the Sophora japonica in the West, Berberis thunbergii cv. Atropurpurea pruning balls in the South, and Malus micromalus in the Northeast. On shady slope, 169 plants of Prunus persica are surrounded by Armeniaca vulgaris in the West, Salix matsudana in the South, and garden road in the North . Volatile collections were conducted from 2 trees of Prunus persica on sunny and shady slope on 29 April 2016, respectively. Two fully blooming floral branches of each tree were sampled. Food grade polythene bag  was used to include the branches with inlet and outlet glass tube with absorbent carbasus . QC-1S Atmospheric sampling instrument  was used to generate the air flow of 1.5 L/min filtered with activated carbon. Each collection was done 10 min. Volatiles analysis were conducted by Turbo Matrix 50 ATD  and Clarus GC /Clarus 600 T MS   with Turb oMass 5.4.2 recognition pattern were used for chemical analysis of collected volatiles. Carrier gas velocity was 2.0 ml/min. First stage thermal desorption temperature  was 260˚C, keeping for 10 min. Cold trap temperature in the first stage thermal desorption process was −30˚C. From first stage to second stage thermal desorption process heat-up speed was 40˚C/s. The second stage thermal desorption temperature was 300˚C, maintaining for 5 min. Thermal desorption mode was import diversion, general dry for 10 min. Desorption flow rate 25.00 ml/min, import diversion flow rate 10.00 ml/min, export diversion flow rate 20.00 ml/min. Chromatographic separation condition that chromatographic column was Elite-5 MS capillary column . Temperature was programmed from 40˚C  to 180˚C at 6˚C/min , then increased to 270˚C at 15˚C/min that hold 3 min. Mass spectrometric detection condition was 220˚C ion source temperature, 250˚C interface temperature and 29 – 600 amu m/z scan range. Constituents were identified by peak area normalization method that calculate relative percentage content of each volatile compound from Prunus persica flower in comparison with standard spetral library NIST08 and combined with the retention time and related chemical experience, according to ATD-GC/MS get TIC effective peak.