[Metabolomics analysis of Lycium barbarum leaves after Aceria pallida infestation].

Aceria pallida is one of the most common pests in the main production areas of Lycium barbarum in China. The mite mainly feeds on foliage, leading to local tissue deformation and formation of massive galls, which seriously affects the growth and yield of L. barbarum. However, little is known about the influence of galling organisms on plant primary and secondary metabolism. In order to compare the metabolites differences between healthy and the mite infested leaves of wolfberry, and provide a scientific basis for the development and utilization of the galled leaves, L. barbarum seedlings were infested with A. pallida artificially in the laboratory, the metabolites of L. barbarum leaves were determined by LC-MS/MS. Our results showed that the leaves were rich in amino acids and flavonoid compounds. A total of 204 compounds from 16 classes were detected in L. barbarum leaves based on LC-MS/MS. The primary metabolites are mainly amino acids, and the secondary metabolites are mainly organic acids and flavonoids. The content of the metabolite in the leaves of L. barbarum was significantly affected by the mite, 30 metabolites such as flavonoids and phenylpropanoids were significantly changed, 21 metabolites were up-regulated and 9 metabolites were down-regulated significantly. There were 8 compounds which has pharmacological and biological activity, such as eriodictyol, isorhamnetin-3-O-neohesperidoside and scopoletin up-regulated significantly. Based on the above findings, we suggest that the galled leaves of L.barbarum have a potential to be developed in the future.

[Ultrastructure observations on antennal sensilla of Heortia vitessoides the most serious pest of Aquilaria sinensis].

Heortia vitessoides is the most serious pest of Aquilaria sinensis,which is an economically important evergreen tree native to China and is the principal source of Chinese agarwood. In severe infestations,the insects completely eat up the leaves of A. sinensis,causing severe economic losses. In a more recent study,we found that the antennal sensilla of adult play important roles in the host location,mating and oviposition of H. vitessoides. Here,the external morphology of the antennal sensilla of H. vitessoides were examined using scanning electron microscopy. The result showed that the antennae of both sexes of H. vitessoides were filiform in shape,which consist of the scape,pedicel and about 64 segments of flagellomeres. Eight morphological sensilla types were recorded in both sexes,including sensilla trichodea,sensilla chaetica,sensilla basiconica,sensilla coeloconica,sensilla styloconica,sensilla auricillica,sensilla squamiformia and böhm bristle. Major differences were recorded in the distribution and quantity of different sensilla types in each segment of antenna. The sensillas are almost confined to the ventral and lateral surfaces rather than the back side of antennae. Antennal flagella contained the most sensilla while the scape and pedicel segments only contained böhm bristles and sensilla squamiformias. Sensilla trichodea Ⅲ were only found on male antennae. These results are discussed in relation to the possible roles of the sensilla types in the host location,mating and oviposition selection behavior of H. vitessoides.

[Overwintering status and species identification of gall mite in major wolfberry production areas].

To identify the wolfberry gall mite species and their overwintering situation in different wolfberry production areas and to provide a scientific basis for their prevention and control. We investigate the phoresy overwintering situation of gall mite, and identify the species of gall mite. Our results showed that the phoretic gall mite was Aceria pallida in all the production areas. The phoresy of gall mite was a common occurrence in different production areas, and the phoresy probability was above 75%. The phoretic behavior of wolfberry gall mite occurred commonly in the other production areas. The control of this mite in different production areas should pay attention to their phoretic behavior.

[Variance analysis on polysaccharide,total flavonoids and total phenols of Lycium barbarum leaves from different production areas].

To compare the polysaccharides( LBLP),total flavonoids( LBLTF) and total phenols( LBLTP) of Lycium barbarum leaves from 14 production areas from Ningxia,Qinghai and Gansu at different harvesting time. Our results showed that there were significant difference of LBLP,LBLTF and LBLTP among different production areas. The LBLP collected from Xiangride township production area,Dulan,Qinghai was as high as 15. 02%. The LBLTF collected from Dagele township production area,Geermu,Qinghai was as high as28. 63%. The LBLTP collected from Keluke township production area,Delingha,Qinghai was as high as 16. 7%. There were also significant difference of these 3 components between different harvest periods. The average LBLP in May( 10. 20%) was significantly higher than that in June( 8. 49%). However,the average LBLTF( 17. 71%) and LBLTP( 12. 77%) in June was significantly higher than that in May( 14. 15%,11. 19%). The LBLTF and LBLTP have a positive correlation with the altitude of production areas,but there was no significant association between LBLP and altitude. The LBLP,LBLTF and LBLTP in different production areas and harvesting time was significant difference,the results provide some references for furthermore development and utilization of L. barbarum leaves.

[Threshold temperature and effective accumulative temperature of Periplaneta Americana].

Periplaneta americana is an important medicinal insect. A series of new drugs developed from it have remarkable clinical effects and are in great demand in the market. Because of unclear biology, the quality and yield of P. americana are affected. Understanding the developmental threshold temperature and effective accumulated temperature of P. americana can provide theoretical basis for standardized culture of P.americana. Under climate chamber, the threshold temperature and effective accumulated temperature for egg development of P. americana to were determined through effective accumulated temperature law. The threshold temperature was （15.8±0.71）°C, the effective accumulated temperature was 415.8±38.05 degree days. A model of the relationship between temperature and developmental rates was established.

[Evaluation of efficiency of traps baited with frequency trembler grid lamps and trap plants for control Heortia vitessoides infectwed in Aquilaria sinensis].

Heortia vitessoides has been a serious defoliating pest of Aquilaria sinensis forests in recent years.The adults displayed strong tropism to the frequency trembler grid lamps and the nectar source plants.The favorite nectar source plants of H.vitessoides adults as the trap plants and the frequency trembler grid lamps in the integrated management of H.vitessoides were studied in the adult eclosion period through both the laboratory and field.The results showed that Kuhnia rosmarnifolia and Santalum album plants showed strong attraction to the H.vitessoides adults, with significant differences among the different nectar source plants.K.rosmarnifolia and S.album as trap plants with board type of planting area to total planting area of 5%-10%, and the frequency trembler grid lamps trapped significantly more adults of H.vitessoides. These results suggested that the frequency trembler grid lamps and trap plants could play an important role in the integrated management of the pest H.vitessoides of A.sinensis.

[Residue degradation dynamics of spinetoram in wolfberry].

The safety interval period and residue dynamics of two main components (XDE-175-J and XDE-175-L) of spinetoram in wolfberry were measured. Field experiment design and sampling method were carried out according to the "Guideline on pesticide residue trials". The wolfberry samples were extracted with acetonitrile by ultrasonic, and analyzed by UPLC-MS/MS. The wolfberry was sprayed with 6% spinetoram suspension concentrate (SC) at recommended dosage (1 500 times) and doubling dosage (750 times) (one time) at fructescence of wolfberry. The half-lives of spinetoram residue under recommended dosage treatment was 3.65-4.25 d, and all the fresh and dried fruits conform to first order kinetics equation. The dissipation rate was over 95% in fresh and dried fruits at 14 d after application. In conclusion, spinetoram belongs to the easily degradable pesticide type.

[Dissipation dynamics of spirodiclofen in wolfberry fruit].

The dissipation of spirodiclofen in fresh fruit and dry fruit of wolfberry was detected in this study to provide a reference for its safe application.According to Pesticide Residue Test Criteria of China, the open-field experiment was conducted in Zhongning courty of Ningxia province, and the dissipation of spirodiclofen was studied by acetonitrile extraction and HPLC-MS/MS detection. The results showed that the half-lives of spirodiclofen in fresh wolfberry fruit and dry wolfberry fruit were 6.9-11.2 days and 8.5-10.4 days, respectively. Spirodiclofen belongs to the easily degradable pesticide type. According to the maximum residue limits (0.5 mg•kg⁻¹) of spirodiclofen of EU for wolfberry, after recommended dosage being sprayed for once, fresh wolfberry fruit was safe to eat after 5 days, and dry wolfberry fruit was safe to eat after 21 days.

[Preliminary studies on critical control point of traceability system in wolfberry].

As a traditional Chinese medicine, wolfberry (Lycium barbarum) has a long cultivation history and a good industrial development foundation. With the development of wolfberry production, the expansion of cultivation area and the increased attention of governments and consumers on food safety, the quality and safety requirement of wolfberry is higher demanded. The quality tracing and traceability system of production entire processes is the important technology tools to protect the wolfberry safety, and to maintain sustained and healthy development of the wolfberry industry. Thus, this article analyzed the wolfberry quality management from the actual situation, the safety hazard sources were discussed according to the HACCP (hazard analysis and critical control point) and GAP (good agricultural practice for Chinese crude drugs), and to provide a reference for the traceability system of wolfberry.

[Investigation of production status in major wolfberry producing areas of China and some suggestions].

To investigate the production status and the safety influence factors of wolfberry in China. We investigated the detailed factors which affect the quality safe of wolfberry in the periods of July-August 2013 and July-September 2009. The factors include fertilizing patterns, the used pesticide and preliminary process wolfberry. The factors were discussed according to the results of investigation, and suggestions were proposed for the management and production departments of wolfberry.

Female sex pheromone blends and male response of the legume pod borer, Maruca vitrata (Lepidoptera: Crambidae), in two populations of mainland China.

The legume pod borer, Maruca vitrata (Lepidoptera: Crambidae; syn. M. testulalis), is a serious pantropical insect pest of grain legumes. Comparative studies of M. vitrata female sex pheromone components in two different geographic populations in China, Wuhan and Huazhou, confirmed that (E,E)-10,12-hexadecadienal (E10,E12-16:Ald) and (E)-10-hexadecenal (E10-16:Ald) were present in variable ratios in all pheromone gland extracts of both populations. (E,E)-10,12-hexadecadienol (E10,E12-16:OH) was always detected in minor amounts using polar DB-WAX columns, but was never detected using medium-polar DB-17 columns for the two populations. E10-16:OH was not found in any of the M. vitrata sex pheromone gland extracts. The average ratios of E10-16:Ald, E10,E12-16:Ald, and E10,E12-16:OH in the pheromone gland extracts of populations from Wuhan and Huazhou were 79.5:100:12.1 and 10.3:100:0.7, respectively. Electrophysiological testing suggested that E10,E12-16:Ald elicited the highest male electroantennography (EAG) response, followed by E10,E12-16:OH and E10-16:Ald. Field-trapping tests with single synthetic sex pheromone lures showed that E10,E12-16:OH alone could not attract males, whereas E10,E12-16:Ald or E10-16:Ald alone attracted few males. Wuhan and Huazhou males were most attracted by lures containing E10-16:Ald + E10,E12-16:Ald + E10,E12-16:OH in doses of (80 + 100 + 10) microg and (10 + 100 + 10) microg, respectively, per vial. Males could discriminate between the blends that were most attractive to their own geographic population and those that were most attractive to the reference population. Our findings suggest that geographic variation exists in the sex pheromone system of M. vitrata in China. The results are discussed with regard to the mechanisms underlying the sex pheromone variation.

[Effects of bionic glue on experimental population dynamics of Paratrioza sinica and its natural enemies].

OBJECTIVE: To find out whether and how the newly invented technique-bionic glue affects the main pest of wolf berry-Paratrioza sinica and its natural enemies Tamarixia lyciumi and Chrysopa septempunctata. METHOD: Spraying bionic glue in field when wolf berry just geminated, investigated the adults and nymphs of P. sinica and it's natural enemies: adults of T. lyciumi and eggs of C. septempunctata. RESULT AND CONCLUSION: Bionic glue can significantly reduce the population number of P. sinica, but with little impacts on its natural enemies of T. lyciumi and C. septempunctata, and more experiments are need for further research.

[Screening for virulence strains of Metarhizium against Dorysthenes hydropicus pascoes].

OBJECTIVE: The aim of the present study was to screen the Metarhizium strains with high virulence against the larvae of Dorysthenes hydropicus, a serious pest of Citrus grandis. METHOD: Thirty six strains of Metarhiziums were isolated from the soil of C. grandis GAP base and collected from other institutions, and the pathogenicity of these strains against 1st instar larvae of D. hydropicus was detected at concentration of 1 x 10(8) conidia/g. The high violence strains against D. hydropicus were cultivated in sabouraud dextrose yeast medium at first, then transfer to rice grain. And the sporulations of these violent strains against D. hydropicus were detected. RESULT: Twenty-eight strains showed virulence against D. hydropicus by preliminary study, and 7 strains of them were collected for further study, 6 of the 7 showed high virulence, the highest cadaver rate was higher than 74%. The conidia production of strain 1 and strain 4 were 2.35 +/- 0. 25 (1 x 10(9) conidia/g), 2.21 +/- 0.27 (1 x 10(9) conidia/g), respectively, showed significantly higher than other strains. CONCLUSION: Strain 1 and strain 4 of the 36 Metarhiziums strains showed high virulence against D. hydropicus, and the highest sporulation ability, so they have a best application prospect.

Secondary Rhinaria Contribute to Major Sexual Dimorphism of Antennae in the Aphid Semiaphis heraclei (Takahashi).

Sexual generation is an important generation in the life cycle of host-alternating aphids, and its population size determines the intensity of the peak in the next spring. Although male trapping techniques based on olfactory stimuli have been successfully established in the field, the biological basis of olfactory perception in males is unclear. In this study, we compared the morphology of antennae and the types, sizes, numbers, and distribution of sensilla between males and sexual females in the host-alternating aphid Semiaphis heraclei (Hemiptera: Aphididae). We found that flagellum length differentiation contributed to the majority of the sexual dimorphism of antennae. Most sensillum types or subtypes, including trichoid sensilla subtype I, campaniform sensilla, and primary rhinaria subtypes I and II, were enlarged in males. In addition, males bore more trichoid sensilla subtype I than sexual females. In particular, secondary rhinaria were present in males only and could not be detected in sexual females. These results revealed the structural basis of male olfactory perception. Our findings provide insight into the mechanism underlying chemical communication between sexual aphids and could thus be useful for pest control.

[Analysis of volatile compounds of inflorescence by GC-MS from Cistanche deserticola].

OBJECTIVE: To study the volatile compounds from inflorescence of Cistanche deserticola and provide basis for its utilization and seed breeding. METHODS: The volatile compounds were collected by dynamic headspace adsorption and analyzed by gas chromatography-mass spectrometry (GC-MS). RESULTS: Forty volatile components were identified in inflorescence of Cistanche deserticola from squaring period to full-bloom period. The main components in buds of Cistanche deserticola were hydrocarbons and green leaf volatiles in squaring period. Some components were characteristic in buds and disappeared or decreased in flowers. The relative contents of some components gradually increased with the buds blooming. And some components only emerged in flowers of Cistanche deserticola. The higher content of esters and aromatics were found in flowers, which were significantly increased in comparison with the volatile compounds from buds. CONCLUSION: The volatile compounds from inflorescence of Cistanche deserticola were complex, consisting of various compositions and significantly different with buds blooming.

Morphology and distribution of sensilla on the antennae and ovipositor of the Hymenopteran parasitoid Pseudotorymus jaapiellae Yang et Chen.

Pseudotorymus jaapiellae is an important ectoparasitoid of the larvae of Gephyraulus lycantha, a serious gall-forming pest that devastates wolfberry, Lycium barbarum, in Northwest China. To provide requisite background for our ongoing research on the mechanisms of P. jaapiellae's host location and subsequent oviposition, we used scanning electron microscopy to describe the external morphologies and distributions of sensilla on their antennae and ovipositors. The geniculate antennae of both male and female P. jaapiellae were each composed of a scape with a basal radiculum, a pedicel, an anellum and a flagellum. We identified nine morphological sensilla types on the antennae of both sexes, including three sensilla trichodea (ST), one sensillum basiconicum (SB), two sensilla chaetica (SCh), one sensillum placodeum (SP), one sensillum coeloconicum (SCo), and one sensillum campaniformia (SCa). Females had significantly more ST I and SP than males had, but males had more ST III than did females. We observed six types of sensilla on the ovipositor, including three ST, one SB, and two SCa. ST II, ST IV and SB II were on the sheath, whereas ST V and the SCa were on the stylus. Finally, the possible biological functions of these sensilla were discussed according to their morphology and ultrastructure. These results provide an important basis for further study on chemical communication between P. jaapiellae and their host, and contribute to the development of a biological control program for G. lycantha, using the parasitoid, P. jaapiellae.

Peach volatile emission and attractiveness of different host plant volatiles blends to Cydia molesta in adjacent peach and pear orchards.

The oriental fruit moth (OFM), Cydia (= Grapholitha) molesta, is a highly damaging pest; peaches are its primary host, and pears serve as post-peach secondary hosts during the late season in China. We collected volatiles from detached peach shoots and fruits, and identified them with gas chromatography-mass spectrometry (GC-MS). Antennally active compounds were identified by gas chromatography-electroantennogram detection (GC-EAD), and these were further tested in the laboratory and field. We detected consistent electroantennographic activity was for ten compounds. Significantly more C. molesta females were caught with a mixture of female EAD-active compounds identified from the detached matured peach fruits (nonanal, butyl acetate, 3-methylbutyl acetate, hexyl acetate, (Z)-3-hexenyl acetate, linalool and farnesene) than other mixtures mimicking the volatile profile from detached matured fruits or shoots. We identified a new GC-EAD active mixture from intact peach shoots composed of nonanal, (Z)-3-hexenyl acetate, (E)-ß-ocimene, and 6-methyl-5-hepten-2-one. In the field test, the background odour of orchards could affect trap catches, and two peach-derived blends together with two previously known pear-derived blends were proven to be able to monitor the seasonal OFM population dispersal in adjacent orchards. These host plant blends will be effective for further designing candidate attractants for season-long C. molesta population dynamic monitoring.

Sample Preparation Method of Scanning and Transmission Electron Microscope for the Appendages of Woodboring Beetle.

This report described sample preparation methods that scanning and transmission electron microscope observations, demonstrated by preparing appendages of the woodboring beetle, Chlorophorus caragana Xie & Wang (2012), for both types of electron microscopy. The scanning electron microscopy (SEM) sample preparation protocol was based on sample chemical fixation, dehydration in a series of ethanol baths, drying, and sputter-coating. By adding Tween 20 (Polyoxyethylene sorbitan laurate) to the fixative and the wash solution, the insect body surface of woodboring beetle was washed more cleanly in SEM. This study's transmission electron microscopy (TEM) sample preparation involved a series of steps including fixation, ethanol dehydration, embedding in resin, positioning using fluorescence microscopy, sectioning, and staining. Fixative with Tween 20 enabled penetrate the insect body wall of woodboring beetle more easily than it would had been without Tween 20, and subsequently better fixed tissues and organs in the body, thus yielded clear transmission electron microscope observations of insect sensilla ultrastructures. The next step of this preparation was determining the positions of insect sensilla in the sample embedded in the resin block by using fluorescence microscopy to increase the precision of target sensilla positioning. This improved slicing accuracy.

[Effects of root extract of host plants on seed germination and haustorium formation of Cistanche deserticola]. / å¯„ä¸»æ¤ç‰©æ ¹æµ¸ææ¶²å¯¹è‚‰è‹è“‰ç§å­èŒå‘åŠå¸å™¨å½¢æˆçš„å½±å“.

We examined the effects of root extracts of Haloxylon ammodendron and Beta vulgaris in Chenopodiaceae extracted by water and ethanol on seed germination and haustorium formation of Cistanche deserticola by filter paper culture dish method. The results showed that only adding root extract had no effect on seed germination and haustorium formation of C. deserticola. The germination rate of C. deserticola seeds treated by adding 10 mg·kg-1 gibberellin to the root extracted by ethanol was not significantly different from that of the control (GA3), whereas those treated by adding gibberellin to the ethanol extract of two kinds of host root was increased by more than 10 times. The germination rate of C. deserticola seeds in the treatment with adding 1 mg·kg-1 fluridone (FL) to root extract was not significantly different from that in the control with only fluridone, while those in the treatment with B. vulgaris root water extraction was the highest (39.4%). Compared to the treatment of adding gibberellin to the root extract, the germination rate of C. deserticola seeds was only increased. When FL was added to the host root extract, the haustorium was formed on the germination tube, with the formation rate of the ethanol extraction group being the highest (16.2%). Seed germination rate of C. deserticola increased to 52.3% when GA3 and FL were added to the ethanol extract of H. ammodendron, but the formation rate of haustorium was not different from that of FL treatment. Only 6.7% of the seed formation haustorium in the control was significantly lower than that in FL treatment. There were differences in the position and shape of the haustorium of C. deserticola seeds under different treatments. The haustorium produced by adding the extract of the host root mostly appeared at the top of the bud tube, and many papillae raised into claws. The haustorium of FL treatment without adding the extract of the host root mostly appeared at the bottom or the top of the bud tube splitting. The results indicated that ethanol extraction and water extraction could extract the substances that could promote the formation of C. deserticola seeds haustorium from the host root, but did not affect seed germination. GA3 and FL could significantly improve the germination rate of C. deserticola seeds, but the formation of the haustorium was affected by some substances in the host root extract.

[Electrophysiological and behavioral responses of gall midge, Jaapiella sp. to volatiles of host plant Lycium barbarum]. / 枸杞红瘿蚊对寄主植物挥发物的电生理和行为反应.

Plant-derived volatiles are important for guiding herbivorous insects to their host plants. Jaapiella sp. (Diptera: Cecidomyiidae) is the most serious pest of Lycium barbarum. The young flower buds of L. barbarum are attractive to females of Jaapiella sp. for oviposition in the field. In this study, we compared response of Jaapiella sp. to volatiles from flower buds of L. barbarum at different stages, and clarified the crucial semiochemicals attracting Jaapiella sp. to lay eggs. Volatiles from young and mature flower buds were collected using SPME, and then identified and quantified by gas chromatography-mass spectrometry (GC-MS). Both electrophysiological and behavior experiments were conducted to measure the attraction of eight synthetic compounds to females and screen the crucial components based on gas chromatography-electroantennogram detection (GC-EAD) and Y-tube bioassays, respectively. Results showed that qualitative and quantitative diffe-rences between two odor profiles. Consistent electroantennographic response was observed for eight compounds from headspace collections, of which five synthetic compounds (3-carene, camphene, terpinolene, d-limonene, and (+)-pinene) were essential for significant preference or avoidance. Our results indicated that these compounds from L. barbarum flower buds could be effective candidates as oviposition attractant or repellents of Jaapiella sp., which would be helpful in the exploitation of new control agent to Jaapiella sp. in the field.