A transgenic maize event ZD12-6 expressing a Bacillus thuringiensis （Bt） fusion protein CrylAb/Cry2Aj and a modified 5-enolpyruvylshikimate-3-phosphate synthase （EPSPS） protein G10 was characterized and evaluated....A transgenic maize event ZD12-6 expressing a Bacillus thuringiensis （Bt） fusion protein CrylAb/Cry2Aj and a modified 5-enolpyruvylshikimate-3-phosphate synthase （EPSPS） protein G10 was characterized and evaluated. Southern blot analysis indicated that ZD12-6 is a single copy integration event. The insert site was determined to be at chromosome 1 by border sequence analysis. Expression analyses of Bt fusion protein CrylAb/Cry2Aj and the EPSPS protein G10 suggested that they are both expressed stably in different generations. Insect bioassays demonstrated that the transgenic plants are highly resistant to Asian corn borer （Ostnnia furnacalis）, cotton boll worm （Helicoverpa armigera）, and armyworm （Mythimna separata）. This study suggested that ZD12-6 has the potential to be developed into a commercial transgenic line.展开更多
With the rapid development of high-throughput sequencing technology and bioinformatics algorithms, great progress has been made in the field of noncoding RNA(ncRNA) in the last decade. RNA molecules have been regarded...With the rapid development of high-throughput sequencing technology and bioinformatics algorithms, great progress has been made in the field of noncoding RNA(ncRNA) in the last decade. RNA molecules have been regarded only as a messenger between DNA and protein for decades, but now they have new roles in the biological process as ncRNAs. A growing number of ncRNAs have been identified in insects from the RNA-Seq data of small RNA libraries or transcriptomes. ncRNAs have varied regulatory functions at the epigenetic, transcriptional, or post-transcriptional levels, and participate in almost all types of biological processes. Here, we review the research progress of four kinds of ncRNAs, including microRNA(miRNA), Piwiinteracting RNA(piRNA), circular RNA(circRNA), and long noncoding RNA(lncRNA) in insects. The discovery, biogenesis mechanisms, and regulatory functions of these ncRNAs are presented here to provide a comprehensive understanding of insect ncRNAs and to promote the application of ncRNAs in insect pest control.展开更多
The determination of the environmental fate of Bt insecticidal protein released by Bt rice plants in paddy soils is a key issue in its ecological risk assessment. In this study, the impacts of soil water content, pH, ...The determination of the environmental fate of Bt insecticidal protein released by Bt rice plants in paddy soils is a key issue in its ecological risk assessment. In this study, the impacts of soil water content, pH, and temperature on the degradation of CrylAb protein expressed in the leaves of Bt rice KMD2 were studied in the laboratory. Three types of paddy soils were used, i.e., blue clayey paddy soil, pale paddy soil on quaternary red soil, and marine-fluvigenic yellow loamy paddy soil. Ground powders of KMD2 leaf blades were mixed with each type of soil, and degradation dynamics of Cry lAb were measured using enzyme-linked immunosorbent assay （ELISA）. The degradation rate of CrylAb was high at the early experimental stage, but slowed down steadily at middle and later stages, which could be described by exponential equations, with the half-life period of degradation determined as 1.8-4.0 d. The soil water content, pH, and temperature could affect the degradation of CrylAb, but the effects of soil pH and temperature were relatively greater. In general, CrylAb degradations were slower under lower soil pH and temperature conditions, especially for marine-fluvigenic yellow loamy paddy soil.展开更多
Numerous Bt rice lines expressing Cry protein derived from Bacillus thuringiensis Berliner （Bt） have been developed since 1989. However, the potential risks posed by Bt rice on non-target organisms still remain deba...Numerous Bt rice lines expressing Cry protein derived from Bacillus thuringiensis Berliner （Bt） have been developed since 1989. However, the potential risks posed by Bt rice on non-target organisms still remain debate. The white-backed planthopper （WBPH）, Sogatella furcifera （Horváth）, is one of the most economically important insect pests of rice in Asian countries and also one of the main non-target herbivores of transgenic rice. In the current study, impacts of transgenic cry1Ab/vip3H＋epsps rice （G6H1） with both insect and herbicide resistance on WBPH were evaluated to ascertain whether this transgenic rice line had potential risks for this sap-sucking pest under laboratory and ifeld conditions. The laboratory results showed that no signiifcant difference in egg developmental duration, nymphal survival rate and female fecundity was found for WBPH between G6H1 and its non-transgenic isoline （XS110）. However, the development duration of nymphs was signiifcantly shorter and female longevity signiifcantly longer when WBPH fed on G6H1 by comparison with those on its control. To verify the results found in laboratory, a 3-yr ifeld trial was conducted to monitor WBPH population using both the vacuum-suction machine and beat plate methods. Although the seasonal density of WBPH nymphs and total density of nymphs and adults were not signiifcantly affected by transgenic rice regardless of the sampling methods, the seasonal density of WBPH adults in transgenic rice plots was slightly lower than that in the control when using the vacuum-suction machine. Based on these results both from laboratory and ifeld, it is clear that our tested transgenic rice line will not lead higher population of WBPH. However, long-term ifeld experiments to monitor the population dynamics of WPBH at large scale need to be conducted to conifrm the present conclusions in future.展开更多
The specificity of the double-stranded RNA(dsRNA) used in the RNA interference(RNAi) technique is crucial for the success of sequence-specific gene silencing. Currently, RNAi-mediated insect control is a trending rese...The specificity of the double-stranded RNA(dsRNA) used in the RNA interference(RNAi) technique is crucial for the success of sequence-specific gene silencing. Currently, RNAi-mediated insect control is a trending research topic.However, the off-target effects of the dsRNA in RNAi are a major concern. In this study, the ds Hvβ’COPI(coat protein complex I, β’ subunit)-treated and untreated transcriptomes of the 28-spotted potato lady beetle(Henosepilachna vigintioctopunctata) were compared to understand its off-target gene silencing effects. The RNA-seq results revealed that 63 and 44 differentially expressed genes(DEGs) were upregulated and downregulated, respectively, in the ds Hvβ’COPI treated group as compared with the control. Validation of the differential expressions of some selected DEGs via reverse transcription-quantitative PCR(RT-qPCR) analysis confirmed the reliability of the transcriptome analysis results. Further downstream analysis revealed that there were no genes homologous with Hvβ’COPI in H. vigintioctopunctata. Additionally,no genes with a >11 bp continuous match with ds Hvβ’COPI were found in the H. vigintioctopunctata transcriptome. Six genes(Hvcitron, Hvhelicase, Hvtransposase, Hvserine, Hvdynein, and Hv E3 ubiquitin) were selected to examine the offtarget activity of ds Hvβ’COPI based on their potential involvement in various H. vigintioctopunctata metabolic pathways.The severity of silencing these six off-target genes was evaluated by employing RNAi. The RNAi results confirmed the downregulation of the expression of all six genes, although there was no significant lethality. The findings of this study will be helpful in the risk analysis of future RNAi-mediated pest control experiments.展开更多
Insect resistance and glyphosate tolerance have been two of the most important traits in the genetic improvement of various crops. In this study, two Bacillus thuringiensis （Bt） insecticidal genes, CrylAc and Cryllg...Insect resistance and glyphosate tolerance have been two of the most important traits in the genetic improvement of various crops. In this study, two Bacillus thuringiensis （Bt） insecticidal genes, CrylAc and Cryllg, and a modified glyphosate-tolerant 5-enolpyruvylshikimate-3-phosphate synthase （EPSPS） gene （GIO） were combined into a single transferred DNA （T-DNA） fragment and introduced into rice by Agrobacterium-mediated transformation. A transgenic line with single-copy T-DNA insertion named GAI-14 was found to be highly resistant to striped stem borer and rice leaf roller, and tolerant to glyphosate. Analysis of T-DNA border sequence suggested that the transgenes were inserted at the chromosome 3 and appeared to have not interrupted any known or putative genes. A field trial observed no significant difference in the basic agronomic traits between GAI-14 and the recipient rice.展开更多
A recent breakthrough in agricultural biotechnology is the introduction of RNAi-mediated strategies in pest control.However, the off-target effects of RNAi pest control are still not fully understood. Here, we studied...A recent breakthrough in agricultural biotechnology is the introduction of RNAi-mediated strategies in pest control.However, the off-target effects of RNAi pest control are still not fully understood. Here, we studied the off-target effects of two insecticidal siRNAs in both target and non-target insects. The results revealed that off-target effects of insecticidal siRNAs occur widely in both target and non-target insects. We classified the expression-changed genes according to their homology to the siRNA-targeted gene, related KEGG pathways with the siRNA-targeted gene and continuous matches with siRNAs. Surprisingly, the unintended significant changes in gene expression levels did not strictly match with the number of contiguous nucleotides in the siRNAs. As expected, the expression of small portions of the homologous and KEGG-related genes were significantly changed. We calculated the Shannon entropy of the transcriptome profile of the insects after injecting them with insecticidal siRNAs. Though hundreds of genes were affected in their expression levels post siRNAtreatment, the Shannon entropy of the transcriptome remained unchanged, suggesting that the transcriptome expression was balanced. Our results provide evidence that siRNAs cross-reacted with individual genes in non-target species, but did not have significant effects on the integrity of the transcriptome profiles in either target or non-target species on a genomic scale. The metric we proposed can be used to estimate the off-target effects of insecticidal siRNAs, which might be useful for evaluating the safety of RNAi in pest control.展开更多
Innovations in sequencing technology and the development of bioinformatics have allowed for studies of the genomics of many rice pests.At present,draft genomes of rice pests including Nilaparvata lugens,Sogatella furc...Innovations in sequencing technology and the development of bioinformatics have allowed for studies of the genomics of many rice pests.At present,draft genomes of rice pests including Nilaparvata lugens,Sogatella furcifera,Laodelphax striatellus,Sesamia inferens,Chilo suppressalis,Scirpophaga incertulas.展开更多
Following the publication of Xu et al.(2022),an error was identified in Figure 1D.Specifically,the top left panel was inadvertently duplicated during figure preparation.To ensure the accuracy and integrity of our publ...Following the publication of Xu et al.(2022),an error was identified in Figure 1D.Specifically,the top left panel was inadvertently duplicated during figure preparation.To ensure the accuracy and integrity of our published work,we request the publication of a corrigendum with the corrected image.We apologize for this oversight and any confusion it may have caused.The amended figure is provided in the updated Supplementary Materials.展开更多
The domesticated silkworm(Bombyx mori)has evolved a highly efficient nitrogen utilization system to support silk production.The silk glands play a pleiotropic role in sequestering nitrogen resources for silk synthesis...The domesticated silkworm(Bombyx mori)has evolved a highly efficient nitrogen utilization system to support silk production.The silk glands play a pleiotropic role in sequestering nitrogen resources for silk synthesis,mitigating aminoacidemia by assimilating free amino acids,and reallocating nitrogen during metamorphosis through programmed cell death.However,the specific functions of nitrogen metabolism-related genes in this process remain unclear.Using CRISPR/Cas9-based gene editing,mutations were generated in glutamine synthetase(GS),glutamate synthetase(GOGAT),asparagine synthetase(AS),glutamate dehydrogenase(GDH)and glutamate oxaloacetate transaminase 1(GOT1).Disruption of GS,GOGAT,and AS consistently reduced silkworm cocoon and pupal weight and significantly down-regulated silk protein gene transcription,whereas GOT1 mutation had no such effect.GOGAT mutants exhibited abnormally enlarged silk glands,whereas GS and AS mutants showed delayed programmed cell death in the silk glands.In contrast,GOT1 mutants displayed normal silk gland morphology but were consistently smaller.Disruption of GS,GOGAT,and AS led to more extensive transcriptional changes,including altered expression of transcription factors in the silk glands,compared with GOT1 mutants.Both GS and GOGAT mutants exhibited up-regulation of AS and GDH,while only GOGAT mutants displayed elevated AS enzymatic activity,suggesting that GOGAT may compete with AS for glutamine in the silk glands to support silk protein synthesis.AS mutants showed significantly elevated GOT activity and up-regulation of several metabolic pathways,indicating that AS may functionally interact with GOT in regulating both silk gland development and programmed cell death during metamorphosis.展开更多
The cotton mealybug Phenacoccus solenopsis Tinsley(Hemiptera: Pseudococcidae), which is native to North America, emerged as a major invasive insect pest of multiple crops in Asia at the beginning of the 21 st century....The cotton mealybug Phenacoccus solenopsis Tinsley(Hemiptera: Pseudococcidae), which is native to North America, emerged as a major invasive insect pest of multiple crops in Asia at the beginning of the 21 st century. Considering the economic significance of this insect and its rapid worldwide spread, we examined the possible factors driving its invasions. In this paper, we summarize the life history traits of P. solenopsis conceivably related to population development in invaded regions; these traits include its use of diverse host plants, reproductive capacity and mode, adaptation to temperature, response to food shortage, and insecticidal resistance. Then, focusing on the multiple trophic interactions that may promote or hinder invasion, we review the mutualistic relationship of this mealybug with ants and predation and parasitism by natural enemies. Finally, we suggest topics for future research and provide our perspective on the biological invasions of this mealybug. We speculate that specific biological factors associated with this mealybug, particularly its wide host range, high reproductive potential, evolved changes in life history traits, and mutualism with ants have played important roles in its invasions, allowing this pest to become established and rapidly increase its population upon its introduction into new regions.展开更多
The ecological effects of plant-virus-vector interactions on invasion of alien plant viral vectors have been rarely investigated. We examined the transmission of Tomato yellow leaf cur/China virus （TYLCCNV） by the i...The ecological effects of plant-virus-vector interactions on invasion of alien plant viral vectors have been rarely investigated. We examined the transmission of Tomato yellow leaf cur/China virus （TYLCCNV） by the invasive Q biotype and the indigenous ZHJ2 biotype of the whitefly Bemisia tabaci, a plant viral vector, as well as the influence of TYLCCNV-infection of plants on the performance of the two whitefly biotypes. Both whitefly biotypes were able to acquire viruses from infected plants and retained them in their bodies, but were unable to transmit them to either tobacco or tomato plants. However, when the Q biotype fed on tobacco plants infected with TYLCCNV, its fecundity and longevity were increased by 7- and 1-fold, respectively, compared to those of the Q biotype fed on uninfected tobacco plants. When the ZHJ2 biotype fed on virus-infected plants, its fecundity and longevity were increased by only 2- and 0.5-fold, respectively. These data show that the Q biotype acquired higher beneficial effects from TYLCCNV-infection of tobacco plants than the ZHJ2 biotype. Thus, the Q biotype whitefly may have advantages in its invasion and displacement of the indigenous ZHJ2 biotype.展开更多
The brown planthopper, Nilaparvata lugens St?l, has become a major threat in tropical Asian and China since the rice green revolution of the 1960 s. Currently, insecticide application remains the primary choice for co...The brown planthopper, Nilaparvata lugens St?l, has become a major threat in tropical Asian and China since the rice green revolution of the 1960 s. Currently, insecticide application remains the primary choice for controlling this rice insect pest, but heavy use of insecticides poses dangerous risks to beneficial natural enemies and pollinators, and stimulates N. lugens reproductivity, and has caused a resurgence of the pest in the major rice-planting regions throughout Asia. Achieving the long-lasting goal of sustainable management of N. lugens requires understanding of the molecular basis of outbreaks of the pest and the development of environment-friendly pest-control strategies. Here, we review the recent molecular advances in N. lugens research on the aspects of its endosymbionts, virus transmission, insecticide resistance, and interaction between N. lugens and rice plants. We also put forward further research directions that may shed some lights on management of the rice pest.展开更多
Glyphosate-resistant crops have been a huge economic success for genetic engineering. The creating of new glypbosateresistant plants would increase the available choices for planting and lower the price of genetically...Glyphosate-resistant crops have been a huge economic success for genetic engineering. The creating of new glypbosateresistant plants would increase the available choices for planting and lower the price of genetically modified crop seeds. A novel G6 gene from Pseudomonas putida that encoded 5-enolpyruvylshikimate-3-phosphate synthase （EPSPS） was previously isolated. The G6 gene was transfected into rice via Agrobacterium-mediated transformation. The transgenic rice obtained was confirmed by PCR, Southern, and Western blots. The lab experiment and field trials further confirmed that the transgenic rice can survive glyphosate spraying at a dose of 8 g L^-1. In contrast, conventional rice was killed at a weed control glyphosate spray dose of 1 g L^-1. Altogether, the present study showed that the G6 gene works well in rice in vivo for glyphosate-resistance.展开更多
Although the biochemical dissection of parasitoid-host interactions is becoming well characterized, the molecular knowledge concerning them is minimal. In order to understand the molecular bases of the host immune res...Although the biochemical dissection of parasitoid-host interactions is becoming well characterized, the molecular knowledge concerning them is minimal. In order to understand the molecular bases of the host immune response to parasitoid attack, we explored the response of Papilio xuthus parasitized by the endoparasitic wasp Pteromalus puparum using proteomic approach. By examining the differential expression of plasma proteins in the parasitized and unparasitized host pupae by two-dimensional (2D) electrophoresis, 16 proteins were found to vary in relation to parasitization compared with unparasitized control samples. All of them were submitted to identification by mass spectrometry coupled with a database search. The modulated proteins were found to fall into the following functional groups: humoral or cellular immunity, detoxification, energy metabolism, and others. This study contributes insights into the molecular mechanism of the relationships between parasitoids and their host insects.展开更多
Chlorantraniliprole was introduced into China from 2008 as a novel insecticide to control the rice stem borer, Chilo suppressalis （Walker）. Cotesia chilonis （Matsumura） is the major parasitoid of C. suppressalis. ...Chlorantraniliprole was introduced into China from 2008 as a novel insecticide to control the rice stem borer, Chilo suppressalis （Walker）. Cotesia chilonis （Matsumura） is the major parasitoid of C. suppressalis. We collected seven populations of C. suppressalis and two populations of C. chilonis in different locations in China in 2009 to investigate the lethal effects of chlorantraniliprole on them. The populations of C. suppressalis from different regions showed various LD50 values ranging from 2.00 to 18.70 ng per larva. Chlorantraniliprole has negligible acute contact toxicity （LC50500 mg L-1） to C. chilonis and its oral toxicity is also much lower than that of fipronil （2 800-fold difference in LC50）. The results indicated that chlorantraniliprole is a good alterative in rice integrated pest management （IPM） programs. The susceptibility data of C. suppressalis will be useful for monitoring resistance levels in future.展开更多
Cold stress responses help insects to survive under low temperatures that would be lethal otherwise.This phenomenon might contribute to the invasion of some Bemisia tabaci cryptic species from subtropical areas to tem...Cold stress responses help insects to survive under low temperatures that would be lethal otherwise.This phenomenon might contribute to the invasion of some Bemisia tabaci cryptic species from subtropical areas to temperate regions.However,the molecular mechanisms regulating cold stress responses in whitefly are yet unclear.Mitogen-activated protein kinases（MAPKs）which including p38,ERK,and JNK,are well known for their roles in regulating metabolic responses to cold stress in many insects.In this study,we explored the possible roles of the MAPKs in response to low temperature stresses in the Mediterranean cryptic species（the Q-biotype）of the B.tabaci species complex.First,we cloned the p38 and ERK genes from the whitefly cDNA library.Next,we analyzed the activation of MAPKs during cold stress in the Mediterranean cryptic species by immuno-blotting.After cold stress,the level of phospho-p38 increased but no significant change was observed in the phosphorylation of ERK and JNK,thus suggesting that the p38 might be responsible for the defense response to low temperature stress.Furthermore,we demonstrated that：i）3 min chilling at 0°C was sufficient for the activation of p38 MAPK pathway in this whitefly;and ii）the amount of phosphorylated p38 increased significantly in the first 20 min of chilling,reversed by 60 min,and then returned to the original level by 120 min.Taken together,our results suggest that the p38 pathway is important during response to low temperature stress in the Mediterranean cryptic species of the B.tabaci species complex.展开更多
Human serum albumin(HSA) is widely utilized for medical purposes and biochemical research.Transgenic rice has proved to be an attractive bioreactor for mass production of recombinant HSA(rHSA).However,transgene spread...Human serum albumin(HSA) is widely utilized for medical purposes and biochemical research.Transgenic rice has proved to be an attractive bioreactor for mass production of recombinant HSA(rHSA).However,transgene spread is a major environmental and food safety concern for transgenic rice expressing proteins of medical value.This study aimed to develop a selectively terminable transgenic rice line expressing HSA in rice seeds,and a simple process for recovery and purification of rHSA for economical manufacture.An HSA expression cassette was inserted into a T-DNA vector encoding an RNA interference(RNAi) cassette suppressing the CYP81A6 gene.This gene detoxifies the herbicide bentazon and is linked to the 5-enolpyruvylshikimate-3-phosphate synthase(EPSPS) cassette which confers glyphosate tolerance.ANX Sepharose Fast Flow(ANX FF) anion exchange chromatography coupled with Butyl Sepharose High Performance(Butyl HP) hydrophobic interaction chromatography was used to purify rHSA.A transgenic rice line,HSA-84,was obtained with stable expression of rHSA of up to 0.72% of the total dry weight of the dehusked rice seeds.This line also demonstrated high sensitivity to bentazon,and thus could be killed selectively by a spray of bentazon.A two-step chromatography purification scheme was established to purify the rHSA from rice seeds to a purity of 99% with a recovery of 62.4%.Results from mass spectrometry and N-terminus sequencing suggested that the purified rHSA was identical to natural plasma-derived HSA.This study provides an alternative strategy for large-scale production of HSA with a built-in transgene safety control mechanism.展开更多
Plant flowering and breeding characteristics are important for us to understand the reproduction of plant populations. In this paper, we studied the reproduction characteristics of Jatropha curcas in Yuanjiang County ...Plant flowering and breeding characteristics are important for us to understand the reproduction of plant populations. In this paper, we studied the reproduction characteristics of Jatropha curcas in Yuanjiang County （23°36＇1＇4, 101°00＇E）, Yunnan Province. The plant produces flowers in dichasial inflorescences. Normally, the flowers are unisexual, and male and female flowers are produced in the same inflorescence. Only a few male flowers are produced in an inflorescence, and fruits are produced only through pollination between different flowers from the same or different plants. By the treatments of emasculation, bagging and artificial pollination in this experiment, there were few but same fruit set ratios when the inflorescences were emasculated, bagged, or bagged with net, except artificial pollination treatments, which showed that Jatropha curcas could produce fruit through apomixis but not wind pollination. When the inflorescences were unbagged, unemasculated and with free pollination treatments, or bagged, emasculated and with artificial cross-pollination treatments, or unbagged, emasculated and with free pollination treatments, there were many fruits produced. It showed that Jatropha curcas shows outcrossing, is self-compatible, and demanding for pollinators. Normally, the male flowers open first and a few flowers bloom in one day in a raceme. These flowers last a long time in bloom. However, a large number of female flowers open from the third to the fifth day, with some female flowers opening first in a few raceme. This shows a tendency to promote xenogamy and minimize geitonogamy.展开更多
The oriental armyworm,Mythimna separata(Lepidoptera:Noctuidae)is a harmful pest which causes heavy loss in agricultural crops.Sex identification of M.separata is very important for understanding the biology and manage...The oriental armyworm,Mythimna separata(Lepidoptera:Noctuidae)is a harmful pest which causes heavy loss in agricultural crops.Sex identification of M.separata is very important for understanding the biology and management of this pest.Development of fast and convenient methods to identify M.separata female and male is the necessary prerequisite for con ducting gender-related studies.In this study,new methods to differentiate sex of M.separata at pupal and adult stages were investigated.Our results showed that posterior abdominal segments containing genital organs which are morphological traits to differentiate female and male.Genital openings are important characters for the female and male pupae differentiation.For female pupa,it locates on the ventral side of eighth and ninth abdominal segments and appears as a visible longitudinal suture;for male pupa,it locates on the ventral side of the ninth abdominal segment and appears as a short slit surrounded by a pair of visible semicircular lumps.Bristle numbers are another indicator for sex differentiation of adults.Each female has three-bristled frenulum and each male has one bristled-frenulum near the base of hind wing of each side of moth body.We confirmed the two sex differentiation methods manually by dissecting genitalia of adults and found their genger was as same as what we examined the sex of pupae and adults using the morphological methods.展开更多
基金Project supported by the Fundamental Research Funds for the Central Universities(No.2017FZA6011)the National Key Transgenic Research Projects(No.2016ZX08010003)of China
文摘A transgenic maize event ZD12-6 expressing a Bacillus thuringiensis （Bt） fusion protein CrylAb/Cry2Aj and a modified 5-enolpyruvylshikimate-3-phosphate synthase （EPSPS） protein G10 was characterized and evaluated. Southern blot analysis indicated that ZD12-6 is a single copy integration event. The insert site was determined to be at chromosome 1 by border sequence analysis. Expression analyses of Bt fusion protein CrylAb/Cry2Aj and the EPSPS protein G10 suggested that they are both expressed stably in different generations. Insect bioassays demonstrated that the transgenic plants are highly resistant to Asian corn borer （Ostnnia furnacalis）, cotton boll worm （Helicoverpa armigera）, and armyworm （Mythimna separata）. This study suggested that ZD12-6 has the potential to be developed into a commercial transgenic line.
基金supported by the National Key Research and Development Program(2017YFD0200900,2017YFD0200904,2017YFC1200602 and 2016YFC1200600)the National Natural Science Foundation of China(31772238,31760514 and 31701785)
文摘With the rapid development of high-throughput sequencing technology and bioinformatics algorithms, great progress has been made in the field of noncoding RNA(ncRNA) in the last decade. RNA molecules have been regarded only as a messenger between DNA and protein for decades, but now they have new roles in the biological process as ncRNAs. A growing number of ncRNAs have been identified in insects from the RNA-Seq data of small RNA libraries or transcriptomes. ncRNAs have varied regulatory functions at the epigenetic, transcriptional, or post-transcriptional levels, and participate in almost all types of biological processes. Here, we review the research progress of four kinds of ncRNAs, including microRNA(miRNA), Piwiinteracting RNA(piRNA), circular RNA(circRNA), and long noncoding RNA(lncRNA) in insects. The discovery, biogenesis mechanisms, and regulatory functions of these ncRNAs are presented here to provide a comprehensive understanding of insect ncRNAs and to promote the application of ncRNAs in insect pest control.
基金This study was financially supported by the National Natural Science Foundation of China （30070500） to Cheng Jiaan.
文摘The determination of the environmental fate of Bt insecticidal protein released by Bt rice plants in paddy soils is a key issue in its ecological risk assessment. In this study, the impacts of soil water content, pH, and temperature on the degradation of CrylAb protein expressed in the leaves of Bt rice KMD2 were studied in the laboratory. Three types of paddy soils were used, i.e., blue clayey paddy soil, pale paddy soil on quaternary red soil, and marine-fluvigenic yellow loamy paddy soil. Ground powders of KMD2 leaf blades were mixed with each type of soil, and degradation dynamics of Cry lAb were measured using enzyme-linked immunosorbent assay （ELISA）. The degradation rate of CrylAb was high at the early experimental stage, but slowed down steadily at middle and later stages, which could be described by exponential equations, with the half-life period of degradation determined as 1.8-4.0 d. The soil water content, pH, and temperature could affect the degradation of CrylAb, but the effects of soil pH and temperature were relatively greater. In general, CrylAb degradations were slower under lower soil pH and temperature conditions, especially for marine-fluvigenic yellow loamy paddy soil.
基金provided by the Special Research Projects for Developing Transgenic Plants, China (2013ZX08011-001)China National Science Fund for Innovative Research Group of Biological Control (31021003)the National Basic Research Progarm of China (973 Program, 2007CB109202)
文摘Numerous Bt rice lines expressing Cry protein derived from Bacillus thuringiensis Berliner （Bt） have been developed since 1989. However, the potential risks posed by Bt rice on non-target organisms still remain debate. The white-backed planthopper （WBPH）, Sogatella furcifera （Horváth）, is one of the most economically important insect pests of rice in Asian countries and also one of the main non-target herbivores of transgenic rice. In the current study, impacts of transgenic cry1Ab/vip3H＋epsps rice （G6H1） with both insect and herbicide resistance on WBPH were evaluated to ascertain whether this transgenic rice line had potential risks for this sap-sucking pest under laboratory and ifeld conditions. The laboratory results showed that no signiifcant difference in egg developmental duration, nymphal survival rate and female fecundity was found for WBPH between G6H1 and its non-transgenic isoline （XS110）. However, the development duration of nymphs was signiifcantly shorter and female longevity signiifcantly longer when WBPH fed on G6H1 by comparison with those on its control. To verify the results found in laboratory, a 3-yr ifeld trial was conducted to monitor WBPH population using both the vacuum-suction machine and beat plate methods. Although the seasonal density of WBPH nymphs and total density of nymphs and adults were not signiifcantly affected by transgenic rice regardless of the sampling methods, the seasonal density of WBPH adults in transgenic rice plots was slightly lower than that in the control when using the vacuum-suction machine. Based on these results both from laboratory and ifeld, it is clear that our tested transgenic rice line will not lead higher population of WBPH. However, long-term ifeld experiments to monitor the population dynamics of WPBH at large scale need to be conducted to conifrm the present conclusions in future.
基金supported by the National Natural Science Foundation of China(31972269 and 32172500)the National Key R&D Program of China(2017YFD0200900)。
文摘The specificity of the double-stranded RNA(dsRNA) used in the RNA interference(RNAi) technique is crucial for the success of sequence-specific gene silencing. Currently, RNAi-mediated insect control is a trending research topic.However, the off-target effects of the dsRNA in RNAi are a major concern. In this study, the ds Hvβ’COPI(coat protein complex I, β’ subunit)-treated and untreated transcriptomes of the 28-spotted potato lady beetle(Henosepilachna vigintioctopunctata) were compared to understand its off-target gene silencing effects. The RNA-seq results revealed that 63 and 44 differentially expressed genes(DEGs) were upregulated and downregulated, respectively, in the ds Hvβ’COPI treated group as compared with the control. Validation of the differential expressions of some selected DEGs via reverse transcription-quantitative PCR(RT-qPCR) analysis confirmed the reliability of the transcriptome analysis results. Further downstream analysis revealed that there were no genes homologous with Hvβ’COPI in H. vigintioctopunctata. Additionally,no genes with a >11 bp continuous match with ds Hvβ’COPI were found in the H. vigintioctopunctata transcriptome. Six genes(Hvcitron, Hvhelicase, Hvtransposase, Hvserine, Hvdynein, and Hv E3 ubiquitin) were selected to examine the offtarget activity of ds Hvβ’COPI based on their potential involvement in various H. vigintioctopunctata metabolic pathways.The severity of silencing these six off-target genes was evaluated by employing RNAi. The RNAi results confirmed the downregulation of the expression of all six genes, although there was no significant lethality. The findings of this study will be helpful in the risk analysis of future RNAi-mediated pest control experiments.
基金supported by the National Natural Science Foundation of China(No.31321063)
文摘Insect resistance and glyphosate tolerance have been two of the most important traits in the genetic improvement of various crops. In this study, two Bacillus thuringiensis （Bt） insecticidal genes, CrylAc and Cryllg, and a modified glyphosate-tolerant 5-enolpyruvylshikimate-3-phosphate synthase （EPSPS） gene （GIO） were combined into a single transferred DNA （T-DNA） fragment and introduced into rice by Agrobacterium-mediated transformation. A transgenic line with single-copy T-DNA insertion named GAI-14 was found to be highly resistant to striped stem borer and rice leaf roller, and tolerant to glyphosate. Analysis of T-DNA border sequence suggested that the transgenes were inserted at the chromosome 3 and appeared to have not interrupted any known or putative genes. A field trial observed no significant difference in the basic agronomic traits between GAI-14 and the recipient rice.
基金funded by grants from the National Science and Technology Major Project of China(2016ZX08011002)。
文摘A recent breakthrough in agricultural biotechnology is the introduction of RNAi-mediated strategies in pest control.However, the off-target effects of RNAi pest control are still not fully understood. Here, we studied the off-target effects of two insecticidal siRNAs in both target and non-target insects. The results revealed that off-target effects of insecticidal siRNAs occur widely in both target and non-target insects. We classified the expression-changed genes according to their homology to the siRNA-targeted gene, related KEGG pathways with the siRNA-targeted gene and continuous matches with siRNAs. Surprisingly, the unintended significant changes in gene expression levels did not strictly match with the number of contiguous nucleotides in the siRNAs. As expected, the expression of small portions of the homologous and KEGG-related genes were significantly changed. We calculated the Shannon entropy of the transcriptome profile of the insects after injecting them with insecticidal siRNAs. Though hundreds of genes were affected in their expression levels post siRNAtreatment, the Shannon entropy of the transcriptome remained unchanged, suggesting that the transcriptome expression was balanced. Our results provide evidence that siRNAs cross-reacted with individual genes in non-target species, but did not have significant effects on the integrity of the transcriptome profiles in either target or non-target species on a genomic scale. The metric we proposed can be used to estimate the off-target effects of insecticidal siRNAs, which might be useful for evaluating the safety of RNAi in pest control.
基金supported by the Zhejiang Provincial Key Research and Development Plan, China(Grant Nos.2020C02001 and 2022C02034)the National Natural Science Foundation of China(Grant No.31672022)。
文摘Innovations in sequencing technology and the development of bioinformatics have allowed for studies of the genomics of many rice pests.At present,draft genomes of rice pests including Nilaparvata lugens,Sogatella furcifera,Laodelphax striatellus,Sesamia inferens,Chilo suppressalis,Scirpophaga incertulas.
文摘Following the publication of Xu et al.(2022),an error was identified in Figure 1D.Specifically,the top left panel was inadvertently duplicated during figure preparation.To ensure the accuracy and integrity of our published work,we request the publication of a corrigendum with the corrected image.We apologize for this oversight and any confusion it may have caused.The amended figure is provided in the updated Supplementary Materials.
基金supported by the National Natural Science Foundation of China(32270458,32070411,2023A1515010657)State Key Laboratory of Resource Insects(SKLSGB-ORP202209)。
文摘The domesticated silkworm(Bombyx mori)has evolved a highly efficient nitrogen utilization system to support silk production.The silk glands play a pleiotropic role in sequestering nitrogen resources for silk synthesis,mitigating aminoacidemia by assimilating free amino acids,and reallocating nitrogen during metamorphosis through programmed cell death.However,the specific functions of nitrogen metabolism-related genes in this process remain unclear.Using CRISPR/Cas9-based gene editing,mutations were generated in glutamine synthetase(GS),glutamate synthetase(GOGAT),asparagine synthetase(AS),glutamate dehydrogenase(GDH)and glutamate oxaloacetate transaminase 1(GOT1).Disruption of GS,GOGAT,and AS consistently reduced silkworm cocoon and pupal weight and significantly down-regulated silk protein gene transcription,whereas GOT1 mutation had no such effect.GOGAT mutants exhibited abnormally enlarged silk glands,whereas GS and AS mutants showed delayed programmed cell death in the silk glands.In contrast,GOT1 mutants displayed normal silk gland morphology but were consistently smaller.Disruption of GS,GOGAT,and AS led to more extensive transcriptional changes,including altered expression of transcription factors in the silk glands,compared with GOT1 mutants.Both GS and GOGAT mutants exhibited up-regulation of AS and GDH,while only GOGAT mutants displayed elevated AS enzymatic activity,suggesting that GOGAT may compete with AS for glutamine in the silk glands to support silk protein synthesis.AS mutants showed significantly elevated GOT activity and up-regulation of several metabolic pathways,indicating that AS may functionally interact with GOT in regulating both silk gland development and programmed cell death during metamorphosis.
基金financially supported by the National Key Research and Development Program of China(2016YFC1200600)
文摘The cotton mealybug Phenacoccus solenopsis Tinsley(Hemiptera: Pseudococcidae), which is native to North America, emerged as a major invasive insect pest of multiple crops in Asia at the beginning of the 21 st century. Considering the economic significance of this insect and its rapid worldwide spread, we examined the possible factors driving its invasions. In this paper, we summarize the life history traits of P. solenopsis conceivably related to population development in invaded regions; these traits include its use of diverse host plants, reproductive capacity and mode, adaptation to temperature, response to food shortage, and insecticidal resistance. Then, focusing on the multiple trophic interactions that may promote or hinder invasion, we review the mutualistic relationship of this mealybug with ants and predation and parasitism by natural enemies. Finally, we suggest topics for future research and provide our perspective on the biological invasions of this mealybug. We speculate that specific biological factors associated with this mealybug, particularly its wide host range, high reproductive potential, evolved changes in life history traits, and mutualism with ants have played important roles in its invasions, allowing this pest to become established and rapidly increase its population upon its introduction into new regions.
基金supported by the National Natural Science Foundation of China (No.30730061)the National Basic Research Program (973) of China (No.2009CB119203)the Zhejiang Provincial Key Agricultural Project (No.2007C12045),China
文摘The ecological effects of plant-virus-vector interactions on invasion of alien plant viral vectors have been rarely investigated. We examined the transmission of Tomato yellow leaf cur/China virus （TYLCCNV） by the invasive Q biotype and the indigenous ZHJ2 biotype of the whitefly Bemisia tabaci, a plant viral vector, as well as the influence of TYLCCNV-infection of plants on the performance of the two whitefly biotypes. Both whitefly biotypes were able to acquire viruses from infected plants and retained them in their bodies, but were unable to transmit them to either tobacco or tomato plants. However, when the Q biotype fed on tobacco plants infected with TYLCCNV, its fecundity and longevity were increased by 7- and 1-fold, respectively, compared to those of the Q biotype fed on uninfected tobacco plants. When the ZHJ2 biotype fed on virus-infected plants, its fecundity and longevity were increased by only 2- and 0.5-fold, respectively. These data show that the Q biotype acquired higher beneficial effects from TYLCCNV-infection of tobacco plants than the ZHJ2 biotype. Thus, the Q biotype whitefly may have advantages in its invasion and displacement of the indigenous ZHJ2 biotype.
基金supported by the National Natural Science Foundation of China(31672025,31471765 and 31630057)
文摘The brown planthopper, Nilaparvata lugens St?l, has become a major threat in tropical Asian and China since the rice green revolution of the 1960 s. Currently, insecticide application remains the primary choice for controlling this rice insect pest, but heavy use of insecticides poses dangerous risks to beneficial natural enemies and pollinators, and stimulates N. lugens reproductivity, and has caused a resurgence of the pest in the major rice-planting regions throughout Asia. Achieving the long-lasting goal of sustainable management of N. lugens requires understanding of the molecular basis of outbreaks of the pest and the development of environment-friendly pest-control strategies. Here, we review the recent molecular advances in N. lugens research on the aspects of its endosymbionts, virus transmission, insecticide resistance, and interaction between N. lugens and rice plants. We also put forward further research directions that may shed some lights on management of the rice pest.
文摘Glyphosate-resistant crops have been a huge economic success for genetic engineering. The creating of new glypbosateresistant plants would increase the available choices for planting and lower the price of genetically modified crop seeds. A novel G6 gene from Pseudomonas putida that encoded 5-enolpyruvylshikimate-3-phosphate synthase （EPSPS） was previously isolated. The G6 gene was transfected into rice via Agrobacterium-mediated transformation. The transgenic rice obtained was confirmed by PCR, Southern, and Western blots. The lab experiment and field trials further confirmed that the transgenic rice can survive glyphosate spraying at a dose of 8 g L^-1. In contrast, conventional rice was killed at a weed control glyphosate spray dose of 1 g L^-1. Altogether, the present study showed that the G6 gene works well in rice in vivo for glyphosate-resistance.
基金Project supported by the National Basic Research Program (973) of China (No. 2006CB102005)the National Natural Science Foundation of China (Nos. 30571251 and 30170626)+1 种基金the Program for New Cen-tury Excellent Talents in University of the Ministry of Education of China (No. NCET-05-0513)the Innovation Research Team Program of the Ministry of Education of China (No. IRT0535)
文摘Although the biochemical dissection of parasitoid-host interactions is becoming well characterized, the molecular knowledge concerning them is minimal. In order to understand the molecular bases of the host immune response to parasitoid attack, we explored the response of Papilio xuthus parasitized by the endoparasitic wasp Pteromalus puparum using proteomic approach. By examining the differential expression of plasma proteins in the parasitized and unparasitized host pupae by two-dimensional (2D) electrophoresis, 16 proteins were found to vary in relation to parasitization compared with unparasitized control samples. All of them were submitted to identification by mass spectrometry coupled with a database search. The modulated proteins were found to fall into the following functional groups: humoral or cellular immunity, detoxification, energy metabolism, and others. This study contributes insights into the molecular mechanism of the relationships between parasitoids and their host insects.
基金supports from the National Basic Research Program of China (973 Program, 2007CB109202)the Special Agricultural Research Projects for Public Welfare, China (200803004)
文摘Chlorantraniliprole was introduced into China from 2008 as a novel insecticide to control the rice stem borer, Chilo suppressalis （Walker）. Cotesia chilonis （Matsumura） is the major parasitoid of C. suppressalis. We collected seven populations of C. suppressalis and two populations of C. chilonis in different locations in China in 2009 to investigate the lethal effects of chlorantraniliprole on them. The populations of C. suppressalis from different regions showed various LD50 values ranging from 2.00 to 18.70 ng per larva. Chlorantraniliprole has negligible acute contact toxicity （LC50500 mg L-1） to C. chilonis and its oral toxicity is also much lower than that of fipronil （2 800-fold difference in LC50）. The results indicated that chlorantraniliprole is a good alterative in rice integrated pest management （IPM） programs. The susceptibility data of C. suppressalis will be useful for monitoring resistance levels in future.
基金supported by the National Natural Science Foundation of China(30730061)the National Basic Research Program of China(2009CB119203)
文摘Cold stress responses help insects to survive under low temperatures that would be lethal otherwise.This phenomenon might contribute to the invasion of some Bemisia tabaci cryptic species from subtropical areas to temperate regions.However,the molecular mechanisms regulating cold stress responses in whitefly are yet unclear.Mitogen-activated protein kinases（MAPKs）which including p38,ERK,and JNK,are well known for their roles in regulating metabolic responses to cold stress in many insects.In this study,we explored the possible roles of the MAPKs in response to low temperature stresses in the Mediterranean cryptic species（the Q-biotype）of the B.tabaci species complex.First,we cloned the p38 and ERK genes from the whitefly cDNA library.Next,we analyzed the activation of MAPKs during cold stress in the Mediterranean cryptic species by immuno-blotting.After cold stress,the level of phospho-p38 increased but no significant change was observed in the phosphorylation of ERK and JNK,thus suggesting that the p38 might be responsible for the defense response to low temperature stress.Furthermore,we demonstrated that：i）3 min chilling at 0°C was sufficient for the activation of p38 MAPK pathway in this whitefly;and ii）the amount of phosphorylated p38 increased significantly in the first 20 min of chilling,reversed by 60 min,and then returned to the original level by 120 min.Taken together,our results suggest that the p38 pathway is important during response to low temperature stress in the Mediterranean cryptic species of the B.tabaci species complex.
基金Project(No.2011ZX08010-003)supported by the Ministry of Agriculture of China
文摘Human serum albumin(HSA) is widely utilized for medical purposes and biochemical research.Transgenic rice has proved to be an attractive bioreactor for mass production of recombinant HSA(rHSA).However,transgene spread is a major environmental and food safety concern for transgenic rice expressing proteins of medical value.This study aimed to develop a selectively terminable transgenic rice line expressing HSA in rice seeds,and a simple process for recovery and purification of rHSA for economical manufacture.An HSA expression cassette was inserted into a T-DNA vector encoding an RNA interference(RNAi) cassette suppressing the CYP81A6 gene.This gene detoxifies the herbicide bentazon and is linked to the 5-enolpyruvylshikimate-3-phosphate synthase(EPSPS) cassette which confers glyphosate tolerance.ANX Sepharose Fast Flow(ANX FF) anion exchange chromatography coupled with Butyl Sepharose High Performance(Butyl HP) hydrophobic interaction chromatography was used to purify rHSA.A transgenic rice line,HSA-84,was obtained with stable expression of rHSA of up to 0.72% of the total dry weight of the dehusked rice seeds.This line also demonstrated high sensitivity to bentazon,and thus could be killed selectively by a spray of bentazon.A two-step chromatography purification scheme was established to purify the rHSA from rice seeds to a purity of 99% with a recovery of 62.4%.Results from mass spectrometry and N-terminus sequencing suggested that the purified rHSA was identical to natural plasma-derived HSA.This study provides an alternative strategy for large-scale production of HSA with a built-in transgene safety control mechanism.
文摘Plant flowering and breeding characteristics are important for us to understand the reproduction of plant populations. In this paper, we studied the reproduction characteristics of Jatropha curcas in Yuanjiang County （23°36＇1＇4, 101°00＇E）, Yunnan Province. The plant produces flowers in dichasial inflorescences. Normally, the flowers are unisexual, and male and female flowers are produced in the same inflorescence. Only a few male flowers are produced in an inflorescence, and fruits are produced only through pollination between different flowers from the same or different plants. By the treatments of emasculation, bagging and artificial pollination in this experiment, there were few but same fruit set ratios when the inflorescences were emasculated, bagged, or bagged with net, except artificial pollination treatments, which showed that Jatropha curcas could produce fruit through apomixis but not wind pollination. When the inflorescences were unbagged, unemasculated and with free pollination treatments, or bagged, emasculated and with artificial cross-pollination treatments, or unbagged, emasculated and with free pollination treatments, there were many fruits produced. It showed that Jatropha curcas shows outcrossing, is self-compatible, and demanding for pollinators. Normally, the male flowers open first and a few flowers bloom in one day in a raceme. These flowers last a long time in bloom. However, a large number of female flowers open from the third to the fifth day, with some female flowers opening first in a few raceme. This shows a tendency to promote xenogamy and minimize geitonogamy.
基金This research was financially supported by the National Natural Science Foundation of China(31601691 and 31272089).The authors thank Dr.Xue Shuang(Anyang In stitute of Tech no logy,Henan Province,China)for technical assistanee in dissecting genitalia.
文摘The oriental armyworm,Mythimna separata(Lepidoptera:Noctuidae)is a harmful pest which causes heavy loss in agricultural crops.Sex identification of M.separata is very important for understanding the biology and management of this pest.Development of fast and convenient methods to identify M.separata female and male is the necessary prerequisite for con ducting gender-related studies.In this study,new methods to differentiate sex of M.separata at pupal and adult stages were investigated.Our results showed that posterior abdominal segments containing genital organs which are morphological traits to differentiate female and male.Genital openings are important characters for the female and male pupae differentiation.For female pupa,it locates on the ventral side of eighth and ninth abdominal segments and appears as a visible longitudinal suture;for male pupa,it locates on the ventral side of the ninth abdominal segment and appears as a short slit surrounded by a pair of visible semicircular lumps.Bristle numbers are another indicator for sex differentiation of adults.Each female has three-bristled frenulum and each male has one bristled-frenulum near the base of hind wing of each side of moth body.We confirmed the two sex differentiation methods manually by dissecting genitalia of adults and found their genger was as same as what we examined the sex of pupae and adults using the morphological methods.
