Apolygus lucorum（Meyer-Dür.） and Erythroneura apicalis（Nawa） are important pests that affect the quality and the yield of grapevine and cause huge economic losses. This paper focuses on the selection of effec...Apolygus lucorum（Meyer-Dür.） and Erythroneura apicalis（Nawa） are important pests that affect the quality and the yield of grapevine and cause huge economic losses. This paper focuses on the selection of effective botanical pesticides to control A. lucorum and E. apicalis. This experiment explores the effect of several botanical pesticides for A. lucorum and E. apicalis, including the 0.5% veratrine, the0.6% Oxygen·Lactone agent, the 5% natural pyrethrin, the composite neem pesticide, the rotenone and the composite nicotine. The 0.5% veratrine shows a stable control efficacy, which is higher than 60% in Chengdu, while the composite nicotine shows the highest efficacy against A. lucorum, which is above 70%. In Yinchuan,the 0.5% veratrine shows the highest efficacy, against the first generation adults and the second generation larvae of E. apicalis, while the 5% natural pyrethrin shows 100% control efficacy against E. apicalis in Nanjiang. The 0.5% veratrine and the composite neem could be used as effective pesticides to control A. lucorum and the 5% natural pyrethrin can be used to control E. apicalis. They could be widely used in the production of pollution-free grapes.展开更多
[Objective] The aim was to analyze effect of climate change on occurrence degree of cotton Lugus lucorum in coastal areas of Jiangsu Province, in order to provide authentic references for better prevention. [Method] S...[Objective] The aim was to analyze effect of climate change on occurrence degree of cotton Lugus lucorum in coastal areas of Jiangsu Province, in order to provide authentic references for better prevention. [Method] Statistical analysis was conducted on Information about occurrence degree of cotton Lugus lucorum in Binhai County in Jiangsu Province from 1985 to 2007 under guidance of Integral regression method. [Result] Because of climate change, cotton Lugus lucorum occurrence de- grees differed in different generations and growth stages in Binhai County. Tempera- ture and humidity constituted the main climate factors. In general, climate change hindered occurrence degree from increasing. We could make use of the period when occurrence degree of cotton Lugus lucorum is prone to significant increase and choose some periods, such as early April, late June and middle August, to improve prevention effect. In addition, attention should be paid to comprehensive measures, such as adjustment of industrial structure, which will reduce pesticide and minimize cotton Lugus lucorum damage and adverse impact from prevention measures. [Conclusion] The results provided authentic references for cotton Lugus lucorum preven- tion in Jiangsu coastal areas.展开更多
Artemisia annua is an important preferred host of the mirid bug Apolygus lucorum in autumn.Volatiles emitted from A.annua attract A.Iucorum.Volatile artemisinic acid of A.annua is a precursor of artemisinin that has b...Artemisia annua is an important preferred host of the mirid bug Apolygus lucorum in autumn.Volatiles emitted from A.annua attract A.Iucorum.Volatile artemisinic acid of A.annua is a precursor of artemisinin that has been widely investigated in the Chinese herbal medicine field.However,little is known at this point about the biological roles of artemisinic acid in regulating the behavioral trends of A.lucorum.In this study,we collected volatiles from A.annua at the seedling stage by using headspace solid phase microextraction(HS-SPME).Gas chromatography-mass spectrometry(GC-MS) analysis showed that approximately 11.03±6.00 and 238.25±121.67 ng hartemisinic acid were detected in volatile samples and milled samples,respectively.Subsequently,a key gene for artemisinic acid synthesis,the cytochrome P450 gene cyp71 av1,was expressed in engineered Saccharomyces cerevisiae to catalyze the production of artemisinic acid.After the addition of exogenous artemisinic alcohol or artemisinic aldehyde,artemisinic acid was identified as the product of the expressed gene.In electroantennogram(EAG) recordings,3-day-old adult A.lucorum showed significant electrophysiological responses to artemisinic alcohol,artemisinic aldehyde and artemisinic acid.Furthermore,3-day-old female bugs were significantly attracted by artemisinic acid and artemisinic alcohol at a concentration of 10 mmol L,whereas 3-day-old male bugs were attracted significantly by 10 mmol Lartemisinic acid and artemisinic aldehyde.We propose that artemisinic acid and its precursors could be used as potential attractant components for the design of novel integrated pest management strategies to control A.lucorum.展开更多
Background Apolygus lucorum is a worldwide omnivorous pest damaging a range of crops and causing great economic losses.Symbiotic bacteria living in insects play a key role in the nutrition,physiology,and behavior of h...Background Apolygus lucorum is a worldwide omnivorous pest damaging a range of crops and causing great economic losses.Symbiotic bacteria living in insects play a key role in the nutrition,physiology,and behavior of hosts.Here,we present an experiment using Illumina HiSeq sequencing targeting the V3–V4 regions of bacteria’s 16S rRNA throughout the entire life cycle of A.lucorum.Results The first and second instar nymphs have the largest alpha diversity compared with other life stages of the insect.Bacterial phyla Proteobacteria(72.29%),Firmicutes(15.24%),Actinobacteria(7.76%)exhibit the largest relative abundance in all developmental stages.Erwinia(23.97%)and Lactococcus(10.62%)are the two genera with the high-est relative abundance.The relative abundance of Erwinia in the nymph stage is significantly greater than the adult stage,and the relative abundance of Lactococcus in 6-day-old and 9-day-old adult females is higher compared with adult males.Conclusions These results reveal that microbial community composition and relative abundance shift dynamically at different life stages,implying that different bacterial phyla and genera may have specific roles in specific life stages such as metabolism,nutrition absorption,detoxification,and reproduction.This study reveals for the first time the community composition and ecological dynamics of symbiotic bacteria throughout the life stages of A.lucorum,and thus may provide insight to new strategies for pest control.展开更多
Plants reshape their transcriptomes, proteomes and metabolomes in response to insect damage. In this study, we used suppression subtractive hybridization to investigate the transcriptomes of two cotton varieties （CCR...Plants reshape their transcriptomes, proteomes and metabolomes in response to insect damage. In this study, we used suppression subtractive hybridization to investigate the transcriptomes of two cotton varieties （CCRI41 and CCRI23） under Apolygus lucorum damage. From the CCRI23 libraries we obtained 92 transcripts and from the CCRI41 libraries we obtained 96 transcripts. 26 and 63 of the transcripts from CCRI23 and CCRI41, respectively, had known functions. Using reverse transcription PCR, we detected expression proifle of genes with known functions. Ultimately, we identiifed eight signiifcantly regulated genes, including one downregulated and four upregulated genes from the CCRI41 libraries, and one downregulated and two upregulated genes from the CCRI23 libraries. Only the gene encoding the polyphenol oxidase （PPO） is involved in plant defense against insect herbivores, and the others are related to improving tolerance to insect damage. Quantitative real-time PCR was used to study changes in expression levels during A. lucorum damage in CCRI23 and CCRI41. Signiifcantly regulated genes from CCRI23 showed a response in CCRI23 but not response in CCRI41. Similarly, signiifcantly regulated genes from CCRI41 showed a response in CCRI41 but not response in CCRI23. The results showed that, among transcriptomes of cotton varieties, there are different responses to A. lucorum damage.展开更多
In recent years, Apolygus htcorum has become the main pest of winter jujube. It sucks juice of young parts of winter jujube and causes falling of buds and flowers, leaf perforations, deformities and fall- off of fruit...In recent years, Apolygus htcorum has become the main pest of winter jujube. It sucks juice of young parts of winter jujube and causes falling of buds and flowers, leaf perforations, deformities and fall- off of fruits, leading to serious economic loss. Considering the occurrence characteristics and control problems, the distribution, occurrence regularity, damage characteristics and outbreak reasons of A. lucorum are overviewed and analyzed in this paper, and the pollution-free control measures are also put forward.展开更多
The widespread planting of genetically engineered cotton producing the CrylAc toxin has led to significantly reduced pesticide applications since 1997. However, conse- quently, the number of green mirid bugs （GMB）, ...The widespread planting of genetically engineered cotton producing the CrylAc toxin has led to significantly reduced pesticide applications since 1997. However, conse- quently, the number of green mirid bugs （GMB）, Apolygus lucorum （Meyer-Diir） has in- creased. So far the GMB, instead of the cotton bollworm Helicoverpa armigera （Hiibner）, has become the major pest in the transgenic Bt cotton field and has influenced cotton yield. Disproportionately, only a few studies on GMB at molecular level have been re- ported. Libraries from both third instar nymphs and adults were sequenced using Illumina technology, producing more than 106 million short reads and assembled into 63 029 uni- genes of mean length 597 nt and N50 813 nt, ranging from 300 nt to 9771 nt. BLASTx analysis against Nr, Swissprot, GO and COG was performed to annotate these unigenes. As a result, 26 478 unigenes （42.01%） matched to known proteins and 107 immune-related, 320 digestive-related and 53 metamorphosis-related genes were detected in these annotated unigenes. Additionally, we profiled gene expression using mapping based differentially expressed genes （DEGs） strategy between the two developmental stages： nymph and adult. The results demonstrated that thousands of genes were significantly differentially ex- pressed at different developmental stages. The transcriptome and gene expression data provided comprehensive and global gene resources of GMB. This transcriptome would improve our understanding of the molecular mechanisms of various underlying biological characteristics, including development, digestion and immunity in GMB. Therefore, these findings could help elucidate the intrinsic factors of the GMB resurgence, offering novel pest management targets for future transgenic cotton breeding.展开更多
The green mirid bug Apolygus lucorum is an agricultural pest that is known to cause damage to more than 150 plant species. Here, we report the transcriptomes ofA. lucorum at three different developmental stages （the ...The green mirid bug Apolygus lucorum is an agricultural pest that is known to cause damage to more than 150 plant species. Here, we report the transcriptomes ofA. lucorum at three different developmental stages （the second and fifth instar nymphs and adults）. A total of 98,236 unigenes with an average length of 1,335nt was obtained, of which 50,640 were annotated, including those encoding digestive enzymes and cytochrome P450s. Comparisons with cotton bollworm and cotton aphid transcriptomes revealed distinct features ofA. lucorum as a mesophyll feeder. The gene expression dynamics varied during development from young nymphs to adults. The high-quality transcriptome data and the gene expression dynamics reported here provide valuable data for a more comprehensive understanding of the physiology and development of mirid bugs, and for mining targets for their control.展开更多
The complete mitochondrial （mt） genome of the plant bug, Apolygus lucorum, an important cotton pest, has been sequenced and annotated in this study. The entire circular genome is 14 768 bp in size and represents the...The complete mitochondrial （mt） genome of the plant bug, Apolygus lucorum, an important cotton pest, has been sequenced and annotated in this study. The entire circular genome is 14 768 bp in size and represents the smallest in presently known heteropteran mt genomes. The mt genome is encoding for two ribosomal RNA （rRNA） genes, 22 transfer RNA （tRNA） genes, 13 protein coding genes and a control region, and the order, content, codon usage and base organization show similarity to a great extent to the hypothetical ancestral model. All protein coding genes use standard initiation codons ATN. Conventional stop codons TAA and TAG have been assigned to the most protein coding genes; however, COIII, ND4 and ND5 genes show incomplete terminator signal （T）. All tRNA genes possess the typical clover leaf structure, but the dihydrouridine arm of tRNAser（A6N） only forms a simple loop. Secondary structure models of rRNA genes are generally in accordance with the former models, although some differences exist in certain parts. Three intergenic spacers have never been found in sequenced mt genomes of Heteroptera. The phylogenetic study based on protein coding genes is largely congruent with previous phylogenetic work. Both Bayesian inference and maximum likelihood analyses highly support the sister relationship ofA. lucorum and Lygus lineolaris, and Miridae presents a sister position to Anthocoridae.展开更多
Heat shock cognate protein 70 （Hsc70） is a very important stress-resistance protein of insects against environmental stresses. We employed fluorescent real-time quan- titative polymerase chain reaction and Western-b...Heat shock cognate protein 70 （Hsc70） is a very important stress-resistance protein of insects against environmental stresses. We employed fluorescent real-time quan- titative polymerase chain reaction and Western-blot techniques to analyze the transcrip- tional and translational expression profiles of AIHSC70 under extreme temperature （4℃ and 40℃） or 4 pesticide stresses in Apolygus lucorum. The results showed that the expres- sion of AIHSC70 were significantly induced by cyhalothrin or extremely high temperature （40℃） in both transcriptional and translational levels （P 〈 0.05）, while the transcriptional and translational level of AIHSC70 decreased significantly in treatments of chlorpyrifos or extreme cold temperature （4℃） （P 〈 0.05）. Moreover, afterApolygus lucorum treated by imidacloprid or emamectin benzoate, the expression of AIHSC70 was only up-regulated significantly at the transcriptional level （P 〈 0.05）, although obviously up-regulated at the translational level of AIHSC70. Therefore, this study confirmed that the Alhsc70 gene played important roles in response to both temperature and pesticide stresses, especially for cyhalothrin or extremely high temperature （40℃）. In addition, the significant polynomial regression correlations between temperature and the Alhsc70 expression level were shown in all the nymph and adult stages （P 〈 0.01）, indicating temperature was an important factor to affect the relative expression of Alhsc70.展开更多
RNA interference(RNAi)is a powerful tool that post-transcriptionally silences target genes in eukaryotic cells.However,silencing efficacy varies greatly among different insect species.Recently,we met with little succe...RNA interference(RNAi)is a powerful tool that post-transcriptionally silences target genes in eukaryotic cells.However,silencing efficacy varies greatly among different insect species.Recently,we met with little success when attempting to knock down genes in the mirid bug Apolygus lucorum via dsRNA injection.The disappearance of double-stranded RNA(dsRNA)could be a potential factor that restricts RNAi efficiency.Here,we found that dsRNA can be degraded in midgut fluids,and a dsRNase of A.lucorum(AldsRNase)was identified and characterized.Sequence alignment indicated that its 6 key amino acid residues and the Mg2+-binding site were similar to those of other insects’dsRNases.The signal peptide and endonuclease non-specific domain shared high sequence identity with the brown-winged green stinkbug Plautia stali dsRNase.AldsRNase showed high salivary gland and midgut expression and was continuously expressed through the whole life cycle,with peaks at the 4th instar ecdysis in the whole body.The purified AldsRNase protein obtained by heterologously expressed can rapidly degrade dsRNA.When comparing the substrate specificity of AldsRNase,3 specific substrates(dsRNA,small interfering RNA,and dsDNA)were all degraded,and the most efficient degradation is dsRNA.Subsequently,immunofluorescence revealed that AldsRNase was expressed in the cytoplasm of midgut cells.Through cloning and functional study of AldsRNase,the enzyme activity and substrate specificity of the recombinant protein,as well as the subcellular localization of nuclease,the reason for the disappearance of dsRNA was explained,which was useful in improving RNAi efficiency in A.lucorum and related species.展开更多
The green plant bug Apolygus lucorum is a major pest of Bacillus thuringiensis cotton in China.Previously,we reported that chlorpyrifos resistance in a laboratory-selected strain of A.lucorum(BZ-R)is associated with t...The green plant bug Apolygus lucorum is a major pest of Bacillus thuringiensis cotton in China.Previously,we reported that chlorpyrifos resistance in a laboratory-selected strain of A.lucorum(BZ-R)is associated with the homozygosis of an allele in the ace-1 gene encoding an alanine to serine substitution at position 216 of acetylcholinesterase-1.Here we describe the results of crosses between the resistant BZ-R strain(41-fold to chlorpyrifos)and the unselected susceptible BZ-S strain homozygous for the wild type alanine allele at position 216.Resistance to chlorpyrifos was inherited as a semi-dominant trait mainly controlled by a single autosomal gene and co-segregates strongly but not completely with the serine substitution in ace-1.Synergism bioassays and enzyme assays showed that minor contributions to resistance are also made by enhanced cytochrome P450 and carboxylesterase activities.A survey of 25 field populations from five Chinese provinces showed strong positive correlations between 50%lethal concentration against chlorpyrifos and S216 allele and genotype frequencies,although the most tolerant populations still only show 40%-50%S216 allele frequencies.The results above provide important information for designing effective resistance monitoring and management strategies for A.lucorum in China.展开更多
基金Supported by the Special Program for the Construction of Modern Agricultural Industrial Technological System(CARS-30-bc)~~
文摘Apolygus lucorum（Meyer-Dür.） and Erythroneura apicalis（Nawa） are important pests that affect the quality and the yield of grapevine and cause huge economic losses. This paper focuses on the selection of effective botanical pesticides to control A. lucorum and E. apicalis. This experiment explores the effect of several botanical pesticides for A. lucorum and E. apicalis, including the 0.5% veratrine, the0.6% Oxygen·Lactone agent, the 5% natural pyrethrin, the composite neem pesticide, the rotenone and the composite nicotine. The 0.5% veratrine shows a stable control efficacy, which is higher than 60% in Chengdu, while the composite nicotine shows the highest efficacy against A. lucorum, which is above 70%. In Yinchuan,the 0.5% veratrine shows the highest efficacy, against the first generation adults and the second generation larvae of E. apicalis, while the 5% natural pyrethrin shows 100% control efficacy against E. apicalis in Nanjiang. The 0.5% veratrine and the composite neem could be used as effective pesticides to control A. lucorum and the 5% natural pyrethrin can be used to control E. apicalis. They could be widely used in the production of pollution-free grapes.
文摘[Objective] The aim was to analyze effect of climate change on occurrence degree of cotton Lugus lucorum in coastal areas of Jiangsu Province, in order to provide authentic references for better prevention. [Method] Statistical analysis was conducted on Information about occurrence degree of cotton Lugus lucorum in Binhai County in Jiangsu Province from 1985 to 2007 under guidance of Integral regression method. [Result] Because of climate change, cotton Lugus lucorum occurrence de- grees differed in different generations and growth stages in Binhai County. Tempera- ture and humidity constituted the main climate factors. In general, climate change hindered occurrence degree from increasing. We could make use of the period when occurrence degree of cotton Lugus lucorum is prone to significant increase and choose some periods, such as early April, late June and middle August, to improve prevention effect. In addition, attention should be paid to comprehensive measures, such as adjustment of industrial structure, which will reduce pesticide and minimize cotton Lugus lucorum damage and adverse impact from prevention measures. [Conclusion] The results provided authentic references for cotton Lugus lucorum preven- tion in Jiangsu coastal areas.
基金supported by the National Natural Science Foundation of China (31772176 and 31972338)the National Key Research and Development Program of China (2019YFD0300100)
文摘Artemisia annua is an important preferred host of the mirid bug Apolygus lucorum in autumn.Volatiles emitted from A.annua attract A.Iucorum.Volatile artemisinic acid of A.annua is a precursor of artemisinin that has been widely investigated in the Chinese herbal medicine field.However,little is known at this point about the biological roles of artemisinic acid in regulating the behavioral trends of A.lucorum.In this study,we collected volatiles from A.annua at the seedling stage by using headspace solid phase microextraction(HS-SPME).Gas chromatography-mass spectrometry(GC-MS) analysis showed that approximately 11.03±6.00 and 238.25±121.67 ng hartemisinic acid were detected in volatile samples and milled samples,respectively.Subsequently,a key gene for artemisinic acid synthesis,the cytochrome P450 gene cyp71 av1,was expressed in engineered Saccharomyces cerevisiae to catalyze the production of artemisinic acid.After the addition of exogenous artemisinic alcohol or artemisinic aldehyde,artemisinic acid was identified as the product of the expressed gene.In electroantennogram(EAG) recordings,3-day-old adult A.lucorum showed significant electrophysiological responses to artemisinic alcohol,artemisinic aldehyde and artemisinic acid.Furthermore,3-day-old female bugs were significantly attracted by artemisinic acid and artemisinic alcohol at a concentration of 10 mmol L,whereas 3-day-old male bugs were attracted significantly by 10 mmol Lartemisinic acid and artemisinic aldehyde.We propose that artemisinic acid and its precursors could be used as potential attractant components for the design of novel integrated pest management strategies to control A.lucorum.
基金This research was supported by Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences.
文摘Background Apolygus lucorum is a worldwide omnivorous pest damaging a range of crops and causing great economic losses.Symbiotic bacteria living in insects play a key role in the nutrition,physiology,and behavior of hosts.Here,we present an experiment using Illumina HiSeq sequencing targeting the V3–V4 regions of bacteria’s 16S rRNA throughout the entire life cycle of A.lucorum.Results The first and second instar nymphs have the largest alpha diversity compared with other life stages of the insect.Bacterial phyla Proteobacteria(72.29%),Firmicutes(15.24%),Actinobacteria(7.76%)exhibit the largest relative abundance in all developmental stages.Erwinia(23.97%)and Lactococcus(10.62%)are the two genera with the high-est relative abundance.The relative abundance of Erwinia in the nymph stage is significantly greater than the adult stage,and the relative abundance of Lactococcus in 6-day-old and 9-day-old adult females is higher compared with adult males.Conclusions These results reveal that microbial community composition and relative abundance shift dynamically at different life stages,implying that different bacterial phyla and genera may have specific roles in specific life stages such as metabolism,nutrition absorption,detoxification,and reproduction.This study reveals for the first time the community composition and ecological dynamics of symbiotic bacteria throughout the life stages of A.lucorum,and thus may provide insight to new strategies for pest control.
基金supported by the National Natural Science Foundation of China (31201518)
文摘Plants reshape their transcriptomes, proteomes and metabolomes in response to insect damage. In this study, we used suppression subtractive hybridization to investigate the transcriptomes of two cotton varieties （CCRI41 and CCRI23） under Apolygus lucorum damage. From the CCRI23 libraries we obtained 92 transcripts and from the CCRI41 libraries we obtained 96 transcripts. 26 and 63 of the transcripts from CCRI23 and CCRI41, respectively, had known functions. Using reverse transcription PCR, we detected expression proifle of genes with known functions. Ultimately, we identiifed eight signiifcantly regulated genes, including one downregulated and four upregulated genes from the CCRI41 libraries, and one downregulated and two upregulated genes from the CCRI23 libraries. Only the gene encoding the polyphenol oxidase （PPO） is involved in plant defense against insect herbivores, and the others are related to improving tolerance to insect damage. Quantitative real-time PCR was used to study changes in expression levels during A. lucorum damage in CCRI23 and CCRI41. Signiifcantly regulated genes from CCRI23 showed a response in CCRI23 but not response in CCRI41. Similarly, signiifcantly regulated genes from CCRI41 showed a response in CCRI41 but not response in CCRI23. The results showed that, among transcriptomes of cotton varieties, there are different responses to A. lucorum damage.
基金Supported by Special Fund for Agro-scientific Research in the Public Interest ( 201103012)
文摘In recent years, Apolygus htcorum has become the main pest of winter jujube. It sucks juice of young parts of winter jujube and causes falling of buds and flowers, leaf perforations, deformities and fall- off of fruits, leading to serious economic loss. Considering the occurrence characteristics and control problems, the distribution, occurrence regularity, damage characteristics and outbreak reasons of A. lucorum are overviewed and analyzed in this paper, and the pollution-free control measures are also put forward.
基金Acknowledgments We thank Dr. K. S. Shelby for comments and edito- rial assistance on the manuscript. This work was funded by the National Genetically Modified Organisms Breed- ing Major Project （2012ZX08009001）, National Natural Science Foundation of China （31230062） and the Na- tional Basic Research Program of China （973 Program, 2012CBl14104）. The ftmders had no role in study de- sign, data collection and analysis, decision to publish, or preparation of the manuscript.
文摘The widespread planting of genetically engineered cotton producing the CrylAc toxin has led to significantly reduced pesticide applications since 1997. However, conse- quently, the number of green mirid bugs （GMB）, Apolygus lucorum （Meyer-Diir） has in- creased. So far the GMB, instead of the cotton bollworm Helicoverpa armigera （Hiibner）, has become the major pest in the transgenic Bt cotton field and has influenced cotton yield. Disproportionately, only a few studies on GMB at molecular level have been re- ported. Libraries from both third instar nymphs and adults were sequenced using Illumina technology, producing more than 106 million short reads and assembled into 63 029 uni- genes of mean length 597 nt and N50 813 nt, ranging from 300 nt to 9771 nt. BLASTx analysis against Nr, Swissprot, GO and COG was performed to annotate these unigenes. As a result, 26 478 unigenes （42.01%） matched to known proteins and 107 immune-related, 320 digestive-related and 53 metamorphosis-related genes were detected in these annotated unigenes. Additionally, we profiled gene expression using mapping based differentially expressed genes （DEGs） strategy between the two developmental stages： nymph and adult. The results demonstrated that thousands of genes were significantly differentially ex- pressed at different developmental stages. The transcriptome and gene expression data provided comprehensive and global gene resources of GMB. This transcriptome would improve our understanding of the molecular mechanisms of various underlying biological characteristics, including development, digestion and immunity in GMB. Therefore, these findings could help elucidate the intrinsic factors of the GMB resurgence, offering novel pest management targets for future transgenic cotton breeding.
基金supported by the Strategic Priority Research Program of Chinese Academy of Sciences(XDB11030000)the Ministry of Agriculture of China(2014ZX08009001-009)
文摘The green mirid bug Apolygus lucorum is an agricultural pest that is known to cause damage to more than 150 plant species. Here, we report the transcriptomes ofA. lucorum at three different developmental stages （the second and fifth instar nymphs and adults）. A total of 98,236 unigenes with an average length of 1,335nt was obtained, of which 50,640 were annotated, including those encoding digestive enzymes and cytochrome P450s. Comparisons with cotton bollworm and cotton aphid transcriptomes revealed distinct features ofA. lucorum as a mesophyll feeder. The gene expression dynamics varied during development from young nymphs to adults. The high-quality transcriptome data and the gene expression dynamics reported here provide valuable data for a more comprehensive understanding of the physiology and development of mirid bugs, and for mining targets for their control.
基金This research is supported by grants from the Special Fund for Agroscientific Research in the Public Interest （Nos. 201103012, 201103022, 201303024）, National Ba- sic Research Program of China （No. 2013CB 127600）, the Natural Science Foundation of Beijing （No. 6112013）, the special Foundation for Scientific Research （No. 2012FY 111100） and the National Natural Science Foun- dation of China （Nos. 1061160186, 31111140015）.
文摘The complete mitochondrial （mt） genome of the plant bug, Apolygus lucorum, an important cotton pest, has been sequenced and annotated in this study. The entire circular genome is 14 768 bp in size and represents the smallest in presently known heteropteran mt genomes. The mt genome is encoding for two ribosomal RNA （rRNA） genes, 22 transfer RNA （tRNA） genes, 13 protein coding genes and a control region, and the order, content, codon usage and base organization show similarity to a great extent to the hypothetical ancestral model. All protein coding genes use standard initiation codons ATN. Conventional stop codons TAA and TAG have been assigned to the most protein coding genes; however, COIII, ND4 and ND5 genes show incomplete terminator signal （T）. All tRNA genes possess the typical clover leaf structure, but the dihydrouridine arm of tRNAser（A6N） only forms a simple loop. Secondary structure models of rRNA genes are generally in accordance with the former models, although some differences exist in certain parts. Three intergenic spacers have never been found in sequenced mt genomes of Heteroptera. The phylogenetic study based on protein coding genes is largely congruent with previous phylogenetic work. Both Bayesian inference and maximum likelihood analyses highly support the sister relationship ofA. lucorum and Lygus lineolaris, and Miridae presents a sister position to Anthocoridae.
基金Acknowledgments This work was supported by the Natural Science Foun- dation of Jiangsu Province （BK20130717）, the Jiangsu Agriculture Science and Technology Innovation Fund （JASTIF）, CX（14）5020, the National Natural Science Foundation of China （31401753）, the Open Fund in State Key Laboratory for Biology of Plant Diseases and Insect Pests （SKL2012OP03）, the Special Fund for Agro-scientific Research in the Public Interest of China （201103012-04）. We thank Professor Zhong-Yan Gu for providing the pesticide AI for this study.
文摘Heat shock cognate protein 70 （Hsc70） is a very important stress-resistance protein of insects against environmental stresses. We employed fluorescent real-time quan- titative polymerase chain reaction and Western-blot techniques to analyze the transcrip- tional and translational expression profiles of AIHSC70 under extreme temperature （4℃ and 40℃） or 4 pesticide stresses in Apolygus lucorum. The results showed that the expres- sion of AIHSC70 were significantly induced by cyhalothrin or extremely high temperature （40℃） in both transcriptional and translational levels （P 〈 0.05）, while the transcriptional and translational level of AIHSC70 decreased significantly in treatments of chlorpyrifos or extreme cold temperature （4℃） （P 〈 0.05）. Moreover, afterApolygus lucorum treated by imidacloprid or emamectin benzoate, the expression of AIHSC70 was only up-regulated significantly at the transcriptional level （P 〈 0.05）, although obviously up-regulated at the translational level of AIHSC70. Therefore, this study confirmed that the Alhsc70 gene played important roles in response to both temperature and pesticide stresses, especially for cyhalothrin or extremely high temperature （40℃）. In addition, the significant polynomial regression correlations between temperature and the Alhsc70 expression level were shown in all the nymph and adult stages （P 〈 0.01）, indicating temperature was an important factor to affect the relative expression of Alhsc70.
基金This study was supported by Jiangsu Agricultural Science and Technology Innovation Fund[CX(21)3088,CX(22)2038]Jiangsu Province Key R&D Program(Modern Agriculture)Project:Surface Project(BE2021303).
文摘RNA interference(RNAi)is a powerful tool that post-transcriptionally silences target genes in eukaryotic cells.However,silencing efficacy varies greatly among different insect species.Recently,we met with little success when attempting to knock down genes in the mirid bug Apolygus lucorum via dsRNA injection.The disappearance of double-stranded RNA(dsRNA)could be a potential factor that restricts RNAi efficiency.Here,we found that dsRNA can be degraded in midgut fluids,and a dsRNase of A.lucorum(AldsRNase)was identified and characterized.Sequence alignment indicated that its 6 key amino acid residues and the Mg2+-binding site were similar to those of other insects’dsRNases.The signal peptide and endonuclease non-specific domain shared high sequence identity with the brown-winged green stinkbug Plautia stali dsRNase.AldsRNase showed high salivary gland and midgut expression and was continuously expressed through the whole life cycle,with peaks at the 4th instar ecdysis in the whole body.The purified AldsRNase protein obtained by heterologously expressed can rapidly degrade dsRNA.When comparing the substrate specificity of AldsRNase,3 specific substrates(dsRNA,small interfering RNA,and dsDNA)were all degraded,and the most efficient degradation is dsRNA.Subsequently,immunofluorescence revealed that AldsRNase was expressed in the cytoplasm of midgut cells.Through cloning and functional study of AldsRNase,the enzyme activity and substrate specificity of the recombinant protein,as well as the subcellular localization of nuclease,the reason for the disappearance of dsRNA was explained,which was useful in improving RNAi efficiency in A.lucorum and related species.
基金National Key Research Development Program of China(No.2016YFD0200500)。
文摘The green plant bug Apolygus lucorum is a major pest of Bacillus thuringiensis cotton in China.Previously,we reported that chlorpyrifos resistance in a laboratory-selected strain of A.lucorum(BZ-R)is associated with the homozygosis of an allele in the ace-1 gene encoding an alanine to serine substitution at position 216 of acetylcholinesterase-1.Here we describe the results of crosses between the resistant BZ-R strain(41-fold to chlorpyrifos)and the unselected susceptible BZ-S strain homozygous for the wild type alanine allele at position 216.Resistance to chlorpyrifos was inherited as a semi-dominant trait mainly controlled by a single autosomal gene and co-segregates strongly but not completely with the serine substitution in ace-1.Synergism bioassays and enzyme assays showed that minor contributions to resistance are also made by enhanced cytochrome P450 and carboxylesterase activities.A survey of 25 field populations from five Chinese provinces showed strong positive correlations between 50%lethal concentration against chlorpyrifos and S216 allele and genotype frequencies,although the most tolerant populations still only show 40%-50%S216 allele frequencies.The results above provide important information for designing effective resistance monitoring and management strategies for A.lucorum in China.
