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Identification of a novel cytochrome P450 CYP321B1 gene from tobacco cutworm （spodoptera litura） and RNA interference to evaluate its role in commonly used insecticides

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Insect Science 2017年第2期24卷 235-247页

作者： Rui-Long Wang Keyan Zhu-Salzman Scott R. Baerson Xiao-Wei Xin Jun Li Yi-Juan Su Ren-Sen Zeng Key Laboratory of Tropical Agro-Environment Ministry of Agriculture College of Natural Resources and Environment South China Agri- cultural University Guangzhou China College of Life Sciences Fujian Agriculture and Forestry University Fuzhou China Departments of Entomology Texas A&M University Texas United States Department of Agriculture-Agricultural Research Service Natural Products Utilization Research Unit University Mississippi USA Guangdong Engineering Research Center for Modem Eco-agriculture and Circular Agriculture Guangzhou China

Insect cytochrome P450 monooxygenases （CYPs or P450s） play an important role in detoxifying insecticides leading to resistance in insect populations. A polyphagous pest, spodoptera litura, has developed resistance to a wide range of insecticides. In the present study, a novel P450 gene, CYP321B1, was cloned from S. litura. The function of CYP321B1 was assessed using RNA interference （RNAi） and monitoring resistance levels for three commonly used insecticides, including chlorpyrifos, β-cypermethrin and methomyl. The full-length complementary DNA sequence of CYP321B1 is 1814 bp long with an open reading frame of 1 488 bp encoding 495 amino acid residues. Quantitative reverse-transcriptase polymerase chain reaction analyses during larval and pupal develop- ment indicated that CYP321B1 expression was highest in the midgut of fifth-instar larvae, followed by fat body and cuticle. The expression of CYP321B1 in the midgut was up- regulated by chlorpyrifos,β-cypermethrin and methomyl with both lethal concentration at 15% （LC15）（50, 100 and 150 μg/mL, respectively） and 50%（LC50） dosages （100,200 and 300μg/mL, respectively）. Addition of piperonyl butoxide （PBO） significantly increased the toxicity ofchlorpyrifos,β-cypermethrin and methomyl to S. litura, suggesting a marked synergism of the three insecticides with PBO and P450-mediated detoxification. RNAi- mediated silencing of CYP321B1 further increased mortality by 25.6% and 38.9% when the fifth-instar larvae were exposed to chlorpyrifos and β-cypermethrin, respectively, at the LCso dose levels. The results demonstrate that CYP321B1 might play an important role in chlorpyrifos and β-cypermethrin detoxification in S. litura.

关键词： CYP321B1 cytochrome P450 monooxygenases insecticide resistance RNAinterference spodoptera litura

Resistance in hybrid derivatives of Lycopersicon accessions against leaf caterpillar, spodoptera litura Fab.

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Insect Science 2008年第2期15卷 141-146页

作者： N. Muthukumaran V. Selvanarayanan Department of Entomology Faculty of Agriculture Annamalai University Annamalainagar Tamil Nadu India

Four tomato accessions, namely Ac 238, Roma, Seijima Jeisei and Varushanadu Local selected from preliminary screening of 321 accessions and their hybrid derivatives, namely HYtF：（Varushanadu Local × Ac 238）, HY2F1（Varushanadu Local × Roma）, HY3F1 （Ac 238 × Roma）（first generation）, HY1F2, HY2F2 and HY3F2 （second generation） were evaluated for their resistance in comparison with a susceptible check, 1979 against the leaf caterpillar, spodoptera litura Fab. both under field and laboratory conditions at Annamalainagar, Tamil Nadu, India. In the field evaluation, accession Seijima Jeisei and hybrid HY3F1 recorded the minimum larval population. In glasshouse and laboratory studies, Seijima Jeisei was the least preferred by S. litura for both feeding and oviposition. The hybrid HYsF： exerted pronounced antibiosis effect on S. litura. Among the various biophysical and biochemical bases of resistance, contents of reducing sugar and phenol were found negatively correlated with S. litura larval feeding. Among the hybrids, HY3F1 and HY3F2 and the parent Seijima Jeisei recorded higher fruit yield per plant in both seasons. Considerable variation in the resistance traits of the hybrids was observed when compared with the parents.

关键词： hybrid derivatives resistance spodoptera litura tomato accession

Analyses of microRNAs and transcriptomes in the midgut of spodoptera litura feeding on Brassica juncea

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Insect Science 2021年第2期28卷 533-547页

作者： Xiao-Peng Zou Yi-Guang Lin Yong-Jie Cen Kang Ma Bin-Bin Qiu Qi-Li Feng Si-Chun Zheng School of Life Sciences South China Normal UniversityGuangzhouChina

spodoptera litura is a destructive agricultural pest in tropical and subtropical *** the molecular mechanisms of *** adaptation to its preferred host plants may help identify target genes useful for pest *** used high-throughput sequencing to characterize the expression patterns of messenger RNAs(mRNAs)and microRNAs(miRNAs)in the midgut of *** fed on Brassica juncea for 6 h and 48 h.A total of 108 known and 134 novel miRNAs were identified,29 miRNAs and 237 mRNAs were differentially expressed at 6 h of *** feeding,26 miRNAs and 433 mRNAs were differentially expressed at 48 *** the mRNAs,the up-regulated genes were mostly enriched in detoxification enzymes(cytochrome P450,esterase,glutathione^-transferase,uridine diphosphate-glucuronosyl transferase),while the down-regulated genes were mostly enriched in proteinases and immune-related ***,most detoxification enzymes begin to up-regulate at 6 h,while most digestion and immunerelated genes begin to up-or down-regulate at 48 *** and 37 differently expressed transcription factors were identified at 6 h and 48 h,which may regulate the functional *** acquired 136 and 41 miRNA versus mRNA pairs at 6 h and 48 h,*** down-regulated and up-regulated miRNAs were predicted to target detoxification enzymes and proteinases,***-time quantitative polymerase chain reaction of nine randomly selected miRNAs and 28 genes confirmed the results of *** analyses of miRNA and mRNA transcriptomes provides useful information about the molecular mechanisms of *** response to ***.

关键词： Brassica juncea midgut miRNAs RNA-Seq spodoptera litura

Transcription factor CncC potentially regulates the expression of multiple detoxification genes that mediate indoxacarb resistance in spodoptera litura

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Insect Science 2021年第5期28卷 1426-1438页

作者： Li Shi Yao Shi Meng-Fei Liu Ya Zhang Xiao-Lan Liao Hunan Provincial Engineering and Technology Research Center for Bio-pesticide and Formulation Processing College of Plant ProtectionHunan Agricultural UniversityChangshaChina Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests ChangshaChina

The xenobiotic transcription factor cap‘n’collar isoform C(CncC)is considered the central regulator of antioxidant and detoxification *** research indicated that CncC regulates three-phase enzymes responsible for insecticide *** this study,the SlituCncC gene from spodoptera litura was obtained and *** polymerase chain reaction(qPCR)analysis showed that SlituCncC was expressed in all developmental stages and tissues,but was highly expressed in 3rd-and 4th-instar larvae,and in the Malpighian tubule,fat body,and *** addition,SlituCncC was up-regulated and more highly induced with indoxacarb treatment in the indoxacarb-resistant strains compared with the susceptible *** interference-mediated gene silencing of SlituCncC significantly increased mortality of *** when exposed to ***,comparative transcriptome analysis showed that 842 genes were down-regulated and 127 genes were up-regulated in SlituC"ncC knockdown *** analysis indicated that 18 three-phase enzymes were identified in the down-regulated genes,of which seven were associated with indoxacarb resistance in *** analysis confirmed that expression of six of these seven genes was consistent with RNA se-quencing *** six detoxification genes were induced by indoxacarb,and the expression patterns were similar to that of ***,the CneC-Maf binding site was predicted in all six gene *** study indicates that the transcription factor SlituCncC may regulate multiple detoxification genes that mediate indoxacarb resistance in ***.

关键词： spodoptera litura indoxacarb resistance transcription factor detoxification genes RNA interference RNA sequencing

CRISPR/Cas9-mediated targeted gene mutagenesis in spodoptera litura

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Insect Science 2016年第3期23卷 469-477页

作者： Hong-Lun Bi Jun XU An-Jiang Tan Yong-Ping Huang College of Life Science Fujian Agriculture and Forestry University Fuzhou Key Laboratory of Insect Developmental and EvolutionaryBiology Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences ShanghaiChina

Custom-designed nuclease technologies such as the clustered regularly in- terspaced short palindromic repeat （CRISPR）-associated （Cas） system provide attractive genome editing tools for insect functional genetics. The targeted gene mutagenesis me- diated by the CR1SPR/Cas9 system has been achieved in several insect orders including Diptera, Lepidoptera and Coleoptera. However, little success has been reported in agricul- tural pests due to the lack of genomic information and embryonic microinjection techniques in these insect species. Here we report that the CRISPR/Cas9 system induced efficient gene mutagenesis in an important Lepidopteran pest spodoptera litura. We targeted the S. litura Abdominal-A （Slabd-A） gene which is an important embryonic development gene and plays a significant role in determining the identities of the abdominal segments of in- sects. Direct injection of Cas9 messenger RNA and Slabd-A-specific single guide RNA （sgRNA） into S. litura embryos successfully induced the typical abd-A deficient pheno- type, which shows anomalous segmentation and ectopic pigmentation during the larval stage. A polymerase chain reaction-based analysis revealed that the Cas9/sgRNA complex effectively induced a targeted mutagenesis in S. litura. These results demonstrate that the CRISPR/Cas9 system is a powerful tool for genome manipulation in Lepidopteran pests such as S. litura.

关键词： abdominal-A gene CRISPR/Cas9 system genome editing microinjection spodoptera litura

Expression profiles of glutathione S-transferase superfamily in spodoptera litura tolerated to sublethal doses of chlorpyrifos

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Insect Science 2016年第5期23卷 675-687页

作者： Ni Zhang Jia Liu Shu-Na Chen Li-Hua Huang Qi-Li Feng Si-Chun Zheng Laboratory of Developmental and Molecular Entomology Guangdong Provincial Key Laboratory of Biotechnology for Plant DevelopmentSchool of Life Sciences South China Normal University Guangzhou China

Chlorpyrifos （CPF） is a broad-spectrum organophosphate insecticide. Glu- tathione S-transferases （GSTs） in insects are a family of detoxification enzymes and they play critical roles in CPF detoxification. spodoptera litura is one of the most destructive agricultural pests in tropical and subtropical areas in the world. In this study, 37 Slgsts from 46 unique transcripts of gsts in S. litura transcriptome data, including eight pre- viously reported GSTs, were identified and their expression patterns in susceptible and 12-generation-CPF-treated strains were analyzed to understand the roles of these Slgsts in sublethal doses of CPF tolerance. The results indicate that the members of the S. litura GST superfamily could be distinguished into three major groups： one group, includ- ing six cytosolic Slgsts （SIGSTel, SIGSTe3, SIGSTelO, SIGSTe15, SIGSTo2 and SIGSTs5） and two microsomal Slgsts （SIMGSTI-2 and SIMGST1-3）, was directly responsible for CPF induction in both 12-generation-treated and susceptible strains; the second group, including three cytosolic Slgsts （SlGSTe13, SIGSTtl and SIGSTzl） and one microsomal Slgst （SIMGSTI-1）, was induced only in the 12-generation-treated strain; the third group, including eight cytosolic Slgsts （two epsilon, three delta, one omega, one zeta and one un- classified Slgst）, was expressed 1.52-5.15-fold higher in the 12-generation-treated strain than in the susceptible strain.

关键词： chlorpyrifos expression glutathione S-transferase spodoptera litura su-perfamily tolerance

Laboratory evaluation of flight activity of the common cutworm, spodoptera litura （Lepidoptera： Noctuidae）

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Insect Science 2010年第1期17卷 53-59页

作者： Ye-Gou Tu Kong-Ming Wu Fang-Sen Xue Yan-Hui Lu State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences Beijing Institute of Entomology Jiangxi Agricultural University Nanchang China

The flight activity of spodoptera litura in tethered conditions is evaluated using a computer-mediated flight-mill in the laboratory. The 3-4-day-old moths fly longer and farther than those of other ages. Male and female moths exhibit similar flight activity. Mating status does not influence the flight duration and distance of 2-day-old females. However, these two flight parameters with a 6-day-old mated female is significantly lower than that of tmmated ones. The optimum temperature for flight ranged from 16-24℃, whereas the optimum RH ranged from 60%--100%. During 72-h period, the total flight duration and distance of 1-day-old male and female moths were 19.6 h （± 5.8） and 83.3 km （± 28.4）, and 24.0 h （± 7.0） and 105.4 km （± 37.4）, respectively. These results indicate that S. litura has a great potential to undertake long-distance migratory flights.

关键词： spodoptera litura flight activity tethered-flight

Transcriptome and metabolome analysis reveal that oral secretions from Helicoverpa armigera and spodoptera litura influence wound-induced host response in cotton

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The Crop Journal 2020年第6期8卷 929-942页

作者： Huan Si Hongling Liu Yiwen Sun Zhongping Xu Sijia Liang Bo Li Xiao Ding Jianying Li Qiongqiong Wang Lin Sun Xianlong Zhang Shuangxia Jin National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural UniversityWuhan 430070HubeiChina

Cotton(Gossypium hirsutum)is an important fiber crop *** attack causes cotton yield and quality ***,little is known about the mechanism of cotton response to insect *** simulated insect feeding by applying insect oral secretions(OS)to wounds,and combined transcriptome and metabolome analysis to investigate how OS from two major pest species(Helicoverpa armigera and spodoptera litura)affect cotton defense *** found that respectively 12,668 and 13,379 genes were differentially expressed in comparison with wounding *** addition of OS,the jasmonic acid signaling pathway was rapidly and strongly induced,whereas genes involved in salicylic acid biosynthesis were *** constructing a coexpression gene network,we identified a hub gene encoding a leucine-rich repeat receptor kinase that may play an important role in early signal recognition and *** from the two insect species altered the abundance of flavonoid-related compounds in different *** remained in lower concentration after OS application than after wounding alone,suggesting a suppressive effect of OS on cotton defense *** study illustrated transcriptional and metabolic changes of cotton in responding to OS from two chewing insect species,identified potential key response genes,and revealed evidence for OS inhibition of wounding-induced cotton defense response.

关键词： Oral secretions Helicoverpa armigera spodoptera litura Cotton Host plant defense

Cloning and structural characterization of juvenile hormone diol kinase in spodoptera litura

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Insect Science 2016年第6期23卷 819-828页

作者： Bao-Juan Zeng Yuan Lv Li-Li Zhang Li-Hua Huang Qi-Li Feng Guangzhou Key Laboratory of Insect Development Regulation and Application Research School of Life Sciences South China Normal University Guangzhou 510631 China

Juvenile hormone （JH） is one of the key insect hormones that regulate metamorphosis. Juvenile hormone diol kinase （JHDK） is an enzyme involved in JH metabolism and catalyzes JH diol to form a polar end product, JH diol phosphate that has no JH activity. In this study, a JHDK complementary DNA （cDNA） was cloned from spodoptera litura and the structure and expression of the gene was characterized. The cDNA was 714 base pairs in length and encoded a protein of 183 amino acids with a molecular mass of 21 kDa and an isoelectric point of 4.55. Based on the structure, three putative calcium binding motifs and guanosine triphosphate-binding motifs were predicted in the protein. Modeling of the 3-D structure showed that the protein consisted of eight α-helixes linked with loops, with no β-sheets. The gene was expressed in the epidermis, fat body and midgut of fifth and sixth instar larvae. The expression level in the epidermis was lower than in the fat body and midgut. The gene was expressed at higher levels at the early stages than in the later stages of fifth and sixth instar midgut and fat body. The results suggest that this gene may be involved in the regulation of the JH titer in larvae ofS. litura.

关键词： juvenile hormone juvenile hormone diol kinase metabolism spodoptera litura