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Identification of four caspase genes from Spodoptera exigua (Lepidoptera: Noctuidae) and their regulations toward different apoptotic stimulations

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insect Science 2020年第6期27卷 1158-1172页

作者： Huan Yu Zi-Qi Li Yi-Yi Ou-Yang Guo-Hua Huang Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests Hunan Agricultural UniversityChangshaChina College of plant Protection Hunan Agricultural UniversityChangshaChina

Apoptosis plays critical roles in multiple biological processes in multicellular organisms.Caspases are known as important participators and regulators of apoptosis.Here,four novel caspase genes of Spodoptera exigua were cloned and characterized which were designated as SeCasp-1,SeCasp-6,ScCasp-7 and SeCasp-S.Analysis of the putative encoded protein sequences of these SeCasps indicated that SeCasp-1 and ScCasp-7 were possible homologs of executor caspases;SeCasp-8 was a possible homolog of initiator caspases;and SeCasp-6 was a unique caspase of S.exigua that shares low similarity with all the identified insect caspases.Based on baculovirus expression system analyses,SeCasp-1 exhibited similar caspase activity to human caspase-1,-3,-4,-6,-8 and-9;SeCasp-6 presented similar caspase activity to human caspase-2,-3,-4,-6,-8 and-9;SeCasp-7 exhibited similar caspase activity to human caspase-2,-3 and-6;and SeCasp-8 presented similar caspase activity only to human caspase-8.Induction with diiTcrent chemicals revealed that SeCasp-1 showed extreme upregulation after 24 h in the treated fat body cell line(IOZCAS-Spex-II)of S.exigua.Developmental expression analysis revealed that SeCasp-1 was highly transcribed in the larval stages,while SeCasp-6,SeCasp-7,SeCasp-H were down-regulated.The in vivo detection of the relative expression levels of SeCasps in S.eixgua larvae inoculated with different pathogens suggested that SeCasp-J was sensitive to Bacillus thuringiensis infection and that SeCasp-6 was sensitive to baculovirus infection.SeCasp-7 and SeCasp-H showed slight changes under either in vitro chemical apoptosis induction or in vivo pathogen infection.

关键词： apoptosis apoptotic inducing insect caspase SeCasp Spodoptera exigua

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 genes.Previous research indicated that CncC regulates three-phase enzymes responsible for insecticide resistance.In this study,the SlituCncC gene from Spodoptera litura was obtained and characterized.Quantitative 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 midgut.In addition,SlituCncC was up-regulated and more highly induced with indoxacarb treatment in the indoxacarb-resistant strains compared with the susceptible strain.RNA interference-mediated gene silencing of SlituCncC significantly increased mortality of S.liura when exposed to in-doxacarb.Furthermore,comparative transcriptome analysis showed that 842 genes were down-regulated and 127 genes were up-regulated in SlituC"ncC knockdown S.litura.Further analysis indicated that 18 three-phase enzymes were identified in the down-regulated genes,of which seven were associated with indoxacarb resistance in S.litura.qPCR analysis confirmed that expression of six of these seven genes was consistent with RNA se-quencing data.All six detoxification genes were induced by indoxacarb,and the expression patterns were similar to that of SlituCncC.Finally,the CneC-Maf binding site was predicted in all six gene promoters.This study indicates that the transcription factor SlituCncC may regulate multiple detoxification genes that mediate indoxacarb resistance in S.litura.

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

Novel strategies for the biocontrol of noctuid pests(Lepidoptera)based on improving ascovirus infectivity using Bacillus thuringiensis

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insect Science 2021年第5期28卷 1452-1467页

作者： Huan Yu Chang-Jin Yang Ni Li Ying Zhao Zhuang-Mei Chen Si-Jia Yi Zi-Qi Li Michael J.Adang Guo-Hua Huang Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests Hunan Agricultural UniversityChangshaHunan410128China College of plant Protection Hunan Agricultural UniversityChangshaHunan410128China Department of Entomology University of GeorgiaAthensGA30602USA

ldentifying novel biocontrol agents and developing new strategies are urgent goals in insect pest biocontrol.Ascoviruses are potential competent insect viruses that may be developed into bioinsecticides,but this aim is impeded by their poor oral infectivity.To improve the per os infectivity of ascovirus,Bacillus thuringiensis kurstaki(Btk)was employed as a helper to damage the midgut of lepidopteran larvae(Helicoverpa armigera,Mythimna separata,Spodoptera frugiperda,and S.litura)in formulations with Heliothis virescens ascovirus isolates(HvAV-3h and HvAV-3j).Btk and ascovirus mixtures(Btk/HvAV-3h and B1k/HvAV-3j)were fed to insect larvae(3rd instar).With the exception of S.frugiperda larvae,which exhibited low mortality after ingesting Btk,the larvae of the other tested species showed three types of response to feeding on the formulas:type I,the tested larvae(H.armigera)were killed by Btk infection so quickly that insufficient time and resources remained for ascoviral invasion;type II,both Btk and the ascovirus were depleted by their competition,such that neither was successfully released or colonized the tissue;type II,Btk was eliminated by the ascovirus,and the ascovirus achieved systemic infection in the tested larvae.The feeding of Btk/ascovirus formulas led to a great reduction in larval diet consumption and resulted in a significant decrease in the emergence rate of H.armigera,M.separata,and S.litura larvae,which suggested that the formulas exerted marked oral control effects on both the contemporary individuals and the next generation of these tested pest species.

关键词： ascovirus Bacillus thuringiensis formulation Heliothis virescens ascovirus 3h(HvAV-3h) Heliothis virescens ascovirus 3j(HvAV-3j) per os infectivity

Potassium sulphate induces resistance of rice against the rootknot nematode Meloidogyne graminicola

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Journal of Integrative Agriculture 2022年第11期21卷 3263-3277页

作者： LIU Mao-Yan PENG De-liang SU Wen XIANG Chao JIAN Jin-zhuo ZHAO Jie PENG Huan LIU Shi-ming KONG Ling-an DAI Liang-ying HUANG Wen-kun LIU Jing key laboratory for biology and control of plant diseases and insect pests College of Plant ProtectionHunan Agricultural UniversityChangsha 410128P.R.China State key laboratory for biology of plant diseases and insect pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193P.R.China School of Agricultural Science Xichang UniversityXichang 615013P.R.China

Potassium(K),an important nutrient element,can improve the stress resistance/tolerance of crops.The application of K in resisting plant-parasitic nematodes shows that the K treatment can reduce the occurrence of nematode diseases and increase crop yield.However,data on K_(2)SO_(4)induced rice resistance against the root-knot nematode Meloidogyne graminicola are still lacking.In this work,K_(2)SO_(4)treatment reduced galls and nematodes in rice plants and delayed the development of nematodes.Rather than affecting the attractiveness of roots to nematodes and the morphological phenotype of giant cells at feeding sites,such an effect is achieved by rapidly priming hydrogen peroxide(H_(2)O_(2))accumulation and increasing callose deposition.Meanwhile,galls and nematodes in rice roots were more in the potassium channel OsAKT11 and transporter OsHAK5 gene-deficient plants than in wild-type,while the K_(2)SO_(4)-induced resistance showed weaker in the defective plants.In addition,during the process of nematode infection,the expression of jasmonic acid(JA)/ethylene(ET)/brassinolide(BR)signaling pathway-related genes and pathogenesis-related(PR)genes OsPR1 a/OsPR1 b was up-regulated in rice after K_(2)SO_(4)treatment.In conclusion,K_(2)SO_(4)induced rice resistance against M.graminicola.The mechanism of inducing resistance was to prime the basal defense and required the participation of the K^(+)channel and transporter in rice.These laid a foundation for further study on the mechanism of rice defense against nematodes and the rational use of potassium fertilizer on improving rice resistance against nematodes in the field.

关键词： rice Meloidogyne graminicola potassium sulphate induced resistance H_(2)O_(2) callose potassium channel and transporter

Recognition of the inducible,secretory small protein OsSSP1 by the membrane receptor OsSSR1 and the co-receptor OsBAK1 confers rice resistance to the blast fungus

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Molecular plant 2024年第5期17卷 807-823页

作者： Tianfeng Zhao Shijie Ma Ziying Kong Haimiao Zhang Yi Wang Junzhe Wang Jiazong Liu Wanzhen Feng Tong Liu Chunyan Liu Suochen Liang Shilin Lu Xinyu Li Haipeng Zhao Chongchong Lu Muhammad Zunair Latif Ziyi Yin Yang Li Xinhua Ding State key laboratory of Crop biology Shandong Provincial Key Laboratory for Biology of Vegetable Diseases and Insect PestsCollege of Plant ProtectionShandong Agricultural UniversityTaian 271018ShandongChina College of Breeding and Multiplication(Sanya Institute of Breeding and Multiplication) Hainan UniversitySanya 572024HainanChina

The plant apoplast,which serves as the frontline battleground for long-term host–pathogen interactions,harbors a wealth of disease resistance resources.However,the identification of the disease resistance proteins in the apoplast is relatively lacking.In this study,we identified and characterized the rice secretory protein OsSSP1(Oryza sativa secretory small protein 1).OsSSP1 can be secreted into the plant apoplast,and either in vitro treatment of recombinant OsSSP1 or overexpression of OsSSP1 in rice could trigger plant immune response.The expression of OsSSP1 is suppressed significantly during Magnaporthe oryzae infection in the susceptible rice variety Taibei 309,and OsSSP1-overexpressing lines all show strong resistance to M.oryzae.Combining the knockout and overexpression results,we found that OsSSP1 positively regulates plant immunity in response to fungal infection.Moreover,the recognition and immune response triggered by OsSSP1 depend on an uncharacterized transmembrane OsSSR1(secretory small protein receptor 1)and the key co-receptor OsBAK1,since most of the induced immune response and resistance are lost in the absence of OsSSR1 or OsBAK1.Intriguingly,the OsSSP1 protein is relatively stable and can still induce plant resistance after 1 week of storage in the open environment,and exogenous OsSSP1 treatment for a 2-week period did not affect rice yield.Collectively,our study reveals that OsSSP1 can be secreted into the apoplast and percepted by OsSSR1 and OsBAK1 during fungal infection,thereby triggering the immune response to enhance plant resistance to M.oryzae.These findings provide novel resources and potential strategies for crop breeding and disease control.

关键词： rice plant apoplast OsssP1 receptor plant resistance Magnaporthe oryzae

Genome analysis of Heliothis virescens ascovirus 3h isolated from China

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Virologica Sinica 2017年第2期32卷 147-154页

作者： Guo-Hua Huang Dian-Hai Hou Manli Wang Xiao-Wen Cheng Zhihong Hu State key laboratory of Virology Microorganisms & Viruses Culture Collection CenterWuhan Institute of VirologyChinese Academy of Sciences Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests Institute of VirologyHunan Agricultural University Department of Microbiology 32 Pearson HallMiami University

No ascovirus isolated from China has been sequenced so far. Therefore, in this study, we aimed to sequence the genome of Heliothis virescens ascovirus 3h(Hv AV-3h) using the 454 pyrosequencing technology. The genome was found to be 190,519-bp long with a G+C content of 45.5%. We also found that it encodes 185 hypothetical open reading frames(ORFs) along with at least 50 amino acids, including 181 ORFs found in other ascoviruses and 4 unique ORFs. Gene-parity plots and phylogenetic analysis revealed a close relationship between Hv AV-3h and three other Hv AV-3a strains and a distant relationship with Spodoptera frugiperda ascovirus 1a(Sf AV-1a), Trichoplusia ni ascovirus 6a(Tn AV-6a), and Diadromus pulchellus ascovirus 4a(Dp AV-4a). Among the 185 potential genes encoded by the genome, 44 core genes were found in all the sequenced ascoviruses. In addition, 25 genes were found to be conserved in all ascoviruses except Dp AV-4a.In the Hv AV-3h genome, 24 baculovirus repeat ORFs(bros) were present, and the typical homologous repeat regions(hrs) were absent. This study supplies information important for understanding the conservation and functions of ascovirus genes as well as the variety of ascoviral genomes.

关键词： Heliothis virescens ascovirus 3h (HvAV-3h ) 染色体组织兄弟基因种系发生的关系

3H-31, A Non-structural Protein of Heliothis virescens ascovirus 3h,Inhibits the Host Larval Cathepsin and Chitinase Activities

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Virologica Sinica 2021年第5期36卷 1036-1051页

作者： Huan Yu Yi-Yi Ou-Yang Chang-Jin Yang Ni Li Madoka Nakai Guo-Hua Huang Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests Hunan Agricultural UniversityChangsha 410128China College of plant Protection Hunan Agricultural UniversityChangsha 410128China Tokyo University of Agriculture and Technology SaiwaiFuchuTokyo 183-8509Japan

3h-31 of Heliothis virescens ascovirus 3h(Hv AV-3h)is a highly conserved gene of ascoviruses.As an early gene of Hv AV-3h,3h-31 codes for a non-structural protein(3H-31)of Hv AV-3h.In the study,3h-31 was initially transcribed and expressed at 3 h post-infection(hpi)in the infected Spodoptera exigua fat body cells(Se FB).3h-31 was further inserted into the bacmid of Autographa californica nucleopolyhedrovirus(Ac MNPV)to generate an infectious baculovirus(Ac MNPV-31).In vivo experiments showed that budded virus production and viral DNA replication decreased with the expression of 3H-31,and lucent tubular structures were found around the virogenic stroma in the Ac MNPV-31-infected Se FB cells.In vivo,both LD50and LD90values of Ac MNPV-31 were significantly higher than those of the wild-type Ac MNPV(Ac MNPV-wt)in third instar S.exigua larvae.An interesting finding was that the liquefaction of the larvae killed by the infection of Ac MNPV-31 was delayed.Chitinase and cathepsin activities of Ac MNPV-31-infected larvae were significantly lower than those of Ac MNPV-wt-infected larvae.The possible regulatory function of the chitinase and cathepsin for 3H-31 was further confirmed by RNAi,which showed that larval cathepsin activity was significantly upregulated,but chitinase activity was not significantly changed due to the RNAi of 3h-31.Based on the obtained results,we assumed that the function of 3H-31 was associated with the inhibition of host larval chitinase and cathepsin activities,so as to restrain the hosts in their larval stages.

关键词： Cathepsin Chitinase Heliothis virescens ascovirus 3h(HvAV-3h) Recombinant virus 3h-31

Systematic identification and functional analysis of long noncoding RNAs involved in indoxacarb resistance in Spodoptera litura

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insect Science 2022年第6期29卷 1721-1736页

作者： Li Shi Wen-Lin Li Hai-Xin Zeng Yao Shi 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 andinsectpests ChangshaChina

Long noncoding RNAs(lncRNAs)are noncoding transcripts that are more than 200 nucleotides long.They play essential roles in regulating a variety of biological processes in many species,including insects,and some lncRNAs have been found to be associated with insecticide resistance.However,the characteristics and biological func-tions of lncRNAs involved in indoxacarb resistance are unknown in Spodoptera litura.We performed RNA sequencing in the SS,InRS,and FInRS of S.litura and identified 11978 lncRNAs,including 3136 intergenic lncRNAs,7393 intronic lncRNAs,and 1449 anti-sense lncRNAs.Compared with the SS,51 lncRNAs were upregulated and 134 lncRNAs were downregulated in the two resistant strains,and 908 differentially expressed mRNAs were predicted as the target genes of the 185 differentially expressed lncRNAs.Further analysis showed that 112 of differentially expressed lncRNAs may be associated with in-doxacarb resistance by regulating the expression of 14 P450s,seven CCEs,one GST,six UGTs,five ABC transporters,and 24 cuticle protein genes,and 79 of differentially ex-pressed lncRNAs may regulate the expression of 14 detoxification genes and 19 cuticle protein genes to participate in indoxacarb resistance by sponging 10 microRNAs.Interest-ingly,47 of differentially expressed lncRNAs may mediate indoxacarb resistance through both lncRNA-mRNA and lncRNA-miRNA-mRNA regulatory pathways.Furthermore,quantitative PCR,RNA interference,and indoxacarb bioassay analyses indicated that over-expressed LNC_004867 and LNC_006576 were involved in indoxacarb resistance.This study provides comprehensive information for lncRNAs of S.litura,and presents evidence that lncRNAs havekey roles in conferring insecticide resistance in S.litura.

关键词： indoxacarb resistance long noncoding RNAs regulatory pathways RNA sequencing Spodopteralitura

Change of 3 Kinds of Enzyme Activity and Soluble Sugar in 3 Chinese Cabbage Varieties with Different Resistance Level During the Process of Plasmodiophora brassicae Infecting Their Roots

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Agricultural Science & Technology 2015年第2期16卷 205-208页

作者： Zuohua REN Hu ZHOU Xiaoli HUANG Qiong SONG Minjie LIU Erming LIU College of plant Protection Hunan Agricultural University Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests College of Information Science and Technology Hunan Agricultural University

There are different levels of resistance in Chinese cabbage varieties against clubroot.The content of catalase(CAT),peroxidase(POD),phenylalanin ammonia-lyase(PAL),Super Oxide Dismutase(SOD)and soluble sugar activities of 3cultivars with different clubroot-resistant levelswere detected after inoculation in this study.The results suggest that the changing rates of CAT,PAL and SOD contents of resistance cultivars after inoculation were higher than those of susceptible cultivars(Baigengbaiye>Zaobaicaitai>Huangjinxiaobaicai);the POD activities of resistance cultivars reached a peak on an earlier day than those of susceptible cultivars;the increasing rate of soluble sugar of susceptible cultivars was higher than that of the resistance cultivars,and ended with two peaks.Therefore,the four enzymes(CAT,POD,PAL and SOD)and soluble sugar may be used as physiological and biochemical reference indexes for the resistance identification to clubroot after inoculation with P.brassicae.

关键词：抗性品种可溶性糖大白菜酶活性甘蓝中国 POD活性苯丙氨酸解氨酶