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In Vitro Infectious Risk Assessment of Heliothis virescens ascovirus 3j(HvAV-3j) toward Non-target Vertebrate Cells

Virologica Sinica 2019年第4期34卷 423-433页

作者： Huan Yu Yi-Yi Ou-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 41012& China College of Plant Protection Hunan Agricultural UniversityChangsha 41012& China Tokyo University of Agriculture and Technology SaiwaiFuchuTokyo 183-8509Japan

As specific pathogens of noctuid pests,including Spodoptera exigua,S.litura,Helicoverpa armigera,and Mythimna separata,ascoviruses are suitable for the development of bioinsecticides.In this study,the infectivity of Heliothis virescens ascovirus 3j(HvAV-3j)on insect and mammalian cells was evaluated.HvAV-3j infection induced drastic morphological changes in Sf9,HzAMl,SeFB,and HaFB cells,including swelling and detachmen Notably,the latter phenomena did not occur in HvAV-3j-inoculated mammalian cells(HEK293,7402,HePG2,PK15,ST,and TM3).MTT assays indicated that HvAV-3j inhibited the growth of host insect cells from the 6th hpi,but no effects were detected in the HvAV-3j-inoculated mammalian cells.Furthermore,viral DNA replication,gene transcription,and protein expression were investigated,and the results consistently suggested that HvAV-3j viruses were not able to replicate their genomic DNA,transcribe,or express their proteins in the non-target vertebrate cells.The HvAV-3j genes were only transcribed and expressed in the four insect cell lines.These results indicated that HvAV-3j was infectious to cells derived from S.frugiperda,S.exigua,H.armigera,and H.zea but not to cells derived from human,pig,and mouse,suggesting that ascoviruses are safe to nontarget vertebrate cells.

关键词： Infectious risk Heliothis virescens ascovirus 3j(HvAV-3j) Mammalian cells Insect cells

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Genome Analysis of Dasineura jujubifolia Toursvirus 2,A Novel ascovirus

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Virologica Sinica 2020年第2期35卷 134-142页

作者： Jun Wang Minglu Yang Haibing Xiao Guo-Hua Huang Fei Deng Zhihong Hu State Key Laboratory of Virology and National Virus Resource Center Wuhan Institute of VirologyChinese Academy of SciencesWuhan 430071China Tarim University Alar 843300China Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests Institute of VirologyHunan Agricultural UniversityChangsha 410128China

So far,ascoviruses have only been identified from Lepidoptera host insects and their transmission vectors-endoparasitic wasps.Here,we reported the first finding of a complete novel ascovirus genome from a Diptera insect,Dasineura jujubifolia.Initially,sequence fragments with homology to ascoviruses were incidentally identified during metagenomic sequencing of the mitochondria of D.jujubifolia(Cecidomyiidae,Diptera)which is a major pest on Ziziphus jujuba.Then a full circular viral genome was assembled from the metagenomic data,which has an A+T percentage of 74%and contains 142,600 bp with 141 open reading frames(ORFs).Among the 141 ORFs,37 were conserved in all sequenced ascoviruses(core genes)including proteins predicted to participate in DNA replication,gene transcription,protein modification,virus assembly,lipid metabolism and apoptosis.Multi-gene families including those encode for baculovirus repeated open reading frames(BROs),myristylated membrane proteins,RING/U-box E3 ubiquitin ligases,and ATP-binding cassette(ABC)transporters were found in the virus genome.Phylogenetic analysis showed that the newly identified virus belongs to genus Toursvirus of Ascoviridae,and is therefore named as Dasineura jujubifolia toursvirus 2(DjTV-2a).The virus becomes the second reported species of the genus after Diadromus pulchellus toursvirus 1(DpTV-1a).The genome arrangement of DjTV-2a is quite different from that of DpTV-1a,suggesting these two viruses separated in an early time of evolution.The results suggest that the ascoviruses may infect a much broader range of hosts than our previous knowledge,and shed lights on the evolution of ascoviruses and particularly on that of the toursviruses.

关键词： Dasineura jujubifolia toursvirus 2(DjTV-2a) ascovirus Toursvirus

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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

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