两种林业昆虫杆状病毒PCR快速检测技术及其应用的研究
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摘要
林业昆虫病毒作为一种高效无害专一性强的杀虫剂,现在已经在森林保护中起到了很大的作用。但其在林业上的使用率还不够广泛,主要是因为病毒防治害虫时潜伏期较化学杀虫剂长,没有后者那么立竿见影的效果,使得使用者对病毒治虫有质疑。现今对昆虫病毒防治效果的研究还仅仅停留在对病毒的形态观测阶段,本实验旨在建立几种以昆虫病毒核酸为基础的检测技术,从而填补昆虫病毒检测在分子生物学方面的空白,同时应用这些技术检测在昆虫体内低水平存在的病毒,为昆虫病毒的垂直传播及防治效果监测提供微观的、确凿的分子生物学证据,进而论证应用昆虫病毒防治林业害虫的可行性,确保森林生态保持稳定。
根据HcNPV pe38基因设计两对引物,利用PCR法检测HcNPV基因组DNA,分别扩增出长为994和614 bp的片段,经测序确定为HcNPV pe38基因片段。应用两对引物分别检测了美国白蛾病虫的总DNA和获得的病毒多角体,其检测最低量分别为1 fg病虫总DNA和3~4 OBs/mL。用不同浓度的病毒感染试虫后,不同时间取其血淋巴用PCR方法检测。结果表明,接毒量为3.53×109 OBs/mL时,36 h后可检出病毒DNA;接毒量为3~4 OBs/mL时,120 h后可检出。病毒可检出时间随接毒浓度的递减而延后。
从秦皇岛地区施用过HcNPV的林地和未施用病毒防虫的林地采集美国白蛾野外幼虫,取其血淋巴做模板进行PCR扩增,结果十分明显,说明之前建立的方法完全可以应用于实践中,同时说明HcNPV对该地区的美国白蛾种群具有持续控制作用。
根据ClanGV egt基因设计两对引物,利用PCR方法检测ClanGV基因组DNA,分别扩增出401和491bp的片段,经测序确定为ClanGV egt基因片段。应用两对引物检测ClanGV病毒悬液,亦可成功扩增出目的基因片段,说明这两对特异性较高,可以用其检测ClanGV的存在,从而为昆虫病毒的野外检测奠定基础。
Forest insect virus as a highly efficient sound-specific insecticide, is played a significant role in the forest protection now. But it is not be used popularly, mainly because virus has a long latent period, its reaction are slower than chemical pesticides, and these questioned the users. This experiment aimed at establishing several insect virus nucleic acid-based detection technologies, which fill the blank of the insect virus detection in the molecular biology. Applied the technologies to detect low concentration of OB suspension can provide some evidence for vertical transmission and reaction effect of insect virus, it also can demonstrate the feasibility of controlling pest with insect virus.
Two pairs of specific primers were used for the detection of the Hyphantria cunea nucleopolyhedrovirus (HcNPV) by PCR. The two pairs of primers were both designed based on gene pe38 of HcNPV genome. The sizes of amplified fragments were 994 and 614 bp, respectively. The two pairs of primers were further used to detect the amount of viral DNA in diseased larvae or polyhedron of HcNPV. The minimum amount of detection could be as less as 1 fg of total DNA or 3?4 OBs/mL of polyhedron. The amount of viral DNA in hemolymph sampled from the larvae infected by different concentrations of polyhedron at different time points was also investigated, and it was found that the lower the concentration of polyhedron used, the later the viral DNA will be detected.
Larvae of Hyphantria cunea were collected from Qinhuangdao. Their hemolymph is detected by PCR, the effect is also obviously. The result shows that the PCR-based method can be applied to practice, and HcNPV has a continuous control to Hyphantria cunea.
Two pairs of specific primers were used for the detection of the Clostera anachoreta Granulovirus (ClanGV) by PCR. The two pairs of primers were both designed based on gene egt of ClanGV genome. The sizes of amplified fragments were 401 and 491 bp, respectively. The two pairs of primers were used to detect the genome DNA and OBs of ClanGV successfully.
根据HcNPV pe38基因设计两对引物,利用PCR法检测HcNPV基因组DNA,分别扩增出长为994和614 bp的片段,经测序确定为HcNPV pe38基因片段。应用两对引物分别检测了美国白蛾病虫的总DNA和获得的病毒多角体,其检测最低量分别为1 fg病虫总DNA和3~4 OBs/mL。用不同浓度的病毒感染试虫后,不同时间取其血淋巴用PCR方法检测。结果表明,接毒量为3.53×109 OBs/mL时,36 h后可检出病毒DNA;接毒量为3~4 OBs/mL时,120 h后可检出。病毒可检出时间随接毒浓度的递减而延后。
从秦皇岛地区施用过HcNPV的林地和未施用病毒防虫的林地采集美国白蛾野外幼虫,取其血淋巴做模板进行PCR扩增,结果十分明显,说明之前建立的方法完全可以应用于实践中,同时说明HcNPV对该地区的美国白蛾种群具有持续控制作用。
根据ClanGV egt基因设计两对引物,利用PCR方法检测ClanGV基因组DNA,分别扩增出401和491bp的片段,经测序确定为ClanGV egt基因片段。应用两对引物检测ClanGV病毒悬液,亦可成功扩增出目的基因片段,说明这两对特异性较高,可以用其检测ClanGV的存在,从而为昆虫病毒的野外检测奠定基础。
Forest insect virus as a highly efficient sound-specific insecticide, is played a significant role in the forest protection now. But it is not be used popularly, mainly because virus has a long latent period, its reaction are slower than chemical pesticides, and these questioned the users. This experiment aimed at establishing several insect virus nucleic acid-based detection technologies, which fill the blank of the insect virus detection in the molecular biology. Applied the technologies to detect low concentration of OB suspension can provide some evidence for vertical transmission and reaction effect of insect virus, it also can demonstrate the feasibility of controlling pest with insect virus.
Two pairs of specific primers were used for the detection of the Hyphantria cunea nucleopolyhedrovirus (HcNPV) by PCR. The two pairs of primers were both designed based on gene pe38 of HcNPV genome. The sizes of amplified fragments were 994 and 614 bp, respectively. The two pairs of primers were further used to detect the amount of viral DNA in diseased larvae or polyhedron of HcNPV. The minimum amount of detection could be as less as 1 fg of total DNA or 3?4 OBs/mL of polyhedron. The amount of viral DNA in hemolymph sampled from the larvae infected by different concentrations of polyhedron at different time points was also investigated, and it was found that the lower the concentration of polyhedron used, the later the viral DNA will be detected.
Larvae of Hyphantria cunea were collected from Qinhuangdao. Their hemolymph is detected by PCR, the effect is also obviously. The result shows that the PCR-based method can be applied to practice, and HcNPV has a continuous control to Hyphantria cunea.
Two pairs of specific primers were used for the detection of the Clostera anachoreta Granulovirus (ClanGV) by PCR. The two pairs of primers were both designed based on gene egt of ClanGV genome. The sizes of amplified fragments were 401 and 491 bp, respectively. The two pairs of primers were used to detect the genome DNA and OBs of ClanGV successfully.
引文
J.萨姆布鲁克,D. W.拉塞尔著.分子克隆实验指南黄培堂等译第三版北京:科学出版社,2002
曹广力,薛仁宇,朱越雄等.美国白蛾核型多角体病毒p35基因的克隆及序列分析昆虫学报,2002,45(6):711-716
曹广力,薛仁宇,朱越雄等.美国白蛾核型多角体病毒超氧化物歧化酶基因的序列分析和表达微生物学报,2001,41(2):173-180
曹晓红,唐英,郭惠.艾滋病病毒的核酸检测法渝西学院学报(自然科学版),2002年l2月,1(4):40-46
陈新文,胡志红.中国棉铃虫单粒包埋核型多角体病毒多角体蛋白基因的序列分析中国病毒学,1997,12(4):346-353
戴冠群,陈庆雄等.鸟类传播昆虫病毒的观察病毒学杂志,1986,1(4):56-64.
段彦丽.美国白蛾NPV与Bt混合致病机理及其对寄主种群持续控制作用,中国林业科学研究院博士论文,2008,1?2
范凌云,胡志红,Just M Vlak等.棉铃虫单核衣壳核多角体病毒解螺旋酶基因的序列分析中国病毒学,2001,16:344-349
龚和.我国昆虫分子生物学研究的回顾和展望昆虫知识,2000,37(1):32-37
龚岷,梁布锋,李暐东等.油桐尺蠖核型多角体病毒DNA聚合酶基因的克隆和序列分析中国病毒学,2000,15:59-65
巩红霞,任永宏,巩强.应用PCR-限制性内切酶技术快速检测副结核分枝杆菌畜牧兽医杂志,2006,25(4):25?27
贡成良,小林淳,宫岛成寿.美国白蛾核型多角体病毒几丁质酶基因核苷酸序列研究病毒学报,1999,15(3):260-269
贡成良,小林淳,宫岛成寿等.HcNPV半胱氨酸蛋白酶基因的核苷酸序列研究生物化学与生物物理学报,1998,30(3):307-310
贡成良,小林淳,金伟等.HcNPV半胱氨酸蛋白酶、几丁质酶基因失活分析生物化学与生物物理学报,2000,32(2):187-191
洪华珠,杨红.杀虫微生物学纲要武汉:武汉师范大学出版社,1997,32-82
洪健,周雪平.ICTV第八次报告的最新病毒分类系统中国病毒学,2006年1月,21(1):84-96
洪靖君,段家龙,彭辉银.昆虫杆状病毒若干基因的研究进展中国病毒学,2002年8月,17 (3):270-275
胡远扬.昆虫病毒研究的回顾与展望中国病毒学,2004年6月,19(3):303-308
李辉,汤历,陈其津.昆虫病毒的研究进展安徽农学通报,2007,13(13):150-152
李家荣.我国有害生物综合治理(IPM)的进展植物医生,1999年4月,12(2):2-3
李荣森.我国微生物防治研究与微生物农药产业化的近展中国病毒学,2000,15(SI):1-14
梁革梅,谭维嘉,郭予远.人工饲养棉铃虫技术的改进植物保护,1999,(2):15-17
刘岱岳,居蜀生.美国白蛾病毒的繁殖与飞机喷施生物防治通报,1986,2(2):74
刘高强,余晓丹.昆虫杆状病毒分子生物学及其应用研究的新进展安徽农业科学,2006,34(9):1748?1750
刘祖强,杨复华,齐义鹏等.杆状病毒的垂直传播及绿色荧光蛋白在棉铃虫幼虫中的表达昆虫学报,2001,44(1):1-9
吕鸿声.昆虫病毒与昆虫病毒病北京:科学出版社,1982
彭建新.杆状病毒分子生物学武汉:华中师范大学出版社,2000,108?109
彭建新,陈曲侯.杆状病毒DNA复制研究的某些进展病毒学报,1995,11(1):93?97
蒲蛰龙.昆虫病理学广州:广东科技出版社,1994
乔鲁芹.美国白蛾核型多角体病毒及其感病寄主检测方法的研究北京:中国林业科学研究院博士论文,2007,58?59
裘卫等.应用昆虫杆状防治害虫生物防治通报,1987,3(1):39-43
曲良建,张永安,王玉珠等.PCR法证实棉铃虫核型多角体病毒对棉铃虫经卵和蛹的垂直传播中国生物防治,2005,21(1):45?48
邵碧英.植物病毒分子检测方法概述植物检疫,2002,16(6):377-379.
沈兴家,张志芳,李奕仁.昆虫杆状病毒egt基因研究进展中国蚕业,2004年5月,25(2):4-7
盛晔.斜纹夜蛾核型多角体病毒日本株ie0、ie1基因和25k基因结构和功能的初步研究苏州大学硕士学位论文,2007,2-7
苏志坚,庞义,余健秀等.污染寄主卵面的斜纹夜蛾核多角体病毒PCR检测及其消除农业生物技术学报,2001,9(2):119?122
孙彤.生物防治在有害生物综合治理上的应用现状及前景辽宁林业科技,2002年,(4):36-38
孙修炼,胡志红,彭辉银.杆状病毒生态学研究进展中国病毒学,2006,12(2):200-206
汤承,索化夷,岳华等.SYBR Green I实时定量PCR快速检测鸭瘟病毒的研究中国动物检疫,2006,23(9):25-28
汤显春,彭辉银,张小霞等.分月扇舟蛾颗粒体病毒的形态和限制性内切酶图谱分析华中农业大学学报,2004,(6):616-619
田克恭.多聚酶链反应在实验动物病毒检测中的应用中国兽医杂志,1997,23(8):56-57
王根,吴祥甫等.中国棉铃虫核型多角体DNA聚合酶基因的克隆和序列分析中国病毒学, 1998,13(1):77-82
王华林,胡志红,孙修炼等.棉铃虫核多角体病毒细胞调亡抑制基因iap3的序列分析中国病毒学,2000,15(杀虫微生物专刊):43-49
王娜,马雅军,王喆之等.PCR相关技术方法在植物病害检测中的应用(综述)上海农业学报, 2004,20 (4):112-115
王庆礼,吴刚.论中国林业现状及可持续发展战略中国人口·资源与环境,1997年l2月,7(4):38-43
王文兵,张志芳,何家禄等.杆状病毒基因组DNA复制相关基因的研究进展生物化学与生物物理进展,2000,27(3):257?261
王新生,郭复明,刘炳龙等.杨扇舟蛾综合防治技术初探安徽林业科技,2004,(1):17
魏建荣,王传珍,杨隽等.美国白蛾卵块及幼虫网幕空间格局研究林业科学研究,2004,17(4):500～50
徐培渝,Jerlin Walke. PCR和限制性内切酶法快速检测C57BL/6J ob小鼠基因型癌变?畸变?突变,2000,12(3):143?145
阎庆生,庞义,农广等.斜纹夜蛾核多角体病毒egt基因的克隆和部分序列分析中山大学学报(自然科学版),1998年5月,37(3):125-127
杨苗苗,李孟楼,曲良建等.南华松叶蜂病毒DnNPV的发现及其毒力测试林业科学,2007,43(7): 138?141
杨忠岐.中国寄生于美国白蛾的啮小蜂一新属一新种昆虫分类学报,1989,11(1-2):117-130
杨忠岐,张永安.重大外来入侵害虫--美国白蛾生物防治技术研究昆虫知识,2007,44(4):465-471
姚东伟,郑宇,张光星.病毒杀虫剂的研究现状北方园艺,2008,(3):69-72
叶恭银,朱俊庆,胡萃.茶尺蠖核型多角体病毒对宿主种群的控制作用植物保护学报,1994,21(3):231-237
张传溪,胡萃,吴祥甫等.茶尺蠖核型多角体病毒polh基因核苷酸序列及其在Baculoviridae中的地位中国生物化学与分子生物学报,1999,(15):365-371
张春龙,纪晓明,潘淑琴等.颗粒体病毒大面积防治杨扇舟蛾试验吉林林业科技,1992年12月,(6):33-35
张康普,黄维正,王平等.杨扇舟蛾天敌种类调查中国森林病虫,2003年3月,22(2):12-14
张小霞,梁振普,彭辉银等.杨扇舟蛾颗粒体病毒gra基因的分析中国病毒学,2005年12月,20(6):660-663
张小霞,张忠信,丁清泉.棉铃虫核多角体病毒iap2基因的克隆和序列分析中国病毒学,2001,(16):338-343
张永安,王贵成,居来提.新疆和田地区利用AciNPV病毒防治春尺蠖试验报告新疆林业,2002,(2):20-21
张永安,曲良建,王玉珠.利用昆虫病毒恢复和构建稳定森林生态系统的理论与实践中国农业科技导报,2007,9(5):45-52
张宗炳,曹骥.害虫防治:策略与方法北京:科学出版社,1999
赵耘,秦玉明,张广川等.RT-PCR和酶切方法区分猪瘟疫苗毒与野毒的研究微生物学通报,2006,33(3):82-87
Ahrens C H, Russell R L, Funk CJ, et al. The sequence of Orgyia pseudotsugata nuclear polyhedrosis virus genome. Virology, 1997, 229: 381-399
Arif B M. A brief journey with insect viruses with emphasis on baculoviruses. Journal of Invertebrate Pathology, 2005, 89: 39?45
Beisner B E, Myers J H. Population density and transmission of virus in experimental populations of the western tent caterpillar (Lepidoptera:Lasiocampidae). Environ Entomol, 1999, 28: 1107-1113
Black B C, B rennan L A, Dierks P M, et al. Commercialization of baculoviral insecticides in baculoviruses.Edited by Lois, K .Miller. Plenum Press. New York.1997
Blissard G., G.. Rohrmann. Baculovirus diversity and molecular biology. Annu Rev. Entmo1, 1990, 35: 127-155
Bonning B C, Hammock B D , Miller L K. Development and potential of genetically engineered insecticides. Biotechnol Genel Eng Rev, 1992, 10:455-489
Bud H M,Kelly D C. The DNA Contained by Nuclear Polyhedrosis Viruses Isolated from Four Spodoptera spp. (Lepidoptera, Noctuidae); Genome Size and Configuration Assessed by Electron Microscopy. Journal of General Virology, 1977, 37: 135-143
Burden J Griffiths C M, Cory J S, et a1. Vertical transmission of sublethal granuloviru infection in the Indian meal moth, Plodia interpunctella. Mol Ecol, 2002, 11: 547-555
Charpentier G, Desmarteaux D, Bourassa JP et al. Utilization of the polymerase chain reaction in the diagnosis of nuclear polyhedrosis virus infections of gypsy moth (Lymantria dispar, Lep., Lymantriidae) populations. J. Appl. Ent., 2003, 127: 405?412
Crozier R H, Crozier Y C. The cytochrome b and ATPase genes of honeybee mitochondrial DNA. Molecular Biology and Evolution, 1992, 9(3): 474-482
Dwyer G, Elkinton J S, Buonaccorsi J P. Host heterogeneity in susceptibility and disease dynamics: Tests of a mathematical model. Am Nat, 1997, 150:685-707
Freemont P S, Hanson I M, Trowsdale J. A novel cysteine– rich sequence motif. Cell, 1991, 64: 483-484
Furuta M., Schrader J O, Schrader H S, et a1.Chlorella virus PBCV-1 encodes a homolog of the bacteriophage T4 UV damage repair gene denV.Appl Environ Microbiol, 1997, 63:1551-1556
Fuxa J R, Richter A R.Quantification of soil-to-plant transport of recombinant nucleopolyhedrovirus: effects of soil type and moisture, air currents, and precipitation.Appl Environ Microbiol, 2001, 67:5166-5170
G. V. P, Reddy, M. Manjunatha. Laboratory and field studies on the integrated pest management of Helicoverpa armigera (Huèbner) in cotton, based on pheromone trap catch threshold level. J. Appl. Ent, 2000, (124): 213-221
Gomi S, Majima K, Maeda S, et al. Sequence analysis of the genome of Bombyx mori nuclear polyhedrosis virus. J Gen Virol, 1999, 80: 1323-1337
Hang X, Linda A G. Purification of Autographa califotnica nucleopolyhedrovirus DNA polymerase from infected insect cells. J Gen Virol, 1999, 80: 2519 - 2526
Herniou E. A., et al. The genenome sequence and evolution of baculoviruses. Annu. Rev. Entomol, 2003, 48: 211-234
Huang J, Levin D B. Identification, transcription and sequence analysis of the Spodoptera littoralis nucleopolyhedrovirus DNA polymerase gene. Arch Virol, 2001, 146: 303-326
Hunter-Fujita F R, Entwistle P F, Evans H F, et a1. Insect viruses and pest management.John Wiley and Sons, 1998
Hyung-Hoan Lee, Hyun-Jae Lee, Kwan-Hee Yoo. Restriction map of the Hyphantria cunea nuclear polyhedrosis virus. J. of Kor. Soc. of Virology, 1990, 20(2): 145-153
Ijkel W F, Van Strien E A, Heldens J GM, et al. Sequence and organization of the Spodoptera exigua multicapsid nucleopolyhedrovirus genome. J Gen Virol, 1999, 80: 3289-3304
Ikeda M, Shikata M, Shirata N, Chaeychomsri S, Kobayashi M. Gene organization and complete sequence of the Hyphantria cunea nucleopolyhedrovirus genome. J.Gen.Virol., 2006, 87(Pt 9): 2549?2562
Kaupp. W. J., Ebing.P.M. horseradish peroxidase labelled probes and enhanced chemilumin escence to detect baculoviruses in gypsy moth and easten spruce budworm larvae. J.Virol.Methods, 1993, 44:89-98
Keating S.T, Burand J.P., Elkinton J. S. DNA hybridization assay for detection of gypsy moth nuclear polyhedrosis virus in infected gypsy moth( Lymantria dispar L.) larvae. Appl. Environ.Microbiol, 1989, 55:2749-2754
Knell R J, Begon M, Thompson D J. Transmission of Plodia interpunctella granulosis virus does not conform to the mass action model. J Anim Ecol, 1998, 67: 592-599
Kukan B. Vertical transmission of nucleopolyhedrovirus in insects. J.Invertebr.Pathol., 1999, 74:149?155
Kukan B., Myers J.H. DNA hybridization assay for detection of nuclear polyhedrsis virus in tent caterpillars. J.Invertebr.Pathol, 1995, 66:231-236
Kuzio J, Pearson M N, Harwood S H, et al. Sequence and analysis of the genome of a baculovirus pathogenic for Lymantria dispar. Virology, 1999, 253: 17 - 34
Maguire T, Harrison P, Hyink O, et al. The inhibitors of apoptosis of Epiphyas post vittana nucleopolyhedrovirus. J Gen Virol , 2000, 81: 2803-2811
Maruniak J E, Brown S E, L.Knudson D. Physical maps of SfMNPV baculovirus DNA and its genomic variants. Virology, 1984, 136(1): 221-234
Moscardi F. Assessment of the application of baculoviruses for control of Lepidoptera. Annu Rev Entomol, 1999, 44:257-289
Pringle C R. The universal system of virus taxonomy, updated to include the new proposals ratified by the International Committee on Taxonomy of Viruses during 1998. Archives of Virology, 1999, 144(2):421-429.
S. Ignacimuthu, S. Jayaraj .Biotechnology and insect pest management. *A report on the two-day National Symposium on Biotechnology and Insect Pest anagement conducted at the Entomology Research Institute, Loyola College, Chennai during 2–3 February 2006. CURRENT SCIENCE, VOL. 90, NO. 7, 10 APRIL 2006
Smith I R, Crook N E. In vivo isolation of baculovirus genotypes. Virology, 1988, 166(1): 240-244
Stiles B, Himmerich S. Autographa californica NPV Isolates: Restriction Endonuclease Analysis and Comparative Biological Activity. Journal of Invertebrate Pathology, 1998, 72(2): 174-177
Summers M D, Anderson D L. Characterization of deoxyribonucleic acid isolated from the granulosis viruses of the cabbage looper, Trichoplusia ni and the fall armyworm, Spodoptera frugiperda. Virology, 1972, 50(2):459-471
Summers M D, Anderson D L. Granulosis virus deoxyribonucleic acid: a closed, double-stranded molecule. Journal of Virology, 1972, 9: 710-713
Sun X L, van der Werf, W.Bianchi F J J A, et a1. Modelling biological control with wild-type and genetically modified baculoviruses in the Helicoverpa armigera-coton system. Ecol model, 2006, in press
T. Hukuhara, H. Namura. Distribution of a Nuelear-PolyhedrosisVirus of the fall Webworm, Hyphantria cunea, in soil. Journal of Invertebrate Pathology, 1972, 19: 308-316
Van Regenmorte M H V, Fauquet C M, Bishop D H L, et a1. Virus Taxonomy Seventh Report of the International Committee on Taxonomy of Virus.New York, San Diego: Academic Press, 2000
Zhou M Z, Sun X C, Hu Z H (周明哲,孙新城,胡志红) et a1.SOD Enhances Infectivity of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus against H. armigra larvae. Virologica Sinica(中国病毒学), 2004, 18:506-507
Zhou M Z, Sun X L, Sun X C, et a1. Horizontal and vertical transmission of wild-type and recombinant
Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus. J Invertebr Pathol, 2005, 89: l65-175
http://www.hebly.gov.cn/showarticle.php?id=5307(美国白蛾在秦皇岛的发生)
http://www.zcny.gov.cn/Article/bzh/200807/1453.htm(农药危害情况)
http://baike.baidu.com/view/93663.htm(美国白蛾简介)
http://baike.baidu.com/view/1919392.htm(杨扇舟蛾简介)
曹广力,薛仁宇,朱越雄等.美国白蛾核型多角体病毒p35基因的克隆及序列分析昆虫学报,2002,45(6):711-716
曹广力,薛仁宇,朱越雄等.美国白蛾核型多角体病毒超氧化物歧化酶基因的序列分析和表达微生物学报,2001,41(2):173-180
曹晓红,唐英,郭惠.艾滋病病毒的核酸检测法渝西学院学报(自然科学版),2002年l2月,1(4):40-46
陈新文,胡志红.中国棉铃虫单粒包埋核型多角体病毒多角体蛋白基因的序列分析中国病毒学,1997,12(4):346-353
戴冠群,陈庆雄等.鸟类传播昆虫病毒的观察病毒学杂志,1986,1(4):56-64.
段彦丽.美国白蛾NPV与Bt混合致病机理及其对寄主种群持续控制作用,中国林业科学研究院博士论文,2008,1?2
范凌云,胡志红,Just M Vlak等.棉铃虫单核衣壳核多角体病毒解螺旋酶基因的序列分析中国病毒学,2001,16:344-349
龚和.我国昆虫分子生物学研究的回顾和展望昆虫知识,2000,37(1):32-37
龚岷,梁布锋,李暐东等.油桐尺蠖核型多角体病毒DNA聚合酶基因的克隆和序列分析中国病毒学,2000,15:59-65
巩红霞,任永宏,巩强.应用PCR-限制性内切酶技术快速检测副结核分枝杆菌畜牧兽医杂志,2006,25(4):25?27
贡成良,小林淳,宫岛成寿.美国白蛾核型多角体病毒几丁质酶基因核苷酸序列研究病毒学报,1999,15(3):260-269
贡成良,小林淳,宫岛成寿等.HcNPV半胱氨酸蛋白酶基因的核苷酸序列研究生物化学与生物物理学报,1998,30(3):307-310
贡成良,小林淳,金伟等.HcNPV半胱氨酸蛋白酶、几丁质酶基因失活分析生物化学与生物物理学报,2000,32(2):187-191
洪华珠,杨红.杀虫微生物学纲要武汉:武汉师范大学出版社,1997,32-82
洪健,周雪平.ICTV第八次报告的最新病毒分类系统中国病毒学,2006年1月,21(1):84-96
洪靖君,段家龙,彭辉银.昆虫杆状病毒若干基因的研究进展中国病毒学,2002年8月,17 (3):270-275
胡远扬.昆虫病毒研究的回顾与展望中国病毒学,2004年6月,19(3):303-308
李辉,汤历,陈其津.昆虫病毒的研究进展安徽农学通报,2007,13(13):150-152
李家荣.我国有害生物综合治理(IPM)的进展植物医生,1999年4月,12(2):2-3
李荣森.我国微生物防治研究与微生物农药产业化的近展中国病毒学,2000,15(SI):1-14
梁革梅,谭维嘉,郭予远.人工饲养棉铃虫技术的改进植物保护,1999,(2):15-17
刘岱岳,居蜀生.美国白蛾病毒的繁殖与飞机喷施生物防治通报,1986,2(2):74
刘高强,余晓丹.昆虫杆状病毒分子生物学及其应用研究的新进展安徽农业科学,2006,34(9):1748?1750
刘祖强,杨复华,齐义鹏等.杆状病毒的垂直传播及绿色荧光蛋白在棉铃虫幼虫中的表达昆虫学报,2001,44(1):1-9
吕鸿声.昆虫病毒与昆虫病毒病北京:科学出版社,1982
彭建新.杆状病毒分子生物学武汉:华中师范大学出版社,2000,108?109
彭建新,陈曲侯.杆状病毒DNA复制研究的某些进展病毒学报,1995,11(1):93?97
蒲蛰龙.昆虫病理学广州:广东科技出版社,1994
乔鲁芹.美国白蛾核型多角体病毒及其感病寄主检测方法的研究北京:中国林业科学研究院博士论文,2007,58?59
裘卫等.应用昆虫杆状防治害虫生物防治通报,1987,3(1):39-43
曲良建,张永安,王玉珠等.PCR法证实棉铃虫核型多角体病毒对棉铃虫经卵和蛹的垂直传播中国生物防治,2005,21(1):45?48
邵碧英.植物病毒分子检测方法概述植物检疫,2002,16(6):377-379.
沈兴家,张志芳,李奕仁.昆虫杆状病毒egt基因研究进展中国蚕业,2004年5月,25(2):4-7
盛晔.斜纹夜蛾核型多角体病毒日本株ie0、ie1基因和25k基因结构和功能的初步研究苏州大学硕士学位论文,2007,2-7
苏志坚,庞义,余健秀等.污染寄主卵面的斜纹夜蛾核多角体病毒PCR检测及其消除农业生物技术学报,2001,9(2):119?122
孙彤.生物防治在有害生物综合治理上的应用现状及前景辽宁林业科技,2002年,(4):36-38
孙修炼,胡志红,彭辉银.杆状病毒生态学研究进展中国病毒学,2006,12(2):200-206
汤承,索化夷,岳华等.SYBR Green I实时定量PCR快速检测鸭瘟病毒的研究中国动物检疫,2006,23(9):25-28
汤显春,彭辉银,张小霞等.分月扇舟蛾颗粒体病毒的形态和限制性内切酶图谱分析华中农业大学学报,2004,(6):616-619
田克恭.多聚酶链反应在实验动物病毒检测中的应用中国兽医杂志,1997,23(8):56-57
王根,吴祥甫等.中国棉铃虫核型多角体DNA聚合酶基因的克隆和序列分析中国病毒学, 1998,13(1):77-82
王华林,胡志红,孙修炼等.棉铃虫核多角体病毒细胞调亡抑制基因iap3的序列分析中国病毒学,2000,15(杀虫微生物专刊):43-49
王娜,马雅军,王喆之等.PCR相关技术方法在植物病害检测中的应用(综述)上海农业学报, 2004,20 (4):112-115
王庆礼,吴刚.论中国林业现状及可持续发展战略中国人口·资源与环境,1997年l2月,7(4):38-43
王文兵,张志芳,何家禄等.杆状病毒基因组DNA复制相关基因的研究进展生物化学与生物物理进展,2000,27(3):257?261
王新生,郭复明,刘炳龙等.杨扇舟蛾综合防治技术初探安徽林业科技,2004,(1):17
魏建荣,王传珍,杨隽等.美国白蛾卵块及幼虫网幕空间格局研究林业科学研究,2004,17(4):500～50
徐培渝,Jerlin Walke. PCR和限制性内切酶法快速检测C57BL/6J ob小鼠基因型癌变?畸变?突变,2000,12(3):143?145
阎庆生,庞义,农广等.斜纹夜蛾核多角体病毒egt基因的克隆和部分序列分析中山大学学报(自然科学版),1998年5月,37(3):125-127
杨苗苗,李孟楼,曲良建等.南华松叶蜂病毒DnNPV的发现及其毒力测试林业科学,2007,43(7): 138?141
杨忠岐.中国寄生于美国白蛾的啮小蜂一新属一新种昆虫分类学报,1989,11(1-2):117-130
杨忠岐,张永安.重大外来入侵害虫--美国白蛾生物防治技术研究昆虫知识,2007,44(4):465-471
姚东伟,郑宇,张光星.病毒杀虫剂的研究现状北方园艺,2008,(3):69-72
叶恭银,朱俊庆,胡萃.茶尺蠖核型多角体病毒对宿主种群的控制作用植物保护学报,1994,21(3):231-237
张传溪,胡萃,吴祥甫等.茶尺蠖核型多角体病毒polh基因核苷酸序列及其在Baculoviridae中的地位中国生物化学与分子生物学报,1999,(15):365-371
张春龙,纪晓明,潘淑琴等.颗粒体病毒大面积防治杨扇舟蛾试验吉林林业科技,1992年12月,(6):33-35
张康普,黄维正,王平等.杨扇舟蛾天敌种类调查中国森林病虫,2003年3月,22(2):12-14
张小霞,梁振普,彭辉银等.杨扇舟蛾颗粒体病毒gra基因的分析中国病毒学,2005年12月,20(6):660-663
张小霞,张忠信,丁清泉.棉铃虫核多角体病毒iap2基因的克隆和序列分析中国病毒学,2001,(16):338-343
张永安,王贵成,居来提.新疆和田地区利用AciNPV病毒防治春尺蠖试验报告新疆林业,2002,(2):20-21
张永安,曲良建,王玉珠.利用昆虫病毒恢复和构建稳定森林生态系统的理论与实践中国农业科技导报,2007,9(5):45-52
张宗炳,曹骥.害虫防治:策略与方法北京:科学出版社,1999
赵耘,秦玉明,张广川等.RT-PCR和酶切方法区分猪瘟疫苗毒与野毒的研究微生物学通报,2006,33(3):82-87
Ahrens C H, Russell R L, Funk CJ, et al. The sequence of Orgyia pseudotsugata nuclear polyhedrosis virus genome. Virology, 1997, 229: 381-399
Arif B M. A brief journey with insect viruses with emphasis on baculoviruses. Journal of Invertebrate Pathology, 2005, 89: 39?45
Beisner B E, Myers J H. Population density and transmission of virus in experimental populations of the western tent caterpillar (Lepidoptera:Lasiocampidae). Environ Entomol, 1999, 28: 1107-1113
Black B C, B rennan L A, Dierks P M, et al. Commercialization of baculoviral insecticides in baculoviruses.Edited by Lois, K .Miller. Plenum Press. New York.1997
Blissard G., G.. Rohrmann. Baculovirus diversity and molecular biology. Annu Rev. Entmo1, 1990, 35: 127-155
Bonning B C, Hammock B D , Miller L K. Development and potential of genetically engineered insecticides. Biotechnol Genel Eng Rev, 1992, 10:455-489
Bud H M,Kelly D C. The DNA Contained by Nuclear Polyhedrosis Viruses Isolated from Four Spodoptera spp. (Lepidoptera, Noctuidae); Genome Size and Configuration Assessed by Electron Microscopy. Journal of General Virology, 1977, 37: 135-143
Burden J Griffiths C M, Cory J S, et a1. Vertical transmission of sublethal granuloviru infection in the Indian meal moth, Plodia interpunctella. Mol Ecol, 2002, 11: 547-555
Charpentier G, Desmarteaux D, Bourassa JP et al. Utilization of the polymerase chain reaction in the diagnosis of nuclear polyhedrosis virus infections of gypsy moth (Lymantria dispar, Lep., Lymantriidae) populations. J. Appl. Ent., 2003, 127: 405?412
Crozier R H, Crozier Y C. The cytochrome b and ATPase genes of honeybee mitochondrial DNA. Molecular Biology and Evolution, 1992, 9(3): 474-482
Dwyer G, Elkinton J S, Buonaccorsi J P. Host heterogeneity in susceptibility and disease dynamics: Tests of a mathematical model. Am Nat, 1997, 150:685-707
Freemont P S, Hanson I M, Trowsdale J. A novel cysteine– rich sequence motif. Cell, 1991, 64: 483-484
Furuta M., Schrader J O, Schrader H S, et a1.Chlorella virus PBCV-1 encodes a homolog of the bacteriophage T4 UV damage repair gene denV.Appl Environ Microbiol, 1997, 63:1551-1556
Fuxa J R, Richter A R.Quantification of soil-to-plant transport of recombinant nucleopolyhedrovirus: effects of soil type and moisture, air currents, and precipitation.Appl Environ Microbiol, 2001, 67:5166-5170
G. V. P, Reddy, M. Manjunatha. Laboratory and field studies on the integrated pest management of Helicoverpa armigera (Huèbner) in cotton, based on pheromone trap catch threshold level. J. Appl. Ent, 2000, (124): 213-221
Gomi S, Majima K, Maeda S, et al. Sequence analysis of the genome of Bombyx mori nuclear polyhedrosis virus. J Gen Virol, 1999, 80: 1323-1337
Hang X, Linda A G. Purification of Autographa califotnica nucleopolyhedrovirus DNA polymerase from infected insect cells. J Gen Virol, 1999, 80: 2519 - 2526
Herniou E. A., et al. The genenome sequence and evolution of baculoviruses. Annu. Rev. Entomol, 2003, 48: 211-234
Huang J, Levin D B. Identification, transcription and sequence analysis of the Spodoptera littoralis nucleopolyhedrovirus DNA polymerase gene. Arch Virol, 2001, 146: 303-326
Hunter-Fujita F R, Entwistle P F, Evans H F, et a1. Insect viruses and pest management.John Wiley and Sons, 1998
Hyung-Hoan Lee, Hyun-Jae Lee, Kwan-Hee Yoo. Restriction map of the Hyphantria cunea nuclear polyhedrosis virus. J. of Kor. Soc. of Virology, 1990, 20(2): 145-153
Ijkel W F, Van Strien E A, Heldens J GM, et al. Sequence and organization of the Spodoptera exigua multicapsid nucleopolyhedrovirus genome. J Gen Virol, 1999, 80: 3289-3304
Ikeda M, Shikata M, Shirata N, Chaeychomsri S, Kobayashi M. Gene organization and complete sequence of the Hyphantria cunea nucleopolyhedrovirus genome. J.Gen.Virol., 2006, 87(Pt 9): 2549?2562
Kaupp. W. J., Ebing.P.M. horseradish peroxidase labelled probes and enhanced chemilumin escence to detect baculoviruses in gypsy moth and easten spruce budworm larvae. J.Virol.Methods, 1993, 44:89-98
Keating S.T, Burand J.P., Elkinton J. S. DNA hybridization assay for detection of gypsy moth nuclear polyhedrosis virus in infected gypsy moth( Lymantria dispar L.) larvae. Appl. Environ.Microbiol, 1989, 55:2749-2754
Knell R J, Begon M, Thompson D J. Transmission of Plodia interpunctella granulosis virus does not conform to the mass action model. J Anim Ecol, 1998, 67: 592-599
Kukan B. Vertical transmission of nucleopolyhedrovirus in insects. J.Invertebr.Pathol., 1999, 74:149?155
Kukan B., Myers J.H. DNA hybridization assay for detection of nuclear polyhedrsis virus in tent caterpillars. J.Invertebr.Pathol, 1995, 66:231-236
Kuzio J, Pearson M N, Harwood S H, et al. Sequence and analysis of the genome of a baculovirus pathogenic for Lymantria dispar. Virology, 1999, 253: 17 - 34
Maguire T, Harrison P, Hyink O, et al. The inhibitors of apoptosis of Epiphyas post vittana nucleopolyhedrovirus. J Gen Virol , 2000, 81: 2803-2811
Maruniak J E, Brown S E, L.Knudson D. Physical maps of SfMNPV baculovirus DNA and its genomic variants. Virology, 1984, 136(1): 221-234
Moscardi F. Assessment of the application of baculoviruses for control of Lepidoptera. Annu Rev Entomol, 1999, 44:257-289
Pringle C R. The universal system of virus taxonomy, updated to include the new proposals ratified by the International Committee on Taxonomy of Viruses during 1998. Archives of Virology, 1999, 144(2):421-429.
S. Ignacimuthu, S. Jayaraj .Biotechnology and insect pest management. *A report on the two-day National Symposium on Biotechnology and Insect Pest anagement conducted at the Entomology Research Institute, Loyola College, Chennai during 2–3 February 2006. CURRENT SCIENCE, VOL. 90, NO. 7, 10 APRIL 2006
Smith I R, Crook N E. In vivo isolation of baculovirus genotypes. Virology, 1988, 166(1): 240-244
Stiles B, Himmerich S. Autographa californica NPV Isolates: Restriction Endonuclease Analysis and Comparative Biological Activity. Journal of Invertebrate Pathology, 1998, 72(2): 174-177
Summers M D, Anderson D L. Characterization of deoxyribonucleic acid isolated from the granulosis viruses of the cabbage looper, Trichoplusia ni and the fall armyworm, Spodoptera frugiperda. Virology, 1972, 50(2):459-471
Summers M D, Anderson D L. Granulosis virus deoxyribonucleic acid: a closed, double-stranded molecule. Journal of Virology, 1972, 9: 710-713
Sun X L, van der Werf, W.Bianchi F J J A, et a1. Modelling biological control with wild-type and genetically modified baculoviruses in the Helicoverpa armigera-coton system. Ecol model, 2006, in press
T. Hukuhara, H. Namura. Distribution of a Nuelear-PolyhedrosisVirus of the fall Webworm, Hyphantria cunea, in soil. Journal of Invertebrate Pathology, 1972, 19: 308-316
Van Regenmorte M H V, Fauquet C M, Bishop D H L, et a1. Virus Taxonomy Seventh Report of the International Committee on Taxonomy of Virus.New York, San Diego: Academic Press, 2000
Zhou M Z, Sun X C, Hu Z H (周明哲,孙新城,胡志红) et a1.SOD Enhances Infectivity of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus against H. armigra larvae. Virologica Sinica(中国病毒学), 2004, 18:506-507
Zhou M Z, Sun X L, Sun X C, et a1. Horizontal and vertical transmission of wild-type and recombinant
Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus. J Invertebr Pathol, 2005, 89: l65-175
http://www.hebly.gov.cn/showarticle.php?id=5307(美国白蛾在秦皇岛的发生)
http://www.zcny.gov.cn/Article/bzh/200807/1453.htm(农药危害情况)
http://baike.baidu.com/view/93663.htm(美国白蛾简介)
http://baike.baidu.com/view/1919392.htm(杨扇舟蛾简介)