Application of PCR Method for Detecting of the EoNPV in Ectropis oblique and Its Biological Activity

ZHANG Yong-an; ZHONG Guo-li; HOU Yu-xia; GONG Yu-mei; QU Liang-jian; WANG Yu-zhu

Volume 21 Issue 4

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Article Navigation > Forest Research > 2008 > 21(4): 451-455

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Application of PCR Method for Detecting of the EoNPV in Ectropis oblique and Its Biological Activity

1.
College of Science, China Agricultural University, Beijing　100094, China
2.
Research Institute ofForest Ecology, Environment and Protection, CAF, Beijing　100091, China
3.
Science and TechnologyDevelopment Centre, State Forestry Administration, Beijing　100714, China
4.
Research Institute ofForest Ecology, Environment and Protection, CAF, Beijing　100091, China

Received Date: 2008-03-19

Abstract

The polymerase chain reaction ( PCR) method for detecting Ectropis obliqua multip le nuclear polyhedrosisviruswas established with self2designed p rimers according to the sequence of polyhedron gene of EoNPV, andEoNPV could be detected from the offsp ring, the data showed that the method would allow the detection of about 1fg, thismethod could be used to identify EoNPV in field p ractices. The method was simp le with good rep roducibilityand selectivity, and high sensitivity, so deserves to be popularized. An infectivity test of EoNPV against the secondinstar larvae of Ectropis oblique was carried out in our laboratory, and LC50 of EoNPV was determined as 2. 2 ×104P IB·mL -1 , the larva was very sensitive to EoNPV, and it had a higher death rate with higher EoNPVconcentration, and the death time p resented positive correlation with the concentration of EoNPV.
- EoNPV,
- Ectropis obliqua,
- PCR detection,
- biological activity

References

[1]	殷坤山,陈华才,唐美君,等. 茶尺蠖病毒杀虫剂田间使用技术的研究[J]. 中国病毒学, 2003, 18 (5) : 492 - 495
[2]	殷坤山,陈华才. 喷施茶尺蠖病毒杀虫剂对茶叶品质的影响[J]. 中国茶叶, 2002, 24 (4) : 5
[3]	曾明森,王庆森,吴光远. 高效氯氟氰菊酯、苦参总碱和高效Bt防治茶尺蠖田间药效试验结果[J]. 茶叶科学技术, 2005 ( 2) :13 - 14
[4]	吴晓晶,胡　萃. 茶尺蠖核型多角体病毒的大量增殖研究[J].浙江农业大学学报, 1987, 13 (2) : 150 - 157
[5]	叶恭银, 朱俊庆. 高温条件下茶尺蠖核型多角体病毒对茶尺蠖繁殖的影响[J]. 中国病毒学, 1992, 7 (3) : 283 - 288
[6]	叶恭银, 朱俊庆,胡　萃. 茶尺蠖核型多角体病毒对宿主幼虫食物摄取与利用的影响[J]. 浙江农业大学学报, 1995, 21 ( 1) :77 - 80
[7]	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[J]. 植物保护学报, 1994, 21 (3) : 231 - 237
[8]	胡　萃, 朱俊庆,叶恭银. 茶尺蠖NPV对茶尺蠖幼虫取食及生长发育的影响[J]. 浙江农业大学学报, 1990, 16 (2) : 113 - 118
[9]	孔庆瑚,何　玲,陈云英. 茶尺蠖核型多角体病毒毒性试验[J].科技通报, 1990, 6 (1) : 32 - 35
[10]	陈棣华,洪北边,赵烨烽. 茶尺蠖核型多角体病毒研究[J]. 生物防治通报, 1989, 5 (4) : 168 - 172
[11]	陈棣华,孙继山. 茶尺蠖多角体病毒的精细结构及发生[J]. 电子显微学报, 1992, 5: 373 - 374
[12]	谭业平, 陶　涛. 昆虫病毒学[M]. 武汉: 武汉出版社, 1991
[13]	陈棣华,赵怀宇,张立人. 三种昆虫核型多角体病毒DNA的限制性内切酶酶解分析[J]. 病毒学报, 1987, 3 (1) : 69 - 75
[14]	张益民. 三种茶树害虫核型多角体病毒蛋白性质的初步研究[J]. 微生物学通报, 1986, 13 (5) : 198 - 200
[15]	赵怀宇, 陈棣华,张立人. 茶尺蠖核型多角体病毒的血清学研究[J]. 病毒学杂志, 1986, 1 (3) : 52 - 59
[16]	Ma X C, Xu H J, Mei J T, et al. Genome sequence and organization of a nucleopolyhedrovirus that infects the tea looper caterp illar,Ectropis oblique [J]. Microbiol, 2006, 44 (1) : 77 - 82
[17]	HughesD S, Possee R D, Ling IA. Activation and detection of alatent baculovirus resemblingM am estra brassicae nuclearpolyhedrosis virus in M. brassicae insect [J]. Virology, 1993, 194:608 - 615
[18]	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[J]. 中国生物防治, 2005,21 (1) : 45 - 48
[19]	贾春生. 利用sp ss软件计算杀虫剂的LC50 [J]. 昆虫知识,2006, 43 (3) : 414 - 417
[20]	郭　郛, 忻介六. 昆虫学实验技术[M]. 北京:科学出版社,1988, 359 - 360
[21]	叶恭银,胡　萃. 温度对茶尺蠖核型多角体病毒增殖动态的影响[J]. 病毒学杂志, 1990, 4 (4) : 397 - 403
[22]	陈亦根,黄明度. 茶尺蠖生物防治进展[J]. 昆虫天敌, 2001, 23(4) : 181 - 184
[23]	肖能文,谭济才. 无公害杀虫剂防治茶尺蠖试验[J]. 茶叶通讯, 2001 (4) : 17 - 19
[24]	雷元胜,夏良胜,石　涛. 4 000 IV /mg Bt颗粒剂防治茶尺蠖药效对比试验[J]. 茶业通报, 2002, 24 (2) : 26
[25]	张益民,王学兰,张世敏. 茶尺蠖核型多体病毒超微结构的初步研究[J]. 科学通报, 1985, (24) : 1918 - 1920
[26]	刘　军, 兰萍章. 用细胞遗传学方法检测茶尺蠖核型多角体病毒的安全性[J]. 生物防治通报, 1988, 4 (1) : 38 - 44
[27]	叶恭银,胡　萃. 茶尺蠖感染核型多角体病毒后病死时间分布的数学模拟[J]. 生态学报, 1994, 14 (2) : 196 - 200
[28]	陈棣华, 钟卫洲,栗陶生. 灰茶尺蛾核型多角体病毒的精细结构及生物学特性研究[J]. 微生物学通报, 1991, 18 ( 6 ) : 321- 323
[29]	叶恭银,胡　萃. 适用于高温季节的茶尺蠖核型多角体病毒制剂的筛选[J]. 植物保护学报, 1991, 18 (3) : 215 - 220
[30]	叶恭银,胡　萃, 洪　健. 温度对茶尺蠖核型多角体病毒形态发生的影响[J]. 浙江农业大学学报, 1991, 17 (2) : 185 - 191
[31]	周晓峰,李　劲,陈海雷. 油茶尺蠖核型多角体病毒毒力的生物测定[J]. 植物保护, 1990, 16 (4) : 25 - 26
[32]	甘运凯,刘年翠,黎国强. 应用混合病毒防治茶园害虫[J]. 湖南微生物学通讯, 1990 (1) : 16 - 17

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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Application of PCR Method for Detecting of the EoNPV in Ectropis oblique and Its Biological Activity

1. College of Science, China Agricultural University, Beijing　100094, China

2. Research Institute ofForest Ecology, Environment and Protection, CAF, Beijing　100091, China

3. Science and TechnologyDevelopment Centre, State Forestry Administration, Beijing　100714, China

4. Research Institute ofForest Ecology, Environment and Protection, CAF, Beijing　100091, China

Received Date: 2008-03-19

Abstract: The polymerase chain reaction ( PCR) method for detecting Ectropis obliqua multip le nuclear polyhedrosisviruswas established with self2designed p rimers according to the sequence of polyhedron gene of EoNPV, andEoNPV could be detected from the offsp ring, the data showed that the method would allow the detection of about 1fg, thismethod could be used to identify EoNPV in field p ractices. The method was simp le with good rep roducibilityand selectivity, and high sensitivity, so deserves to be popularized. An infectivity test of EoNPV against the secondinstar larvae of Ectropis oblique was carried out in our laboratory, and LC50 of EoNPV was determined as 2. 2 ×104P IB·mL -1 , the larva was very sensitive to EoNPV, and it had a higher death rate with higher EoNPVconcentration, and the death time p resented positive correlation with the concentration of EoNPV.

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Reference (32)

[1]	殷坤山,陈华才,唐美君,等. 茶尺蠖病毒杀虫剂田间使用技术的研究[J]. 中国病毒学, 2003, 18 (5) : 492 - 495
[2]	殷坤山,陈华才. 喷施茶尺蠖病毒杀虫剂对茶叶品质的影响[J]. 中国茶叶, 2002, 24 (4) : 5
[3]	曾明森,王庆森,吴光远. 高效氯氟氰菊酯、苦参总碱和高效Bt防治茶尺蠖田间药效试验结果[J]. 茶叶科学技术, 2005 ( 2) :13 - 14
[4]	吴晓晶,胡　萃. 茶尺蠖核型多角体病毒的大量增殖研究[J].浙江农业大学学报, 1987, 13 (2) : 150 - 157
[5]	叶恭银, 朱俊庆. 高温条件下茶尺蠖核型多角体病毒对茶尺蠖繁殖的影响[J]. 中国病毒学, 1992, 7 (3) : 283 - 288
[6]	叶恭银, 朱俊庆,胡　萃. 茶尺蠖核型多角体病毒对宿主幼虫食物摄取与利用的影响[J]. 浙江农业大学学报, 1995, 21 ( 1) :77 - 80
[7]	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[J]. 植物保护学报, 1994, 21 (3) : 231 - 237
[8]	胡　萃, 朱俊庆,叶恭银. 茶尺蠖NPV对茶尺蠖幼虫取食及生长发育的影响[J]. 浙江农业大学学报, 1990, 16 (2) : 113 - 118
[9]	孔庆瑚,何　玲,陈云英. 茶尺蠖核型多角体病毒毒性试验[J].科技通报, 1990, 6 (1) : 32 - 35
[10]	陈棣华,洪北边,赵烨烽. 茶尺蠖核型多角体病毒研究[J]. 生物防治通报, 1989, 5 (4) : 168 - 172
[11]	陈棣华,孙继山. 茶尺蠖多角体病毒的精细结构及发生[J]. 电子显微学报, 1992, 5: 373 - 374
[12]	谭业平, 陶　涛. 昆虫病毒学[M]. 武汉: 武汉出版社, 1991
[13]	陈棣华,赵怀宇,张立人. 三种昆虫核型多角体病毒DNA的限制性内切酶酶解分析[J]. 病毒学报, 1987, 3 (1) : 69 - 75
[14]	张益民. 三种茶树害虫核型多角体病毒蛋白性质的初步研究[J]. 微生物学通报, 1986, 13 (5) : 198 - 200
[15]	赵怀宇, 陈棣华,张立人. 茶尺蠖核型多角体病毒的血清学研究[J]. 病毒学杂志, 1986, 1 (3) : 52 - 59
[16]	Ma X C, Xu H J, Mei J T, et al. Genome sequence and organization of a nucleopolyhedrovirus that infects the tea looper caterp illar,Ectropis oblique [J]. Microbiol, 2006, 44 (1) : 77 - 82
[17]	HughesD S, Possee R D, Ling IA. Activation and detection of alatent baculovirus resemblingM am estra brassicae nuclearpolyhedrosis virus in M. brassicae insect [J]. Virology, 1993, 194:608 - 615
[18]	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[J]. 中国生物防治, 2005,21 (1) : 45 - 48
[19]	贾春生. 利用sp ss软件计算杀虫剂的LC50 [J]. 昆虫知识,2006, 43 (3) : 414 - 417
[20]	郭　郛, 忻介六. 昆虫学实验技术[M]. 北京:科学出版社,1988, 359 - 360
[21]	叶恭银,胡　萃. 温度对茶尺蠖核型多角体病毒增殖动态的影响[J]. 病毒学杂志, 1990, 4 (4) : 397 - 403
[22]	陈亦根,黄明度. 茶尺蠖生物防治进展[J]. 昆虫天敌, 2001, 23(4) : 181 - 184
[23]	肖能文,谭济才. 无公害杀虫剂防治茶尺蠖试验[J]. 茶叶通讯, 2001 (4) : 17 - 19
[24]	雷元胜,夏良胜,石　涛. 4 000 IV /mg Bt颗粒剂防治茶尺蠖药效对比试验[J]. 茶业通报, 2002, 24 (2) : 26
[25]	张益民,王学兰,张世敏. 茶尺蠖核型多体病毒超微结构的初步研究[J]. 科学通报, 1985, (24) : 1918 - 1920
[26]	刘　军, 兰萍章. 用细胞遗传学方法检测茶尺蠖核型多角体病毒的安全性[J]. 生物防治通报, 1988, 4 (1) : 38 - 44
[27]	叶恭银,胡　萃. 茶尺蠖感染核型多角体病毒后病死时间分布的数学模拟[J]. 生态学报, 1994, 14 (2) : 196 - 200
[28]	陈棣华, 钟卫洲,栗陶生. 灰茶尺蛾核型多角体病毒的精细结构及生物学特性研究[J]. 微生物学通报, 1991, 18 ( 6 ) : 321- 323
[29]	叶恭银,胡　萃. 适用于高温季节的茶尺蠖核型多角体病毒制剂的筛选[J]. 植物保护学报, 1991, 18 (3) : 215 - 220
[30]	叶恭银,胡　萃, 洪　健. 温度对茶尺蠖核型多角体病毒形态发生的影响[J]. 浙江农业大学学报, 1991, 17 (2) : 185 - 191
[31]	周晓峰,李　劲,陈海雷. 油茶尺蠖核型多角体病毒毒力的生物测定[J]. 植物保护, 1990, 16 (4) : 25 - 26
[32]	甘运凯,刘年翠,黎国强. 应用混合病毒防治茶园害虫[J]. 湖南微生物学通讯, 1990 (1) : 16 - 17

Application of PCR Method for Detecting of the EoNPV in Ectropis oblique and Its Biological Activity

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Application of PCR Method for Detecting of the EoNPV in Ectropis oblique and Its Biological Activity

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