转聚合cry1A.105、cry2Ab2和cp4epsps基因抗虫耐除草剂玉米的田间抗性评价
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摘要
为明确转聚合cry1A.105、cry2Ab2和cp4epsps基因的抗虫耐除草剂玉米的抗虫性和对除草剂的耐受性,本文通过田间人工接虫,以食叶级别、存活幼虫数、蛀孔数、隧道长度和雌穗被害级别等分别作为评价玉米心叶期和穗期的抗虫性标准,评价了转聚合cry1A.105、cry2Ab2和cp4epsps基因,具有抗虫耐除草剂复合性状的玉米品种‘ND6708YGRR’和‘ND6515YGRR’及同型非转基因‘ND6708’、‘ND6515’和当地‘郑单958’品种对亚洲玉米螟、黏虫和棉铃虫的抗虫效果;通过喷施除草剂后调查玉米植株的存活率,评价上述品种对除草剂的耐受性。田间试验结果表明:‘ND6708YGRR’和‘ND6515YGRR’两个转基因玉米品种对亚洲玉米螟、黏虫及棉铃虫的抗性水平均为高抗,且在4叶期和8叶期喷施草甘膦后其受害率均为0,而同型对照品种和当地对照品种喷施同剂量草甘膦后受害率为100%。综上,聚合cry1A.105、cry2Ab2和cp4epsps基因,具有抗虫耐除草剂复合性状的玉米品种‘ND6708YGRR’和‘ND6515YGRR’对亚洲玉米螟、黏虫和棉铃虫具有较强的抗虫效果,对草甘膦具有较好的耐受性。此研究可为将来建立以转Bt基因玉米为主要手段的综合防治技术体系提供依据。
To quantify resistance against insect pests and tolerance to glyphosate of transgenic corn lines expressing cry1A.105+cry2Ab2+cp4epsps, the leaf feeding rating, larval survival rate, the number of borer holes and the tunnel length of corn lines/hybrids ‘ND6708 YGRR' and ‘ND6515 YGRR' containing its conventional non-transgenic cultivars ‘ND6708' and ‘ND6515', and ‘Zhengdan 958', were evaluated by artificial infestations of the larvae of Ostrinia furnacalis(Guenée), Mythimna separata(Walker) and Helicoverpa armigera(Hübner) in field trials, respectively. Also, the mortality of plants after spraying herbicides was investigated to evaluate their tolerance to glyphosate. The results showed that corn lines/hybrids ‘ND6708 YGRR' and ‘ND6515 YGRR' expressing cry1A.105+cry2Ab2+cp4epsps were highly resistant to O. furnacalis, M. separata and H. armigera, and also performed excellent tolerance to glyphosate after spraying glyphosate with recommended dose at the 4 and 8 leaves stage, respectively. In contrast, the mortalities of conventional non-transgenic cultivars were 100% after spraying the same dose of glyphosate. These results demonstrate that transgenic corn lines expressing cry1A.105+cry2Ab2+cp4epsps were resistant to O. furnacalis, M. separata and H. armigera, and tolerant to glyphosate. This study will help to construct the strategy of planting transgenic Bt corn for controlling the Lepidopteran pests as one of main tactics in the future.
To quantify resistance against insect pests and tolerance to glyphosate of transgenic corn lines expressing cry1A.105+cry2Ab2+cp4epsps, the leaf feeding rating, larval survival rate, the number of borer holes and the tunnel length of corn lines/hybrids ‘ND6708 YGRR' and ‘ND6515 YGRR' containing its conventional non-transgenic cultivars ‘ND6708' and ‘ND6515', and ‘Zhengdan 958', were evaluated by artificial infestations of the larvae of Ostrinia furnacalis(Guenée), Mythimna separata(Walker) and Helicoverpa armigera(Hübner) in field trials, respectively. Also, the mortality of plants after spraying herbicides was investigated to evaluate their tolerance to glyphosate. The results showed that corn lines/hybrids ‘ND6708 YGRR' and ‘ND6515 YGRR' expressing cry1A.105+cry2Ab2+cp4epsps were highly resistant to O. furnacalis, M. separata and H. armigera, and also performed excellent tolerance to glyphosate after spraying glyphosate with recommended dose at the 4 and 8 leaves stage, respectively. In contrast, the mortalities of conventional non-transgenic cultivars were 100% after spraying the same dose of glyphosate. These results demonstrate that transgenic corn lines expressing cry1A.105+cry2Ab2+cp4epsps were resistant to O. furnacalis, M. separata and H. armigera, and tolerant to glyphosate. This study will help to construct the strategy of planting transgenic Bt corn for controlling the Lepidopteran pests as one of main tactics in the future.
引文
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[2] JAMES C. Global status of commercialized biotech/GM crops[R]. ISAAA Briefs: Brief 52, Ithaca, NY, 2016.
[3] HUTCHISON W D, BURKNESS E C, MITCHELL P D, et al. Areawide suppression of European corn borer with Bt maize reaps savings to non-Bt maize growers [J]. Science, 2010, 330(6001): 222-225.
[4] DIVELY G P, VENUGOPAL P D, BEAN D, et al. Regional pest suppression associated with widespread Bt maize adoption benefits vegetable growers [J]. Proceedings of the National Academy of Sciences of the United States of America, 2018, 115(13):3320-3325.
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[7] 王振营,鲁新,何康来,等.我国研究亚洲玉米螟历史、现状与展望[J].沈阳农业大学学报,2000,31(5):402-412.
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[10] 曾娟,姜玉英,刘杰.2012年黏虫暴发特点分析与监测预警建议[J].植物保护,2013,39(2):117-121.
[11] 程登发,赵中华.我国部分地区粘虫暴发原因分析与对策建议[J].种子科技,2016(10):89-90.
[12] 王振营,何康来,文丽萍,等.第四代棉铃虫卵在华北夏玉米田的时空分布[J].中国农业科学,2001,34(2):153-156.
[13] FAROMóS G P, ANDREADIS S S, DE LA POZA M, et al. Comparative assessment of the field-susceptibility of Sesamia nonagrioides to the Cry1Ab toxin in areas with different adoption rates of Bt maize and in Bt-free areas [J]. Crop Protection, 2011, 30: 902-906.
[14] PéREZ-HEDO M, REITER D, LóPEZ C, et al. Processing of the maize Bt toxin in the gut of Mythimna unipuncta caterpillars [J]. Entomologia Experimentalis et Applicata, 2013, 148: 56-64.
[15] GONZáLEZ-CABRERA J, GARCíA M, HERNáNDEZ-CRESPO P, et al. Resistance to Bt maize in Mythimna unipuncta (Lepidoptera: Noctuidae) is mediated by alteration in Cry1Ab protein activation [J]. Insect Biochemistry and Molecular Biology, 2013, 43: 635-643.
[16] HE Kanglai, WANG Zhenying, ZHOU Dayong, et al. Evaluation of transgenic Bt corn for resistance to the Asian corn borer (Lepidoptera: Pyralidae)[J]. Journal of Economic Entomology, 2003, 96(3):935-940.
[17] 王振营,王冬妍,何康来,等.转Bt基因玉米对粘虫的室内杀虫效果评价[J].植物保护学报,2005,32(2):153-157.
[18] 常雪艳,何康来,王振营,等.转Bt基因玉米对棉铃虫的抗性评价[J].植物保护学报,2006,33(4):374-378.
[19] 常雪,常雪艳,何康来,等.转cry1Ab基因玉米对粘虫的抗性评价[J].植物保护学报,2007,34(3):225-228.
[20] 中华人民共和国农业行业标准.NT/T 1248.5-2006:玉米抗病虫性鉴定技术规范第5部分:玉米抗玉米螟鉴定技术规范[S].北京:中国农业出版社,2006.
[21] 中华人民共和国农业部.农业部953号公告-10.1-2007:转基因植物及其产品环境安全检测:抗虫玉米第1部分:抗虫性[S].北京:中国农业出版社,2007.
[22] 中华人民共和国国家标准.GB/T 17980.42-2000:田间药效试验准则(一)除草剂防治玉米地杂草[S].北京:中国标准出版社,2005.
[23] KOZIEL M G, BELAND G L, BOWMAN C, et al. Field performance of elite transgenic maize plants expressing an insecticidal protein derived from Bacillus thuringiensis [J]. Bio-Technology, 1993, 11(2): 194-200.
[24] SIEBERT M W, NOLTING S P, HENDRIX W, et al. Evaluation of corn hybrids expressing Cry1F, Cry1A. 105, Cry2Ab2, Cry34Ab1/Cry35Ab1, and Cry3Bb1 against southern United States insect pests [J]. Journal of Economic Entomology, 2012, 105(5): 1825-1834.
[25] 王振营,何康来,石洁,等.桃蛀螟在玉米上危害加重原因与控制对策[J].植物保护,2006,32(2):67-69.
[26] 石洁,王振营,何康来.黄淮海地区夏玉米病虫害发生趋势与原因分析[J].植物保护,2005,31(5):63-65.
[2] JAMES C. Global status of commercialized biotech/GM crops[R]. ISAAA Briefs: Brief 52, Ithaca, NY, 2016.
[3] HUTCHISON W D, BURKNESS E C, MITCHELL P D, et al. Areawide suppression of European corn borer with Bt maize reaps savings to non-Bt maize growers [J]. Science, 2010, 330(6001): 222-225.
[4] DIVELY G P, VENUGOPAL P D, BEAN D, et al. Regional pest suppression associated with widespread Bt maize adoption benefits vegetable growers [J]. Proceedings of the National Academy of Sciences of the United States of America, 2018, 115(13):3320-3325.
[5] USEPA (United States Environmental Protection Agency), Current and previously section 3 PIP registrations [R]. http://www. epa. gov/pesticides/biopesticides/pips/piplist. htm, 2009.
[6] ZHAO Jianzhou, CAO Jun, COLLINS H L, et al. Concurrent use of transgenic plants expressing a single and two Bacillus thuringiensis genes speeds insect adaptation to pyramided plants [J]. Proceedings of the National Academy of Sciences of the United States of America, 2005, 102: 8426-8430.
[7] 王振营,鲁新,何康来,等.我国研究亚洲玉米螟历史、现状与展望[J].沈阳农业大学学报,2000,31(5):402-412.
[8] 江幸福,张蕾,程云霞,等.我国粘虫发生危害新特点及趋势分析[J].应用昆虫学报,2014,51(4):1444-1449.
[9] 中国农业科学院植物保护研究所,中国植物保护学会.中国农作物病虫害上册[M].北京:中国农业出版社,2014:673-682.
[10] 曾娟,姜玉英,刘杰.2012年黏虫暴发特点分析与监测预警建议[J].植物保护,2013,39(2):117-121.
[11] 程登发,赵中华.我国部分地区粘虫暴发原因分析与对策建议[J].种子科技,2016(10):89-90.
[12] 王振营,何康来,文丽萍,等.第四代棉铃虫卵在华北夏玉米田的时空分布[J].中国农业科学,2001,34(2):153-156.
[13] FAROMóS G P, ANDREADIS S S, DE LA POZA M, et al. Comparative assessment of the field-susceptibility of Sesamia nonagrioides to the Cry1Ab toxin in areas with different adoption rates of Bt maize and in Bt-free areas [J]. Crop Protection, 2011, 30: 902-906.
[14] PéREZ-HEDO M, REITER D, LóPEZ C, et al. Processing of the maize Bt toxin in the gut of Mythimna unipuncta caterpillars [J]. Entomologia Experimentalis et Applicata, 2013, 148: 56-64.
[15] GONZáLEZ-CABRERA J, GARCíA M, HERNáNDEZ-CRESPO P, et al. Resistance to Bt maize in Mythimna unipuncta (Lepidoptera: Noctuidae) is mediated by alteration in Cry1Ab protein activation [J]. Insect Biochemistry and Molecular Biology, 2013, 43: 635-643.
[16] HE Kanglai, WANG Zhenying, ZHOU Dayong, et al. Evaluation of transgenic Bt corn for resistance to the Asian corn borer (Lepidoptera: Pyralidae)[J]. Journal of Economic Entomology, 2003, 96(3):935-940.
[17] 王振营,王冬妍,何康来,等.转Bt基因玉米对粘虫的室内杀虫效果评价[J].植物保护学报,2005,32(2):153-157.
[18] 常雪艳,何康来,王振营,等.转Bt基因玉米对棉铃虫的抗性评价[J].植物保护学报,2006,33(4):374-378.
[19] 常雪,常雪艳,何康来,等.转cry1Ab基因玉米对粘虫的抗性评价[J].植物保护学报,2007,34(3):225-228.
[20] 中华人民共和国农业行业标准.NT/T 1248.5-2006:玉米抗病虫性鉴定技术规范第5部分:玉米抗玉米螟鉴定技术规范[S].北京:中国农业出版社,2006.
[21] 中华人民共和国农业部.农业部953号公告-10.1-2007:转基因植物及其产品环境安全检测:抗虫玉米第1部分:抗虫性[S].北京:中国农业出版社,2007.
[22] 中华人民共和国国家标准.GB/T 17980.42-2000:田间药效试验准则(一)除草剂防治玉米地杂草[S].北京:中国标准出版社,2005.
[23] KOZIEL M G, BELAND G L, BOWMAN C, et al. Field performance of elite transgenic maize plants expressing an insecticidal protein derived from Bacillus thuringiensis [J]. Bio-Technology, 1993, 11(2): 194-200.
[24] SIEBERT M W, NOLTING S P, HENDRIX W, et al. Evaluation of corn hybrids expressing Cry1F, Cry1A. 105, Cry2Ab2, Cry34Ab1/Cry35Ab1, and Cry3Bb1 against southern United States insect pests [J]. Journal of Economic Entomology, 2012, 105(5): 1825-1834.
[25] 王振营,何康来,石洁,等.桃蛀螟在玉米上危害加重原因与控制对策[J].植物保护,2006,32(2):67-69.
[26] 石洁,王振营,何康来.黄淮海地区夏玉米病虫害发生趋势与原因分析[J].植物保护,2005,31(5):63-65.