转Cry1Ac/1Ab基因棉花对棉铃虫的控制效果及对甜菜夜蛾和斜纹夜蛾生长发育的影响
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摘要
为评估转Cry1Ac/1Ab基因棉花对棉铃虫Helicoverpa armigera的抗性及对非靶标害虫甜菜夜蛾Spodoptera exigua和斜纹夜蛾S.litura生长发育的影响,采用室内生测法测定其对棉铃虫的抗性及对甜菜夜蛾和斜纹夜蛾不同龄期幼虫存活率、营养代谢及中肠酶活性的影响。结果表明,转Cry1Ac/1Ab基因棉花对棉铃虫第2代幼虫的抗性程度最高,幼虫校正死亡率达91.33%,但对甜菜夜蛾和斜纹夜蛾的抗性程度较低,幼虫校正死亡率分别为15.33%和13.33%。甜菜夜蛾和斜纹夜蛾各龄期幼虫取食转Cry1Ac/1Ab基因棉花后,其存活率与取食常规棉花对照无显著差异;甜菜夜蛾对转Cry1Ac/1Ab基因棉花叶片的相对取食量、近似消化率分别为16.68和93.12%,均高于取食常规棉花对照的10.72和92.00%,但差异不显著,而斜纹夜蛾取食转Cry1Ac/1Ab基因棉花后的各项营养指标均低于取食常规棉花对照,差异也不显著。取食转Cry1Ac/1Ab基因棉花后,甜菜夜蛾的过氧化物酶活性为0.02 U/mg prot,显著低于取食常规棉花的0.05 U/mg prot;斜纹夜蛾的酸性磷酸酶活性为0.15 U/mg prot,高于取食常规棉花的0.10 U/mg prot,但差异不显著,其它中肠酶活性均低于对照,亦无显著差异。
To assess the resistance of transgenic cotton to cotton bollworm Helicoverpa armigera(Hübner) and its effect on growth and development of Spodoptera exigua and S. litura, the larval survival rate, nutrition and energy metabolism levels and the midgut enzyme activity were tested with laboratory bioassay methods. The corrected mortality of bollworms reared on transgenic Cry1Ac/1Ab cotton peaked at 91.33% in 2 nd generation bollworms, indicating that this variety had high resistance to the2 nd generation cotton bollworm. However, its resistance to S. exigua and S. litura was low, in which the corrected larval mortalities of these two pests fed on transgenic Cry1Ac/1Ab cotton were 15.33% and13.33%, respectively. The survival rates of S. exigua and S. litura larvae in different developmental stages fed on transgenic cotton were not significantly different from that on the conventional cotton. The relative feeding rate and approximate digestion rate of S. exigua fed on transgenic cotton were 16.68 and93.12%, higher than that on the conventional cotton(10.72 and 92.00%), but not statistically significant.The nutrition index of S. litura was lower than that fed on conventional cotton, but the results were not statistically significant. When fed with transgenic Cry1Ac/1Ab cotton, the peroxidase activity of S. exigua was 0.02 U/mg prot, significantly lower than that when fed on conventional cotton(0.05 U/mg prot), and the acid phosphatase activity of S. litura was only 0.15 U/mg prot, higher than that when fed on conventional cotton(0.10 U/mg prot), but the activities of other midgut enzymes were not significantly different from the control.
To assess the resistance of transgenic cotton to cotton bollworm Helicoverpa armigera(Hübner) and its effect on growth and development of Spodoptera exigua and S. litura, the larval survival rate, nutrition and energy metabolism levels and the midgut enzyme activity were tested with laboratory bioassay methods. The corrected mortality of bollworms reared on transgenic Cry1Ac/1Ab cotton peaked at 91.33% in 2 nd generation bollworms, indicating that this variety had high resistance to the2 nd generation cotton bollworm. However, its resistance to S. exigua and S. litura was low, in which the corrected larval mortalities of these two pests fed on transgenic Cry1Ac/1Ab cotton were 15.33% and13.33%, respectively. The survival rates of S. exigua and S. litura larvae in different developmental stages fed on transgenic cotton were not significantly different from that on the conventional cotton. The relative feeding rate and approximate digestion rate of S. exigua fed on transgenic cotton were 16.68 and93.12%, higher than that on the conventional cotton(10.72 and 92.00%), but not statistically significant.The nutrition index of S. litura was lower than that fed on conventional cotton, but the results were not statistically significant. When fed with transgenic Cry1Ac/1Ab cotton, the peroxidase activity of S. exigua was 0.02 U/mg prot, significantly lower than that when fed on conventional cotton(0.05 U/mg prot), and the acid phosphatase activity of S. litura was only 0.15 U/mg prot, higher than that when fed on conventional cotton(0.10 U/mg prot), but the activities of other midgut enzymes were not significantly different from the control.
引文
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Guo JY,Gang W,Wan FH.2010.Activities of digestive and detoxification enzymes in multiple generations of beet armyworm,Spodoptera exigua(Hübner),in response to transgenic Bt cotton.Journal of Pest Science,83(4):453-460
Guo WJ,Lu CY,Xiong YQ,Ma HJ,Li SQ.2012.Effects of transgenic cry1Ab/cry1Ac rice on the activities of three protective enzymes in larvae of Sesamia inferens(Lepidoptera:Noctuidae).Acta Entomologica Sinica,55(8):958-963(in Chinese)[郭文娟,陆驰宇,熊应强,马鹤娟,李绍勤.2012.转cry1Ab/cry1Ac基因水稻对大螟幼虫体内三种保护酶活性的影响.昆虫学报,55(8):958-963]
Hu QJ,Zhou FC,Yang YZ,Yang AM,Zhang JJ,He J,Zhou JH,Shao JZ.2014.Effects of transgenic Bt-Cry1Ac cotton on activities of Bemisia tabaci enzymes.Chinese Journal of Eco-Agriculture,22(9):1079-1085(in Chinese)[胡其靖,周福才,杨益众,杨爱民,张建军,何菁,周建华,邵久之.2014.转Bt-Cry1Ac基因棉花对烟粉虱体内几种酶活力的影响.中国生态农业学报,22(9):1079-1085]
Huang DL,Liu HQ,Jiang SX.2006.Effects of Zhongmiansuo 45 and Zhongmiansuo 41 on experimental population of Spodoptera litura.Journal of Plant Protection,33(1):1-5(in Chinese)[黄东林,刘汉勤,蒋思霞.2006.转双价基因抗虫棉对斜纹夜蛾实验种群的影响.植物保护学报,33(1):1-5]
Jiang Q,Su HH,Yang YZ.2013.Relationship between midgut proteinases,detoxification enzymes,protective enzymes in insect and the resistance to Bt.Journal of Environmental Entomology,35(1):95-101(in Chinese)[蒋晴,苏宏华,杨益众.2013.昆虫对Bt毒素的抗性与中肠蛋白酶、解毒酶及保护酶系活性的关系.环境昆虫学报,35(1):95-101]
Lalitha C,Muralikrishna T,Sravani S,Devaki K.2012.In vitro evaluation of native Bacillus thuringiensis isolates against II instar Spodoptera litura(Fabricius).Annals of Plant Protection Sciences,20(1):91-97
Li HF,Liu X,Qu Q,Deng ZY,Lu ZX,Chen WX,Xu HL,Zhu Z.2002.Yield and insect resistance analysis of transgenic maize(Zea mays L.)with transgenic insect resistant fusion gene(cry1Ac3-cpti).Progress in Natural Science,12(1):37-40(in Chinese)[李慧芬,刘翔,曲强,邓朝阳,路子显,陈宛新,徐鸿林,朱祯.2002.转抗虫融合基因(cry1Ac3-cpti)玉米(Zea mays L.)植株的获得及其抗虫性分析.自然科学进展,12(1):37-40]
Li ZY.2012.Studies on the effects of transgenic Cry1Ab+Cry1Ac rice against key insect pests in paddy fields.Master Thesis.Nanjing:Nanjing Agricultural University(in Chinese)[李志毅.2012.转Cry1Ab/Cry1Ac融合基因抗虫水稻对田间主要昆虫种群的影响研究.硕士学位论文.南京:南京农业大学]
Luo JY,Cui JJ,Zhang S,Xin HJ,Wang CY.2013.Effects of transgenic Cry1Ac plus Cry2Ab cotton on pest survival,growth and enzymes activity of three kinds of pests.Cotton Science,25(1):68-72(in Chinese)[雒珺瑜,崔金杰,张帅,辛惠江,王春义.2013.转Cry1Ac+Cry2Ab抗虫棉对3种害虫的存活、生长发育及中肠相关酶活性的影响.棉花学报,25(1):68-72]
Sivasupramaniam S,Moar WJ,Ruschke LG,Osborn JA,Jiang C,Sebaugh JL,Brown GR,Shappley ZW,Oppenhuizen ME,Mullins JW,et al.2008.Toxicity and characterization of cotton expressing Bacillus thuringiensis Cry1Ac and Cry2Ab2 proteins for control of lepidopteran pests.Journal of Economic Entomology,101(2):546-554
Su HH,Li D,Fan N,Yang YZ.2010.Insect resistance of different tissues of transgenic cotton to Spodoptera exigua(Hübner).Agricultural Science&Technology,11(8):140-144
Wan P,Wu KM,Huang MS,Yu DZ,Wu JP.2008.Population dynamics of Spodoptera litura(Lepidoptera:Noctuidae)on Bt cotton in the Yangtze River Valley of China.Environmental Entomology,37(4):1043-1048
Wu G,Harris MK,Guo JY,Wan FH.2009.Response of multiple generations of beet armyworm,Spodoptera exigua(Hübner),feeding on transgenic Bt cotton.Journal of Applied Entomology,133(2):90-100
Wu KM,Li WD,Feng HQ,Guo YY.2002.Seasonal abundance of the mirids,Lygus lucorum and Adelphocoris spp.(Hemiptera:Miridae)on Bt cotton in northern China.Crop Protection,21(10):997-1002
Wu KM,Xu G,Guo YY.2001.Seasonal population dynamics of white fly adults on cotton in northern China.Plant Protection,27(2):14-15(in Chinese)[吴孔明,徐广,郭予元.2001.华北北部地区棉田烟粉虱成虫季节性动态.植物保护,27(2):14-15]
Xia JY,Cui JJ,Chang RQ.2000.Study on the resistance of transgenic cotton against Spodoptera exigua.China Cotton,27(9):10-11(in Chinese)[夏敬源,崔金杰,常蕊芹.2000.转基因抗虫棉对甜菜夜蛾的抗性研究.中国棉花,27(9):10-11]
Xu D,Guo JM,Cong SB,Wu HH,Huang MS,Wang Q,Wan P.2016.The effectiveness of genetically modified stacked cotton against four species of Lepidoptera and expression analysis of Bt proteins.Cotton Science 28(5):478-484(in Chinese)[许冬,郭建明,丛胜波,武怀恒,黄民松,王强,万鹏.2016.复合性状转基因棉花杀虫蛋白表达量及对4种鳞翅目害虫的抗性.棉花学报,28(5):478-484]
Zhang JH,Guo JY,Wan FH,Xia JY.2006.Effects of transgenic Bt cotton on the susceptibility to insecticides and detoxification enzyme activities of cotton aphid(Aphis gossypii Glover).Acta Entomologica Sinica,49(6):938-943(in Chinese)[张炬红,郭建英,万方浩,夏敬源.2006.转Bt基因抗虫棉对棉蚜的杀虫剂敏感性及解毒酶系的影响.昆虫学报,49(6):938-943]
Zhang SY,Li DM,Xie BY.2004.Effects of Bt toxic protein on development and activities of several relative enzymes in Helicoverpa armigera.Entomological Knowledge,41(6):536-540(in Chinese)[张少燕,李典谟,谢宝瑜.2004.Bt毒蛋白对棉铃虫的生长发育和相关酶活性的影响.昆虫知识,41(6):536-540]
Zhang XL,Chen P,Chen CF,Yang YZ.2007.Effect of the transgenic Bt cotton on laboratory population increasing of the beet armyworm Spodoptera exigua Hübner.Journal of Plant Protection,34(4):391-395(in Chinese)[张小丽,陈萍,陈翠芳,杨益众.2007.转Bt基因抗虫棉对甜菜夜蛾实验种群增长的影响.植物保护学报,34(4):391-395]
Zheng XL,Wang P,Wang XP,Lei CL.2010.Damage,occurrence and control of Spodoptera exigua on transgenic cottons.Plant Protection,36(3):34-38(in Chinese)[郑霞林,王攀,王小平,雷朝亮.2010.转基因棉甜菜夜蛾的为害现状、暴发成因及防治现状.植物保护,36(3):34-38]
Zhu JH,Zhang FP,Ren HG.2005.Development and nutrition of Prodenia litura on four food plants.Chinese Bulletin of Entomology,42(6):643-646(in Chinese)[朱俊洪,张方平,任洪刚.2005.四种食料植物对斜纹夜蛾生长发育及营养指标的影响.昆虫知识,42(6):643-646]
Chen ZH.1987.Quantitative measurement and calculation of insect nutrition index.Entomological Knowledge,24(5):299-301(in Chinese)[陈志辉.1987.昆虫营养指标的定量测量与计算.昆虫知识,24(5):299-301]
Govindan K,Gunasekaran K,Kuttalam S,Aiswariya KK.2010.Laboratory evaluation of transgenic Bt cotton and non Bt cotton plant parts against third instar larvae of Spodoptera litura(Fab.)(Noctuidae:Lepidoptera).Journal of Biopesticides,3(2):432-436
Guo JY,Gang W,Wan FH.2010.Activities of digestive and detoxification enzymes in multiple generations of beet armyworm,Spodoptera exigua(Hübner),in response to transgenic Bt cotton.Journal of Pest Science,83(4):453-460
Guo WJ,Lu CY,Xiong YQ,Ma HJ,Li SQ.2012.Effects of transgenic cry1Ab/cry1Ac rice on the activities of three protective enzymes in larvae of Sesamia inferens(Lepidoptera:Noctuidae).Acta Entomologica Sinica,55(8):958-963(in Chinese)[郭文娟,陆驰宇,熊应强,马鹤娟,李绍勤.2012.转cry1Ab/cry1Ac基因水稻对大螟幼虫体内三种保护酶活性的影响.昆虫学报,55(8):958-963]
Hu QJ,Zhou FC,Yang YZ,Yang AM,Zhang JJ,He J,Zhou JH,Shao JZ.2014.Effects of transgenic Bt-Cry1Ac cotton on activities of Bemisia tabaci enzymes.Chinese Journal of Eco-Agriculture,22(9):1079-1085(in Chinese)[胡其靖,周福才,杨益众,杨爱民,张建军,何菁,周建华,邵久之.2014.转Bt-Cry1Ac基因棉花对烟粉虱体内几种酶活力的影响.中国生态农业学报,22(9):1079-1085]
Huang DL,Liu HQ,Jiang SX.2006.Effects of Zhongmiansuo 45 and Zhongmiansuo 41 on experimental population of Spodoptera litura.Journal of Plant Protection,33(1):1-5(in Chinese)[黄东林,刘汉勤,蒋思霞.2006.转双价基因抗虫棉对斜纹夜蛾实验种群的影响.植物保护学报,33(1):1-5]
Jiang Q,Su HH,Yang YZ.2013.Relationship between midgut proteinases,detoxification enzymes,protective enzymes in insect and the resistance to Bt.Journal of Environmental Entomology,35(1):95-101(in Chinese)[蒋晴,苏宏华,杨益众.2013.昆虫对Bt毒素的抗性与中肠蛋白酶、解毒酶及保护酶系活性的关系.环境昆虫学报,35(1):95-101]
Lalitha C,Muralikrishna T,Sravani S,Devaki K.2012.In vitro evaluation of native Bacillus thuringiensis isolates against II instar Spodoptera litura(Fabricius).Annals of Plant Protection Sciences,20(1):91-97
Li HF,Liu X,Qu Q,Deng ZY,Lu ZX,Chen WX,Xu HL,Zhu Z.2002.Yield and insect resistance analysis of transgenic maize(Zea mays L.)with transgenic insect resistant fusion gene(cry1Ac3-cpti).Progress in Natural Science,12(1):37-40(in Chinese)[李慧芬,刘翔,曲强,邓朝阳,路子显,陈宛新,徐鸿林,朱祯.2002.转抗虫融合基因(cry1Ac3-cpti)玉米(Zea mays L.)植株的获得及其抗虫性分析.自然科学进展,12(1):37-40]
Li ZY.2012.Studies on the effects of transgenic Cry1Ab+Cry1Ac rice against key insect pests in paddy fields.Master Thesis.Nanjing:Nanjing Agricultural University(in Chinese)[李志毅.2012.转Cry1Ab/Cry1Ac融合基因抗虫水稻对田间主要昆虫种群的影响研究.硕士学位论文.南京:南京农业大学]
Luo JY,Cui JJ,Zhang S,Xin HJ,Wang CY.2013.Effects of transgenic Cry1Ac plus Cry2Ab cotton on pest survival,growth and enzymes activity of three kinds of pests.Cotton Science,25(1):68-72(in Chinese)[雒珺瑜,崔金杰,张帅,辛惠江,王春义.2013.转Cry1Ac+Cry2Ab抗虫棉对3种害虫的存活、生长发育及中肠相关酶活性的影响.棉花学报,25(1):68-72]
Sivasupramaniam S,Moar WJ,Ruschke LG,Osborn JA,Jiang C,Sebaugh JL,Brown GR,Shappley ZW,Oppenhuizen ME,Mullins JW,et al.2008.Toxicity and characterization of cotton expressing Bacillus thuringiensis Cry1Ac and Cry2Ab2 proteins for control of lepidopteran pests.Journal of Economic Entomology,101(2):546-554
Su HH,Li D,Fan N,Yang YZ.2010.Insect resistance of different tissues of transgenic cotton to Spodoptera exigua(Hübner).Agricultural Science&Technology,11(8):140-144
Wan P,Wu KM,Huang MS,Yu DZ,Wu JP.2008.Population dynamics of Spodoptera litura(Lepidoptera:Noctuidae)on Bt cotton in the Yangtze River Valley of China.Environmental Entomology,37(4):1043-1048
Wu G,Harris MK,Guo JY,Wan FH.2009.Response of multiple generations of beet armyworm,Spodoptera exigua(Hübner),feeding on transgenic Bt cotton.Journal of Applied Entomology,133(2):90-100
Wu KM,Li WD,Feng HQ,Guo YY.2002.Seasonal abundance of the mirids,Lygus lucorum and Adelphocoris spp.(Hemiptera:Miridae)on Bt cotton in northern China.Crop Protection,21(10):997-1002
Wu KM,Xu G,Guo YY.2001.Seasonal population dynamics of white fly adults on cotton in northern China.Plant Protection,27(2):14-15(in Chinese)[吴孔明,徐广,郭予元.2001.华北北部地区棉田烟粉虱成虫季节性动态.植物保护,27(2):14-15]
Xia JY,Cui JJ,Chang RQ.2000.Study on the resistance of transgenic cotton against Spodoptera exigua.China Cotton,27(9):10-11(in Chinese)[夏敬源,崔金杰,常蕊芹.2000.转基因抗虫棉对甜菜夜蛾的抗性研究.中国棉花,27(9):10-11]
Xu D,Guo JM,Cong SB,Wu HH,Huang MS,Wang Q,Wan P.2016.The effectiveness of genetically modified stacked cotton against four species of Lepidoptera and expression analysis of Bt proteins.Cotton Science 28(5):478-484(in Chinese)[许冬,郭建明,丛胜波,武怀恒,黄民松,王强,万鹏.2016.复合性状转基因棉花杀虫蛋白表达量及对4种鳞翅目害虫的抗性.棉花学报,28(5):478-484]
Zhang JH,Guo JY,Wan FH,Xia JY.2006.Effects of transgenic Bt cotton on the susceptibility to insecticides and detoxification enzyme activities of cotton aphid(Aphis gossypii Glover).Acta Entomologica Sinica,49(6):938-943(in Chinese)[张炬红,郭建英,万方浩,夏敬源.2006.转Bt基因抗虫棉对棉蚜的杀虫剂敏感性及解毒酶系的影响.昆虫学报,49(6):938-943]
Zhang SY,Li DM,Xie BY.2004.Effects of Bt toxic protein on development and activities of several relative enzymes in Helicoverpa armigera.Entomological Knowledge,41(6):536-540(in Chinese)[张少燕,李典谟,谢宝瑜.2004.Bt毒蛋白对棉铃虫的生长发育和相关酶活性的影响.昆虫知识,41(6):536-540]
Zhang XL,Chen P,Chen CF,Yang YZ.2007.Effect of the transgenic Bt cotton on laboratory population increasing of the beet armyworm Spodoptera exigua Hübner.Journal of Plant Protection,34(4):391-395(in Chinese)[张小丽,陈萍,陈翠芳,杨益众.2007.转Bt基因抗虫棉对甜菜夜蛾实验种群增长的影响.植物保护学报,34(4):391-395]
Zheng XL,Wang P,Wang XP,Lei CL.2010.Damage,occurrence and control of Spodoptera exigua on transgenic cottons.Plant Protection,36(3):34-38(in Chinese)[郑霞林,王攀,王小平,雷朝亮.2010.转基因棉甜菜夜蛾的为害现状、暴发成因及防治现状.植物保护,36(3):34-38]
Zhu JH,Zhang FP,Ren HG.2005.Development and nutrition of Prodenia litura on four food plants.Chinese Bulletin of Entomology,42(6):643-646(in Chinese)[朱俊洪,张方平,任洪刚.2005.四种食料植物对斜纹夜蛾生长发育及营养指标的影响.昆虫知识,42(6):643-646]