转Bt基因抗虫玉米HGK60的农艺性状分析

doi:10.13560/j.cnki.biotech.bull.1985.2018-0219

生物技术通报 ›› 2018, Vol. 34 ›› Issue (7): 92-100.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0219

转Bt基因抗虫玉米HGK60的农艺性状分析

梁海生^{1, 2}, 李梦桃^{2, 4}, 李圣彦², 汪海², 张杰³, 郎志宏²

1. 西南科技大学生命科学与工程学院,绵阳 621010;
2. 中国农业科学院生物技术研究所,北京100081;
3. 中国农业科学院植物保护研究所/植物病虫害生物学国家重点实验室,北京 100193;
4. 东北农业大学生命科学学院,哈尔滨 150030

收稿日期:2018-03-16 出版日期:2018-07-26 发布日期:2018-08-01
作者简介:梁海生,男,硕士研究生,研究方向：植物分子生物学与基因工程;E-mail：lhs1549631002@163.com
基金资助:
转基因生物新品种培育重大专项（2016ZX08003-001）

Agronomic Traits Analysis of Transgenic Bt cry1Ah Maize HGK60 Line

LIANG Hai-sheng^{1, 2}, LI Meng-tao^{2, 4}, LI Sheng-yan², WANG Hai², ZHANG Jie³, LANG Zhi-hong²

1. Southwest University of Science and Technology,School of Life Science and Engineering,Mianyang 621010;
2. Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081;
3. Institute of Plant Protection,Chinese Academy of Agricultural Sciences/State Key Laboratory for Biology of Plant Diseases and Insect Pests,Beijing 100193;
4. Northeast Agricultural University,School of Life Science,Harbin 150030

Received:2018-03-16 Published:2018-07-26 Online:2018-08-01

摘要/Abstract

摘要： 为了确定转Bt cry1Ah抗虫玉米HGK60的自交系及其杂交后代外源基因的遗传表达稳定性和农艺性状,通过实时荧光定量PCR和ELISA分析外源基因的遗传表达稳定性,通过室内外生测和田间性状考量分析农艺性状。荧光定量PCR结果表明Bt cry1Ah基因在玉米的不同组织中可以正常转录,但RNA表达水平存在一定的差异;ELISA结果表明在转基因植株的不同发育时期、不同器官中Cry1Ah的蛋白表达量顺序：雄穗>叶片>苞叶>籽粒>花丝>穗轴。两地连续三代的田间及室内抗虫性检测结果表明HGK60抗虫玉米对亚洲玉米螟均表现出很好的抗性。性状考量结果表明HGK60抗虫玉米与受体材料对照比较,种子发芽率、生育期、穗行数、穗长、千粒重等农艺性状均无显著差异。通过多年多点田间试验和分子检测结果证明HGK60转基因抗虫玉米中外源基因稳定的遗传和表达,对亚洲玉米螟有很好的抗性,农艺性状与对照材料无显著差异。HGK60转基因抗虫玉米对亚洲玉米螟的良好抗性使其具有很好的产业化应用前景。

关键词: cry1Ah, 转基因玉米, 遗传稳定性, 农艺性状

Abstract: To determine the genetic expression stability and agronomic traits of the inbred lines of Bt cry1Ah-resistant maize HGK60 and its hybrid progenies,we analyzed the genetic expression stability of the exogenous genes by fluorescence quantitative PCR and ELISA,both bioassay indoor and outdoor. Analysis of agronomic traits were carried out by bioassay and agronomic traits considerations. The result of fluorescent quantitative RT-PCR showed that Bt cry1Ah could be transcribed normally in different plant tissues,and the expression level of RNA was different. The results of ELISA showed that the protein expression of Cry1Ah in different organs at different developmental stages：tassel > leaf > husk > grain > filament > cob. Through two locations and three consecutive generations of bioassay indoor and outdoor,the HGK60 line showed high resistant to Asian corn borer. Compared with the recipient inbred and hybrid lines,seed germination rate,growth period,ear number,spike length,thousand grains weight and other agronomic traits considerations indicated are not significantly different. Through experiments and molecular tests tracking over many years,the genetic and expression stability of the exogenous genes in HGK60 transgenic maize has been proven to be very resistant to Asian corn borer,and there is no significant difference between agronomic traits and control materials. The good resistance of HGK60 transgenic insect-resistant corn to Asian corn borer proved that HGK60 transgenic insect-resistant maize has good commercial application prospect.

Key words: cry1Ah gene, transgenic maize, genetic stability, agronomic traits

梁海生, 李梦桃, 李圣彦, 汪海, 张杰, 郎志宏. 转Bt基因抗虫玉米HGK60的农艺性状分析[J]. 生物技术通报, 2018, 34(7): 92-100.

LIANG Hai-sheng, LI Meng-tao, LI Sheng-yan, WANG Hai, ZHANG Jie, LANG Zhi-hong. Agronomic Traits Analysis of Transgenic Bt cry1Ah Maize HGK60 Line[J]. Biotechnology Bulletin, 2018, 34(7): 92-100.

参考文献

[1] 齐涛. 中国玉米国际竞争力研究[D]. 杨凌:西北农林科技大学, 2011.
[2] 沈平, 章秋艳, 林友华, 等. 推进我国转基因玉米产业化的思考[J]. 中国生物工程杂志, 2016, 36(04):24-29.
[3] 康岭生, 王玉民, 姜昱, 等. 转Bt基因玉米的抗螟性及产量分析[J]. 玉米科学, 2009, 17(1):62-64, 70.
[4] 陈化榜. 美国转基因玉米的生产概况和发展趋势[J]. 玉米科学, 2008, 16(3):1-3.
[5] Brookes G, Barfoot P.Environmental impacts of genetically modified(GM)crop use 1996-2015:Impacts on pesticide use and carbon emissions[C]. GM Crops & Food, 2017, 8(2):117-147.
[6] Mungai NW, Motavalli PP, Nelson KA, et al.Differences in yields, residue composition and n mineralization dynamics of Bt and Non-Bt maize[J]. Nutrient Cycling in Agroecosystems, 2005, 73(1):101-109.
[7] Dillehay BL, Roth GW, Calvin DD, et al.Performance of Bt corn hybrids, their near isolines, and leading corn hybrids in pennsylvania and maryland[J]. Agronomy Journal, 2004, 96(3):818-824.
[8] Pellegrino E, Bedini S, Nuti M, et al.Impact of genetically engineered maize on agronomic, environmental and toxicological traits:a meta-analysis of 21 years of field data[J]. Scientific Reports, 2018, 8(1):2045-2322.
[9] Wang X, Zhang X, Yang JT, et al.Effect on transcriptome and metabolome of stacked transgenic maize containing insecticidal cry and glyphosate tolerance epsps genes[J]. The Plant J, 2018, 93(6):1007-1016.
[10] Xue J, Liang G, Crickmore N, et al.Cloning and characterization of a novel Cry1A toxin from Bacillus thuringiensis with high toxicity to the Asian corn borer and other lepidopteran insects[J]. FEMS microbiology letters, 2008, 280(1):95-101.
[11] Xue J, Zhou ZS, Song FP, et al.Identification of the minimal active fragment of the Cry1Ah toxin[J]. Biotechnology letters, 2011, 33(3):531-537.
[12] Li XY, Lang ZH, Zhang J, et al.Acquisition of insect-resistant transgenic maize harboring a truncated cry1Ah gene via agrobacterium-mediated transformation[J]. Journal of Integrative Agriculture, 2014, 13(5):937-944.
[13] 戴军, 李秀影, 朱莉, 等. 转Bt cry1Ah基因抗虫玉米的分子检测及农艺性状分析[J]. 生物技术通报, 2014(5):62-68.
[14] 刘柱. 可变盐单胞菌中草甘膦抗性EPSP合酶新基因克隆、大肠杆菌表达及其抗性机制的研究[D]. 雅安:四川大学, 2004.
[15] 宋苗, 汪海, 张杰, 等. 转Bt cry1Ah基因抗虫玉米对亚洲玉米螟、棉铃虫和黏虫的抗性评价[J]. 生物技术通报, 2016, 32(6):69-75.
[16] Trtikova M, Wikmark OG, Zemp N, et al.Transgene expression and Bt protein content in transgenic Bt maize(MON810)under optimal and stressful environmental conditions[J]. PLoS One, 2015, 10(4):e0123011.
[17] 吴乃虎. 基因工程原理(下册)[M]. 北京:科学出版社, 2001:278-290.
[18] Paz J, Pla M, Papazova N, et al.Stability of the MON 810 transgene in maize[J]. Plant Molecular Biology, 2010, 74(6):563-571.
[19] Makarevitch I, Stupar RM, Iniguez AL, et al.Natural variation for alleles under epigenetic control by the maize chromomethylase Zmet2[J]. Genetics, 2007, 177(2):749-760.
[20] Sigman MJ, Slotkin RK.The first rule of plant transposable element silencing:location, location, location[J]. The Plant Cell, 2016, 28(2):304-313.
[21] Oka R, Zicola J, Weber B, et al.Genome-wide mapping of transcriptional enhancer candidates using DNA and chromatin features in maize[J]. Genome Biology, 2017, 18(1):137.
[22] Liu HJ, Luo X, Niu LY, et al.Distant eQTLs and non-coding sequences play critical roles in regulating gene expression and quantitative trait variation in maize[J]. Molecular Plant, 2017, 10(3):414-426.
[23] 刘月娥. 玉米对区域光、温、水资源变化的响应研究[D]. 北京:中国农业科学院, 2013.
[24] 康领生, 姜志磊, 刘洋, 等. 转基因玉米SW12-859的抗螟性及农艺性状评价[J]. 玉米科学, 2017, 25(5):45-48.
[25] 王延锋. 转Bt基因抗虫玉米田间试验与遗传稳定性分析[D]. 哈尔滨:东北农业大学, 2010.

转Bt基因抗虫玉米HGK60的农艺性状分析

Agronomic Traits Analysis of Transgenic Bt cry1Ah Maize HGK60 Line

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	范昕琦, 王海燕, 陈静, 张晓娟, 郭琦, 梁笃, 周福平, 聂萌恩, 张一中, 柳青山. EMS诱变对高粱成苗及M₁主要农艺性状的影响[J]. 生物技术通报, 2023, 39(7): 173-184.
[2]	朱少喜, 金肇阳, 葛建镕, 王蕊, 王凤格, 路运才. 基于KASP平台的转基因玉米高通量特异性检测方法[J]. 生物技术通报, 2023, 39(6): 133-140.
[3]	卢振万, 李雪琪, 黄金光, 周焕斌. 利用胞嘧啶碱基编辑技术创制耐草甘膦水稻[J]. 生物技术通报, 2023, 39(2): 63-69.
[4]	李圣彦, 李香银, 李鹏程, 张明俊, 张杰, 郎志宏. 转基因玉米2HVB5的性状鉴定及遗传稳定性分析[J]. 生物技术通报, 2023, 39(1): 21-30.
[5]	李鹏程, 张明俊, 王银晓, 李香银, 李圣彦, 郎志宏. 转基因玉米HGK60在不同遗传背景下抗虫性鉴定及农艺性状分析[J]. 生物技术通报, 2023, 39(1): 40-47.
[6]	王翠翠, 傅达奇. 泛素-蛋白酶体系统影响植物农艺性状的研究进展[J]. 生物技术通报, 2023, 39(1): 72-83.
[7]	杨青青, 唐家琪, 张昌泉, 高继平, 刘巧泉. KASP标记技术在主要农作物中的应用及展望[J]. 生物技术通报, 2022, 38(4): 58-71.
[8]	焦悦, 韩宇, 杨桥, 黄耀辉, 安吉翠, 杨亚洲, 叶纪明. 全球转基因玉米商业化发展态势概述及启示[J]. 生物技术通报, 2021, 37(4): 164-176.
[9]	温洪涛, 李夏莹, 杨洋, 陈子言, 丁一佳, 张秀杰, 张瑞英. 玉米转基因成分筛查策略[J]. 生物技术通报, 2020, 36(5): 39-47.
[10]	王颢潜, 肖芳, 杨蕾, 缪青梅, 张旭冬, 张秀杰. 转基因玉米双抗12-5转化体特异性PCR方法验证结果分析[J]. 生物技术通报, 2020, 36(5): 48-55.
[11]	杨镇州, 刘刚, 许丽. 基于RNAi技术的转基因玉米逆转录数字PCR检测方法[J]. 生物技术通报, 2020, 36(5): 56-63.
[12]	李葱葱, 谢苹, 董立明, 夏蔚, 兰青阔, 闫伟, 龙丽坤, 李飞武. 抗虫耐除草剂玉米GH5112E-117C定性PCR检测方法[J]. 生物技术通报, 2020, 36(5): 64-67.
[13]	张海淼, 李洋, 刘海峰, 孔令广, 丁新华. 水稻重要农艺性状调控基因及其育种利用研究进展[J]. 生物技术通报, 2020, 36(12): 155-169.
[14]	李爱国, 李积铭, 李和平, 刘桂华, 宋聪敏, 武军艳. 适宜河北省旱寒区的冬油菜品种的筛选[J]. 生物技术通报, 2020, 36(1): 95-100.
[15]	王翠云, 刘艳, 刘允军. 外源基因在转基因玉米中的整合位点分析[J]. 生物技术通报, 2019, 35(3): 1-5.