Agronomic Traits Analysis of Transgenic Bt cry1Ah Maize HGK60 Line

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

Abstract

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

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.

References

[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.