Insect Resistance Identification and Agronomy Traits Analysis of Transgenic Maize HGK60 with Different Genetic Backgrounds

doi:10.13560/j.cnki.biotech.bull.1985.2022-1227

Abstract

Abstract:

In order to study the genetic stability and insect resistance of transgenic maize HGK60 under different genetic backgrounds, HGK60 transgenic maize harboring Bt cry1Ah gene was used as donor, and the cry1Ah gene was introduced into inbred line Zheng 58, Chang 7-2, lx05-4 and lx03-2 by successive backcrossing respectively, and transgenic maize inbred line HGK60-Zheng 58, HGK60-Chang 7-2, HGK60-lx03-2, and HGK60-lx05-4 were acquired. The hybrid line HGK60-zhengdan 958（HGK60-Zheng 58 with HGK60-Chang 7-2）and HGK60-Ludan 9066（HGK60-lx03-2 with HGK60-lx05-4）were obtained by crossing. The foreign cry1Ah gene was confirmed to be transferred into the maize with different genetic background by event-special PCR detection. The expressions of Cry1Ah proteins in the transgenic maize leaves of the inbred and hybrid lines under different genetic backgrounds were insignificantly different by ELISA detection. Bioassay in field and in laboratory showed that the transgenic maize with different genetic backgrounds had same high resistance to Asia corn borer, and the mortality rate reached 100% at 4 d after indoor inoculation. The agronomic traits of the transgenic maize HGK60 with different genetic backgrounds demonstrated that there was no significant difference in agronomic traits between the transgenic maize and the non-transgenic control maize, and transgenic maize HGK60 may be used for the breeding of insect-resistant maize varieties.

Key words: transgenic maize, cry1Ah gene, different genetic background, insect resistance, agronomic traits

LI Peng-cheng, ZHANG Ming-jun, WANG Yin-xiao, LI Xiang-yin, LI Sheng-yan, LANG Zhi-hong. Insect Resistance Identification and Agronomy Traits Analysis of Transgenic Maize HGK60 with Different Genetic Backgrounds[J]. Biotechnology Bulletin, 2023, 39(1): 40-47.

Figures/Tables 5

References 19

[1]	刘杰, 李天娇, 姜玉英, 等. 2020年我国玉米主要病虫害发生特点[J]. 中国植保导刊, 2021, 41(8): 30-35.
	Liu J, Li TJ, Jiang YY, et al. Occurrence characteristics of main maize diseases and insect pests in China in 2020[J]. China Plant Prot, 2021, 41(8): 30-35.
[2]	国际农业生物技术应用服务组织. 2019年全球生物技术/转基因作物商业化发展态势[J]. 中国生物工程杂志, 2021, 41(1): 114-119.
	International Service for the Acquisition of Agri-biotech Applications. Development trend of global biotechnology/GM crop commercialization in 2019[J]. China Biotechnol, 2021, 41(1): 114-119.
[3]	Xie W, Ali T, Cui Q, et al. Economic impacts of commercializing insect-resistant GM maize in China[J]. China Agric Econ Rev, 2017, 9(3): 340-354.
[4]	梁晋刚, 张旭冬, 毕研哲, 等. 转基因抗虫玉米发展现状与展望[J]. 中国生物工程杂志, 2021, 41(6): 98-104.
	Liang JG, Zhang XD, Bi YZ, et al. Development status and prospect of genetically modified insect-resistant maize[J]. China Biotechnol, 2021, 41(6): 98-104.
[5]	Li YH, Hallerman EM, Wu KM, et al. Insect-resistant genetically engineered crops in China: development, application, and prospects for use[J]. Annu Rev Entomol, 2020, 65: 273-292. doi: 10.1146/annurev-ento-011019-025039 pmid: 31594412
[6]	Xue J, Liang GM, 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 Microbiol Lett, 2008, 280(1): 95-101. doi: 10.1111/j.1574-6968.2007.01053.x URL
[7]	Xue J, Zhou Z, Song F, et al. Identification of the minimal active fragment of the Cry1Ah toxin[J]. Biotechnol Lett, 2011, 33(3): 531-537. doi: 10.1007/s10529-010-0452-0 pmid: 21046197
[8]	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.
	Dai J, Li XY, Zhu L, et al. Molecular detection and agronomic traits analysis of insect-resistant transgenic maize harboring Bt cry1Ah gene[J]. Biotechnol Bull, 2014(5): 62-68.
[9]	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. doi: 10.13560/j.cnki.biotech.bull.1985.2016.06.011
	Song M, Wang H, Zhang J, et al. Resistance evaluation of Bt cry1Ah-transgenic maize to Asian corn borer, cotton bollworm and oriental armyworm[J]. Biotechnol Bull, 2016, 32(6): 69-75.
[10]	梁海生, 李梦桃, 李圣彦, 等. 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. doi: 10.13560/j.cnki.biotech.bull.1985.2018-0219
	Liang HS, Li MT, Li SY, et al. Agronomic traits analysis of transgenic Bt cry1Ah maize HGK60 line[J]. Biotechnol Bull, 2018, 34(7): 92-100.
[11]	卢美贞, 崔海瑞, 姚艳玲, 等. 影响苏云金芽孢杆菌基因在转基因植物中表达的因素[J]. 细胞生物学杂志, 2005, 27(5): 509-513.
	Lu MZ, Cui HR, Yao YL, et al. Factors affecting expression of Bacillus thuringiensis genes in transgenic plants[J]. Chin J Coll Biol, 2005, 27(5): 509-513.
[12]	杨宙, 吕再萍, 黄仁良, 等. Bt蛋白在水稻不同遗传背景下的含量及对抗虫性的影响[J]. 植物保护学报, 2017, 44(6): 1017-1023.
	Yang Z, Lü ZP, Huang RL, et al. Content of Bt protein in different backgrounds of rice and its effect on insect resistance[J]. J Plant Prot, 2017, 44(6): 1017-1023.
[13]	王冬妍, 王振营, 何康来, 等. Bt玉米杀虫蛋白含量的时空表达及对亚洲玉米螟的杀虫效果[J]. 中国农业科学, 2004, 37(8): 1155-1159.
	Wang DY, Wang ZY, He KL, et al. Temporal and spatial expression of CrylAb toxin in transgenic Bt corn and its effects on Asian corn borer, Ostrinia furnacalis(guenee)[J]. Sci Agric Sin, 2004, 37(8): 1155-1159.
[14]	刘晓贝, 白树雄, 王振营, 等. 转cry1Ab/cry1Ac基因玉米Cry1Ab/Cry1Ac融合蛋白表达及对亚洲玉米螟的室内杀虫效果[J]. 昆虫学报, 2020, 63(10): 1201-1206.
	Liu XB, Bai SX, Wang ZY, et al. Expression of Cry1Ab/Cry1Ac fusion protein in the transgenic cry1Ab/cry1Ac maize and its control efficacy against the Asian corn borer, Ostrinia furnacalis(Lepidoptera: Crambidae), in the laboratory[J]. Acta Entomol Sin, 2020, 63(10): 1201-1206.
[15]	林敏. 农业生物育种技术的发展历程及产业化对策[J]. 生物技术进展, 2021, 11(4): 405-417.
	Lin M. The development course and industrialization countermeasure of agricultural biological breeding technology[J]. Curr Biotechnol, 2021, 11(4): 405-417.
[16]	Venkatesh TV, Cook K, Liu B, et al. Compositional differences between near-isogenic GM and conventional maize hybrids are associated with backcrossing practices in conventional breeding[J]. Plant Biotechnol J, 2015, 13(2): 200-210. doi: 10.1111/pbi.12248 pmid: 25196222
[17]	夏兰芹, 王远, 郭三堆. 外源基因在转基因植物中的表达与稳定性[J]. 生物技术通报, 2000(3): 8-12.
	Xia LQ, Wang Y, Guo SD. The stability of the expression of foreign genes in transgenic plants[J]. Biotechnol Inf, 2000(3): 8-12.
[18]	孙国清. 不同抗虫棉品种杀虫蛋白表达研究及影响抗虫性分析[D]. 乌鲁木齐: 新疆农业大学, 2010.
	Sun GQ. Study on expression and influencing factor of Bt insecticidal protein in transgenic insect-resistant cotton[D]. Urumqi: Xinjiang Agricultural University, 2010.
[19]	王培, 何康来, 王振营, 等. 转cry1Ac玉米对亚洲玉米螟的抗性评价[J]. 植物保护学报, 2012, 39(5): 395-400.
	Wang P, He KL, Wang ZY, et al. Evaluating transgenic cry1Ac maize for resistance to Ostrinia furnacalis(Guenée)[J]. J Plant Prot, 2012, 39(5): 395-400.

玉米材料 Corn material	2020年海南三亚 Sanya, Hainan in 2020		2021年河北廊坊 Langfang, Hebei in 2021		2021年海南三亚 Sanya, Hainan in 2022
玉米材料 Corn material	3 d死亡率 3-day mortality rate/%	4 d死亡率 4-day mortality rate/%	3 d死亡率 3-day mortality rate/%	4 d死亡率 4-day mortality rate/%	3 d死亡率 3-day mortality rate/%	4 d死亡率 4-day mortality rate/%
HGK60-郑58	100.00±0.00	-	100.00±0.00	-	91.67±4.17	100.00±0.00
HGK60-昌7-2	100.00±0.00	-	100.00±0.00	-	93.06±2.41	100.00±0.00
HGK60-lx03-2	100.00±0.00	-	100.00±0.00	-	97.22±2.41	100.00±0.00
HGK60-lx05-4	100.00±0.00	-	100.00±0.00	-	94.44±2.41	100.00±0.00
HGK60-郑单958	100.00±0.00	-	100.00±0.00	-	94.44±2.41	100.00±0.00
HGK60-鲁单9066	100.00±0.00	-	100.00±0.00	-	98.61±2.41	100.00±0.00
非转基因玉米-郑58	1.39±2.41	1.39±2.41	0.00±0.00	0.00±0.00	5.56±2.41	5.56±2.41

玉米材料 Corn material	2020年海南三亚 Sanya, Hainan in 2020		2021年河北廊坊 Langfang, Hebei in 2021		2021年海南三亚 Sanya, Hainan in 2022
玉米材料 Corn material	3 d死亡率 3-day mortality rate/%	4 d死亡率 4-day mortality rate/%	3 d死亡率 3-day mortality rate/%	4 d死亡率 4-day mortality rate/%	3 d死亡率 3-day mortality rate/%	4 d死亡率 4-day mortality rate/%
HGK60-郑58	100.00±0.00	-	100.00±0.00	-	91.67±4.17	100.00±0.00
HGK60-昌7-2	100.00±0.00	-	100.00±0.00	-	93.06±2.41	100.00±0.00
HGK60-lx03-2	100.00±0.00	-	100.00±0.00	-	97.22±2.41	100.00±0.00
HGK60-lx05-4	100.00±0.00	-	100.00±0.00	-	94.44±2.41	100.00±0.00
HGK60-郑单958	100.00±0.00	-	100.00±0.00	-	94.44±2.41	100.00±0.00
HGK60-鲁单9066	100.00±0.00	-	100.00±0.00	-	98.61±2.41	100.00±0.00
非转基因玉米-郑58	1.39±2.41	1.39±2.41	0.00±0.00	0.00±0.00	5.56±2.41	5.56±2.41

年代地点 Year and location	转基因玉米 Transgenic maize	抗性等级 Resistance level	非转基因玉米 Non-transgenic maize	抗性等级 Resistance level
2020年海南三亚	HGK60-郑58	1.34±0.38a	郑58	7.03±0.23b
	HGK60-昌7-2	1.31±0.57a	昌7-2	6.34±0.23b
	HGK60-lx03-2	1.48±0.35a	lx03-2	6.48±0.78b
	HGK60-lx05-4	1.72±0.31a	lx05-4	6.95±0.49b
	HGK60-郑单958	1.23±0.94a	郑单958	6.67±0.69b
	HGK60-鲁单9066	1.28±0.46a	鲁单9066	6.89±0.87b
2021年河北廊坊	HGK60-郑58	1.39±0.45a	郑58	7.56±0.49b
	HGK60-昌7-2	1.78±0.30a	昌7-2	6.63±0.71b
	HGK60-lx03-2	1.48±0.45a	lx03-2	6.93±0.59b
	HGK60-lx05-4	1.42±0.40a	lx05-4	7.01±0.52b
	HGK60-郑单958	1.31±0.29a	郑单958	6.85±0.27b
	HGK60-鲁单9066	1.37±0.65a	鲁单9066	6.42±0.68b
2021年海南三亚	HGK60-郑58	1.21±0.71a	郑58	6.21±0.18b
	HGK60-昌7-2	1.51±0.54a	昌7-2	6.78±0.56b
	HGK60-lx03-2	1.63±0.34a	lx03-2	6.97±0.72b
	HGK60-lx05-4	1.51±0.11a	lx05-4	7.31±0.67b
	HGK60-郑单958	1.18±0.54a	郑单958	6.41±0.93b
	HGK60-鲁单9066	1.23±0.43a	鲁单9066	6.56±0.67b

年代地点 Year and location	转基因玉米 Transgenic maize	抗性等级 Resistance level	非转基因玉米 Non-transgenic maize	抗性等级 Resistance level
2020年海南三亚	HGK60-郑58	1.34±0.38a	郑58	7.03±0.23b
	HGK60-昌7-2	1.31±0.57a	昌7-2	6.34±0.23b
	HGK60-lx03-2	1.48±0.35a	lx03-2	6.48±0.78b
	HGK60-lx05-4	1.72±0.31a	lx05-4	6.95±0.49b
	HGK60-郑单958	1.23±0.94a	郑单958	6.67±0.69b
	HGK60-鲁单9066	1.28±0.46a	鲁单9066	6.89±0.87b
2021年河北廊坊	HGK60-郑58	1.39±0.45a	郑58	7.56±0.49b
	HGK60-昌7-2	1.78±0.30a	昌7-2	6.63±0.71b
	HGK60-lx03-2	1.48±0.45a	lx03-2	6.93±0.59b
	HGK60-lx05-4	1.42±0.40a	lx05-4	7.01±0.52b
	HGK60-郑单958	1.31±0.29a	郑单958	6.85±0.27b
	HGK60-鲁单9066	1.37±0.65a	鲁单9066	6.42±0.68b
2021年海南三亚	HGK60-郑58	1.21±0.71a	郑58	6.21±0.18b
	HGK60-昌7-2	1.51±0.54a	昌7-2	6.78±0.56b
	HGK60-lx03-2	1.63±0.34a	lx03-2	6.97±0.72b
	HGK60-lx05-4	1.51±0.11a	lx05-4	7.31±0.67b
	HGK60-郑单958	1.18±0.54a	郑单958	6.41±0.93b
	HGK60-鲁单9066	1.23±0.43a	鲁单9066	6.56±0.67b

年代地点 Year and location	农艺性状 Agronomy traits	HGK60-郑单958 HGK60-zhengdan 958	郑单958 Zhengdan 958	HGK60-鲁单9066 HGK60-Ludan 9066	鲁单9066 Ludan 9066
2020年海南三亚	株高	190.34±12.31a	193.13±11.71a	223.97±10.13a	228.14±12.15a
	穗位高	90.44±5.87a	92.15±6.87a	101.08±7.81a	103.78±11.57a
	穗长	16.01±0.85a	15.95±1.06a	14.91±1.75a	14.13±1.48a
	穗粗	4.43±0.25a	4.77±0.22a	3.78±0.28a	3.78±0.22a
	穗行数	13.95±1.45a	14.98±1.25a	12.87±1.01a	12.00±0.91a
	秃尖长	0.42±0.31a	0.44±0.33a	0.44±0.25a	0.46±0.37a
	行粒数	38.64±3.43a	38.21±2.23a	39.84±4.75a	38.85±4.52a
	百粒重	27.13±2.22a	26.24±3.24a	22.41±1.83a	23.22±2.33a
	单株产量	152.74±7.13a	150.82±12.81a	112.65±13.04a	115.84±12.91a
2021年河北廊坊	株高	248.04±10.87a	250.72±10.96a	294.77±11.14a	288.28±12.64a
	穗位高	119.51±9.04a	120.61±9.15a	133.34±7.47a	137.18±9.92a
	穗长	19.81±1.22a	18.94±1.41a	21.64±2.33a	22.05±0.87a
	穗粗	4.85±0.32a	4.81±0.26a	4.65±0.24a	4.65±0.16a
	穗行数	15.62±1.27a	15.81±1.47a	13.76±1.21a	13.85±1.56a
	秃尖长	0.56±0.36a	0.65±0.46a	0.47±0.47a	0.75±0.56a
	行粒数	39.71±3.15a	39.63±3.96a	41.35±6.76a	42.86±2.55a
	百粒重	34.85±1.34a	35.23±1.55a	29.51±1.16a	29.91±1.35a
	单株产量	188.51±12.35a	191.61±13.26a	161.57±10.1a	164.26±12.63a
2021年海南三亚	株高	187.34±11.23a	188.24±13.42a	224.01±11.24a	226.89±11.26a
	穗位高	88.43±4.67a	90.10±5.65a	99.08±6.19a	101.68±12.68a
	穗长	15.93±0.45a	16.02±1.23a	14.92±2.03a	14.61±1.39a
	穗粗	4.35±0.21a	4.43±0.18a	3.56±0.30a	3.61±0.32a
	穗行数	14.01±1.14a	14.83±1.34a	12.73±1.34a	12.34±0.56a
	秃尖长	0.40±0.24a	0.45±0.38a	0.45±0.34a	0.46±0.35a
	行粒数	37.94±2.54a	38.45±3.12a	38.45±4.67a	39.05±4.52a
	百粒重	26.63±1.39a	25.93±2.18a	23.02±1.75a	24.01±2.61a
	单株产量	155.53±6.19a	153.54±11.67a	113.43±12.07a	114.45±13.45a