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

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

摘要/Abstract

摘要：

为研究转基因玉米HGK60在不同遗传背景下遗传稳定性和抗虫效果，利用转Bt cry1Ah基因的转基因玉米HGK60为供体，通过回交转育的方式将cry1Ah基因分别导入玉米自交系郑58、昌7-2、lx05-4、lx03-2，获得转基因玉米自交系HGK60-郑58、HGK60-昌7-2、HGK60-lx03-2、HGK60-lx05-4，并杂交获得HGK60-郑单958（HGK60-郑58 × HGK60-昌7-2）和HGK60-鲁单9066（HGK60-lx05-4 × HGK60-lx03-2），转化体特异性PCR证明cry1Ah基因已转入不同遗传背景玉米中，ELISA检测不同遗传背景转基因玉米叶片中Cry1Ah蛋白表达情况，结果表明在不同遗传背景玉米自交系和杂交种中Cry1Ah蛋白表达没有显著差异；田间人工接虫和室内玉米螟抗虫性鉴定结果表明，不同遗传背景的转基因玉米高抗玉米螟，室内接虫后4 d幼虫死亡率达到100%；对不同遗传背景转基因玉米HGK60进行农艺性状分析，结果显示与受体对照玉米相比，两者之间农艺性状没有显著差异，转基因玉米HGK60可用于抗虫玉米品种的选育。

关键词: 转基因玉米, cry1Ah基因, 不同遗传背景, 抗虫性, 农艺性状

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

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.

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.

图/表 5

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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