转基因玉米2HVB5的性状鉴定及遗传稳定性分析

doi:10.13560/j.cnki.biotech.bull.1985.2022-0536

摘要/Abstract

摘要：

为了明确转基因玉米2HVB5的目标性状及遗传稳定性，对回交转育郑58的BC₅S₁、BC₅S₂代转基因玉米2HVB5分别进行了Southern blot、ELISA、室内和田间生物活性测定、靶标除草剂草铵膦耐受性分析及农艺性状调查。结果显示，2HVB5中目的基因cry2Ah-vp和bar都是以单拷贝的形式整合到玉米基因组并稳定遗传，Cry2Ah-vp和PAT蛋白在2HVB5植株的不同时期、不同组织部位均有表达，其中在叶片中的表达量相对较高，分别达到2-3.5 μg/g FW（鲜重）和8-17 μg/g FW（鲜重）。室内生物活性检测结果表明，2HVB5转基因玉米对东方粘虫和棉铃虫有很高的抗性，接虫后4-5 d幼虫死亡率达100%，对草地贪夜蛾有明显的体重抑制。田间抗虫性鉴定结果也表明，2HVB5转基因玉米对东方粘虫和棉铃虫均达到高抗水平，平均抗性级别分别为1.19-1.29和0.60-0.73。2HVB5转基因玉米可耐受田间使用中剂量4倍量的草铵膦，农艺性状与对照郑58相比无显著差异。转基因玉米2HVB5遗传稳定，高抗虫耐除草剂，可用于玉米害虫尤其是夜蛾科害虫的防治，具有产业化应用前景。

关键词: 抗虫耐除草剂, 转基因玉米, 性状鉴定, 遗传稳定性

Abstract:

In order to confirm the target traits and genetic stability of transgenic maize 2HVB5，Southern blot, ELISA, bioassay in laboratory and field, target herbicide-tolerance analysis and agronomic traits investigation were carried out on BC₅S₁ and BC₅S₂ transgenic maize 2HVB5 plants which had been backcrossed to Zheng 58. The results showed that the target genes cry2Ah-vp and bar were integrated into maize genome in the form of single copy and steady inheritance in 2HVB5 plants. Cry2Ah-VP and PAT proteins were expressed in different stages and different tissues in 2HVB5 plants, and the expressions in the leaves were relatively high, reaching 2-3.5 μg/g fresh weight（FW）and 8-17 μg/g FW respectively. The bioassay results in laboratory showed that 2HVB5 plants had high resistance to armyworm（Mythimna separate）and cotton bollworm（Helicoverpa armigera）， the corrected mortality rates of armyworm and cotton bollworm larvae reached 100% on 4-5 d, and the 2HVB5 plants presented a significant weight inhibiton to fall armyworm（Spodoptera frugiperda）. The bioassay results in field also showed that 2HVB5 plants had high resistances to armyworm and cotton bollworm, the average resistance indexes were 1.19-1.29 and 0.60-0.73， respectively. The transgenic maize 2HVB5 tolerated 4 times the recommended medium dose for glufosinate application in field, and there were no significant differences in agronomic traits compared with the control Zheng 58. These indicate that transgenic maize 2HVB5 is of genetic stability and high insect resistance and herbicide resistance. It can be used for the control of corn pests, especially Noctuidae pests, and has a prospect of commercial application.

Key words: insect resistance and herbicide tolerance, transgenic maize, trait identification, genetic stability

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.

LI Sheng-yan, LI Xiang-yin, LI Peng-cheng, ZHANG Ming-jun, ZHANG Jie, LANG Zhi-hong. Identification of Target Traits and Genetic Stability of Transgenic Maize 2HVB5[J]. Biotechnology Bulletin, 2023, 39(1): 21-30.

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代别 Generation	试验材料 Sample	食叶级别 Grade of leaf damage	抗性类型 Type of resistance
BC₅S₁	2HVB5	1.19±0.40a	高抗
BC₅S₁	Z58	7.10±0.94b	感
BC₅S₂	2HVB5	1.29±0.56a	高抗
BC₅S₂	Z58	7.05±1.24b	感

代别 Generation	试验材料 Sample	食叶级别 Grade of leaf damage	抗性类型 Type of resistance
BC₅S₁	2HVB5	1.19±0.40a	高抗
BC₅S₁	Z58	7.10±0.94b	感
BC₅S₂	2HVB5	1.29±0.56a	高抗
BC₅S₂	Z58	7.05±1.24b	感

代别 Generation	试验材料 Samples	食穗级别 Grade of ear damage	抗性类型 Type of resistance
BC₅S₁	2HVB5	0.60±0.56a	高抗
BC₅S₁	Z58	3.20±1.73b	中抗
BC₅S₂	2HVB5	0.73±0.45a	高抗
BC₅S₂	Z58	3.97±1.47b	中抗

代别 Generation	试验材料 Samples	食穗级别 Grade of ear damage	抗性类型 Type of resistance
BC₅S₁	2HVB5	0.60±0.56a	高抗
BC₅S₁	Z58	3.20±1.73b	中抗
BC₅S₂	2HVB5	0.73±0.45a	高抗
BC₅S₂	Z58	3.97±1.47b	中抗

代别 Generation	材料 Samples	草铵膦处理 Glufosinate treatment/（g·hm^-2）	成苗率Planting percent/%			植株高度Plant height/cm			受害率Damage percent/%			单株产量 Yield per plant/g
代别 Generation	材料 Samples	草铵膦处理 Glufosinate treatment/（g·hm^-2）	1周 1 week	2周 2 weeks	4周 4 weeks	1周 1 week	2周 2 weeks	4周 4 weeks	1周 1 week	2周 2 weeks	4周 4 weeks	单株产量 Yield per plant/g
BC₅S₁	Z58	0	100	100	100	46.3±1.38a	54.95±1.47a	113.45±1.47a	0	0	0	106.7±2.32a
	2HVB5	0	100	100	100	46.65±1.14a	55.7±1.34a	113.6±1.5a	0	0	0	105.7±2.62a
	Z58	600	0	0	0	0	0	0	100	100	100	0
	2HVB5	600	100	100	100	46.5±1.15a	55.05±1.19a	113.15±1.95a	0	0	0	106.6±2.68a
	Z58	1 200	0	0	0	0	0	0	100	100	100	0
	2HVB5	1 200	100	100	100	46.35±0.99a	55±1.56a	112.5±2.14a	0	0	0	105±2.70a
	Z58	2 400	0	0	0	0	0	0	100	100	100	0
	2HVB5	2 400	100	100	100	47.1±1.07a	55±1.65a	113.5±2.48a	0	0	0	106.7±2.45a
BC₅S₂	Z58	0	100	100	100	46.05±1.32a	53.1±1.62a	112.45±1.7a	0	0	0	106.4±2.56a
	2HVB5	0	100	100	100	45.25±1.07a	53.95±1.43a	112.55±2.04a	0	0	0	106.6±2.95a
	Z58	600	0	0	0	0	0	0	100	100	100	0
	2HVB5	600	100	100	100	46±1.21a	53±1.62a	112.7±2.05a	0	0	0	105.15±2.16a
	Z58	1 200	0	0	0	0	0	0	100	100	100	0
	2HVB5	1 200	100	100	100	45.85±1.39a	53.5±1.54a	112.1±1.62a	0	0	0	106.5±2.89a
	Z58	2 400	0	0	0	0	0	0	100	100	100	0
	2HVB5	2 400	100	100	100	45.65±1.35a	53.45±1.76a	112.95±1.54a	0	0	0	106.45±2.5a