Identification of Target Traits and Genetic Stability of Transgenic Maize 2HVB5

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

Abstract

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

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.

Figures/Tables 8

References 27

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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