抗虫与耐草甘膦转基因棉花株系的抗性评价与遗传稳定分析

doi:10.3864/j.issn.0578-1752.2025.12.002

Abstract

Abstract:

【Objective】Cotton bollworm (Helicoverpa armigera) and field weeds are major constraints to high-yield cotton production. Existing varieties with single traits (insect resistance or herbicide tolerance) fail to meet the demands of efficient cultivation. Developing transgenic cotton varieties with combined insect resistance and glyphosate tolerance will provide high-efficiency germplasm resources for stress-resistant cotton breeding.【Method】The insect-resistant fusion gene cry1Ac-vip3Da and glyphosate-tolerant gene g10-epsps were introduced into cotton R15 through Agrobacterium-mediated method, regenerated transgenic plants were screened via PCR, positive lines underwent multi-generation self-pollination to achieve homozygosity, and stable lines with superior resistance were selected. The expression of target genes in different tissues of transgenic lines was analyzed using qRT-PCR and ELISA. Bioactivity assays and glyphosate tolerance tests were conducted to evaluate the genetic stability of insect resistance and herbicide tolerance across generations (T₄-T₆). Agronomic traits of transgenic lines were comprehensively assessed. 【Result】Eight positive transgenic lines with dual resistance were identified through PCR screening, and CA-6, CA-7 and CA-17 lines exhibited higher resistance. qRT-PCR revealed high expression of cry1Ac-vip3Da and g10-epsps in all tissues of these lines, and expression levels varied significantly among tissues. ELISA analysis demonstrated significant differences in Cry1Ac-Vip3Da and G10-EPSPS protein content across tissues of the three transgenic lines, with the highest levels observed in leaves. Protein accumulation gradually decreased during the developmental stages (from the four-leaf stage to boll-opening stage), but remained stable across T₄-T₆ generations. Bioactivity assays and glyphosate tolerance tests demonstrated that three transgenic cotton lines (T₄-T₆ generations) exhibited corrected mortality rates of 65.12%-82.75%, tolerated glyphosate at over four times the recommended dosage, and showed no attenuation of resistance across generations. There were no significant differences in plant height, number of fruit branches, number of bells per plant, bell weight, lint percentage, seed cotton yield, and lint cotton yield between transgenic lines and R15.【Conclusion】The exogenous genes cry1Ac-vip3Da and g10-epsps were stably inherited across generations in transgenic lines CA-6, CA-7, and CA-17, conferring dual insect resistance and glyphosate tolerance without compromising agronomic performance.

Key words: cry1Ac-vip3Da, g10-epsps, transgenic cotton lines, insect resistance, glyphosate tolerance, stable genetics

WEN Jin, NING YanFang, QIN Xin, LIU Yuan, ZHANG XiaoLing, ZHU YongHong, TIAN ShiMin, MA YanBin. Resistance Evaluation and Genetic Stability Analysis of Insect- Resistant and Glyphosate-Tolerant Transgenic Cotton Lines[J].Scientia Agricultura Sinica, 2025, 58(12): 2291-2302.

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References 46

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引物名称 Primer name	引物序列 Primer sequence (5′-3′)	用途 Usage
g10-epsps-F	CTCTTCTTCCTGACGGACTC	PCR
g10-epsps-R	GACTTTCTGATGATGTGGTGTGC
cry1Ac-vip3Da-F	CCATCTCACGCCATACTTCC
cry1Ac-vip3Da-R	CACCTTGATTGTCCCTGGTC
Histone3-QF	GGTGGTGTGAAGAAGCCTCAT	qRT-PCR
Histone3-QR	AATTTCACGAACAAGCCTCTGGAA
cry1Ac-vip3Da-QF	GAGGTACGAGGTGACTGCCAA
cry1Ac-vip3Da-QR	TCCTGTACTCAGCCTCACTGCT
g10-epsps-QF	CTCCCTTATCGGTTTCTCTGAAG
g10-epsps-QR	TGTAGTTCTTGGATGGCTGTGC

表型参数 Phenotypic parameters	R15	CA-6			CA-7			CA-17
表型参数 Phenotypic parameters	R15	T₄	T₅	T₆	T₄	T₅	T₆	T₄	T₅	T₆
叶片受害率 Leaf damage rate (%）	100.00	10.14±0.35b	11.68±0.85a	10.93±0.84ab	3.76±0.28e	3.97±0.33e	3.56±0.56e	5.38±0.55d	6.54±0.63c	6.38±0.35cd
成活率 Survival rate (%)	0.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00	100.00

材料 Material	株高 Plant height (cm)	果枝数 Fruit branches	单株铃数 Bolls number	铃重 Boll weight (g)	籽棉产量 Seed cotton yield (kg·hm^-2)	皮棉产量 Lint yield (kg·hm^-2)	衣分 Lint percentage (%)
R15	98.33±9.01	13.56±2.13	25.8±1.8	5.60±0.34	4231.56	1760.33	41.60±2.12
CA-6	100.43±4.51	14.10±1.93	25.6±3.2	5.90±0.65	4236.26	1745.34	41.20±2.06
CA-7	99.27±4.95	14.00±2.21	28.4±2.1*	5.40±1.01	4396.89	1798.33	40.90±2.25
CA-17	96.17±5.92	13.30±1.49	24.1±1.9	5.70±0.74	4191.50	1680.79	40.10±2.40

Resistance Evaluation and Genetic Stability Analysis of Insect- Resistant and Glyphosate-Tolerant Transgenic Cotton Lines

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[2]	KOU Jiang-Tao, SHI Shang-Li, HU Gui-Xin, ZHOU Wan-Hai, YAO Tuo. Photosynthetic Physiology of Odontothrips Damaged Medicago sativa [J]. Scientia Agricultura Sinica, 2013, 46(12): 2459-2470.
[3]	,,,,,. Inheritance Analysis and Mapping QTLs Related to Cotton Worm Resistance in Soybeans [J]. Scientia Agricultura Sinica, 2005, 38(07): 1369-1372 .