中国农业科学2025，Vol.58Issue(12)：2291-2302,12.DOI:10.3864/j.issn.0578-1752.2025.12.002
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
Resistance Evaluation and Genetic Stability Analysis of Insect-Resistant and Glyphosate-Tolerant Transgenic Cotton Lines
摘要
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.关键词
cry1Ac-vip3Da/g10-epsps/转基因棉花/抗虫性/草甘膦耐受性/稳定遗传Key words
cry1Ac-vip3Da/g10-epsps/transgenic cotton lines/insect resistance/glyphosate tolerance/stable genetics引用本文复制引用
文晋,宁艳芳,秦欣,刘圆,张晓玲,朱永红,田时敏,马燕斌..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[J].中国农业科学,2025,58(12):2291-2302,12.基金项目
山西省应用基础研究面上项目(202103021224145)、山西省基础研究计划(202403021222085)、山西省高等学校科技创新项目(2023L050)、山西省重点研发计划(202302140601004) （202103021224145）
