棉花学报2025，Vol.37Issue(5)：429-438,10.DOI:10.11963/cs20250046
棉花茎尖遗传转化体系优化研究
Optimization of the genetic transformation system of cotton shoot tip
摘要
Abstract
[Objective]The study aims at investigating the optimal transformation system to enhance genetic transformation efficiency of cotton shoot tips.[Methods]The shoot tips of Baimian 1 seeds were used as the explant.Different concentrations of ethylenediaminetetra-acetic acid(EDTA),various water bath treatment times,and different concentrations of bacterial suspension were set during the infection stage.During the co-culture stage,six exogenous additives(lanthanum chloride,EDTA,salicylic acid,thidiazuron,zeatin,and silver nitrate)at varying concentrations were introduced.During the screening stage,the concentrations of spectinomycin and ratios of various plant growth regulators(kinetin,naphthalene acetic acid,6-benzylaminopurine,indole butyric acid,and 2,4-dichlorophenoxyacetic acid)were adjusted.The effects of these treatments on the transient transformation efficiency,germination rate,seedling survival rate,and positive rate of cotton shoot tips were analyzed.[Results]During the infection stage,adjusting the OD600 of bacterial suspension to 0.7 and pre-treating shoot tips with 100 μmol·L-1 EDTA in a water bath for 15 min can significantly improve the transient transformation efficiency(28.09％).During the co-culture phase,adding 1.5 mg·L-1 thidiazuron obtained the highest transient transformation efficiency(41.47％)and a relatively high germination rate(32.79％).In the screening phase,adding 200 mg·L-1 spectinomycin resulted in a higher seedling survival rate(32.44％)and the highest positive rate(19.97％);1 mg·L-1 6-benzylaminopurine+0.1 mg·L-1 naphthalene acetic acid treatment yielded a significantly higher germination rate(34.73％)than other plant growth regulator combinations.Under the optimized conditions described above,the transformation efficiency of cotton shoot tips reached 7.62％,with a positive rate of 18.52％.[Conclusion]The cotton genetic transformation system has been optimized,providing a reference for further improving the transformation efficiency of cotton shoot tips.关键词
棉花/遗传转化/茎尖分生组织/转化效率/阳性率/外源添加物/植物生长调节剂Key words
cotton/genetic transformation/shoot tip meristem/transformation efficiency/positive rate/exogenous additives/plant growth regulators引用本文复制引用
葛桓羽,左东云,王巧连,何嫚,于锐,程海亮,陈全家,吕丽敏,宋国立..棉花茎尖遗传转化体系优化研究[J].棉花学报,2025,37(5):429-438,10.基金项目
国家重点研发计划(2024YFD1200305) （2024YFD1200305）
三亚中国农业科学院国家南繁研究院"南繁专项"(YBXM2546) （YBXM2546）
