核农学报2017，Vol.31Issue(10)：1881-1888,8.DOI:10.11869/j.issn.100-8551.2017.10.1881
农杆菌介导陆地棉棕彩选1号茎尖遗传转化
Agrobacterium-mediated Genetic Transformation of Stem Apexes of Brown Fiber Cotton Zongcaixuan 1
摘要
Abstract
In order to establish an efficient genetic transformation system for stem apexes of brown cotton,Zongcaixuan 1 that belongs to upland cotton was used as materials for genetic transformation using Agrobacterium-mediated method.In this paper,the shoot tips from 7-day-old seedlings of Zongcaixuan 1 were selected as receptors,and co-culture time,co-culture temperature,bacteria OD600 and infection time were optimized by orthogonal test.In addition,the effects of the size of the shoot tip and the vacuum infiltration on the transformation efficiency were also studied,and the genetic transformation system of stem apexes from brown cotton was initially established.The results showed that the stem apexes with 0.5 cm hypocotyl were suitable for transformation,and the suitable conditions for Agrobacterium-mediated transformation were Agro bacteria concentration of OD600 =1.0,infection time for 20 min,co-culture time for 72 h and co-culture temporature for 24℃.Moreover,vacuum osmotic treatment was helpful to the occurrence of resistant seedlings.At first,plantlets were screened on the medium containing 100 mg· L-1 kanamycin,and 102 resistant plants were obtained.Subsequently,these plants were further detected by PCR and Southern blot,and in total 24 transgenic plants were obtained with a single copy gene insertion.The transformation rate was 0.4 ％.Furthermore,the optimization conditions for Agrobacterium-mediated genetic transformation of stem apexes of Zongcaixuan 1 were established in this study,which laid a foundation for the genetic transformation of brown cotton.关键词
棕色棉/茎尖/农杆菌/遗传转化Key words
brown cotton/stem apex/Agrobacterium tumefaciens/genetic transformation引用本文复制引用
高俊山,许磊,陈文,沈知临,蔡永萍,林毅..农杆菌介导陆地棉棕彩选1号茎尖遗传转化[J].核农学报,2017,31(10):1881-1888,8.基金项目
国家自然科学基金(31672497),安徽省“115”产业创新团队(皖人才办[2011]2号) （31672497）
