数字农科院2.0

Rapid And Convenient Transformation Of C.otton (Gossypium Hirsutum L.) U sing In Planta Shoot Apex Via Glyphosate Selection

文献类型： 外文期刊

作者： Wang, Kevin Yueju;Guo Wen-Fang;Wang Nan;Liu De-Hu;Li Gang-Qiang;Li Jun

作者机构：

关键词： shoot apex transformation; CP4-EPSPS; glyphosate; in planta; transgenic cotton

期刊名称： JOURNAL OF INTEGRATIVE AGRICULTURE

ISSN： 2095-3119

年卷期： 2018 年 17 卷 10 期

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摘要： Cotton plants are recalcitrant with regards to transformation and induced regeneration. In the present study, 5-enolpyruvylshikimate-3-phosphate (EPSPS), a glyphosate resistant gene from the bacterium Agrobacterium sp. strain CP4, was introduced into an elite Bt transgenic cotton cultivar with a modified technique involving in planta Agrobacteriummediated transformation of shoot apex. Primary transformants were initially screened using a 0.26% glyphosate spray and subsequently by PCR analysis. Five out of 4000 transformants from T-1 seeds were obtained resulting in an in planta transformation rate of 0.125%. Four homozygous lines were produced by continuous self-fertilization and both PCR-based selection and glyphosate resistance. Transgene insertion was analyzed by Southern blot analysis. Gene transcription and protein expression levels in the transgenic cotton lines were further investigated by RT-PCR, Western blot, and ELISA methods. Transgenic T-3 plants were resistant to as much as 0.4% of glyphosate treatments in field trials. Our results indicate that the cotton shoot apex transformation technique which is both tissue-culture and genotype-independent would enable the exploitation of transgene technology in different cotton cultivars. Since this method does not require sterile conditions, the use of specialized growth media or the application of plant hormones, it can be conducted under the greenhouse condition.

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