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中国农学通报 ›› 2011, Vol. 27 ›› Issue (12): 255-260.

所属专题: 生物技术 棉花

• 生物技术科学 • 上一篇    下一篇

过量表达棉花GhACO2基因增强拟南芥抗逆性研究

张萍 王斐 孙辉 李鸿彬   

  • 收稿日期:2011-01-24 修回日期:2011-02-23 出版日期:2011-05-25 发布日期:2011-05-25
  • 基金资助:

    农业部转基因生物新品种培育重大专项;兵团博士基金项目;石河子大学高层次人才项目

Overexpression of a Cotton GhACO2 Gene Enhance the Stress Tolerance of Ababidopsis Thaliana

  • Received:2011-01-24 Revised:2011-02-23 Online:2011-05-25 Published:2011-05-25

摘要:

乙烯在植物生长发育过程中起到非常重要的作用,ACO基因对于细胞内乙烯的合成起到关键的作用,然而乙烯在植物响应非生物胁迫方面的功能有很大的未知性。研究通过PCR方法从棉花基因组中克隆获得GhACO2基因,构建了植物过量表达载体p35S::GhACO2,通过花序侵染法转化拟南芥,用卡那霉素对转化植株进行初步筛选,进一步对阳性植株进行PCR和GUS组织化学分析检测,结果表明,在转基因拟南芥叶片中有很强的GUS活性,茎和根有微量表达;GhACO2基因已经整合到拟南芥基因组中,成功获得转基因拟南芥。经过纯合筛选后获得转基因T2代拟南芥植株,利用NaCl和PEG6000对T2代植株进行盐胁迫和干旱胁迫处理,结果显示,与野生型拟南芥相比,转GhACO2基因拟南芥对盐胁迫和干旱胁迫的耐受性显著性的增强。研究结果为进一步探讨GhACO2的生物学功能和利用基因工程技术进行转基因育种质奠定了基础。

关键词: 时间结构, 时间结构

Abstract:

Ethylene signaling plays important roles in multiple processes of plant growth and development, ACO gene is the key factor regulating biosynthesis of ethylene, however, functions of ACO gene and ethylene in abiotic stress responses remain largely unknown. Here, we cloned a cotton GhACO2 gene from cotton fiber cDNA and constructed a plant overexpression vector p35S::GhACO2. Genetic transformation of Arabidopsis was performed by floral dip method, successive selection were completed with Kanamycin and further PCR and GUS histochemical detection of transgenic arabidopsis plants. The results of selection showed that GUS enzyme of transgenic arabidopsis plants was expressed greatly in leaf and little in stem and root, GhACO2 gene was integrated in the Arabidopsis genome and transgenic positive plants were obtained successfully. The T2 generation homozygous transgenic plants were obtained by selfing, and were used to undergo salt-stress and drought-stress treatment by NaCl and PEG6000. The results indicated that, with the comparison of Col wide type plants, stress tolerances of transgenic plants were enhanced significantly. All these results establish the basis of researching biological functions of GhACO2 further and transgenic breeding by Genetic engineering technology.