高分子量麦谷蛋白亚基基因1Bx14转化小麦

 【目的】利用基因枪将优质高分子量麦谷蛋白亚基基因1Bx14导入普通小麦栽培品种绵阳19，为利用优质基因进行小麦品质改良奠定基础。【方法】构建小麦优质高分子量麦谷蛋白亚基基因1Bx14的胚乳特异性植物表达载体，利用基因枪将其转入不含该亚基的小麦品种绵阳19幼胚愈伤组织中，经潮霉素抗性筛选、PCR检测后，阳性植株进行PCR-Southern和Southern杂交验证，利用SDS-PAGE分析目的基因在转基因后代籽粒中的表达。【结果】从转化的652块愈伤组织中共获得6株转基因阳性植株，转化效率0.92% 。转化后代种子分析表明，1Bx14在部分转基因后代种子中表达，但同时也导致部分内源高分子量麦谷蛋白亚基基因不同程度的沉默。【结论】本研究成功地将小麦高分子量麦谷蛋白亚基基因1Bx14导入普通小麦绵阳19中，并在部分后代种子中得到了表达。

Transformation of Wheat with High Molecular Weight Glutenin Subunit Gene 1Bx14

【Objective】 The objective of this study is to use genetic transformation of high quality HMW-GS gene 1Bx14 to improve wheat bread-making quality. 【Method】 This investigation, based on the clone of HMW-GS gene 1Bx14 and construction of plant expression vectors, transformed the immature embryo of Mianyang19 by microprojectile bombardment. After selection with hygromycin and tested by PCR, the positive progenies were confirmed by PCR-Southern and Southern blotting. The compositions of high molecular weight glutenin subunit were analyzed by SDS-PAGE.【Result】Six transformed plantlet was obtained from 652 transformed callus, yielding a transformation frequency of 0.92%. The HMW-GS gene 1Bx14 was expressed in some seeds of transgenic plants with the expression of other subunits suppressed in varying degrees. 【Conclusion】The high molecular weight glutenin subunit gene 1Bx14 was transformed into wheat Mianyang 19 and was expressed in some transgenic progenies.