农杆菌介导AGPL基因转化玉米的研究

Study on Transformation of AGPL Gene Into Maize Mediated by Agrobacterium Tumefaciens

随着市场对工业用玉米淀粉需求的日益增加,培育高淀粉玉米新种质新品种目前成为解决这一问题的关键。本试验针对这一问题开展工作,成功克隆了水稻胚乳特异型启动子RP5和玉米合成关键酶基因AGPL,利用中间载体pGM-T-RP5和pGM-T-AGPL,将RP5和AGPL定向连接到表达载体pCAMBIA3301上,构建表达载体pRP5-AGPL。以东北骨干玉米自交系Y9812幼胚为受体材料,通过农杆菌介导法,获得玉米再生植株,经PCR分子检测,共有17株呈阳性,转化率达到4.4%。本研究成功地将目的基因和启动子转到玉米自交系Y9812中,并建立了一套稳定的玉米遗传转化体系,为今后转化玉米淀粉合成相关基因奠定坚实基础。

With the increasing of requirements for starch in modern industry,breeding of high starch is the key to solve this problem.In this study,a plant expression vector was successfully constructed which contained the rice endosperm specific promoter RP5 and the key gene AGPL in starch biosynthesis.Moreover,immature embryos of inbred line W9812 widely used in the Northeast were used as recipient materials and transformed mediated by Agrobacterium Tumefaciens.Regenerated plants were obtained and a total of 17 plants were positive by PCR molecular detection,transformation ratio was 4.4%.This study established a stable genetic transformation system and laid the foundation on the transformation of starch biosynthesis genes for future research.