转反义蜡质基因水稻亲本后代性状分析

Analysis on the characters of progenies of the rice transformed with anti-sense waxy gene

以单质粒转化的粳稻广粳一号和双质粒共转化的籼稻01早5202所获得的转反义蜡质基因水稻后代为供试材料,通过潮霉素抗性、PCR检测等手段分析了外源基因的遗传分离特性,同时,还分析了转基因材料的直链淀粉含量、waxy蛋白含量的变化特性。结果表明,无论是采用标记基因(hpt)与目的基因(Anti-sense waxy)连锁的单质粒转化,还是采用双质粒共转化,其供试的后代植株材料都发生了外源基因分离现象,且转基因植株材料的直链淀粉含量都有所下降,有些单株的直链淀粉含量已降至10%(质量百分比)以下,远低于对照(其直链淀粉含量为22.04%);SDS-PAGE检测结果显示,供试的转基因材料的waxy蛋白的含量与对应的直链淀粉含量呈正相关性。

The segregation of exogenous genes was studied by hygromycin-resistant and PCR experiments in the transgenic rice （Oryza sativa L. subsp, japonica and indica） with anti-sense waxy gene, meanwhile the change of amylose and waxy protein contents in progenies of transgenic rice was analyzed. The results showed no matter the rice Guangjing No.1（O. Sativa L. subsp, japonica） were transformed by p13w4 plasmid carrying anti-sense waxy gene and hygrornycin-resistant gene, or in the rice 01Z5202 （O. sativa L. subsp, indica） were co-transformed by p13w8 plasmid carrying anti-sense waxy gene and p1300 plasmid carrying hygrornycin-resistant gene, the target gene（s） had been segregated in the progenies; the content of amylose of the transgenic plants was lower than those in non-transgenic ones, and the content of amylose in some of transgenic plants was less than 10.0% （of the weight of grain）, which was much lower than those in the control （about 22.04%）; and the analysis with SDS-PAGE showed the content of the waxy protein are positively correlated with the content of amylose in the tested transgenic rice materials.