基于PCR和蛋白检测分析多基因转化甘蔗的遗传稳定性

发布时间：2018-07-25 14:07

【摘要】：甘蔗是重要的糖料作物,也是重要的能源作物。由于复杂的遗传背景、较低的可育性和缺乏优异种质资源等因素给甘蔗品种改良带来了很大的困难,通过转基因技术为甘蔗品种改良提供了有效的途径。而转基因植物的遗传稳定性是植物转基因育种过程中一项重要的研究课题。本研究通过一次多基因遗传转化,同时将四个目的基因导入甘蔗品种新台糖22号中。通过PCR检测,对转基因甘蔗T_0、T_1和T_2代的遗传稳定性进行分析;通过外源蛋白快速试纸条检测,对转基因植株四个外源基因中的基因(bar抗除草剂基因和cry1Ab抗虫基因)蛋白表达进行检测。证明一次四基因遗传转化的甘蔗转基因株系,在T_0代出现了部分基因丢失的现象,且T_0代甘蔗的主茎和分蘖可能为不同的转基因事件。但如果是四个基因均能完整整合到基因组中的株系,则四个基因可以在T_0、T_1和T_2代中得到稳定遗传,也可以稳定的检测得到外源基因蛋白的表达。
[Abstract]:Sugarcane is an important sugar crop as well as an important energy crop. Because of the complex genetic background, low fertility and lack of excellent germplasm resources, it is very difficult to improve sugarcane varieties. Transgenic technology provides an effective way for sugarcane variety improvement. Genetic stability of transgenic plants is an important research topic in the process of plant transgenic breeding. In this study, four target genes were introduced into sugarcane variety Xintaotang 22 by a polygenic genetic transformation. The genetic stability of transgenic sugarcane T _ 0 / T _ 1 and T _ s _ 2 generations was analyzed by PCR, and the foreign protein rapid test strip was used to detect the genetic stability of the transgenic sugarcane. The protein expression of four foreign genes (bar herbicide resistant gene and cry1Ab insect-resistant gene) were detected. It was proved that partial gene loss occurred in the first generation of transgenic sugarcane lines transformed by four genes, and the main stem and tiller of sugarcane in T _ S _ 0 generation might be different transgenic events. However, if the four genes can be integrated into the genome completely, the four genes can be stably inherited in the two generations of T _ (0) T _ (1) and T _ (th), and the expression of exogenous gene proteins can also be detected stably.
【作者单位】：中国热带农业科学院甘蔗研究中心中国热带农业科学院热带生物技术研究所农业部热带作物生物学与遗传资源利用重点实验室;
【基金】：海南省自然科学基金项目(20163122) 国家自然科学基金课题(31371687)共同资助
【分类号】：Q943.2;S566.1

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