摘要
以GUS染色为阳性的抗性胚状体继代再生植株的叶片为材料,通过PCR分析结合GUS染色对橡胶树转基因植株进行初期的阳性鉴定。结果表明,在59份转基因再生植株中,两对特异引物的PCR扩增结果与GUS染色检测结果一致,其中37株仍然表现为阳性,而22株却检测不到目的基因的存在以及GUS基因的表达,表现为外源基因的丢失。因此,初步推测低选择压造成了早期阳性胚状体在长期的连续继代培养再生植株的过程中发生了外源基因的丢失现象,并结合现有的研究提出了初步的解决措施。
The regeneration plant leaves derived from positive embryos with GUS staining were used,and the positive transfomants were judged by PCR and GUS staining.The results revealed that PCR analysis and GUS staining were identical among 59 transformants,with 37 transformants positive 22 transformants negtive confirmed by PCR and GUS,indicating that the genes were lost.So we speculated that the reason of gene lose was a result of low selection pressure in the long-term and consecutive subculture of positive embryos,and here the preliminary solutions to gene lose are proposed in transformation of Hevea.
出处
《热带作物学报》
CSCD
北大核心
2013年第4期591-595,共5页
Chinese Journal of Tropical Crops
基金
国家自然科学基金项目(No.30960322)
国家自然科学基金项目(No.31200503)
现代农业产业技术体系建设专项(No.NYCYTX-34)
关键词
橡胶树
转基因检测
稳定性
基因丢失
继代
Hevea brasiliensis
Transgenic detection
Stability
Gene lose
Subculture
