用叶片检测转基因水稻对潮霉素反应的可靠性研究  

Reliability study on the response of HPT gene in transgenic rice to hygromycin by leaf assay

作　　者：张洪 马骏 胡国成[1] 斯华敏[1] 付亚萍[1] 戴良英[2] 孙宗修[1] 

机构地区：[1]中国水稻研究所水稻生物学国家重点实验室,浙江杭州310006 [2]湖南农业大学生物安全科技学院,湖南长沙410128

出　　处：《浙江农业学报》2005年第6期341-345,共5页Acta Agriculturae Zhejiangensis

基　　金：国家973计划项目(G199901161);国家863计划项目(2002AA2Z1001);国家自然科学基金项目(30470933)资助

摘　　要：水稻的转基因研究一般以潮霉素磷酸转移酶(HyglomychlPhosphtobamfemse,6P6)基因HPT作为选择标记,因此潮霉素抗性检测的可靠性至关重要。该项研究以中花11和日本晴的T1,T2和T6代转基因水稻为材料,研究了苗期、减数分裂的完全伸展叶和乳熟期不同部位的叶片对潮霉素的抗性反应,同步提取相应叶片的DNA进行HPT基因的PCR扩增,两者符合率达到97.04%。结果表明,利用叶片对潮霉素的反应检测转基因水稻的方法适用于不同世代、不同生育期和不同部位的叶片,具有很高的可行性和可靠性。HyglomychlPhosphtobamfemse gene （HPT） is widely used for the selectable marker gene in rice transformation. This research is focus on determination of method reliability for response of HPT gene in transgenic rice to hygromycin by leaf assay. Full expanded leaves at seedling and meiosis stages and leaves from different parts of plants at milk stage, as well as leaves from different generation of transgenic rice （ T1, T2 and T6 ） from Oryza sativa L. subsp. Japonica, cv. Zbonghua 11 and Nipponbare were used as for both leaf assaying and DNA extraction. The response of leaves to hygromycin and PCR amplification of a HPT fragment were made at the same time. The result consistency from these two methods appreached 97.04 %. The result showed that this method was suitable for different generations, development stages and parts of plants of transgenic rice, and had high feasibility and rehabihty.

关 键 词：转基因水稻 潮霉素 检测 

分 类 号：Q343.1[生物学—遗传学]