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Position: Home > Articles > Identification and application of new selectable marker genes for rice nuclear transformation Journal of Huazhong Agricultural University 2015,34 (1) 1-8

水稻新型核转化筛选标记基因的鉴定及应用

作  者:
张娟娟;林拥军;周菲
单  位:
华中农业大学作物遗传改良国家重点实验室/华中农业大学国家植物基因研究中心(武汉)
关键词:
水稻;转基因;筛选标记基因;D-氨基酸;生物安全
摘  要:
将人工合成的来自Escherichia coli的dsdA基因与来自Schizosaccharomyces pombe的dao1基因,通过农杆菌侵染方式转化到水稻中花11中。对转化植株进行分子生物学检测表明,目的基因整合到了水稻核基因组中,其中获得转化dsdA的单拷贝植株7个,转化dao1的单拷贝植株6个。通过对单拷贝转基因家系表达量检测及T0代种子萌发测验发现,dao1基因在2个(编号6,7)转基因家系中表达量显著,T0代种子在含D-Ala的培养基上,具有明显抗性。dsdA基因在所有的转基因家系中表达量相对偏低,在含D-Ser的培养基上,T0代种子有一定抗性。
译  名:
Identification and application of new selectable marker genes for rice nuclear transformation
作  者:
ZHANG Juan-juan;LIN Yong-jun;ZHOU Fei;National Key Laboratory of Crop Genetic Improvement/National Center of Plant Gene Research (Wuhan),Huazhong Agricultural University;
关键词:
rice;;transgene;;selectable marker genes;;D-amino acid;;biosafety
摘  要:
Two new selectable marker gene dsdAand dao1 from Escherichia coli and Schizosaccharomyces were synthesized by codon-optimization,and then transformed into japonica rice cultivar zhonghua 11 through Agrobacterium mediated gene transformation.Southern hybridization of the DNA transformants showed that 7single-copy plants of dsdAgene and 6single-copy plants of dao1 gene were obtained.Results of detecting expression levels of the single-copy transformants and the germination texts of T0 generation seeds showed that there were two single-copy gene transformants(numbered as 6and 7)of dao1 gene,indicating high expression levels and consistent with seed germination testing on D-Ala.In all single-copy transformants of dsdAgene,the expression levels of dsdAwere relatively low.Test of seed germination showed that there was the resistance to D-Ser.This study initially explored the application of dsdA,dao1 as new selectable marker genes for rice nuclear transformation,and will lay the foundation for its extensive applications.

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