Generation of marker-free, bacterial blight-resistant transgenic sterile line and hybrid rice with Xa21
Generation of marker-free, bacterial blight-resistant transgenic sterile line and hybrid rice with Xa21
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DOI：
10.1111/j.1439-0523.2011.01847.x
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发表时间：
2011-08-01
期刊：
影响因子：
2
通讯作者：
Zhai, Wenxue
中科院分区：
文献类型：
--
作者：
Gao, Lifen;Xia, Zhihui;Zhai, Wenxue
水稻白叶枯病是由黄单胞菌水稻致病变种引起的。Xoo是商业化杂交水稻生产中的主要生产限制因素，因为大多数用于杂交生产的亲本系对Xoo敏感。本研究利用农杆菌介导的双右边界载体pDRBXa 21，将克隆的抗稻瘟病基因Xa 21导入籼型三系杂交稻保持系“D 62 B”。分子鉴定和抗性分析表明，Xa 21已整合到转基因植株D 62 B的基因组中，并在T2代获得了纯合、单拷贝、无标记的保持系D 62 B-Xa 21。然后将D 62 B-Xa 21与“D 62 A”回交以产生纯合的转基因不育系D 62 A-Xa 21。经花粉碘-碘化钾（I-2/KI）染色和田间观察，Xa 21的导入对不育系的育性及其它农艺性状无明显影响。逆转录-聚合酶链反应显示Xa 21基因在转基因株系中正常表达。以D 62 A-Xa 21为亲本配制的杂交组合可遗传对BB的高抗。
Bacterial blight (BB) of rice caused by Xanthomonas oryzae pv. oryzae (Xoo) is a major production constraint in commercial hybrid rice production because most of the parental lines used in hybrid production are susceptible to Xoo. In this study, the cloned BB resistance gene Xa21 was transferred into 'D62B', a maintainer line of three-line indica hybrid rice using the double right-border vector pDRBXa21 through an Agrobacterium tumefaciens-mediated system. Molecular and resistance analysis revealed that Xa21 was integrated into the genome of transgenic 'D62B', and the homozygous, single-copy and marker-free maintainer line D62B-Xa21 was obtained in the T2 generation. D62B-Xa21 was then backcrossed with 'D62A' to produce a homozygous transgenic sterile line D62A-Xa21. The introduction of Xa21 did not cause any obvious alteration to sterility and other agronomic traits of the sterile line through iodine-potassium iodide (I-2/KI) staining of pollen grains and field observation. Reverse transcriptase-polymerase chain reaction revealed a normal expression pattern of the Xa21 gene in the transgenic lines. The hybrid combinations derived from D62A-Xa21 could inherit high resistance to BB.