中国水稻科学

中国水稻科学

• 研究报告 • 上一篇    下一篇

分子标记辅助选择与花药培养相结合快速聚合水稻白叶枯病抗性基因

王兴春; 杨长登*; 李西明; 马良勇   

  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
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2004-01-10 发布日期:2004-01-10

Rapid Pyramiding of Resistance Genes to Bacterial Blight in Rice by Using the Integrated Technique of Marker Assisted-Selection and Anther Culture

WANG Xing-chun; YANG Chang-deng*; LI Xi-ming; MA Liang-yong   

  1. （State Key Laboratory of Rice Biology; China National Rice Research Institute; Hangzhou 310006; China）
  • Received:1900-01-01 Revised:1900-01-01 Online:2004-01-10 Published:2004-01-10

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被引次数

116

摘要： 以受微效多基因控制白叶枯病抗性的五丰占2号和主基因[i]xa5[/i]控制的IRBB5为材料，应用分子标记辅助选择、花药培养和回交技术研究了主基因与微效多基因聚合的途径。经查阅GeneBank中RG556的序列，利用在线引物设计软件Primer 3设计了一条新的R引物：5′CCAGACACCACTGCACATTC3′，克服了原引物Tm值相差太大，扩增效率低的缺点，使PCR扩增效率提高了近10倍。采用病斑长度与五丰占2号相近且携有xa5基因的五丰占2号[sup]2[/sup]/IRBB5 B[sub]1[/sub]F[sub]1[/sub]植株进行花药培养，获得30株二倍体植株，其中16株携有[i]xa5[/i]基因。携有[i]xa5[/i]基因的植株的平均病斑长度仅0.1 cm，对白叶枯病的抗性明显强于双亲。成功地聚合了白叶枯病抗性主基因和微效多基因。从配组到获得主基因和微效多基因聚合材料仅用两年时间，比常规育种方法缩短1~2年。

关键词: 白叶枯病, 抗性基因, 分子标记辅助选择, 花药培养, 基因聚合

Abstract: WFZ (Wufengzhan 2) and IRBB5, with minor-polygene and a major gene xa5 resistant to bacterial blight respectively, were used as parental lines to pyramid minor-polygene and major genes by using marker assisted-selection, anther culture and backcross. The new piece of R primer, 5′CCAGACACCACTGCACATTC 3′, was designed according to the sequence of RG556 in GeneBank using Primer 3, the new R primer has a similar Tm to the F primer. The PCR efficiency of amplification was greatly improved, about ten times higher than those of the old ones. The individual of B1F1 derived from WFZ 2/IRBB5 showing similar lesion length to WFZ and with gene xa5 were selected for anther culture, sixteen of 30 diploid lines with xa5 incorporated were obtained. These lines showed a higher level of resistance than both of their parents with only 0.1 cm lesion in length. The result showed that the minor-polygene and the major gene were successfully pyramided. It took only two years from the cross to the pyramid, about 1-2 years less than that of the traditional breeding method.

Key words: bacterial blight, resistance gene, maker assisted-selection, anther culture, gene pyramiding