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为在水稻育种中快速与高效利用稻瘟病抗性基因Pi25,本文利用该基因不同等位基因编码区序列差异开发了4套CAPS标记(CAP1/HincII、CAP3/BglII、CAP3/NdeI和CAP3/Hpy99I),并利用169份稻种资源、98个重组自交系(RIL)以及217个水稻转基因后代,对4套标记的准确性和选择效果进行了验证。结果表明,4套标记均能准确地检测Pi25/pi25座位。其中,标记CAP1/HincII和CAP3/Hpy99I特异性识别并酶切显性等位基因,而标记CAP3/BglII和CAP3/NdeI特异性识别并酶切隐性等位基因。利用稻瘟病菌株JS001-20接种RIL与转基因材料,抗性表现与标记检测的结果完全一致,表明该CAPS标记准确可靠。分析稻种资源后发现,Pi25基因频率较低(1.2%),说明该基因在我国水稻稻瘟病抗性育种中还没有被充分利用。本文的研究结果特别是开发的2对识别并酶切显性等位基因的CAPS标记可用于分子标记辅助选择,改良我国早籼稻的稻瘟病抗性。
In order to rapidly and efficiently use rice blast resistance gene Pi25 in rice breeding, four sets of CAPS markers (CAP1 / HincII, CAP3 / BglII, CAP3 / NdeI and CAP3 / Hpy99I) were developed using sequence differences of the different alleles of the gene ), And the accuracy and the selection effect of the four sets of markers were verified by using 169 rice seeds, 98 recombinant inbred lines (RIL) and 217 transgenic rice offspring. The results showed that 4 sets of markers can accurately detect Pi25 / pi25 seat. Among them, the markers CAP1 / HincII and CAP3 / Hpy99I specifically recognized and cleaved the dominant alleles while the markers CAP3 / BglII and CAP3 / NdeI specifically recognized and cleaved the recessive alleles. Inoculation of RIL and transgenic materials with rice blast strain JS001-20 was consistent with the results of marker detection, indicating that the CAPS marker was accurate and reliable. Analysis of the rice germplasm resources showed that the frequency of Pi25 gene was low (1.2%), which indicated that the gene was not fully utilized in rice blast resistance breeding in China. The results of this study, in particular, the two pairs of CAPS markers developed and identified as dominant alleles, can be used for molecular marker-assisted selection to improve rice blast resistance in early indica rice.