作　　者： ; ; ; ; (陈存贤）;

机构地区： 广东省农业科学院

出　　处： 《分子植物育种》 2005年第6期821-828,共8页

摘　　要： 本文是在先期研究工作的基础上,利用从柑桔与枳属间杂种的基因组DNA所获得的类受体激酶(RLK)的候选抗病基因序列,重新设计引物,对柑桔抗溃疡病材料和感病材料开展以PCR扩增为基础的对比分析,其中一对引物‘19h16/DdeI’的扩增产物经限制性内切酶DdeI酶切,揭示了抗性材料(IchangPape-da,Meiwakumquat,MaruhikumquatandNagamikumquat)和感病材料(FlyingDragon,ValenciaorangeandPalestinesweetlime)之间的多态性差异,同时,在C.Ichangensis的自交F1代和Palestinesweetlime×IchangPapeda的杂交F1代群体中也见明显的多态性差异;对这些F1代个体进行溃疡病病原[Xanthomonasax-onopodispv.Citri(Xac)]接种作抗病性表型鉴定,结果表明,该分子标记‘19h16/DdeI’与柑桔溃疡病抗性密切相关;经纯化测序的结果进一步证明,抗性材料PCR扩增产物具有完全一致的序列和DdeI酶切位点,但感病材料缺少这一DdeI酶切位点;染色体步行(primerwalking)在BAC文库的对应单克隆中获得了一个完整的抗病基因序列,与以前获得的2个抗病基因序列‘17o6RLKP’和‘26m19RLKP’相比,‘19h16RLKP’也具有Xa21抗病基因蛋白的所有特征,包括含有一个信号肽、同样数目的亮氨酸重复序列、跨膜域和激酶域等。基于该序列的开放读码框,发展了特异性更强可靠性更高的另一分子标记,在今后的研究工作中具有较大的应用潜力。 By utilizing the receptor-like kinase （RLK） RGC sequences previously obtained from the genomic DNA of an intergeneric Citrus and Poncirus hybrid, PCR amplification with one pair of new-designed primers was found to reveal polymorphism between the theoretical resistant species （Ichang Papeda, Meiwa kumquat,Maruhi kumquat and Nagami kumquat） and susceptible species （Flying Dragon, Valencia orange and Palestine sweet lime ） and among the self-crossing population of C. Ichangensis and F1 population of Palestine sweet lime×Ichang Papeda （PI family）, followed by restriction of amplified products with DdeI. The phenotype evaluation by inoculation with citrus canker [Xanthomonas axonopodis pv. Citri （Xac）] confirmed that the markerpurified and sequenced. The results confirmed that the resistance species （Kumquat and Ichang Papeda） have identical fragment sequences and DdeI restriction site, but the susceptible species lacked the DdeI site. After sequencing the BAC clone insert by primer walking, an entire gene sequence was obtained. Comparing with the previously obtained two RLK sequences 17o6RLKP and 26m19RLKP, 19hl6RLKP contained all the features of the rice Xa21 protein, including a signal peptide, the same number of leucine-rich-repeats, and transmembrane and kinase domains. Based on the ORFs of the sequence, one more specific and reliable marker was developed,which could be of great potential in future application.

关 键 词： 柑桔 溃疡病 抗性基因 类受体激酶 分子标记

领　　域： [农业科学] [农业科学] [农业科学] [农业科学]