广亲和基因S5的克隆为水稻品种亲和特性的鉴定提供了分子学依据。利用PCR技术对水稻S5位点中籼、粳序列间存在差异的区段进行扩增，并结合芹菜核酸内切酶CEL Ⅰ对其进行酶切鉴定，从而区分S5不同的基因型。研究结果表明该方法能准确鉴定不同水稻品种中S5位点的籼粳特性，并能发现新的差异序列。
Cloning of the major locus S5， a widecompatibility gene has laid a molecular basis for identification of the widecompatibility varieties(WCV). According to the difference in S5 locus sequence among indica, japonica and WCV, PCR was conducted and celery endonuclease CEL Ⅰ,which cleaves DNA with high specificity at sites of basesubstitution mismatch, was used to digest PCR products so as to distinguish various S5 DNA fragments. The results showed that the method is accurate in indicajaponica classification. At the same time, a new genotype on the S5 locus was found.
\[1\]张桂权, 卢永根, 张华, 等. 栽培稻(Oryza sativa)杂种不育性的遗传研究： Ⅳ.F1花粉不育性的基因型. 遗传学报, 1994, 21: 3441.
\[2\]严长杰, 梁国华, 顾世梁, 等. 典型籼粳杂种不育性的分子标记分析及其遗传基础. 遗传学报, 2003, 30: 267276.
\[3\]高勇, 滕利生, 申宗坦. 有关三系籼粳杂交稻的广亲和测验种的探讨. 中国水稻科学, 1995, 9(1): 4952.
\[4\]朱旭东, 王建林, 熊振民, 等. 水稻亚种间杂种优势利用中广亲和性的研究. 中国水稻科学, 1994, 8: 211216.
\[5\]Ikihashi H, Araki H. Variety screening of compatibility types revealed in F1 fertility of distant cross in rice. Jpn J Breed, 1984, 34: 304313.
\[6\]Ikihashi H, Araki H. Genetics of F1 sterility in remote crosses of rice. Rice Genet, 1986: 119130.
\[7\]Chen J J, Ding J H, Zhang Q F, et al. A triallelic system of S5 is a major regulator of the reproductive barrier and compatibility of indicajaponica hybrids in rice. PNAS, 2008, 105(32): 1143611441.
\[8\]Fischer S G, Lerman L S. Lengthindependent separation of DNA restriction fragments in twodimensional gel electrophoresis. Cell, 1979, 16(1): 191200.
\[9\]Rosenbaum V, Riesner D. Temperaturegradient gel electrophoresis: Thermodynamic analysis of nucleic acids and proteins in purified form and in cellular extracts．Biophys Chem, 1987, 26(2/3): 235246．
\[10\]Orita M, Wahana H, Kanazawa H, et al．Detection of polymorphisms of human DNA by gel electrophoresis as singlestrand conformation polymorphisms．PNAS, 1989, 86(8): 27662770.
\[11\]Tournier I, Raux G, Di Fiore F, et al. Analysis of the allelespecific expression of the mismatch repair gene MLH1 using a simple DHPLCBased method. Hum Mutat, 2004, 23(4): 379384.
\[12\]Fukuen S, Fukuda T, Maune H, et al. Novel detection assay by PCRRFLP and frequency of the CYP3A5 SNPS, CYP3A5 and 6 in a Japanese population. Pharmacogenetics, 2002, 12(4): 331334.
\[13\]Twito T, Weigend S, Blum S, et al. Biodiversity of 20 chicken breeds assessed by SNPs located in gene regions. Cytogenet Genome Res, 2007, 117: 319326.
\[14\]Wallace R B, Shaffer J, Murphy R F. Hybridization of synthetic oligodeoxyribonucleotides to ФX 174 DNA:The effect of single base pair mismatch. Nucl Acids Res, 1979, 6(11): 35433558.
\[15\]Gresham D, Ruderfer D M, Pratt S C. Genomewide detection of polymorphisms at nucleotide resolution with a single DNA microarray. Science, 2006, 11(5769): 19321936.
\[16\]Ren J. Highthroughput singlestrand conformation polymorphism analysis by capillary electrophoresis. Biomedical Sci Appl, 2000, 741(2): 115128.
\[17\]Nordfors L, Jansson M, Sandberg G. Largescale genotyping of single nucleotide polymorphisms by pyrosequencingtrade mark and validation against the 5′nuclease assay. Hum Mutat, 2002, 19(4): 395401.
\[18\]吴海滨, 朱汝财, 赵德刚. TILLING技术的原理与方法述评. 分子植物育种, 2004, 2(4): 574580.
\[19\]李建春, 李锡香, 宋明. EcoTILLING技术及其应用. 植物遗传资源学报, 2007, 8(1): 123126.
\[20\]Bradley J, Chris B, Luca C, et al. Mismatch cleavage by singlestrand specific Nucleases. Nucleic Acids Res, 2004, 32(8): 26322641.
\[21\]Pimkin M, Caretti E, Canutescu A, et al. Recombinant nucleases CEL Ⅰ from celery and SP Ⅰ from spinach for mutation detection. BMC Biotechnol, 2007, 7: 29.
\[22\]韩锁义, 杨玛丽, 盖钧镒, 等. CEL Ⅰ酶的粗提取及其活性检测. 遗传, 2006, 28(9): 11121116.
\[23\]林英, 陈冬妹, 赵萍, 等. CEL Ⅰ酶的粗纯化及活性分析. 农业生物技术学报, 2007, 15(6): 10061011.
