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首页> 外文期刊>Euphytica >Identification and characterization of RAPD and SCAR markers linked to anthracnose resistance gene in sorghum [Sorghum bicolor (L.) Moench]
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Identification and characterization of RAPD and SCAR markers linked to anthracnose resistance gene in sorghum [Sorghum bicolor (L.) Moench]

机译：与高粱炭疽病抗性基因相关的RAPD和SCAR标记的鉴定与鉴定[高粱]

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Anthracnose, one of the destructive foliar diseases of sorghum growing in warm humid regions, is incited by the fungus Colletotrichum graminicola.The inheritance of anthracnose resistance was studied using the parental cultivars of Sorghum bicolor (L.) Moench, HC 136 (susceptible to anthracnose) and G 73 (anthracnose resistant). The F1 and F2 plants were inoculated with the local isolates of C. graminicola cultures. The F2 plants showed a segregation ratio of 3 (susceptible): 1(resistant) indicating that the locus for resistance to anthracnose in sorghum accession G 73 segregates as a recessive trait in a cross to susceptible cultivar HC 136. RAPD (random amplified polymorphic DNA) marker OPJ 011437 was identified as marker closely linked to anthracnose resistance gene in sorghum by bulked segregant analysis of HC 136 × G73 derived recombinant inbred lines (RILs) of sorghum. A total of 84 random decamer primers were used to screen polymorphism among the parental genotypes. Among these, only 24 primers were polymorphic. On bulked segregant analysis, primer OPJ 01 amplified a 1437 bp fragment only in resistant parent G 73 and resistant bulk. The marker OPJ 011437 was cloned and sequenced. The sequence of RAPD marker OPJ 011437 was used to generate specific markers called sequence characterized amplified regions (SCARs). A pair of SCAR markers SCJ 01-1 and SCJ 01-2 was developed using Mac Vector program. SCAR amplification of resistant and susceptible parents along with their respective bulks and RILs confirmed that SCAR marker SCJ 01 is at the same loci as that of RAPD marker OPJ 011437 and hence, is linked to anthracnose resistance gene. Resistant parent G 73 and resistant bulk amplified single specific band on PCR amplification using SCAR primer pairs. The RAPD marker OPJ 011437 was mapped at a distance of 3.26 cM apart from the locus governing anthracnose resistance on the sorghum genetic map by the segregation analysis of the RILs. Using BLAST program, it was found that the marker showed 100 per cent alignment with the contig{_}3966 located on the longer arm of chromosome 8 of sorghum genome. Therefore, these identified RAPD and SCAR markers can be used in the resistance-breeding program of sorghum anthracnose by marker-assisted selection.
机译：炭疽菌是一种在温暖潮湿地区生长的高粱的破坏性叶面疾病之一，受到真菌Colletotrichum graminicola的激发。炭疽病抗性的遗传是利用双色高粱Moench，HC 136（易患炭疽病）的亲本品种研究的。 ）和G 73（耐炭疽病）。 F1 和F2 植物分别接种了当地的C. graminicola培养物。 F2 植物的分离比为3（敏感）：1（抗性），这表明高粱登录号G 73中对炭疽病的抗性位点在与易感品种HC 136的杂交中作为隐性性状分离。RAPD（通过HC 136×G73衍生的高粱重组近交系的大体分离分析，鉴定出OPJ 011437 标记为与高粱炭疽病抗性基因密切相关的标记。总共84个随机十聚体引物用于筛选亲本基因型之间的多态性。其中，只有24个引物是多态的。在大量分离物分析中，引物OPJ 01仅在抗性亲本G 73和抗性主体中扩增了1437 bp片段。标记OPJ 011437 被克隆并测序。 RAPD标记OPJ 011437 的序列用于产生称为序列特征扩增区域（SCAR）的特异性标记。使用Mac Vector程序开发了一对SCAR标记SCJ 01-1和SCJ 01-2。抗性和易感亲本的SCAR扩增以及它们各自的体积和RIL证实了SCAR标记SCJ 01与RAPD标记OPJ 011437 位于相同的基因座，因此与炭疽病抗性基因相关。使用SCAR引物对进行PCR扩增时，抗性亲本G 73和抗性体扩增单个特异性条带。通过对RIL的分离分析，将RAPD标记OPJ 011437 映射到高粱遗传图谱上与控制炭疽病抗性的位点相距3.26 cM的位置。使用BLAST程序，发现该标记与位于高粱基因组8号染色体较长臂上的contig {_} 3966有100％对齐。因此，这些鉴定出的RAPD和SCAR标记可以通过标记辅助选择用于高粱炭疽病的抗性育种程序。

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