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用于油菜B基因组渐渗系的RGA和RAPD衍生SCAR标记,该渐渗系赋予油菜对黑胫病的抗性

RGA- and RAPD-derived SCAR markers for a Brassica B-genome introgression conferring resistance to blackleg in oilseed rape.

作者信息

Saal B, Struss D

机构信息

Institut für Pflanzengenetik und Kulturpflanzenforschung, Gatersleben, Germany.

出版信息

Theor Appl Genet. 2005 Jul;111(2):281-90. doi: 10.1007/s00122-005-2022-8. Epub 2005 May 11.

Abstract

An introgression derived from the B genome of Brassica juncea in spring-type oilseed rape (B. napus) conferring recessively inherited cotyledon resistance against several pathotypes of the blackleg fungus Leptosphaeria maculans was mapped using PCR-based molecular markers. Resistance-associated B-genome-specific randomly amplified (RAPD) and resistance gene analog (RGA) DNA polymorphisms were converted into three sequence-specific markers (SCARs; B5-1520, C5-1000, RGALm). The flanking sequence of the RGALm locus was determined by genomic walking, leading to a 1,610-bp EcoRV fragment which showed extensive homology to known and putative resistance genes of a cluster on Arabidopsis chromosome 5. Partial sequence analysis of the genomic RAPD segment OPC-05-1700 revealed strong homology to the gibberellin 2-oxidase gene of Arabidopsis. The SCAR markers were analyzed in two segregating populations and were found to be linked in coupling to each other, and in repulsion to the resistance locus. In both populations, markers deviated significantly from a monogenic 3:1 segregation ratio, with plants lacking the markers being more frequent than expected. Although the mode of introgression is yet unknown, the recombinant individuals observed among susceptible progeny suggest homeology between the B-genome-specific segment and its B. napus counterpart. This would offer prospects for reducing the size of the introgression and further fine mapping of the resistance locus.

摘要

利用基于PCR的分子标记,对源自芥菜型油菜(B. juncea)B基因组导入春性油菜(B. napus)中的渐渗系进行了定位,该渐渗系赋予对黑斑病菌(Leptosphaeria maculans)几种致病型的隐性遗传子叶抗性。与抗性相关的B基因组特异性随机扩增多态性DNA(RAPD)和抗性基因类似物(RGA)被转化为三个序列特异性标记(SCARs;B5 - 1520、C5 - 1000、RGALm)。通过基因组步移确定了RGALm位点的侧翼序列,得到一个1610 bp的EcoRV片段,该片段与拟南芥第5号染色体上一个簇的已知和推测抗性基因具有广泛同源性。对基因组RAPD片段OPC - 05 - 1700的部分序列分析显示,其与拟南芥赤霉素2 -氧化酶基因具有高度同源性。在两个分离群体中对SCAR标记进行了分析,发现它们相互呈相引连锁,与抗性位点呈相斥连锁。在两个群体中,标记均显著偏离单基因3:1的分离比例,缺乏标记的植株比预期更为常见。尽管渐渗模式尚不清楚,但在感病后代中观察到的重组个体表明B基因组特异性区段与其甘蓝型油菜对应区段之间存在部分同源性。这将为缩小渐渗系大小和进一步精细定位抗性位点提供前景。

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