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利用来自11个国家的34个水稻白叶枯病菌菌株,以高感品种IR24、含有已克隆的抗白叶枯病基因Xa21的品系C64(IRBB21)、中花11和日本晴4个水稻品种作为参照,对一个含有来自小粒野生稻(Oryza minuta)的抗白叶枯病基因Xa-min(t)的渐渗品系(introgression line)78-15进行了抗谱鉴定。结果表明,Xa-min(t)是一个广谱、高抗的白叶枯病抗性基因,同时,以IR24和78-15为亲本进行杂交,并采用分离群体分组分析法(BSA)、随机引物扩增多态性DNA(RAPD)和序列特异性扩增区(SCAR)筛选与Xa-min(t)连锁的分子标记,共筛选了800个购自Operon公司的随机引物,其中有2个引物,BE05和BE06,可扩增出与Xa-min(t)连锁的RAPD标记BE05_(300)和BE06_(1400)。对这两个DNA片段克隆并测序,根据两端序列设计了两对长度为21～24bp的特异引物扩增供试材料,其中一个RAPD标记可转化成稳定、重复性好的SCAR标记ScBE05_(300),用ScBE05_(300)和BE06_(1400)标记分别对IR24和78-15的杂交F_2代948和719个单株进行的连锁分析表明,这两个标记与Xa-min(t)的连锁距离分别为2.2和3.7cM,并位于Xa-min(t)的同一侧,这些结果为Xa-min(t)的精细物理图谱的构建奠定了基础。
A total of 34 rice bacterial blight isolates from 11 countries were screened for their susceptibility to IR24, IR24 (IRBB21), Zhonghua 11 and Nipponbare , An introgression line 78-15 containing the bacterial blight resistance gene Xa-min (t) from Oryza minuta was identified. The results showed that Xa-min (t) was a broad-spectrum and highly resistant bacterial leaf blight resistance gene. At the same time, IR24 and 78-15 were used as parents for hybridization and segregation population segregation analysis (BSA) Random amplified polymorphic DNA (RAPD) and sequence specific amplification region (SCAR) were used to screen the molecular markers linked to Xa-min (t). A total of 800 random primers purchased from Operon Company were screened, of which 2 Primers, BE05 and BE06, amplified RAPD markers BE05_ (300) and BE06_ (1400) linked to Xa-min (t). The two DNA fragments were cloned and sequenced. Based on the two-terminal sequence, two pairs of specific primers 21 to 24 bp in length were designed for amplification. One RAPD marker was transformed into a stable and reproducible SCAR marker ScBE05_ (300 ). The linkage analysis of 948 and 719 individuals of F_2 generation with IR24 and 78-15 with ScBE05_ (300) and BE06_ (1400) markers showed that the linkage distance between these two markers and Xa-min (t) 2.2 and 3.7 cM, respectively, located on the same side of Xa-min (t), these results provide a basis for the construction of a fine physical map of Xa-min (t).