Kaga A, Ishimoto M
Legume Breeding Laboratory, National Agriculture Research Center, Tsukuba, Ibaraki, Japan.
Mol Gen Genet. 1998 May;258(4):378-84. doi: 10.1007/s004380050744.
Bruchid resistance, controlled by a single dominant gene (Br) in a wild mungbean accession (TC1966), has been incorporated into cultivated mungbean (Vigna radiata). The resistance gene simultaneously confers inhibitory activity against the bean bug, Riptortus clavatus Thunberg (Hemiptera: Alydidae). The resultant isogenic line (BC20 generation) was characterized by the presence of a group of novel cyclopeptide alkaloids, called vignatic acids. A linkage map was constructed for Br and the vignatic acid gene (Va) using restriction fragment length polymorphism (RFLP) markers and a segregating BC20F2 population. By screening resistant and susceptible parental lines with 479 primers, eight randomly amplified polymorphic DNA (RAPD) markers linked to Br were identified and cloned for use as RFLP probes. All eight RAPD-based markers, one mungbean, and four common bean genomic clones were effectively integrated around Br within a 3.7-cM interval. Br was mapped to a 0.7-cM segment between a cluster consisting of six markers and a common bean RFLP marker, Bng110. The six markers are closest to the bruchid resistance gene, approximately 0.2 cM away. The vignatic acid gene, Va, cosegregated with bruchid resistance. However, one individual was identified in the BC20F2 population that retained vignatic acids in spite of its bruchid susceptibility. Consequently, Va was mapped to a single locus at the same position as the cluster of markers and 0.2 cM away from Br. These results suggest that the vignatic acids are not the principal factors responsible for bruchid resistance in V. radiata but will facilitate the use of map-based cloning strategies to isolate the Br gene.
由野生绿豆种质(TC1966)中的一个显性单基因(Br)控制的豆象抗性已被导入栽培绿豆(Vigna radiata)。该抗性基因同时赋予对豆蝽Riptortus clavatus Thunberg(半翅目:缘蝽科)的抑制活性。所得的近等基因系(BC20代)的特征是存在一组名为vignatic acids的新型环肽生物碱。利用限制性片段长度多态性(RFLP)标记和分离的BC20F2群体构建了Br和vignatic酸基因(Va)的连锁图谱。通过用479对引物筛选抗性和感病亲本系,鉴定并克隆了8个与Br连锁的随机扩增多态性DNA(RAPD)标记用作RFLP探针。所有8个基于RAPD的标记、1个绿豆和4个菜豆基因组克隆在3.7厘摩的区间内有效地整合到Br周围。Br被定位在由6个标记组成的簇和一个菜豆RFLP标记Bng110之间的0.7厘摩区段。这6个标记最接近豆象抗性基因,距离约为0.2厘摩。vignatic酸基因Va与豆象抗性共分离。然而,在BC20F2群体中鉴定出一个个体,尽管它对豆象敏感,但仍保留vignatic酸。因此,Va被定位到与标记簇相同位置的一个单一位点,距离Br 0.2厘摩。这些结果表明,vignatic酸不是绿豆对豆象抗性的主要因素,但将有助于利用基于图谱的克隆策略分离Br基因。