Morales Mónica, Orjeda Gisella, Nieto Cristina, van Leeuwen Hans, Monfort Amparo, Charpentier Myriam, Caboche Michel, Arús Pere, Puigdomènech Pere, Aranda Miguel A, Dogimont Catherine, Bendahmane Abdelhafid, Garcia-Mas Jordi
Departament de Genètica Vegetal, Laboratori de Genètica Molecular Vegetal CSIC-IRTA, carretera de Cabrils s/n, 08348 Cabrils (Barcelona), Spain.
Theor Appl Genet. 2005 Sep;111(5):914-22. doi: 10.1007/s00122-005-0019-y. Epub 2005 Oct 18.
Melon necrotic spot virus (MNSV) is a member of the genus Carmovirus, which produces severe yield losses in melon and cucumber crops. The nsv gene is the only known natural source of resistance against MNSV in melon, and confers protection against all widespread strains of this virus. nsv has been previously mapped in melon linkage group 11, in a region spanning 5.9 cM, saturated with RAPD and AFLP markers. To identify the nsv gene by positional cloning, we started construction of a high-resolution map for this locus. On the basis of the two mapping populations, F(2) and BC1, which share the same resistant parent PI 161375 (nsv/nsv), and using more than 3,000 offspring, a high-resolution genetic map has been constructed in the region around the nsv locus, spanning 3.2 cM between CAPS markers M 29 and M 132. The availability of two melon BAC libraries allowed for screening and the identification of new markers closer to the resistance gene, by means of BAC-end sequencing and mapping. We constructed a BAC contig in this region and identified the marker 52 K 20 sp 6, which co-segregates with nsv in 408 F(2) and 2.727 BC1 individuals in both mapping populations. We also identified a single 100 kb BAC that physically contains the resistance gene and covers a genetic distance of 0.73 cM between both BAC ends. These are the basis for the isolation of the nsv recessive-resistance gene.
甜瓜坏死斑病毒(MNSV)是番茄病毒属的成员,可导致甜瓜和黄瓜作物严重减产。nsv基因是甜瓜中已知的唯一对MNSV具有抗性的天然来源,可抵御该病毒的所有广泛流行株系。此前已将nsv基因定位在甜瓜连锁群11中一个跨度为5.9厘摩的区域,该区域富含随机扩增多态性DNA(RAPD)和扩增片段长度多态性(AFLP)标记。为通过定位克隆鉴定nsv基因,我们开始构建该基因座的高分辨率图谱。基于两个共享相同抗性亲本PI 161375(nsv/nsv)的作图群体F(2)和BC1,并使用3000多个后代,在nsv基因座周围区域构建了一个高分辨率遗传图谱,该区域在酶切扩增多态性序列(CAPS)标记M 29和M 132之间跨度为3.2厘摩。两个甜瓜细菌人工染色体(BAC)文库的可用性使得通过BAC末端测序和作图筛选并鉴定出更接近抗性基因的新标记成为可能。我们在该区域构建了一个BAC重叠群,并鉴定出标记52 K 20 sp 6,在两个作图群体的408个F(2)个体和2727个BC1个体中,该标记与nsv共分离。我们还鉴定出一个单一的100 kb BAC,其物理上包含抗性基因,且两个BAC末端之间的遗传距离为0.73厘摩。这些是分离nsv隐性抗性基因的基础。