Melotto M, Coelho M F, Pedrosa-Harand A, Kelly J D, Camargo L E A
Departamento de Fitopatologia, Laboratório de Genética Molecular, ESALQ, Universidade de São Paulo, C.P. 9, 13418-900, Piracicaba, SP, Brazil.
Theor Appl Genet. 2004 Aug;109(4):690-9. doi: 10.1007/s00122-004-1697-6. Epub 2004 Jun 24.
The broadest based resistance to anthracnose of common bean ( Phaseolus vulgaris L.) is conferred by the Co-4 locus. We sequenced a bacterial artificial chromosome clone harboring part of the Co-4 locus of the bean genotype Sprite and assembled a single contig of 106.5 kb for functional annotation. This region contained five copies of the COK-4 gene that encodes for a serine threonine kinase protein previously mapped to the Co-4 locus and 19 novel genes with no similarity to any previously identified genes of common bean. Several putative genes of the Co-4 locus seemed to be expressed as they matched perfectly with bean expressed sequence tags. The expression of the COK-4 genes was assessed by reverse transcription (RT)-PCR, and a single 850-bp cDNA fragment was sequenced and compared with the genomic sequences of the COK-4 homologs. Although the COK-4 cDNA was isolated from a different bean cultivar, it showed high similarity (95%) to the exons of genes BA17 and BA21, suggesting that they were expressed. In a phylogenetic tree including all currently available Pto-like sequences from Phaseolus species, the COK-4 homologs formed a single cluster with the Pto gene, whereas two sequences from P. coccineus and all sequences of P. vulgaris formed two closely related clusters. The Co-4 locus was physically mapped to the short arm of bean chromosome 3, which corresponds to linkage group B8. This study represents a first step in gaining an understanding of the genomic organization of an anthracnose resistance locus of common bean and provides molecular data for comparative analysis with other plant species.
普通菜豆(Phaseolus vulgaris L.)对炭疽病的最广泛抗性由Co-4基因座赋予。我们对包含菜豆基因型Sprite的Co-4基因座部分的细菌人工染色体克隆进行了测序,并组装了一个106.5 kb的单一重叠群用于功能注释。该区域包含五个COK-4基因拷贝,该基因编码一种丝氨酸苏氨酸激酶蛋白,先前已定位到Co-4基因座,还有19个新基因,与普通菜豆中任何先前鉴定的基因均无相似性。Co-4基因座的几个推定基因似乎有表达,因为它们与菜豆表达序列标签完美匹配。通过逆转录(RT)-PCR评估COK-4基因的表达,并对一个850 bp的cDNA片段进行测序,并与COK-4同源基因的基因组序列进行比较。尽管COK-4 cDNA是从不同的菜豆品种中分离出来的,但它与基因BA17和BA21的外显子具有高度相似性(95%),表明它们有表达。在一个包含菜豆属物种所有现有类Pto序列的系统发育树中,COK-4同源基因与Pto基因形成一个单一的簇,而来自多花菜豆的两个序列和普通菜豆的所有序列形成两个密切相关的簇。Co-4基因座被物理定位到菜豆染色体3的短臂上,该短臂对应于连锁群B8。这项研究是了解普通菜豆炭疽病抗性基因座基因组组织的第一步,并为与其他植物物种的比较分析提供了分子数据。