Hinchliffe Doug J, Lu Yingzhi, Potenza Carol, Segupta-Gopalan Champa, Cantrell Roy G, Zhang Jinfa
Department of Agronomy and Horticulture, New Mexico State University, Las Cruces, NM 88003, USA.
Theor Appl Genet. 2005 Apr;110(6):1074-85. doi: 10.1007/s00122-005-1928-5. Epub 2005 Feb 22.
Degenerate primers designed from conserved motifs of known plant resistance gene products were used to amplify genomic DNA sequences from the root-knot nematode (Meloidogyne incognita) resistance genetic source, Upland cotton (Gossypium hirsutum) cultivar Auburn 634 RNR. A total of 165 clones were isolated, and sequence analysis revealed 57 of the clones to be novel nucleotide sequences, many containing the resistance (R)-protein nucleotide-binding site motif. A cluster analysis was performed with resistance gene analogue (RGA) nucleotide sequences isolated in this study, in addition to 99 cotton RGA nucleotide sequences already deposited in GenBank, to generate a phylogenetic tree of cotton R genes. The cotton RGA nucleotide sequences were arranged into 11 groups and 56 sub-groups, based on genetic distances. Multiple sequence alignments were performed on the RGA sequences of each sub-group, and either the consensus sequences or individual RGA sequences were used to design 61 RGA-sequence-tagged site primers. A recombinant inbred line (RIL) population of cultivated tetraploid cotton was genotyped using RGA-specific primers that amplified polymorphic fragments between the two RIL parents. Nine RGA markers were mapped to homeologous chromosomes 12 and 26, based on linkage to existing markers that are located on these chromosomes.
根据已知植物抗性基因产物的保守基序设计的简并引物，用于从抗根结线虫（南方根结线虫）的遗传源陆地棉（陆地棉）品种奥本634 RNR中扩增基因组DNA序列。总共分离出165个克隆，序列分析表明其中57个克隆为新的核苷酸序列，许多含有抗性（R）蛋白核苷酸结合位点基序。除了已存入GenBank的99个棉花RGA核苷酸序列外，还对本研究中分离的抗性基因类似物（RGA）核苷酸序列进行了聚类分析，以生成棉花R基因的系统发育树。根据遗传距离，棉花RGA核苷酸序列被分为11个组和56个亚组。对每个亚组的RGA序列进行了多序列比对，并使用共有序列或单个RGA序列设计了61个RGA序列标签位点引物。利用能扩增两个重组自交系亲本之间多态性片段的RGA特异性引物，对栽培四倍体棉花的重组自交系（RIL）群体进行基因分型。基于与位于这些染色体上的现有标记的连锁关系，将9个RGA标记定位到同源染色体12和26上。
