Tian Yanyan, Fan Longjiang, Thurau Tim, Jung Christian, Cai Daguang
Institute of Crop Science and Plant Breeding, Christian-Albrechts-University of Kiel, Kiel, Germany.
J Mol Evol. 2004 Jan;58(1):40-53. doi: 10.1007/s00239-003-2524-4.
The majority of known plant resistance genes encode proteins with conserved nucleotide-binding sites and leucine-rich repeats (NBS-LRR). Degenerate primers based on conserved NBS-LRR motifs were used to amplify analogues of resistance genes from the dicot sugar beet. Along with a cDNA library screen, the PCR screen identified 27 genomic and 12 expressed NBS-LRR RGAs (nlRGAs) sugar beet clones. The clones were classified into three subfamilies based on nucleotide sequence identity. Sequence analyses suggested that point mutations, such as nucleotide substitutions and insertion/deletions, are probably the primary source of diversity of sugar beet nlRGAs. A phylogenetic analysis revealed an ancestral relationship among sugar beet nlRGAs and resistance genes from various angiosperm species. One group appeared to share the same common ancestor as Prf, Rx, RPP8, and Mi, whereas the second group originated from the ancestral gene from which 12C1, Xa1, and Cre3 arose. The predicted protein products of the nlRGAs isolated in this study are all members of the non-TIR-type resistance gene subfamily and share strong sequence and structural similarities with non-TIR-type resistance proteins. No representatives of the TIR-type RGAs were detected either by PCR amplification using TIR type-specific primers or by in silico screening of more than 16,000 sugar beet ESTs. These findings suggest that TIR type of RGAs is absent from the sugar beet genome. The possible evolutionary loss of TIR type RGAs in the sugar beet is discussed.
大多数已知的植物抗性基因编码具有保守核苷酸结合位点和富含亮氨酸重复序列(NBS-LRR)的蛋白质。基于保守的NBS-LRR基序设计的简并引物被用于从双子叶植物甜菜中扩增抗性基因类似物。通过PCR筛选以及cDNA文库筛选,鉴定出了27个基因组NBS-LRR抗性基因类似物(nlRGAs)和12个表达的甜菜克隆。根据核苷酸序列同一性,这些克隆被分为三个亚家族。序列分析表明,点突变,如核苷酸替换和插入/缺失,可能是甜菜nlRGAs多样性的主要来源。系统发育分析揭示了甜菜nlRGAs与各种被子植物抗性基因之间的祖先关系。一组似乎与Prf、Rx、RPP8和Mi共享同一个共同祖先,而第二组起源于12C1、Xa1和Cre3所源自的祖先基因。本研究中分离出的nlRGAs的预测蛋白质产物均为非TIR型抗性基因亚家族的成员,并且与非TIR型抗性蛋白具有很强的序列和结构相似性。无论是使用TIR型特异性引物进行PCR扩增,还是对超过16,000个甜菜EST进行电子筛选,均未检测到TIR型RGAs的代表序列。这些发现表明甜菜基因组中不存在TIR型RGAs。本文还讨论了甜菜中TIR型RGAs可能的进化丢失情况。