关键词: 黑麦, 75K γ-黑麦碱, 分子鉴定, 转移

Abstract: [Objective] In this study, we aimed to study the genetic phylogenetic relationship of 75K γ-secalin gene in rye(Secale cereale L) and transfer the gene in Chinese rye line Qinling into common wheat. [Method] The experiments were contained of amplification, cloning, sequencing, and sequence analysis of five kinds rye using specific primers secF/secR of 75K γ-secalin subunit. We obtained hybrids of the ordinary wheat(Chinese Spring) as female parent and the rye as male parent. We analyzed 75K γ-secalins protein subunits of hybrids by the SDS-PAGE from the progenies. We observed somatic chromosome number and analyzed in situ hybridization with apical producer. [Results] Sequence analysis showed that the cultivar had typical primary structure of 75K γ-secalin in Qinling(AS156) from China, Wrens(CIse35) from USA, 4041(PI168199) from Turkey, Polko(PI412949) from South Africa, and 278(PI436188) from Chile, respectively. However, they showed a lot variations, including insertion/delection and SNP in repeat region. Moreover, the sequence from AS156 had a deletion of Cys in C-terminal. A clade was clustered by the rye sequences from north latitude of China, USA, and Turkey and the other clade from south latitude of South Africa and Chile. The 75K γ-secalin gene on chromosome 2R in Chinese rye line Qinling was transferred in common wheat line Chinese Spring. A F8 line, WYQ122, with this gene shows good fertility and consistent character in agronomy. This gene in WYQ122 was verified by SDS-PAGE analysis and sequencing data of 75K γ-secalin gene. Cytological observation indicated that WYQ122 has 41 or 42 chromosomes. GISH proved this line containing rye chromosomes. [Conclusion] There was genetic diversity for 75K γ-secalin gene among rye lines. This gene was transferred into common wheat via chromosomal substitution.

Key words: Secale cereale L, 75K γ-secalin, molecular identification, transfer