Keywords
Wheat, ¦¸-Gliadin, Gene Cloning, Sequence Analysis
Abstract
In order to obtain relevant information about the ¦Ø-gliadin gene of the excellent variety Shaan 253, specific primers were designed based on the reported ¦Ø-gliadin gene sequence, and the PCR method was used to amplify the total DNA of the wheat (triticumaestivuml.) variety Shaan 253. Fragments of about 1000 bp were recovered, purified, cloned and sequenced, and five different new omega-gliadin genes were obtained, named gli-s253-1, gli-s253-2, gli-s253-3, gli-s253- 4 and gli-s253-5, the genbank accession numbers are: gq423428, gq423429, gq423430, gq423431 and gq423432 respectively. Sequence analysis shows that: (1) There are four main types of ¦Ø-gliadin gene silencing (q¡ú*, i¡ú*, r¡ú* and w¡ú*), with the first type being the dominant type (accounting for 66.6%) ; (2) From the perspective of the triplet code, the two coding methods of glutamine (q) are most prone to silent mutations (caa¡útaa and cag¡útag); (3) From the perspective of mutation sites, silencing mainly occurs For the first base of the triplet code (accounting for 77.8%), the frequency of mutations in the second base accounted for 33.3%, but no silent mutation was found at the third base; gq423428 had a rare ata ¡ú taa type of silence, and at the same time Variations involving two sites; (4) In terms of the number of gene silent mutations, gq423428 and gq423429 each have up to 6 times, of which 4 sites (349, 373, 478 and 592) are shared by the above 5 records; (5) Judging from the structure of the encoded mature protein, silent mutations are mainly concentrated in the ¦Ø-gliadin repeat region, and no such mutations are found in the signal peptide region and the N-terminal region; (6) Evolutionary analysis shows that ¦Ø in the Triticum family -gliadin/secalin is divided into 3 branches, among which ¦Ø-gliadin derived from wheat is clustered into one branch. The above results show that this study successfully isolated five ¦Ø-gliadin coding genes with rich diversity from Shaan 253, providing a reference for in-depth research on high-quality sources and molecular-assisted breeding.
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Original research done by Li Min, Gao Xiang, Chen Qijiao, Dong Jian, Zhao Wanchun, Li Yanliang, Wang Mingxia, Chen Ruiji, Pang Hongxi, Li Zheqing, Liu Jun
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