|Table of Contents|

Interference Vector Construction of Soybean Translation Elongation Factor GmeEF1A and Soybean Transformation(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2016年06期
Page:
902-910
Research Field:
Publishing date:

Info

Title:
Interference Vector Construction of Soybean Translation Elongation Factor GmeEF1A and Soybean Transformation
Author(s):
LIAO Wen-lin;  LUAN He-xiang;  GAO Le;  DING Xue-ni;  SONG Ying-pei; YIN Jin-long; NIU Hao-peng;  ZHI Hai-jian
（National Center for Soybean Improvement of Nanjing Agricultural University/National Key Laboratory for Crop Genetics and Germplasm Enhancement, Nanjing 210095, China）
Keywords:
Soybean mosaic virus;  GmeEF1A-gene;  RNAi;  GATEWAY technology;  Agrobacterium-mediated soybean transformation
PACS:
-
DOI:
10.11861/j.issn.1000-9841.2016.06.0902
Abstract:
Previous research indicated the interaction between GmeEF1A and P3 protein of soybean mosaic virus, and it is related to SMV replication in soybean. In order to determine the conserved interval of soybean GmeEF1A, we made the nucleotide sequence and amino acid sequence alignment from 5 isoforms of GmeEF1A, cloned the 180 bp interference fragment named GmeEF1Ai. The RNAi vector pB7GWIWG2(II)-eEF1Ai was constructed by GATEWAY technology, and transfered into soybean genotype Tianlong 1 through Agrobacteriummediated system.The sequencing result of GmeEF1Ai fragment in vector matched with the expected sequence completely, and the insert direction was conversed identified by PCR amplification and restriction digestion. We obtained eight transgenic seedlings, seven plants of them were positive detected by PCR, herbicide painting and Liberty Link strip. The data of the fluorescence quantitative PCR showed 4 seedlings contained one copy of bar gene. The results of expression levels of GmeEF1A indicated GmeEF1Ai vector could inhibit the expression of 5 isoforms in different extent. The results of this study provided a material to verify the function of GmeEF1A in SMV resistance and new germplasm resource in soybean mosaic virus- resistance breeding.

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Last Update: 2016-12-08