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Cloning and Expression Analysis of GmGST12 in Soybean(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2015年05期
Page:
782-788
Research Field:
Publishing date:

Info

Title:
Cloning and Expression Analysis of GmGST12 in Soybean
Author(s):
HAN Shao-huai;  LI Jia-jia;  ZHANG Jing-yao;  ZHANG Hao;  LI Yan-wei;  DING Xian-long;  HE Ting-ting;  YANG Shou-ping
Soybean Research Institute, Nanjing Agricultural University/National Center for Soybean Improvement/MOA Key Laboratory for Biology and Genetic Improvement of Soybean(General)/National Key Laboratory of Grop Genetics and Germplasm Enhancement, Nanjing 210095, China
Keywords:
Glycine max;  Cytoplasmic-nuclear male sterility;  Glutathione S-transferases（GSTs）;  Gene cloning;  Expression analysis
PACS:
-
DOI:
10.11861/j.issn.1000-9841.2015.05.0782
Abstract:
The reactive oxygen species (ROS) play an important role in plants which act as intracellular or intercellular signals.But high concentration of ROS might lead to plant male sterility. Glutathione S-transferases（GSTs） are crucial to the detoxification for the cells.Based on comparative transcriptome analysis between the cytoplasmic-nuclear male sterile line NJCMS1A and its maintainer NJCMS1B in Glycine max, a differential expression gene of GmGST12 was identified in our previous research. In the present study, GmGST12 gene was cloned from the flower buds of NJCMS1A and NJCMS1B by homology-based cloning method.The coding DNA sequences (CDSs) of GmGST12 from NJCMS1A and NJCMS1B were 708 bp long with the same nucleotide sequences and coded the glutathione S-transferases protein consisted of 235 amino acids. Phylogenetic analysis showed that the homology between GmGST12 and AtGSTU9 from Arabidopsis thaliana was the highest and the identity of the amino acid sequences was 52%. The tissues expression analysis showed that the mRNA expression level of GmGST12 in flower buds of NJCMS1A was significantly higher than that in flower buds of NJCMS1B, but no difference was found in root, steam and leaf.The result of subcellular localization indicated that GmGST12 was located in the cytoplasm and nucleus.In addition, the plant overexpression vector pCAMBIA3301-GmGST12 was constructed for further transgenic research. All the above results provided the foundation for future study on the molecular mechanism of cytoplasmic-nuclear male sterility in soybean.

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Last Update: 2015-11-07