Isolation of Salt Stress-Related Genes from Aspergillus glaucus CCHA by Random Overexpression in Escherichia coli
The halotolerant fungus Aspergillus glaucus CCHA was isolated from the surface of wild vegetation around a saltern with the salinity range being 0–31%. Here, a full-length cDNA library of A. glaucus under salt stress was constructed to identify genes related to salt tolerance, and one hundred clones...
Saved in:
Main Authors: | , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Wiley
2014-01-01
|
Series: | The Scientific World Journal |
Online Access: | http://dx.doi.org/10.1155/2014/620959 |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
_version_ | 1832551206919077888 |
---|---|
author | Jie Fang Xiaojiao Han Lihua Xie Mingying Liu Guirong Qiao Jing Jiang Renying Zhuo |
author_facet | Jie Fang Xiaojiao Han Lihua Xie Mingying Liu Guirong Qiao Jing Jiang Renying Zhuo |
author_sort | Jie Fang |
collection | DOAJ |
description | The halotolerant fungus Aspergillus glaucus CCHA was isolated from the surface of wild vegetation around a saltern with the salinity range being 0–31%. Here, a full-length cDNA library of A. glaucus under salt stress was constructed to identify genes related to salt tolerance, and one hundred clones were randomly selected for sequencing and bioinformatics analysis. Among these, 82 putative sequences were functionally annotated as being involved in signal transduction, osmolyte synthesis and transport, or regulation of transcription. Subsequently, the cDNA library was transformed into E. coli cells to screen for putative salt stress-related clones. Five putative positive clones were obtained from E. coli cells grown on LB agar containing 1 M NaCl, on which they showed rapid growth compared to the empty vector control line. Analysis of transgenic Arabidopsis thaliana lines overexpressing CCHA-2142 demonstrated that the gene conferred increased salt tolerance to plants as well by protecting the cellular membranes, suppressing the inhibition of chlorophyll biosynthesis. These results highlight the utility of this A. glaucus cDNA library as a tool for isolating and characterizing genes related to salt tolerance. Furthermore, the identified genes can be used for the study of the underlying biology of halotolerance. |
format | Article |
id | doaj-art-a264e4eff91e47c4ac5bb4df9862d61b |
institution | Kabale University |
issn | 2356-6140 1537-744X |
language | English |
publishDate | 2014-01-01 |
publisher | Wiley |
record_format | Article |
series | The Scientific World Journal |
spelling | doaj-art-a264e4eff91e47c4ac5bb4df9862d61b2025-02-03T06:04:42ZengWileyThe Scientific World Journal2356-61401537-744X2014-01-01201410.1155/2014/620959620959Isolation of Salt Stress-Related Genes from Aspergillus glaucus CCHA by Random Overexpression in Escherichia coliJie Fang0Xiaojiao Han1Lihua Xie2Mingying Liu3Guirong Qiao4Jing Jiang5Renying Zhuo6State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, ChinaState Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, ChinaState Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, ChinaState Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, ChinaState Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, ChinaState Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, ChinaState Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, ChinaThe halotolerant fungus Aspergillus glaucus CCHA was isolated from the surface of wild vegetation around a saltern with the salinity range being 0–31%. Here, a full-length cDNA library of A. glaucus under salt stress was constructed to identify genes related to salt tolerance, and one hundred clones were randomly selected for sequencing and bioinformatics analysis. Among these, 82 putative sequences were functionally annotated as being involved in signal transduction, osmolyte synthesis and transport, or regulation of transcription. Subsequently, the cDNA library was transformed into E. coli cells to screen for putative salt stress-related clones. Five putative positive clones were obtained from E. coli cells grown on LB agar containing 1 M NaCl, on which they showed rapid growth compared to the empty vector control line. Analysis of transgenic Arabidopsis thaliana lines overexpressing CCHA-2142 demonstrated that the gene conferred increased salt tolerance to plants as well by protecting the cellular membranes, suppressing the inhibition of chlorophyll biosynthesis. These results highlight the utility of this A. glaucus cDNA library as a tool for isolating and characterizing genes related to salt tolerance. Furthermore, the identified genes can be used for the study of the underlying biology of halotolerance.http://dx.doi.org/10.1155/2014/620959 |
spellingShingle | Jie Fang Xiaojiao Han Lihua Xie Mingying Liu Guirong Qiao Jing Jiang Renying Zhuo Isolation of Salt Stress-Related Genes from Aspergillus glaucus CCHA by Random Overexpression in Escherichia coli The Scientific World Journal |
title | Isolation of Salt Stress-Related Genes from Aspergillus glaucus CCHA by Random Overexpression in Escherichia coli |
title_full | Isolation of Salt Stress-Related Genes from Aspergillus glaucus CCHA by Random Overexpression in Escherichia coli |
title_fullStr | Isolation of Salt Stress-Related Genes from Aspergillus glaucus CCHA by Random Overexpression in Escherichia coli |
title_full_unstemmed | Isolation of Salt Stress-Related Genes from Aspergillus glaucus CCHA by Random Overexpression in Escherichia coli |
title_short | Isolation of Salt Stress-Related Genes from Aspergillus glaucus CCHA by Random Overexpression in Escherichia coli |
title_sort | isolation of salt stress related genes from aspergillus glaucus ccha by random overexpression in escherichia coli |
url | http://dx.doi.org/10.1155/2014/620959 |
work_keys_str_mv | AT jiefang isolationofsaltstressrelatedgenesfromaspergillusglaucuscchabyrandomoverexpressioninescherichiacoli AT xiaojiaohan isolationofsaltstressrelatedgenesfromaspergillusglaucuscchabyrandomoverexpressioninescherichiacoli AT lihuaxie isolationofsaltstressrelatedgenesfromaspergillusglaucuscchabyrandomoverexpressioninescherichiacoli AT mingyingliu isolationofsaltstressrelatedgenesfromaspergillusglaucuscchabyrandomoverexpressioninescherichiacoli AT guirongqiao isolationofsaltstressrelatedgenesfromaspergillusglaucuscchabyrandomoverexpressioninescherichiacoli AT jingjiang isolationofsaltstressrelatedgenesfromaspergillusglaucuscchabyrandomoverexpressioninescherichiacoli AT renyingzhuo isolationofsaltstressrelatedgenesfromaspergillusglaucuscchabyrandomoverexpressioninescherichiacoli |