Construction and Analysis of Suppression Subtractive cDNA Library from Fertile Disk Floret Buds of Zinnia elegans

In order to isolate genes related to anther development and understand molecular basis of male sterility, cDNA library of Zinnia elegans was constructed using suppression subtractive hybridization (SSH) approach. 672 different expressed clones were selected from the fertile disk floret buds. PCR res...

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Main Authors: Rui-hua PANG, Guang-ying MA, Xue-yuan LOU, Man-zhu BAO, Yao-mei YE
Format: Article
Language:English
Published: Elsevier 2012-04-01
Series:Journal of Integrative Agriculture
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2095311912600434
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spelling doaj-554eb7494ebd4428be8df2ab645b9ea12021-06-07T06:47:33ZengElsevierJournal of Integrative Agriculture2095-31192012-04-01114567575Construction and Analysis of Suppression Subtractive cDNA Library from Fertile Disk Floret Buds of Zinnia elegansRui-hua PANG0Guang-ying MA1Xue-yuan LOU2Man-zhu BAO3Yao-mei YE4Key Laboratory of Horticultural Plant Biology, Ministry of Education/College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan 430070, P.R. China; College of Life Sciences, Xinyang Normal University, Xinyang 464000, P.R. ChinaKey Laboratory of Horticultural Plant Biology, Ministry of Education/College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan 430070, P.R. ChinaKey Laboratory of Horticultural Plant Biology, Ministry of Education/College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan 430070, P.R. ChinaKey Laboratory of Horticultural Plant Biology, Ministry of Education/College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan 430070, P.R. ChinaKey Laboratory of Horticultural Plant Biology, Ministry of Education/College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan 430070, P.R. China; Correspondence YE Yao-mei, Tel: +86-27-87286948In order to isolate genes related to anther development and understand molecular basis of male sterility, cDNA library of Zinnia elegans was constructed using suppression subtractive hybridization (SSH) approach. 672 different expressed clones were selected from the fertile disk floret buds. PCR results showed that cDNA inserts were ranged from 100 to 750 bp. 303 positive clones screened by dot-blot hybridization were sequenced. 273 out of 303 sequenced clones produced readable sequences; these sequences represent 87 non-repetitive sequences. The homology alignment showed that 76 expressed sequence tags (ESTs) had functional annotations in GenBank, the other 11 ESTs without any homology to the known gene. In addition, 87 ESTs were divided into 17 groups according to MIPS of Arabidopsis thaliana database. Sequence data from the cDNA library have been deposited with the GenBank under the accession numbers GT067016-GT067085. As an important result in this study, 7 genes related to anther development were isolated. Results from semi-quantitative RT-PCR showed 6 genes were expressed only in disk florets of fertile plants compared with that of male sterile plants. These ESTs obtained provide important clues for further isolation and identification of fertility-related genes in Z. elegans.http://www.sciencedirect.com/science/article/pii/S2095311912600434Zinnia eleganssuppression subtractive hybridization (SSH)male sterilityanther developmentexpressed sequence tags (ESTs)
collection DOAJ
language English
format Article
sources DOAJ
author Rui-hua PANG
Guang-ying MA
Xue-yuan LOU
Man-zhu BAO
Yao-mei YE
spellingShingle Rui-hua PANG
Guang-ying MA
Xue-yuan LOU
Man-zhu BAO
Yao-mei YE
Construction and Analysis of Suppression Subtractive cDNA Library from Fertile Disk Floret Buds of Zinnia elegans
Journal of Integrative Agriculture
Zinnia elegans
suppression subtractive hybridization (SSH)
male sterility
anther development
expressed sequence tags (ESTs)
author_facet Rui-hua PANG
Guang-ying MA
Xue-yuan LOU
Man-zhu BAO
Yao-mei YE
author_sort Rui-hua PANG
title Construction and Analysis of Suppression Subtractive cDNA Library from Fertile Disk Floret Buds of Zinnia elegans
title_short Construction and Analysis of Suppression Subtractive cDNA Library from Fertile Disk Floret Buds of Zinnia elegans
title_full Construction and Analysis of Suppression Subtractive cDNA Library from Fertile Disk Floret Buds of Zinnia elegans
title_fullStr Construction and Analysis of Suppression Subtractive cDNA Library from Fertile Disk Floret Buds of Zinnia elegans
title_full_unstemmed Construction and Analysis of Suppression Subtractive cDNA Library from Fertile Disk Floret Buds of Zinnia elegans
title_sort construction and analysis of suppression subtractive cdna library from fertile disk floret buds of zinnia elegans
publisher Elsevier
series Journal of Integrative Agriculture
issn 2095-3119
publishDate 2012-04-01
description In order to isolate genes related to anther development and understand molecular basis of male sterility, cDNA library of Zinnia elegans was constructed using suppression subtractive hybridization (SSH) approach. 672 different expressed clones were selected from the fertile disk floret buds. PCR results showed that cDNA inserts were ranged from 100 to 750 bp. 303 positive clones screened by dot-blot hybridization were sequenced. 273 out of 303 sequenced clones produced readable sequences; these sequences represent 87 non-repetitive sequences. The homology alignment showed that 76 expressed sequence tags (ESTs) had functional annotations in GenBank, the other 11 ESTs without any homology to the known gene. In addition, 87 ESTs were divided into 17 groups according to MIPS of Arabidopsis thaliana database. Sequence data from the cDNA library have been deposited with the GenBank under the accession numbers GT067016-GT067085. As an important result in this study, 7 genes related to anther development were isolated. Results from semi-quantitative RT-PCR showed 6 genes were expressed only in disk florets of fertile plants compared with that of male sterile plants. These ESTs obtained provide important clues for further isolation and identification of fertility-related genes in Z. elegans.
topic Zinnia elegans
suppression subtractive hybridization (SSH)
male sterility
anther development
expressed sequence tags (ESTs)
url http://www.sciencedirect.com/science/article/pii/S2095311912600434
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