Extraction of total RNA and identification of different genes in mangrove plant involved in its adaptability to the variety of stresses

Nucleic acids extraction from mangroves as woody plants needs lots of practical experience. In the present study, three different methods and the RNeasy plant mini kit were used to extract nucleic acids from mangrove plant. Modified CTAB method provided high integrity and concentration of DNA and RN...

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Main Authors: Sahebi, Mahbob, Musa, Mohamed Hanafi, Abdullah, Siti Nor Akmal, Nejat, Naghmeh, Yusop, Mohd Rafii, Azizi, Parisa
Format: Article
Language:English
Published: University of Agriculture, Faisalabad, Pakistan 2013
Online Access:http://psasir.upm.edu.my/id/eprint/29421/
http://psasir.upm.edu.my/id/eprint/29421/1/29421.pdf
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author Sahebi, Mahbob
Musa, Mohamed Hanafi
Abdullah, Siti Nor Akmal
Nejat, Naghmeh
Yusop, Mohd Rafii
Azizi, Parisa
author_facet Sahebi, Mahbob
Musa, Mohamed Hanafi
Abdullah, Siti Nor Akmal
Nejat, Naghmeh
Yusop, Mohd Rafii
Azizi, Parisa
author_sort Sahebi, Mahbob
building UPM Institutional Repository
collection Online Access
description Nucleic acids extraction from mangroves as woody plants needs lots of practical experience. In the present study, three different methods and the RNeasy plant mini kit were used to extract nucleic acids from mangrove plant. Modified CTAB method provided high integrity and concentration of DNA and RNA, respectively from roots and leaves. This method was successful in extraction of RNA from roots but failed for leaves of mangrove. The CTAB with β-mercaptoethanol increases RNA concentration while it decreases the purity of RNA. High yield of RNA extracted from mangrove leaves was obtained by modifying the SDS method. However, the integrity of RNA decreased. When polyvinylpyrrolidone was used to inhibit polyphenol oxidase activity, the yield and integrity of RNA had improved significantly. Therefore, the objective of this study was to isolate and identify the different genes involved in the adaptation of mangrove plants to different kind of stresses. Two-month old seedlings of the Rhizophoraapiculata were exposed to 450 mM NaCl for 24 hours under hydroponic culture. Then, the expression of dihydrolipoamide dehydrogenase (DLDH) gene was studied using reverse transcriptase-PCR and real-time qRT-PCR.
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publisher University of Agriculture, Faisalabad, Pakistan
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spelling upm-294212016-07-12T09:29:42Z http://psasir.upm.edu.my/id/eprint/29421/ Extraction of total RNA and identification of different genes in mangrove plant involved in its adaptability to the variety of stresses Sahebi, Mahbob Musa, Mohamed Hanafi Abdullah, Siti Nor Akmal Nejat, Naghmeh Yusop, Mohd Rafii Azizi, Parisa Nucleic acids extraction from mangroves as woody plants needs lots of practical experience. In the present study, three different methods and the RNeasy plant mini kit were used to extract nucleic acids from mangrove plant. Modified CTAB method provided high integrity and concentration of DNA and RNA, respectively from roots and leaves. This method was successful in extraction of RNA from roots but failed for leaves of mangrove. The CTAB with β-mercaptoethanol increases RNA concentration while it decreases the purity of RNA. High yield of RNA extracted from mangrove leaves was obtained by modifying the SDS method. However, the integrity of RNA decreased. When polyvinylpyrrolidone was used to inhibit polyphenol oxidase activity, the yield and integrity of RNA had improved significantly. Therefore, the objective of this study was to isolate and identify the different genes involved in the adaptation of mangrove plants to different kind of stresses. Two-month old seedlings of the Rhizophoraapiculata were exposed to 450 mM NaCl for 24 hours under hydroponic culture. Then, the expression of dihydrolipoamide dehydrogenase (DLDH) gene was studied using reverse transcriptase-PCR and real-time qRT-PCR. University of Agriculture, Faisalabad, Pakistan 2013 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/29421/1/29421.pdf Sahebi, Mahbob and Musa, Mohamed Hanafi and Abdullah, Siti Nor Akmal and Nejat, Naghmeh and Yusop, Mohd Rafii and Azizi, Parisa (2013) Extraction of total RNA and identification of different genes in mangrove plant involved in its adaptability to the variety of stresses. Pakistan Journal of Agricultural Sciences, 50 (4). pp. 529-536. ISSN 0552-9034; ESSN: 2076-0906 http://www.pakjas.com.pk/viewpapers.aspx?valv=50&vali=133
spellingShingle Sahebi, Mahbob
Musa, Mohamed Hanafi
Abdullah, Siti Nor Akmal
Nejat, Naghmeh
Yusop, Mohd Rafii
Azizi, Parisa
Extraction of total RNA and identification of different genes in mangrove plant involved in its adaptability to the variety of stresses
title Extraction of total RNA and identification of different genes in mangrove plant involved in its adaptability to the variety of stresses
title_full Extraction of total RNA and identification of different genes in mangrove plant involved in its adaptability to the variety of stresses
title_fullStr Extraction of total RNA and identification of different genes in mangrove plant involved in its adaptability to the variety of stresses
title_full_unstemmed Extraction of total RNA and identification of different genes in mangrove plant involved in its adaptability to the variety of stresses
title_short Extraction of total RNA and identification of different genes in mangrove plant involved in its adaptability to the variety of stresses
title_sort extraction of total rna and identification of different genes in mangrove plant involved in its adaptability to the variety of stresses
url http://psasir.upm.edu.my/id/eprint/29421/
http://psasir.upm.edu.my/id/eprint/29421/
http://psasir.upm.edu.my/id/eprint/29421/1/29421.pdf