Isolation and Expression Analysis of Ethylene Receptor Gene from Oncidium Gower Ramsey Flower

The process of flower senescence is influenced by the plant hormone, ethylene. In Arabidopsis, ethylene perception is controlled by a family of five genes, including ETHYLENE RESPONSE 1 (ETR1), ETHYLENE RESPONSE SENSOR 1 (ERS1), ETHYLENE RESPONSE 2 (ETR2), ETHYLENE RESPONSE SENSOR 2 (ERS2) AND ETHYL...

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Main Author:	Mohamed Bahari, Umikalsum
Format:	Thesis
Language:	English English
Published:	2009
Subjects:	Plants - Effect of ethylene on Plant hormones Oncidium
Online Access:	http://psasir.upm.edu.my/id/eprint/21116/1/IB_2009_20_IR.pdf
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spelling	my-upm-ir.211162013-05-27T08:15:07Z Isolation and Expression Analysis of Ethylene Receptor Gene from Oncidium Gower Ramsey Flower 2009-12 Mohamed Bahari, Umikalsum The process of flower senescence is influenced by the plant hormone, ethylene. In Arabidopsis, ethylene perception is controlled by a family of five genes, including ETHYLENE RESPONSE 1 (ETR1), ETHYLENE RESPONSE SENSOR 1 (ERS1), ETHYLENE RESPONSE 2 (ETR2), ETHYLENE RESPONSE SENSOR 2 (ERS2) AND ETHYLENE-INSENSITIVE 4 (EIN4). They fall into two subfamilies based on their sequence similarities. In flower, it has been reported that similar set of genes are also involved. This study was carried out to isolate the gene that is involved in ethylene signaling from Oncidium Gower Ramsey flower. Total RNA was extracted from self-pollinated Oncidium Gower Ramsey flowers followed with reverse transcriptase-polymerase chain reaction (RT-PCR) using two degenerate primer pairs (F1/R1 and F2/R2). These primers were designed based on the conserved regions of ethylene receptor genes of various plant species. Fragments of the expected size were yielded and sequenced. DNA sequences analysis showed a very high homology to the full length ethylene receptor gene, ETHYLENE RECEPTOR 2 (ER2) (Genebank AF276234) of Oncidium Gower Ramsey. From here, specific primers (F3/R3) were designed for 5’ and 3’ ends of the open reading frame (ORF) region of the ER2 gene in order to clone a similar full length gene. The purified and sequenced PCR fragment had 98% DNA sequence similarity to the ER2 gene. However, the newly amplified fragment was only 1595bp, encoded for 422 amino acids as compared to the 2389bp (631 amino acids) of the full length ER2. Sequence alignment of the two genes indicated two truncated regions between nucleotide number 1015 to 1643 (628bp) and 2013 to 2027 (14bp). These results demonstrated that we have cloned an ethylene gene named ER25, which encodes for a protein that has a missing region in the histidine kinase domain as compared to the sequence of ER protein for a similar orchid hybrid. In addition, expression of the isolated gene was detected by real-time RT-PCR at a very low level in the tested flower tissues, as well as in roots. Our results suggest that ER25 may be involved in the development of different plant tissues Plants - Effect of ethylene on Plant hormones Oncidium 2009-12 Thesis http://psasir.upm.edu.my/id/eprint/21116/ http://psasir.upm.edu.my/id/eprint/21116/1/IB_2009_20_IR.pdf application/pdf en public masters Universiti Putra Malaysia Plants - Effect of ethylene on Plant hormones Oncidium Institute of Bioscience English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
topic	Plants - Effect of ethylene on Plant hormones Oncidium
spellingShingle	Plants - Effect of ethylene on Plant hormones Oncidium Mohamed Bahari, Umikalsum Isolation and Expression Analysis of Ethylene Receptor Gene from Oncidium Gower Ramsey Flower
description	The process of flower senescence is influenced by the plant hormone, ethylene. In Arabidopsis, ethylene perception is controlled by a family of five genes, including ETHYLENE RESPONSE 1 (ETR1), ETHYLENE RESPONSE SENSOR 1 (ERS1), ETHYLENE RESPONSE 2 (ETR2), ETHYLENE RESPONSE SENSOR 2 (ERS2) AND ETHYLENE-INSENSITIVE 4 (EIN4). They fall into two subfamilies based on their sequence similarities. In flower, it has been reported that similar set of genes are also involved. This study was carried out to isolate the gene that is involved in ethylene signaling from Oncidium Gower Ramsey flower. Total RNA was extracted from self-pollinated Oncidium Gower Ramsey flowers followed with reverse transcriptase-polymerase chain reaction (RT-PCR) using two degenerate primer pairs (F1/R1 and F2/R2). These primers were designed based on the conserved regions of ethylene receptor genes of various plant species. Fragments of the expected size were yielded and sequenced. DNA sequences analysis showed a very high homology to the full length ethylene receptor gene, ETHYLENE RECEPTOR 2 (ER2) (Genebank AF276234) of Oncidium Gower Ramsey. From here, specific primers (F3/R3) were designed for 5’ and 3’ ends of the open reading frame (ORF) region of the ER2 gene in order to clone a similar full length gene. The purified and sequenced PCR fragment had 98% DNA sequence similarity to the ER2 gene. However, the newly amplified fragment was only 1595bp, encoded for 422 amino acids as compared to the 2389bp (631 amino acids) of the full length ER2. Sequence alignment of the two genes indicated two truncated regions between nucleotide number 1015 to 1643 (628bp) and 2013 to 2027 (14bp). These results demonstrated that we have cloned an ethylene gene named ER25, which encodes for a protein that has a missing region in the histidine kinase domain as compared to the sequence of ER protein for a similar orchid hybrid. In addition, expression of the isolated gene was detected by real-time RT-PCR at a very low level in the tested flower tissues, as well as in roots. Our results suggest that ER25 may be involved in the development of different plant tissues
format	Thesis
qualification_level	Master's degree
author	Mohamed Bahari, Umikalsum
author_facet	Mohamed Bahari, Umikalsum
author_sort	Mohamed Bahari, Umikalsum
title	Isolation and Expression Analysis of Ethylene Receptor Gene from Oncidium Gower Ramsey Flower
title_short	Isolation and Expression Analysis of Ethylene Receptor Gene from Oncidium Gower Ramsey Flower
title_full	Isolation and Expression Analysis of Ethylene Receptor Gene from Oncidium Gower Ramsey Flower
title_fullStr	Isolation and Expression Analysis of Ethylene Receptor Gene from Oncidium Gower Ramsey Flower
title_full_unstemmed	Isolation and Expression Analysis of Ethylene Receptor Gene from Oncidium Gower Ramsey Flower
title_sort	isolation and expression analysis of ethylene receptor gene from oncidium gower ramsey flower
granting_institution	Universiti Putra Malaysia
granting_department	Institute of Bioscience
publishDate	2009
url	http://psasir.upm.edu.my/id/eprint/21116/1/IB_2009_20_IR.pdf
_version_	1747811481991249920

Isolation and Expression Analysis of Ethylene Receptor Gene from Oncidium Gower Ramsey Flower

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