Isolation And Characterization Of Disease And Stress Related Genes From Muskmelon (Cucumis Melo L.)

Soil salinity and attack by pathogen a re the major abiotic stresses in plant agriculture worldwide. Past efforts to improve plant tolerance to osmotic stress and pathogen attack through breeding and genetic engineering have shown limited success owing to the genetic complexity of stress response...

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Main Author:	Lee, Weng Wah
Format:	Thesis
Language:	English English
Published:	2002
Subjects:	Diseases - Complications and sequelae Diseases - Genetic aspects
Online Access:	http://psasir.upm.edu.my/id/eprint/8472/1/FSMB_2002_10_IR.pdf
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id	my-upm-ir.8472
record_format	uketd_dc
spelling	my-upm-ir.84722024-01-26T06:58:44Z Isolation And Characterization Of Disease And Stress Related Genes From Muskmelon (Cucumis Melo L.) 2002-01 Lee, Weng Wah Soil salinity and attack by pathogen a re the major abiotic stresses in plant agriculture worldwide. Past efforts to improve plant tolerance to osmotic stress and pathogen attack through breeding and genetic engineering have shown limited success owing to the genetic complexity of stress responses. Large-scale partial sequencing of expressed sequence tags (ESTs) towards cataloging and categorizing genetically abiotic stress responses can assist a means for the rapid discovery of stress-specific genes. A combination of two types of abiotic stresses (200mM of NaCI and 14 min of 254nm UV-irradiation) were applied to muskmelon seedlings prior to the isolation of RNA for cDNA library construction. Cold plaque hybridisation using non-induced cDNA as probe to screen the stress-induced cDNA library for stress and disease-related genes was performed. A total of 550 ESTs were generated from a stress-induced cDNA library with a cDNA insert size ranging between 500-2500bp. Classification of the ESTs based on their designated functions showed that the majority of isolated genes were involved in salt tolerance and disease resistance. The results showed that the generation of stress-induced ESTs by partial sequencing of random cDNA clones along with expression analysis are efficient approach to identify isolated genes that a re responsible for salt tolerance and disease resistance on a large scale. We believe our dbEST and the associated DNA materials would be a useful resource to scientists engaged in studies on stress-tolerance and disease resistance. MYB proteins a re known to regulate different branches of flavonoid metabolism in plants in response to wounding, UV irradiation and ethephon treatment. Their role as transcriptional regulators in response to stress treatment and pathogen attack also has been reported (Urao et al., 1993; Abe et al., 1997). A 279bp partial MYB-related protein was also isolated by an RT-PCR method using the degenerate primers. At the same time, the double induced stress-related muskmelon cDNA library of salt and UV irradiation provide a good source to study the gene expression and regulatory of MYB gene in relation to the stress and UV treatment. Due to the high similarity of the partial MYB sequence to the cloning vector, p Bluescript SK(+), screening of the full length MYB gene was unsuccessful. Diseases - Complications and sequelae Diseases - Genetic aspects 2002-01 Thesis http://psasir.upm.edu.my/id/eprint/8472/ http://psasir.upm.edu.my/id/eprint/8472/1/FSMB_2002_10_IR.pdf text en public masters Universiti Putra Malaysia Diseases - Complications and sequelae Diseases - Genetic aspects Food Science and Technology Tan, Siang Hee English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
advisor	Tan, Siang Hee
topic	Diseases - Complications and sequelae Diseases - Genetic aspects
spellingShingle	Diseases - Complications and sequelae Diseases - Genetic aspects Lee, Weng Wah Isolation And Characterization Of Disease And Stress Related Genes From Muskmelon (Cucumis Melo L.)
description	Soil salinity and attack by pathogen a re the major abiotic stresses in plant agriculture worldwide. Past efforts to improve plant tolerance to osmotic stress and pathogen attack through breeding and genetic engineering have shown limited success owing to the genetic complexity of stress responses. Large-scale partial sequencing of expressed sequence tags (ESTs) towards cataloging and categorizing genetically abiotic stress responses can assist a means for the rapid discovery of stress-specific genes. A combination of two types of abiotic stresses (200mM of NaCI and 14 min of 254nm UV-irradiation) were applied to muskmelon seedlings prior to the isolation of RNA for cDNA library construction. Cold plaque hybridisation using non-induced cDNA as probe to screen the stress-induced cDNA library for stress and disease-related genes was performed. A total of 550 ESTs were generated from a stress-induced cDNA library with a cDNA insert size ranging between 500-2500bp. Classification of the ESTs based on their designated functions showed that the majority of isolated genes were involved in salt tolerance and disease resistance. The results showed that the generation of stress-induced ESTs by partial sequencing of random cDNA clones along with expression analysis are efficient approach to identify isolated genes that a re responsible for salt tolerance and disease resistance on a large scale. We believe our dbEST and the associated DNA materials would be a useful resource to scientists engaged in studies on stress-tolerance and disease resistance. MYB proteins a re known to regulate different branches of flavonoid metabolism in plants in response to wounding, UV irradiation and ethephon treatment. Their role as transcriptional regulators in response to stress treatment and pathogen attack also has been reported (Urao et al., 1993; Abe et al., 1997). A 279bp partial MYB-related protein was also isolated by an RT-PCR method using the degenerate primers. At the same time, the double induced stress-related muskmelon cDNA library of salt and UV irradiation provide a good source to study the gene expression and regulatory of MYB gene in relation to the stress and UV treatment. Due to the high similarity of the partial MYB sequence to the cloning vector, p Bluescript SK(+), screening of the full length MYB gene was unsuccessful.
format	Thesis
qualification_level	Master's degree
author	Lee, Weng Wah
author_facet	Lee, Weng Wah
author_sort	Lee, Weng Wah
title	Isolation And Characterization Of Disease And Stress Related Genes From Muskmelon (Cucumis Melo L.)
title_short	Isolation And Characterization Of Disease And Stress Related Genes From Muskmelon (Cucumis Melo L.)
title_full	Isolation And Characterization Of Disease And Stress Related Genes From Muskmelon (Cucumis Melo L.)
title_fullStr	Isolation And Characterization Of Disease And Stress Related Genes From Muskmelon (Cucumis Melo L.)
title_full_unstemmed	Isolation And Characterization Of Disease And Stress Related Genes From Muskmelon (Cucumis Melo L.)
title_sort	isolation and characterization of disease and stress related genes from muskmelon (cucumis melo l.)
granting_institution	Universiti Putra Malaysia
granting_department	Food Science and Technology
publishDate	2002
url	http://psasir.upm.edu.my/id/eprint/8472/1/FSMB_2002_10_IR.pdf
_version_	1794018756077289472

Isolation And Characterization Of Disease And Stress Related Genes From Muskmelon (Cucumis Melo L.)

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