The Cloning, Characterisation and Antisense Cassettes Construction for Some Pigmentation Genes from Oncidium SPP.

Genes that are responsible for color formation within the anthocyanin and carotenoid biosynthesis pathway have been isolated, studied and manipulated to produce novel colors in many plant species. Therefore, in this study, the genes that are involved in flower color formation namely CHS, F3H, DFR...

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Main Author: Manickam, Sugumaran
Format: Thesis
Language:English
English
Published: 2000
Subjects:
Online Access:http://psasir.upm.edu.my/id/eprint/8414/1/FSMB_2000_2_UPMIR.pdf
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spelling my-upm-ir.84142024-01-11T02:49:19Z The Cloning, Characterisation and Antisense Cassettes Construction for Some Pigmentation Genes from Oncidium SPP. 2000-06 Manickam, Sugumaran Genes that are responsible for color formation within the anthocyanin and carotenoid biosynthesis pathway have been isolated, studied and manipulated to produce novel colors in many plant species. Therefore, in this study, the genes that are involved in flower color formation namely CHS, F3H, DFR and PSY will be isolated and studied in the orchid species, Oncidium goldiana. The construction of antisense cassettes will also be carried out for CHS, DFR and PSY. RNA was isolated from various stages of Oncidium go/diana and its purity and concentration was determined. Partial genes of CHS (605bp), F3H (503bp), DFR (415bp) and PSY (543bp) was isolated from flower petals by RT-PCR using degenerate primers. A BLAST search for homology revealed that these sequences had high homology at both the amino and nucleic acid levels with other known plantThe temporal and spatial gene expression studies using semi-quantitative RT -PCR showed that F3H and DFR were floral specific and they had the highest expression at the yellow buds stage. The level of CHS was the highest in partially opened flowers and PSY was expressed at its highest levels in green buds. CHS and PSY were not floral specific as they were also present in leaves. The antisense cassettes were constructed for CHS, DFR and PSY. These vectors were driven by a 35S promoter and Nos 3' terminator sequence. The CHS vector was 13'480bp, the DFR was 13380bp and PSY was 13430bp. These antisense cassettes were suitable for transformation by biolistic gun bombardment or via Agrobacleriunt infection Oncidium. Cloning. 2000-06 Thesis http://psasir.upm.edu.my/id/eprint/8414/ http://psasir.upm.edu.my/id/eprint/8414/1/FSMB_2000_2_UPMIR.pdf text en public masters Universiti Putra Malaysia Oncidium. Cloning. Food Science and Technology Kulaveerasingam, Harikrishna English
institution Universiti Putra Malaysia
collection PSAS Institutional Repository
language English
English
advisor Kulaveerasingam, Harikrishna
topic Oncidium.
Cloning.

spellingShingle Oncidium.
Cloning.

Manickam, Sugumaran
The Cloning, Characterisation and Antisense Cassettes Construction for Some Pigmentation Genes from Oncidium SPP.
description Genes that are responsible for color formation within the anthocyanin and carotenoid biosynthesis pathway have been isolated, studied and manipulated to produce novel colors in many plant species. Therefore, in this study, the genes that are involved in flower color formation namely CHS, F3H, DFR and PSY will be isolated and studied in the orchid species, Oncidium goldiana. The construction of antisense cassettes will also be carried out for CHS, DFR and PSY. RNA was isolated from various stages of Oncidium go/diana and its purity and concentration was determined. Partial genes of CHS (605bp), F3H (503bp), DFR (415bp) and PSY (543bp) was isolated from flower petals by RT-PCR using degenerate primers. A BLAST search for homology revealed that these sequences had high homology at both the amino and nucleic acid levels with other known plantThe temporal and spatial gene expression studies using semi-quantitative RT -PCR showed that F3H and DFR were floral specific and they had the highest expression at the yellow buds stage. The level of CHS was the highest in partially opened flowers and PSY was expressed at its highest levels in green buds. CHS and PSY were not floral specific as they were also present in leaves. The antisense cassettes were constructed for CHS, DFR and PSY. These vectors were driven by a 35S promoter and Nos 3' terminator sequence. The CHS vector was 13'480bp, the DFR was 13380bp and PSY was 13430bp. These antisense cassettes were suitable for transformation by biolistic gun bombardment or via Agrobacleriunt infection
format Thesis
qualification_level Master's degree
author Manickam, Sugumaran
author_facet Manickam, Sugumaran
author_sort Manickam, Sugumaran
title The Cloning, Characterisation and Antisense Cassettes Construction for Some Pigmentation Genes from Oncidium SPP.
title_short The Cloning, Characterisation and Antisense Cassettes Construction for Some Pigmentation Genes from Oncidium SPP.
title_full The Cloning, Characterisation and Antisense Cassettes Construction for Some Pigmentation Genes from Oncidium SPP.
title_fullStr The Cloning, Characterisation and Antisense Cassettes Construction for Some Pigmentation Genes from Oncidium SPP.
title_full_unstemmed The Cloning, Characterisation and Antisense Cassettes Construction for Some Pigmentation Genes from Oncidium SPP.
title_sort cloning, characterisation and antisense cassettes construction for some pigmentation genes from oncidium spp.
granting_institution Universiti Putra Malaysia
granting_department Food Science and Technology
publishDate 2000
url http://psasir.upm.edu.my/id/eprint/8414/1/FSMB_2000_2_UPMIR.pdf
_version_ 1794018744218943488