Detection of oil palm haploids using stomata counting, flow cytometry, microsatellite and genomic in situ hybridization techniques

Haploids are individuals with gametic chromosome (n) and valuable for breeding purposes such as to produce better hybrids and improve parental performances in hybrid combinations. Several methods were applied in order to screen haploid and doubled haploid plants such as stomata counting, flow cytome...

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Main Author:	Zulkifili, Muhammad Azwan
Format:	Thesis
Language:	English
Published:	2016
Subjects:	Oil palm Androgenesis in plants
Online Access:	http://psasir.upm.edu.my/id/eprint/76534/1/FS%202016%2089%20-%20IR.pdf
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id	my-upm-ir.76534
record_format	uketd_dc
spelling	my-upm-ir.765342020-01-30T06:27:21Z Detection of oil palm haploids using stomata counting, flow cytometry, microsatellite and genomic in situ hybridization techniques 2016-12 Zulkifili, Muhammad Azwan Haploids are individuals with gametic chromosome (n) and valuable for breeding purposes such as to produce better hybrids and improve parental performances in hybrid combinations. Several methods were applied in order to screen haploid and doubled haploid plants such as stomata counting, flow cytometric analysis, microsatellite markers analysis and genomic in situ hybridization (GISH). However, the reliable method to screen for oil palm haploid and doubled haploid is yet to be reported. For ploidy level analysis, stomata counting was done by using ANNOVA but showed unreliable result due to factors such as atmosphere, water and disease. For homozygosity analysis, a hypodiploid, a mixed ploidy and two potential haploid seedlings were obtained via flow cytometry. These oil palm variants were screened with 15 microsatellite markers to determine the homozygosity. However, all samples were shown to be heterozygous. For characterization of parental genomes in intergeneric hybrids, sample used derived from wide hybridization technique which involved cross between four dura flowers with Cocos nucifera pollen. Then, GISH was carried out to characterize the parental genomes of these hybrids. The result showed the absence of signals from the hybrid chromosomes which mean that there was no Cocos nucifera genome introgressed into the hybrid. It may be due to the small proportion of Cocos nucifera. These hybrids then were further analysed using 16 specific coconut microsatellite markers. Two primers managed to exhibit the Cocos genome in all the samples. This proved that the microsatellite markers analysis was reliable to characterize the small proportion of Cocos nucifera genome in intergeneric hybrid. In conclusion, this study had showed that microsatellite analysis is the most reliable method to screen for haploid and doubled haploid oil palm. Oil palm Androgenesis in plants 2016-12 Thesis http://psasir.upm.edu.my/id/eprint/76534/ http://psasir.upm.edu.my/id/eprint/76534/1/FS%202016%2089%20-%20IR.pdf text en public masters Universiti Putra Malaysia Oil palm Androgenesis in plants
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English
topic	Oil palm Androgenesis in plants
spellingShingle	Oil palm Androgenesis in plants Zulkifili, Muhammad Azwan Detection of oil palm haploids using stomata counting, flow cytometry, microsatellite and genomic in situ hybridization techniques
description	Haploids are individuals with gametic chromosome (n) and valuable for breeding purposes such as to produce better hybrids and improve parental performances in hybrid combinations. Several methods were applied in order to screen haploid and doubled haploid plants such as stomata counting, flow cytometric analysis, microsatellite markers analysis and genomic in situ hybridization (GISH). However, the reliable method to screen for oil palm haploid and doubled haploid is yet to be reported. For ploidy level analysis, stomata counting was done by using ANNOVA but showed unreliable result due to factors such as atmosphere, water and disease. For homozygosity analysis, a hypodiploid, a mixed ploidy and two potential haploid seedlings were obtained via flow cytometry. These oil palm variants were screened with 15 microsatellite markers to determine the homozygosity. However, all samples were shown to be heterozygous. For characterization of parental genomes in intergeneric hybrids, sample used derived from wide hybridization technique which involved cross between four dura flowers with Cocos nucifera pollen. Then, GISH was carried out to characterize the parental genomes of these hybrids. The result showed the absence of signals from the hybrid chromosomes which mean that there was no Cocos nucifera genome introgressed into the hybrid. It may be due to the small proportion of Cocos nucifera. These hybrids then were further analysed using 16 specific coconut microsatellite markers. Two primers managed to exhibit the Cocos genome in all the samples. This proved that the microsatellite markers analysis was reliable to characterize the small proportion of Cocos nucifera genome in intergeneric hybrid. In conclusion, this study had showed that microsatellite analysis is the most reliable method to screen for haploid and doubled haploid oil palm.
format	Thesis
qualification_level	Master's degree
author	Zulkifili, Muhammad Azwan
author_facet	Zulkifili, Muhammad Azwan
author_sort	Zulkifili, Muhammad Azwan
title	Detection of oil palm haploids using stomata counting, flow cytometry, microsatellite and genomic in situ hybridization techniques
title_short	Detection of oil palm haploids using stomata counting, flow cytometry, microsatellite and genomic in situ hybridization techniques
title_full	Detection of oil palm haploids using stomata counting, flow cytometry, microsatellite and genomic in situ hybridization techniques
title_fullStr	Detection of oil palm haploids using stomata counting, flow cytometry, microsatellite and genomic in situ hybridization techniques
title_full_unstemmed	Detection of oil palm haploids using stomata counting, flow cytometry, microsatellite and genomic in situ hybridization techniques
title_sort	detection of oil palm haploids using stomata counting, flow cytometry, microsatellite and genomic in situ hybridization techniques
granting_institution	Universiti Putra Malaysia
publishDate	2016
url	http://psasir.upm.edu.my/id/eprint/76534/1/FS%202016%2089%20-%20IR.pdf
_version_	1747813168856432640

Detection of oil palm haploids using stomata counting, flow cytometry, microsatellite and genomic in situ hybridization techniques

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