Cover Image

Functional analysis of Barley (Hordeum vulgare L.) Cellulose synthase-like F6 promoter through transgene expression in rice (Oryza sativa L.).

The knowledge of the functional aspects of the promoters are necessary prior to the application of the interested promoters to overexpress transgene in transgenic plants for gene study, improvement of quality traits and biofortification. Currently, there are lacking of characterised endosperm-specif...

Full description

Saved in:
Bibliographic Details
Main Author: Jamahari, Azreena
Format: Thesis
Language:English
Published: 2020
Subjects:
Barley
Online Access:http://psasir.upm.edu.my/id/eprint/99235/1/t%20FSPM%202020%204.pdf
http://psasir.upm.edu.my/id/eprint/99235/
Tags: Add Tag
No Tags, Be the first to tag this record!
id my.upm.eprints.99235
record_format eprints
spelling my.upm.eprints.992352023-04-05T23:53:02Z http://psasir.upm.edu.my/id/eprint/99235/ Functional analysis of Barley (Hordeum vulgare L.) Cellulose synthase-like F6 promoter through transgene expression in rice (Oryza sativa L.). Jamahari, Azreena The knowledge of the functional aspects of the promoters are necessary prior to the application of the interested promoters to overexpress transgene in transgenic plants for gene study, improvement of quality traits and biofortification. Currently, there are lacking of characterised endosperm-specific promoters to produce strong transgene expression in the endosperm tissue of cereal plants at a specific grain development or maturation period. The Cellulose synthase-like F6(CslF6) gene is majorly responsible for the production of beta-glucan in the cereal plants, including barley, oat, wheat and rice. Beta-glucan can be found ubiquitously in the endosperm tissues of barley grains. The HvCslF6 promoter is predicted to drive strong endosperm-specific expression at mid to late grain development stage in transgenic rice, based on previous HvCslF6 gene expression studies. The present study characterised the functional length of HvCslF6 promoter and its tissue-specificity expression pattern through transgene expression in rice. The 2771 bp putative promoter of HvCslF6 gene from Sloop barley was isolated and analysed in-silico. Multiple endosperm-specific elements were identified along the promoter region, suggesting that the promoter may drive endosperm-specific expression pattern. Two transformation vectors, F6Prom1 (2771bpHvCslF6prom::GUS gene) and F6Prom3 (1257bpHvCslF6prom::GUS gene) were successfully constructed and permanently transformed into the Nipponbare rice. The HvCslF6 promoter was functional in transgenic rice as the GUS blue staining was observed in all tested body part of matureT0 plants and remained in the T1 seedlings. The promoter also drove selectively strong expression activity in the endosperm tissue and embryo of the rice grain in comparison to other plant body parts regardless of the promoter lengths. Both GUS histochemical staining and quantitative GUS activity analysis revealed that the expression of the GUS gene driven by 1257 bp HvCslF6 promoter was more potent than that of 2771 bp. The data suggested that the 1257 bp HvCslF6 promoter was sufficient to direct strong transgene expression specifically in the endosperm tissue of the transgenic rice. This verified endosperm-specific promoter will be useful to drive the expression of transgene in rice grain, including beta-glucan synthase, for generating high beta-glucan content rice in the future. Keywords: Endosperm-specific promoter, CslF6 gene, barley, transgenic rice, permanent plant transformation, GUS reporter gene. 2020-11 Thesis NonPeerReviewed text en http://psasir.upm.edu.my/id/eprint/99235/1/t%20FSPM%202020%204.pdf Jamahari, Azreena (2020) Functional analysis of Barley (Hordeum vulgare L.) Cellulose synthase-like F6 promoter through transgene expression in rice (Oryza sativa L.). Masters thesis, Universiti Putra Malaysia. Barley
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
topic Barley
spellingShingle Barley
Jamahari, Azreena
Functional analysis of Barley (Hordeum vulgare L.) Cellulose synthase-like F6 promoter through transgene expression in rice (Oryza sativa L.).
description The knowledge of the functional aspects of the promoters are necessary prior to the application of the interested promoters to overexpress transgene in transgenic plants for gene study, improvement of quality traits and biofortification. Currently, there are lacking of characterised endosperm-specific promoters to produce strong transgene expression in the endosperm tissue of cereal plants at a specific grain development or maturation period. The Cellulose synthase-like F6(CslF6) gene is majorly responsible for the production of beta-glucan in the cereal plants, including barley, oat, wheat and rice. Beta-glucan can be found ubiquitously in the endosperm tissues of barley grains. The HvCslF6 promoter is predicted to drive strong endosperm-specific expression at mid to late grain development stage in transgenic rice, based on previous HvCslF6 gene expression studies. The present study characterised the functional length of HvCslF6 promoter and its tissue-specificity expression pattern through transgene expression in rice. The 2771 bp putative promoter of HvCslF6 gene from Sloop barley was isolated and analysed in-silico. Multiple endosperm-specific elements were identified along the promoter region, suggesting that the promoter may drive endosperm-specific expression pattern. Two transformation vectors, F6Prom1 (2771bpHvCslF6prom::GUS gene) and F6Prom3 (1257bpHvCslF6prom::GUS gene) were successfully constructed and permanently transformed into the Nipponbare rice. The HvCslF6 promoter was functional in transgenic rice as the GUS blue staining was observed in all tested body part of matureT0 plants and remained in the T1 seedlings. The promoter also drove selectively strong expression activity in the endosperm tissue and embryo of the rice grain in comparison to other plant body parts regardless of the promoter lengths. Both GUS histochemical staining and quantitative GUS activity analysis revealed that the expression of the GUS gene driven by 1257 bp HvCslF6 promoter was more potent than that of 2771 bp. The data suggested that the 1257 bp HvCslF6 promoter was sufficient to direct strong transgene expression specifically in the endosperm tissue of the transgenic rice. This verified endosperm-specific promoter will be useful to drive the expression of transgene in rice grain, including beta-glucan synthase, for generating high beta-glucan content rice in the future. Keywords: Endosperm-specific promoter, CslF6 gene, barley, transgenic rice, permanent plant transformation, GUS reporter gene.
format Thesis
author Jamahari, Azreena
author_facet Jamahari, Azreena
author_sort Jamahari, Azreena
title Functional analysis of Barley (Hordeum vulgare L.) Cellulose synthase-like F6 promoter through transgene expression in rice (Oryza sativa L.).
title_short Functional analysis of Barley (Hordeum vulgare L.) Cellulose synthase-like F6 promoter through transgene expression in rice (Oryza sativa L.).
title_full Functional analysis of Barley (Hordeum vulgare L.) Cellulose synthase-like F6 promoter through transgene expression in rice (Oryza sativa L.).
title_fullStr Functional analysis of Barley (Hordeum vulgare L.) Cellulose synthase-like F6 promoter through transgene expression in rice (Oryza sativa L.).
title_full_unstemmed Functional analysis of Barley (Hordeum vulgare L.) Cellulose synthase-like F6 promoter through transgene expression in rice (Oryza sativa L.).
title_sort functional analysis of barley (hordeum vulgare l.) cellulose synthase-like f6 promoter through transgene expression in rice (oryza sativa l.).
publishDate 2020
url http://psasir.upm.edu.my/id/eprint/99235/1/t%20FSPM%202020%204.pdf
http://psasir.upm.edu.my/id/eprint/99235/
_version_ 1762394234335264768
score 13.211869

Similar Items