Manipulating Rab GTPase activity in wheat to alter gluten quality for breadmaking

In the developing endosperm of bread wheat (Triticum aestivum), seed storage proteins are produced on the rough endoplasmic reticulum (ER) and transported to protein bodies, specialised vacuoles for the storage of protein. The important gluten proteins of wheat are transported to the protein bodies...

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Main Author: Tyler, Adam Michael
Format: Thesis (University of Nottingham only)
Language:English
Published: 2012
Online Access:https://eprints.nottingham.ac.uk/12914/
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author Tyler, Adam Michael
author_facet Tyler, Adam Michael
author_sort Tyler, Adam Michael
building Nottingham Research Data Repository
collection Online Access
description In the developing endosperm of bread wheat (Triticum aestivum), seed storage proteins are produced on the rough endoplasmic reticulum (ER) and transported to protein bodies, specialised vacuoles for the storage of protein. The important gluten proteins of wheat are transported to the protein bodies they are stored in by two distinct routes. One route consists of vesicles that bud directly off the ER, while the other involves transport through the Golgi (Arcalis et al, 2004). In plants, the RabD clade mediates ER to Golgi vesicle transport (Batoko et al, 2000). Available sequence information for Rab GTPases in Arabidopsis, rice, Brachypodium and bread wheat was compiled and compared in phenetic trees. Partial genetic sequences were assembled using the first draft of the Chinese Spring wheat genome. A suitable candidate gene from the RabD clade (TaRabD2a) was chosen for down-regulation by RNA interference (RNAi) and an RNAi construct was used to transform wheat plants. Using real time PCR, all four available RabD genes were shown to be knocked down in the developing endosperm of transgenic wheat. The transgenic grain was found to produce flour with significantly altered processing properties when measured by farinograph and extensograph. SE-HPLC found that a smaller proportion of HMW-GS and large LMW-GS are incorporated into the glutenin macropolymer in the transgenic dough. Lower protein content but a similar protein profile on SDS-PAGE was seen in the transgenic grain
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format Thesis (University of Nottingham only)
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institution University of Nottingham Malaysia Campus
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language English
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publishDate 2012
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spelling nottingham-129142025-02-28T11:22:04Z https://eprints.nottingham.ac.uk/12914/ Manipulating Rab GTPase activity in wheat to alter gluten quality for breadmaking Tyler, Adam Michael In the developing endosperm of bread wheat (Triticum aestivum), seed storage proteins are produced on the rough endoplasmic reticulum (ER) and transported to protein bodies, specialised vacuoles for the storage of protein. The important gluten proteins of wheat are transported to the protein bodies they are stored in by two distinct routes. One route consists of vesicles that bud directly off the ER, while the other involves transport through the Golgi (Arcalis et al, 2004). In plants, the RabD clade mediates ER to Golgi vesicle transport (Batoko et al, 2000). Available sequence information for Rab GTPases in Arabidopsis, rice, Brachypodium and bread wheat was compiled and compared in phenetic trees. Partial genetic sequences were assembled using the first draft of the Chinese Spring wheat genome. A suitable candidate gene from the RabD clade (TaRabD2a) was chosen for down-regulation by RNA interference (RNAi) and an RNAi construct was used to transform wheat plants. Using real time PCR, all four available RabD genes were shown to be knocked down in the developing endosperm of transgenic wheat. The transgenic grain was found to produce flour with significantly altered processing properties when measured by farinograph and extensograph. SE-HPLC found that a smaller proportion of HMW-GS and large LMW-GS are incorporated into the glutenin macropolymer in the transgenic dough. Lower protein content but a similar protein profile on SDS-PAGE was seen in the transgenic grain 2012-12-13 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en arr https://eprints.nottingham.ac.uk/12914/1/Thesis_Hardbound_AMT_6.pdf Tyler, Adam Michael (2012) Manipulating Rab GTPase activity in wheat to alter gluten quality for breadmaking. PhD thesis, University of Nottingham.
spellingShingle Tyler, Adam Michael
Manipulating Rab GTPase activity in wheat to alter gluten quality for breadmaking
title Manipulating Rab GTPase activity in wheat to alter gluten quality for breadmaking
title_full Manipulating Rab GTPase activity in wheat to alter gluten quality for breadmaking
title_fullStr Manipulating Rab GTPase activity in wheat to alter gluten quality for breadmaking
title_full_unstemmed Manipulating Rab GTPase activity in wheat to alter gluten quality for breadmaking
title_short Manipulating Rab GTPase activity in wheat to alter gluten quality for breadmaking
title_sort manipulating rab gtpase activity in wheat to alter gluten quality for breadmaking
url https://eprints.nottingham.ac.uk/12914/