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

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Tyler, Adam Michael (2012) Manipulating Rab GTPase activity in wheat to alter gluten quality for breadmaking. PhD thesis, University of Nottingham.

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Abstract

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

Item Type: Thesis (University of Nottingham only) (PhD)
Supervisors: Lycett, G.W.
Lu, C.
Subjects: S Agriculture > SB Plant culture
Q Science > QK Botany > QK710 Plant physiology
Faculties/Schools: UK Campuses > Faculty of Science > School of Biosciences
Item ID: 12914
Depositing User: EP, Services
Date Deposited: 07 May 2013 08:38
Last Modified: 15 Dec 2017 07:37
URI: https://eprints.nottingham.ac.uk/id/eprint/12914

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