The Bowman-Birk family of plant protease inhibitors are serine protease inhibitors that can
simultaneously inhibit trypsin and/or chymotrypsin. Unique features include an
unusually high thermal stability, tolerance towards low pH and resistance towards the
action of proteases. Bowman-Birk protease inhibitors have been implicated in various
biotic and abiotic stress conditions in plants. LRW222 was recently identified by means of
SSH in Puccinia triticina-infected wheat and was found to be homologous to Wali5, a
wound- and aluminium stress-induced Bowman-Birk type protease inhibitor. The aim of this
study was to determine the expression levels and role of the LRW222 protein in Pucciniainfected
wheat.
This study identified and validated stable reference genes for qPCR gene expression
analysis in rust-infected wheat. qPCR is the preferred method to quantify mRNA levels, but
requires validated reference genes for data normalisation. As prescribed by the MIQE
guidelines, the GeNorm-Plus algorithm was used to examine the expression stability of six
candidate reference genes in resistant Avocet Yr1 wheat infected with P. triticina, P.
striiformis and P. graminis f. sp. tritici respectively. These reference genes were used in the
gene expression analysis of LRW222. Results indicated induced expression in both resistant
and susceptible wheat, indicating its possible involvement in the early general defence
response of wheat. Co-immunoprecipitation and mass spectrometry was used in an attempt
to identify LRW222-interacting proteins to shed light on its specific role in plant defence.
Protein identification was inconclusive, but results indicated that under natural conditions,
LRW222 may exist as multimers in wheat.
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:ufs/oai:etd.uovs.ac.za:etd-07162013-151819 |
| Date | 16 July 2013 |
| Creators | Scholtz, Jakobus Johannes |
| Contributors | Dr B Visser |
| Publisher | University of the Free State |
| Source Sets | South African National ETD Portal |
| Language | en-uk |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.uovs.ac.za//theses/available/etd-07162013-151819/restricted/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to University Free State or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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