Probing allocrite preferences of 2 naturally occurring variants of the wheat LR34 ABC transporter

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1139/bcb-2016-0058
AuthorSearch for: 1; Search for: 2; Search for: 1; Search for: 1; Search for: 1; Search for: 1
Name affiliation
  1. National Research Council Canada. Aquatic and Crop Resource Development
  2. National Research Council Canada. Energy, Mining and Environment
FormatText
TypeArticle
Journal titleBiochemistry and Cell Biology
ISSN0829-8211
1208-6002
Volume94
Issue5
Pages459–470
Subjectleaf rust; PDR/ABCG transporter; leaf tip necrosis (LTN); adult plant resistance (APR); transporteur PDR/ABCG; nécrose des extrémités foliaires; résistance des plants adultes
Abstract
For almost a century, the wheat Lr34 gene has conferred durable resistance against fungal rust diseases. While sequence homology predicts a putative ATP binding cassette transporter, the molecules that are transported (allocrites) by the encoded LR34 variants, and any associated mechanism of resistance, remain enigmatic. Here, the in vitro transport characteristics of 2 naturally occurring Lr34 variants (that differ in their ability to mediate disease resistance; Lr34sus and Lr34res) are investigated. Initially, a method to express and purify recombinant LR34Sus and LR34Res pseudo half-molecules from Saccharomyces cerevisiae, is described. Subsequently, a semi-targeted chlorophyll catabolite (CC) extraction from Lr34res-expressing wheat plants was performed based on previous reports highlighting increased accumulation of CCs in Lr34res-expressing flag leaves. Following partial biochemical characterization, this extract was applied to an LR34 in vitro proteoliposome transport assay. Mass spectroscopic analyses of transported metabolites revealed that LR34Sus imported a wheat metabolite of 618 Da and that the LR34Res transporter did not. While the identity of the LR34Sus transported metabolite remains to be confirmed and any allocrites of LR34Res remain to be detected, this work demonstrates that these variants have different allocrite preferences, a finding that may be relevant to the mechanism of disease resistance.
Publication date
PublisherNational Research Council of Canada. Research Press
LanguageEnglish
Peer reviewedYes
NRC numberNRC-ACRD-56297
NPARC number23001057
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Record created2016-12-06
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