H3K4/K9 acetylation and Lr28-mediated expression of six leaf rust responsive genes in wheat (Triticum aestivum)
Development of leaf rust-resistant cultivars is a priority during wheat breeding, since leaf rust causes major losses in yield. Resistance against leaf rust due to Lr genes is partly controlled by epigenetic modifications including histone acetylation that is known to respond to biotic/abiotic stresses. In the present study, enrichment of H3K4ac and H3K9ac in promoters of six defense responsive genes (N-acetyltransferase, WRKY 40, WRKY 70, ASR1, Peroxidase 12 and Sarcosine oxidase) was compared with their expression in a pair of near-isogenic lines (NILs) for the gene Lr28 following inoculation with leaf rust pathotype ’77-5’; ChIP-qPCR was used for this purpose. The proximal and distal promoters of these genes contained a number of motifs that are known to respond to biotic stresses. The enrichment of two acetylation marks changed with passage of time; changes in expression of two of the six genes (N-acetyltransferase and peroxidase12), largely matched with changes in H3K4/H3K9 acetylation patterns of the two promoter regions. For example, enrichment of both the marks matched with higher expression of N-acetyltransferase gene in susceptible NIL and the deacetylation (H3K4ac) largely matched with reduced gene expression in resistant NIL. In peroxidase12, enrichment of H3K4ac and H3K9ac largely matched with higher expression in both the NILs. In the remaining four genes, changes in H3 acetylation did not always match with gene expression levels. This indicated complexity in the regulation of the expression of these remaining four genes, which may be controlled by other epigenetic/genetic regulatory mechanisms that need further analysis.
KeywordsBread wheat Leaf rust resistance Epigenetics Histone modifications Acetylation Chromatin immunoprecipitation (ChIP)
Financial assistance from the Department of Biotechnology (Grant number BT/PR3337/AGR/2/819/2011), Government of India for carrying out this study is gratefully acknowledged. CS, SK and GS were each awarded JRF/SRF by the Department of Biotechnology, Government of India. PKG and HSB were each awarded the position of Senior Scientist by Indian National Science Academy, New Delhi. PKG was also awarded a National Academy of Sciences India (NASI) Senior Scientist Platinum Jubilee Fellowship during the tenure of this research work.
PKG and HSB conceived the experiment. CS conducted most of the experiments including ChIP-qPCR with the help of SK, who provided regular timely suggestions during the course of experimental work. SK also helped CS in preparation of first draft of the manuscript, which was edited and finalized by PKG, HSB and PKS with the help of GS, SK, CS and SR. NJ and KVP raised and provided the plant material.
Compliance with ethical standards
Conflict of interest
All the authors declare that there is no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
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