Drought Stress-Related Physiological Changes and Histone Modifications in Barley Primary Leaves at HSP17 Gene

Abstract

Stress-inducible genes undergo epigenetic modifications under stress conditions. To investigate if HSP17, of which transcripts accumulate in plant cells under stress, is regulated through epigenetic mechanisms under drought stress, 5-day-old barley (Hordeum vulgare cv. Carina) plants were subjected to progressive drought through water withholding for 22 days. Changes in physiological status and expression of HSP17 gene were monitored in primary leaves of control and drought-treated plants every two days. Twelve days after drought started, control and drought-treated plants were analyzed by chromatin-immunoprecipitation using antibodies against three histone modifications (H3K4me3, H3K9ac, and H3K9me2) and H3 itself. Already after four days of drought treatment, stomatal conductance was severely decreased. Thereafter, maximum and quantum yield of photosystem II (PSII), regulated and non-regulated energy dissipation in PSII, and later also chlorophyll content, were affected by drought, indicating the stress-induced onset of senescence. At the 12th day of drought, before leaf water content declined, expression of HSP17 gene was increased two-fold in drought-treated plants compared to the controls. Twelve days of drought caused an increase in H3 and a loss in H3K9me2 not only at HSP17, but also at constitutively transcribed reference genes ACTIN, PROTEIN PHOSPHATASE 2A (pp2A), and at silent regions BM9, CEREBA. In contrast, H3K4me3 showed a specific increase at HSP17 gene at the beginning and the middle part of the coding region, indicating that this mark is critical for the drought-responsive transcription status of a gene.