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    Histone deacetylase 9 interacts with SiHAT3.1 and SiHDA19 to repress dehydration responses through H3K9 deacetylation in foxtail millet
    (Oxford University Press, 2024) Kumar, Verandra; Singh, Babita; Singh, Roshan Kumar; Sharma, Namisha; Muthamilarasan, Mehanathan; Sawant, Samir V; Prasad, Manoj
    Climate change inflicts several stresses on plants, of which dehydration stress severely affects growth and productivity. C4 plants possess better adaptability to dehydration stress; however, the role of epigenetic modifications underlying this trait is unclear. Particularly, the molecular links between histone modifiers and their regulation remain elusive. In this study, genome-wide H3K9 acetylation (H3K9ac) enrichment using ChIP-seq was performed in two foxtail millet cultivars contrastingly differing in dehydration tolerance (IC403579; cv. IC4 – tolerant, and IC480117; cv. IC41 – sensitive). It revealed that a histone deacetylase, SiHDA9, was significantly up-regulated in the sensitive cultivar. Further characterization indicated that SiHDA9 interacts with SiHAT3.1 and SiHDA19 to form a repressor complex. SiHDA9 might be recruited through the SiHAT3.1 recognition sequence onto the upstream of dehydration-responsive genes to decrease H3K9 acetylation levels. The silencing of SiHDA9 resulted in the up-regulation of crucial genes, namely, SiRAB18, SiRAP2.4, SiP5CS2, SiRD22, SiPIP1;4 and SiLHCB2.3, which imparted dehydration tolerance in the sensitive cultivar (IC41). Overall, the study provides mechanistic insights into SiHDA9-mediated regulation of dehydration stress response in foxtail millet.
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    Validation of an allele-specific marker associated with dehydration stress tolerance in a core set of foxtail millet accessions
    (John Wiley & Sons, 2013) Lata, Charu; Prasad, Manoj
    Dehydration responsive element binding (DREB) genes play an important role in improving and enhancing stress tolerance of plants in response to abiotic stresses by interacting with DRE/CRT cis- element present in the promoter region of various stress-responsive genes. Thus, characterization of these genes and development of functional markers for effective alleles is important for crop improvement programmes. Identification of a synonymous single nucleotide polymorphism associated with dehydration tolerance at 558th bp (an A/G transition) in the SiDREB2 gene of foxtail millet (Setaria italica L.) and development of an allele-specific marker (ASM) has previously been reported by us. Here, we report on validation of the ASM in a core set of 170 foxtail millet accessions through single-marker regression analysis. The regression of lipid peroxidation (LP) on the ASM suggested that the SiDREB2 associated trait contributed to 27.18% of the total variation in LP among the 170 accessions indicating it to be a major QTL for dehydration tolerance. Thus, this ASM can be used for marker-aided breeding and allele-mining for dehydration tolerance in foxtail millet.
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    Association of SNP in a novel DREB2-like gene SiDREB2 with stress tolerance in foxtail millet [Setaria italica (L.)]
    (Oxford University Press, 2011) Lata, Charu; Bhutty, Sarita; Bahadur, Ranjit Prasad; Majee, Manoj; Prasad, Manoj
    The DREB genes code for important plant transcription factors involved in the abiotic stress response and signal transduction. Characterization of DREB genes and development of functional markers for effective alleles is important for marker-assisted selection in foxtail millet. Here the characterization of a cDNA (SiDREB2) encoding a putative dehydration-responsive element-binding protein 2 from foxtail millet and the development of an allele-specific marker (ASM) for dehydration tolerance is reported. A cDNA clone (GenBank accession no. GT090998) coding for a putative DREB2 protein was isolated as a differentially expressed gene from a 6 h dehydration stress SSH library. A 5' RACE (rapid amplification of cDNA ends) was carried out to obtain the full-length cDNA, and sequence analysis showed that SiDREB2 encoded a polypeptide of 234 amino acids with a predicted mol. wt of 25.72 kDa and a theoretical pI of 5.14. A theoretical model of the tertiary structure shows that it has a highly conserved GCC-box-binding N-terminal domain, and an acidic C-terminus that acts as an activation domain for transcription. Based on its similarity to AP2 domains, SiDREB2 was classified into the A-2 subgroup of the DREB subfamily. Quantitative real-time PCR analysis showed significant up-regulation of SiDREB2 by dehydration (polyethylene glycol) and salinity (NaCl), while its expression was less affected by other stresses. A synonymous single nucleotide polymorphism (SNP) associated with dehydration tolerance was detected at the 558th base pair (an A/G transition) in the SiDREB2 gene in a core set of 45 foxtail millet accessions used. Based on the identified SNP, three primers were designed to develop an ASM for dehydration tolerance. The ASM produced a 261 bp fragment in all the tolerant accessions and produced no amplification in the sensitive accessions. The use of this ASM might be faster, cheaper, and more reproducible than other SNP genotyping methods, and thus will enable marker-aided breeding of foxtail millet for dehydration tolerance.