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Author: search.filters.author.Lata, CharuSubject: search.filters.subject.dehydration stress

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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.