Đang chuẩn bị liên kết để tải về tài liệu:
Isolation and in silico characterization of disease resistance gene analogues (RGAs) from minor Millets

Minor millets are under-exploited plant species often dubbed as ‘Crops for the future’ as they are best adapted to less fertile area and can survive limited water situations and relatively free from pests and diseases. Degenerate oligo-PCR strategy was employed to isolate resistant gene analogues from minor millets by amplifying samples using NBSLRR gene specific primers. Genomic DNA was isolated from the young leaves of minor millets viz., Foxtail millet (Setaria italica) var. CO 5 and Finger millet (Eleusine corocana) var. CO (Ra) 14 by following CTAB method with minor modifications. PCR amplification of the genomic DNA with degenerate primers, revealed presence of 500bp product in the minor millet samples typical to the size of other reported R-gene analogues. The amplicons TENAIKKM1 sequenced was of size 527 bases and RagiKKM1 was of size 509 bases, the sequences with high similarity to NBS-LRR family are deposited in GenBank, NCBI. In silico characterization of the sequences were done using bioinformatics tools by similarity search, pattern recognition and diversity analysis, which resulted in classifying the genes as those belonging to NBS-LRR class as that of other grass family NBS-LRR kind of proteins. Further, in silico characterization ensured that they belong to Non-TIR family of R genes as evident by the conserved residues in the kinase 1 and kinase2 domains. The diversity among the novel RGAs characterized in this study may lead to mining of R- genes in other under-exploited minor millets. | Isolation and in silico characterization of disease resistance gene analogues (RGAs) from minor Millets