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This study was conducted to describe the role of the chloroplast maturase K (matK) genetic polymorphism in the reciprocal crossing between five barley varieties using several in vitro / in silico tools. Besides, the final consequences of the matK gene polymorphism on its protein structure, function, and interactions were predicted computationally. Five parental varieties were crossed to each other by full reciprocal crossing design, DNA was extracted from seeds and two different primers’ pairs were designed to scan matK gene. Then, polymerase chain reaction - single-stranded conformation polymorphism (PCR-SSCP) were performed. Two distinct haplotypes in both parents and artificial F1 hybrids in the matK gene were observed in both amplified fragments. This finding indicated that the studied gene had no participation in the reciprocal crossing performed. Three SNPs were identified; two of them are non-synonymous (nsSNPs), namely G387V and L459M. The effect of these missense mutations on the matK protein was analyzed by several in silico tools. It was shown that the coding SNP, L459M was predicted to have much more effective consequences on matK protein structure and function. While the I-Mutant 2.0 prediction tool showed a decrease in stability for these two nsSNPs, which may destabilize the protein interactions to some extent. In conclusion, though the observed missense mutations in the matK gene have no suggestive role in the reciprocally crossed barley varieties, they caused dramatic alterations in several matK protein moieties, which may lead to potential subsequent changes in the matK protein-mediated RNA splicing mechanisms. |
