Abstract

Analysis of mitochondrial DNA using amplified fragment length polymorphism markers of isonuclear allocytoplasmic male sterile wheat accessions and their maintainer lines

To produce a good F1 hybrid variety wheat crop, it is necessary to explore novel cytoplasmic male sterility (CMS) lines and their maintainer line. This study aimed to identify cytoplasmic variation in three isonuclear-alloplasmic male sterile lines Aegilops kotschyi (Ae.kots) -90-110, Aegilops ventricosa (Ae.ven) -90-110, and Triticum spelta (T.spelta) -90-110 and their maintainer line, A-90-110, at the molecular level. Mitochondrial DNA (mtDNA) was isolated using a combination of centrifugation and density gradient ultracentrifugation, sucrose sedimentation, lysis with sodium dodecyl sulfate (SDS), potassium proteinase, and phenol/chloroform extraction methods. To detect mtDNA purity, specific primers were designed for nuclear (β-actin) and mitochondrial (COXIII) genes.

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