Abstract

orf256 is a wheat mitochondrial gene associated with cytoplasmic male sterility (CMS) that has different organization in various species. This study exploited the orf256 gene as a mitochondrial DNA marker to study the genetic fingerprint of Triticum and Aegilops species. PCR followed by sequencing of common parts of the orf256 gene were employed to determine the fingerprint and molecular evolution of Triticum and Aegilops species. Although many primer pairs were used, two pairs of orf256 specific primers (5:-94/C: 482, 5:253/C: 482), amplified DNA fragments of 576 bp and 230 bp respectively in all species were tested. A common 500 bp of nine species of Triticum and Aegilops were aligned and showed consistent results with that obtained from other similar chloroplast or nuclear genes. Base alignment showed that there were various numbers of base substitutions in all species compared to S. cereal (Sc) (the outgroup species). Phylogenetic relationship revealed similar locations and proximity on phylogenetic trees established using plastid and nuclear genes. The results of this study open a good route to use unknown function genes of mitochondria in studying the molecular relationships and evolution of wheat and complex plant genomes.

Highlights

  • IntroductionHexaploid bread wheat (Triticum aestivum) is a member of Triticeae tribe, which includes barley (Hordeum vulgare) and rye (Secale cereale) as well as other diploid and tetraploid wheats

  • Hexaploid bread wheat (Triticum aestivum) is a member of Triticeae tribe, which includes barley (Hordeum vulgare) and rye (Secale cereale) as well as other diploid and tetraploid wheats.Meiotic studies indicated that the general evolution of the Triticeae tribe has been defined by divergence at the diploid level from a common diploid ancestor and convergence at the polyploid level involving the diverged diploid genomes [1]

  • Other primer combinations (Table 1) amplified various fragments from different species except Triticum turgidum; we limited the comparison to these two fragments

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Summary

Introduction

Hexaploid bread wheat (Triticum aestivum) is a member of Triticeae tribe, which includes barley (Hordeum vulgare) and rye (Secale cereale) as well as other diploid and tetraploid wheats. Were used in molecular phylogenetic analysis of the Triticum and Aegilops species This included A, D, and S diploids and A genome polyploids using a system based on sequences of large fragments [4,19,20]. T. aestivum, its presence in T. temopheevii, and the presence of a related sequence in rice, (3) the specific transcriptional and translational characteristics of orf256 depending on the source of the nucleus and the relationship to cytoplasmic male sterility, and (4) the lack of a known function for orf256 This gives a good opportunity to follow changes in its sequence, its location, its rearrangement, and its presence or absence in Triticum and Aegilops species. Orf256 was used as a molecular tool to establish a DNA fingerprint and phylogenetic relationship among Triticum and Aegilops species and their evolutionary changes

DNA Primer
Growth of Wheat Shoots
Isolation of DNA
PCR Amplification
Sequencing of the PCR Amplified Fragments
Sequence Alignment and Phylogenetic Relationship
Results and Discussion
Sequencing and Bioinformatic Analysis of PCR Products
Phylogenetic Analysis
Distance Matrix
Conclusions
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