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Isolation and identification of perennial H.villosa centromere repeat sequences

Author: WeiPei
Tutor: RenZhengLong；YangZuJun
School: University of Electronic Science and Technology
Course: Biochemistry and Molecular Biology
Keywords: Centromere Fluorescence in situ hybridization H.villosa Repeat sequences Ty3-gypsy retrotransposons
CLC: S512.1
Type: Master's thesis
Year: 2011
Downloads: 13
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Abstract


H.villosa is Dasypyrum (also known as Haynaldia) wheat Relatives, including two species distributed in the Mediterranean and South-West Asia diploid H.villosa (D. villosum genome for VV) and distributed in North Africa tetraploid perennial H.villosa, (D. breviaristatum, genomic V ~~ bV ~ bV ~ bV ~~ b). H.villosa species has a variety of good traits cultivated wheat needs, such as resistance to powdery mildew, resistance to stem rust and leaf rust, anti-total eclipse of the disease, cold drought and high protein content, as wheat disease resistance, high yield important genetic resources, quality breeding. The from the perennial H.villosa isolated by PCR amplification, the size of the two fragments were repetitive sequences of 920bp and 1003bp, respectively the name pDvC1 and pDvC2. BLAST sequence than found pDvC1 and pDvC2 and Ty3-gypsy-type retrotransposons coat protein coding genes have a high degree of sequence homology. As a probe pDvC1, and pDvC2 sequence, using fluorescence in situ hybridization analysis of the distribution of them in a variety of species of Triticeae chromosome. In situ hybridization showed that in the perennial H.villosa chromosome centromeric region, pDvC1 and pDvC2 strong hybridization signal. With them hybridization durum wheat the - diploid cluster villosa amphidiploid (AABBVV), results showed that in areas all chromosome centromere signals appear, however, in wheat chromosome fluorescence signal intensity near the centromere region significantly weaker than H.villosa chromosome the centromere region near the signal. This shows the obvious amplification Ty3-gypsy-type retrotransposons in the species H.villosa the centromeric region. Thinopyrum intermedium (Thinopyrum intermedium), in situ hybridization, the results found in the middle of Thinopyrum 6 chromosome centromere significant amplification, which we previously thought the H.villosa case of Elytrigia closer relationship results coincide. According to repeat sequences pDvC1, we designed a pair of SCAR primers Mark Triticeae species PCR amplification test. The results show that this pair of primers containing species H. villosa chromosomes can be identified also identified wheat - perennial cluster villosa translocation line in H.villosa forward centromere sequence import. Obtained in this study H.villosa chromosome centromere-specific repetitive sequences have important reference value of the in-depth study of the evolution of repetitive sequences in Triticeae species.

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