Article

Development and application of a genome specific PCR marker for Haynaldia villosa

College of Crop Science, China Agricultural University, Beijing 100094, China.
Acta Genetica Sinica 05/2003; 30(4):350-6.
Source: PubMed

ABSTRACT

Random amplified polymorphic DNA (RAPD) analysis was performed on common wheat Chinese Spring, H. villosa, addition lines of H. villosa chromosome in CS, substitution line 3V of H. villosa chromosome in Triticum aestivum. A genome specific polymorphic DNA segment from H. villosa, OPF02757, was obtained. On the basis of cloning and sequencing of OPF02757, two PCR primers were designed and a genome specific PCR marker for H. villosa was established. The PCR marker including 677 bp was localized on all the seven pairs of H. villosa chromosomes. The result of PCR amplification by the primers indicated that there was a specific band of 677 bp in the materials containing H. villosa Chromosome such as T. aestivum-H. villosa addition, T. aestivum-H. villosa substitution, T. aestivum-H. villosa amphidiploid, T. durum-H. villosa amphidiploid and H. villosum from different accessions, and there was no specific band of 677 bp if the materials did not contain H. villosa chromosome, such as T. aestivum, T. durum, Secale cereale, Hordeum vulgare, Thinopyrum elongatum, Thinopyrum intermedium. Therefore, the PCR maker of 677 bp is specific to H. villosa genome, and could be used as molecular marker for detection of chromosomes of H. villosa in wheat.

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    • "In the present study, we isolated a D. villosum-specific Gypsy-like retrotransposon sequence C1-10 by RAPD. It lacks homologies to OPF02 757 (Liu et al. 2003), indicating that it might be a novel Gypsy-like retrotransposon specific to D. villosum. The C1-10 contains one copy of the motif CAAAA (invert) which was responsible for the breakage–reunion mechanism (Appels et al. 1986) and the basic characteristic of direct repeats, indicating the intra-element recombination contributing to the genome expansion of D. villosum (Yang et al. 2006) (figure 2). "
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    • "If this genome-specific PCR-marker is used prior to DDM analysis, those chromosomal aberrations involving any alien segments inside or outside the interval designated by the two distal markers would be detected easily. However, we have not yet obtained this kind of marker and all the H. villosa genomespecific sequences reported so far are not desirable because of their limited genomic distribution or unstable PCR quality (De Pace et al. 1992; Li et al. 1995; Liu et al. 2003; Zhang et al. 2013). Therefore, exploitation of a new wide-genome-spanning repetitive sequence and its derived stable PCR marker for H. villosa is necessary, which will further increase the efficiency of the DDM strategy. "
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    • "villosa lines (Chen et al. 1996b), to show the presence H. villosa chromatin in sexual hybrids (Zhang et al. 1998), as well as to identify the hybrid calli obtained from asymmetric somatic hybridization (Xia et al. 1998). Moreover, a specific RAPD marker found by Liu et al. (2003) could be used for detection of H. villosa chromatin in other genetic backgrounds. Specific RAPD/SCAR or RAPD markers that can be used for easy screening in wheat for presence/absence of resistance genes have been obtained. "
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