Studies on genome relationship and species-specific PCR marker for Dasypyrum breviaristatum in Triticeae

Abstract

Dasypyrum breviaristatum and nine related species in Triticeae were analyzed using the random amplified polymorphic DNA (RAPD) technique, in order to understand the genetic relationship and to develop species specific markers. The genome relationship dendrogram shows that D. breviaristatum and D. villosum could not be grouped together, indicating that D. breviaristatum was unlikely to be directly derived from D. villosum, while D. breviaristatum was closest to Thinopyrum intermedium, which implied that they might have similar breeding behaviors when introducing their chromatins into wheat. A D. breviaristatum genome specific RAPD product of 1182bp, was cloned and designated as pDb12H. Sequence analysis revealed that pDb12H was strongly homologuos to a long terminal repeat (LTR) Sabrina retrotransposon newly reported in Hordeum. The pDb12H was converted into a PCR based marker, which allows effectively monitoring the D. breviaristatum chromatin introgression into wheat. Fluorescence in situ hybridization (FISH) suggested that pDb12H was specifically hybridized throughout all D. breviaristatum chromosomes arms except for the terminal and centromeric regions, which can be used to characterize wheat -D. breviaristatum chromosome translocation. The genomes repetitive element will also be useful to study gene interactions between the wheat and alien genomes after the polyploidization.

A Novel Wheat-<b><i>Dasypyrum breviaristatum </i></b>Substitution Line with Stripe Rust Resistance

“…The Dasypyrum genome-specific LTR sequence pDb12H can distinguish Dasypyrum chromatin in a wheat background [Yang et al 2006;Liu et al 2009]. A total of 10 mitotic chromosome spreads from D11-5 probed with pDb12H were observed by FISH.…”

“…For FISH analysis, the probe pAs1, containing a 1-kb DNA fragment isolated from Aegilops tauschii cloned in the plasmid pUC8 [Rayburn and Gill, 1986], pSc119.2 [McIntyre et al, 1990], and pHvG38 [Pedersen and Langridge, 1997] with a microsatellite (GAA) n for identifying the B-genome of wheat were generously provided by Dr. B. Friebe, Wheat Genetic and Genomic Resources Center, Kansas State University, USA. The LTR probe pDb12H was used to distinguish the Dasypyrum genome in the wheat background as reported by Yang et al [2006]. Sheared genomic DNA of Chinese Spring wheat (CS, genomes AABBDD, 2n = 42) was used as blocking DNA.…”

“…DNA was extracted from young leaves of D. breviaristatum , TDH-2, TDV-1, D11-5, and CS [Yang et al, 2006]. PCR-based Landmark Unique Gene (PLUG) primers were designed according to Ishikawa et al [2009].…”

Characterization of a genome-specific Gypsy-like retrotransposon sequence and development of a molecular marker specific for Dasypyrum villosum (L.)

“…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). However because the whole sequence of C1-10 as retrotransposon is not known, we can not figure out which Ty3/Gypsy lineage it belongs to.…”

Isolation and chromosomal localization of new MITE-like sequences from Secale

“…FISH is an important method to study the chromosomal organization, composition and evolution (Cuadrado & Schwarzacher 1998;Li et al 2004), while it is an effective assistance to trace alien chromatin (Belyayev 2001;Yang et al 2006). Although many repetitive DNA sequences for rye genome had been found, the majority of them are specific to single chromosome or a location rather than universally detecting rye chromatin (Rogowsky et al 1992;Nagaki et al 1999).…”