Novel expressed sequence tags of an alpine-cold plant species,Gymnadenia conopsea (Orchidaceae)

Gymnadenia conopsea,an alpine Orchidaceae plant,was one of the widely used Tibetan traditional medicines.In this study,we sequenced total 105 expressed sequence tags (ESTs) from a full-length cDNA expression library constructed by the Oligo-capping technique.The further bioinformatic analyses suggested that the 65 represented unique sequences showed high homology to previously identified genes in other plants:30 sequences matched to other uncharacterized expressed sequence tags (ESTs),and 10 sequences showed no good matches to available sequences in DNA databases.Gene ontology annotation by InterProScan indicated that many of these cDNAs (7 percent) have no known molecular functions and may be unique to G.conopsea.Fifty-five ESTs with matched proteins were involved in a series of diverse functions,in which molecular function such as 'binding' (42.9 percent) and 'catalytic activity' (25.0 percent) were the most frequent functions of the cDNAs.This cDNA library provided a critical basis for further investigation of functional genes expression under cold stress in this alpine species.In addition,13 ESTs-based polymerase chain reaction (PCR) primers were designed and can also be used for genotypic identification and for the genetic diversity analysis of G.conopsea and its closely related species.

New PCR based markers allowed to identify Secale chromatin in wheat-Secale africanum introgression lines

The genus of Secale has many agronomically important characters.In order to use the best of this species,markers tracking the rye chromatin incorporated into wheat must be developed.In this study,one rye genome-specific random amplified polymorphic DNA(RAPD)marker was isolated from Secale africanum(Ra genome).Two cloned markers,named OPP13_(1165) and OPP13_(662),were 1165 bp and 662 bp,respectively.Sequence analysis revealed that OPP13_(1165) was highly homologous to a part of a new class of transposon-like gene called the Revolver family,and OPP13_(662) was partially similar to LTR gypsy-like retrotransposon.Fluorescence in situ hybridization(FISH)showed only OPP13_(1165) localized within the whole arms of rye except their terminal regions and no signal was detected on wheat chromosomes,while OPP13_(662) had no hybridization signal detected on wheat and rye genomes.Based on these sequences,two pairs of sequence-characterized amplified region(SCAR)primers were designed,and the resulted SCAR markers were able to target both cultivated and wild Secale species.The FISH patterns and the two SCAR markers should be able to identify and track all wheat-rye translocation lines,especially the S.africanum chromatin.