Abstract:

The alien addition lines of T. aestivum can be used to breed the alien substitution lines and the alien translocation lines, to locate the alien genes on the specific chromosome, and to construct the alien chromosome DNA library. Elymus rectisetus (Nees in Lehm) A. Löve et Connor is the only species known with apomixes in the tribe of Triticeae. Three 2n=44 lines 1026A1, 1057A1 and 1035A2 were derived from [E. rectisetus × Fukuhokomugi (T. aestivum)] BC₂F₂ progeny. Cytogenetic, genomic in situ hybridization (GISH) and RAPD were applied to identify and tag E. rectisetus chromosomes of three lines. Chromosome configurations at PMC MⅠ of three lines were 2n=22Ⅱ. Chromosome configurations of hybrid F₁ between three lines and Fukuhokomugi were 2n=21Ⅱ+1Ⅰ. Chromosome configuration of half diallel cross F₁ between three lines were 2n=21Ⅱ+2Ⅰ. The probe was total genomic DNA of E. rectisetus accession 1050 labbelled with BioNick™ Labeling System and the block DNA was total genomic DNA of Fukuhokomugi. GISH analysis demonstrated that two E. rectisetus chromosome of three lines were detected, respectively. Genomic DNAs was isolated from E. rectisetus accession 1050, Fukuhokomugi and three lines, and was used for PCR amplification with 13 RAPD primers (OPB-08, OPB-14, OPE-09, OPF-05, OPF-09, OPJ-05, OPK-03, OPN-12, OPP-20, OPS-12, OPT-20, OPZ-09 and OPZ-11). OPB-14 and OPE-09 could amplify the E. rectisetus specific DNA bands OPB-14900bp and OPE-09750bp in 1026A1. OPB-14 could amplify the E. rectisetus specific DNA bands OPB-141000bp in 1057A1. The results of cytogenetic and GISH analysis showed that 1026A1, 1057A1 and 1035A2 lines were T. aestivum-E. rectisetus alien disomic addition lines with different E. rectisetus chromosomes. The results of RAPD amplification showed that OPB-14900bp, OPE-09750bp and OPB-141000bp could be used as molecular markers to detect E. rectisetus chromation in alien disomic addition lines 1026A1 and 1057A1, respectively.

Key words: Alien addition lines, Triticum aestivum, Elymus rectisetus, Genomic in situ Hybridization (GISH), RAPD