普通小麦与Elymus rectisetus异附加系的分子细胞遗传学鉴定

Elymus rectisetus (Nees in Lehm) A. Löve et Connor是目前小麦族中发现的唯一的无融合生殖种，为了鉴定和标记从普通小麦与E. rectisetus BC₂F₂衍生后代中选育的2n=44株系1026A1、1057A1和1035A2的外源染色体，应用细胞学、基因组原位杂交和RAPD方法进行了研究。经细胞学鉴定，3个株系花粉母细胞减数分裂中期Ⅰ（PMC MⅠ）染色体构型均为2n=22Ⅱ，与普通小麦Fukuhokomugi杂交F₁的PMC MⅠ染色体构型均为2n =21Ⅱ+1Ⅰ，两两杂交F1的PMC MⅠ染色体构型均为2n=21Ⅱ+2Ⅰ，表明它们是分别附加了1对互不相同外源染色体的普通小麦-E. rectisetus二体异附加系。标记E. rectisetus品系1050的基因组DNA为探针DNA，对3个异附加系进行原位杂交，分别鉴定出附加的1对E. rectisetus染色体。应用13个引物对2个亲本和3个异附加系进行RAPD分析，获得了可分别用于检测1026A1和1057A1中所附加的E. rectisetus染色体遗传物质的分子标记OPB-14_900bp、OPE-09_750bp和OPB-14_1000bp。

Molecular Cytogenetics Identifications on Triticum aestivum-Elymus rectisetus Alien Addition Lines

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.