PCR and sequence analysis of barley chromosome 2H subjected to the gametocidal action of chromosome 2C
Gametocidal (Gc) chromosomes induce various types of chromosomal mutations during gametogenesis in the chromosomes of common wheat and alien chromosomes added to common wheat. However, it is not yet known whether the Gc chromosome causes aberrations at the nucleotide level because mutations caused by Gc chromosomes have been studied only by cytological screening. In order to know whether the Gc chromosome induces point mutations, we conducted PCR analysis and sequencing with the progeny of a common wheat line that is disomic for barley chromosome 2H and monosomic for Gc chromosome 2C. We analyzed 18 2H-specific EST sequences using 81 progeny plants carrying a cytologically normal-appearing 2H chromosome and found no nucleotide changes in the analyzed 1,419 sequences (in total 647,075 bp). During this analysis, we found six plants for which some ESTs could not be PCR amplified, suggesting the presence of chromosomal mutations in these plants. The cytological and PCR analyses of the progeny of the six plants confirmed the occurrence of chromosomal mutations in the parental plants. These results suggested that the Gc chromosome mostly induced chromosomal aberrations, not nucleotide changes, and that the Gc-induced chromosomal mutations in the six plants occurred after fertilization.
KeywordsCommon Wheat Chinese Spring Fraction Length Barley Chromosome Alien Chromosome
Polymerase chain reaction
Expressed sequence tag
Fluorescence in situ hybridization
Genomic in situ hybridization
The aberrant 2H lines developed in this study will be deposited with the National BioResource Project-Wheat. This work was partially supported by the National BioResource Project, MEXT Japan. Contribution number 609 from the Laboratory of Plant Genetics, Graduate School of Agriculture, Kyoto University.