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Drought stress-responsive transcript analysis of wheat-rye translocation line using cDNA-AFLP
Wheat-rye translocation lines are widely used in wheat breeding programmes by reason of biotic stress tolerances. Though there have been a number of researches regarding abiotic stress tolerance, the tolerance of the lines depends on wheat genetic background, not on rye chromosome. Here, we investigated wheat-rye translocation specific transcripts derived from cDNA-AFLP under drought stress, which may help to elucidate the reaction under the stress. ‘OK91G117’ (1BL.1RS translocation) and ‘OK91G144’ (non-translocation) were used as materials, which are near-isolines for 1RS. 25% PEG 6000 was added in culture solution to simulate drought condition and root tissues were sampled at each 0 h, 3 h, 6 h, 12 h, 24 h, and 48 h after PEG treatment for RNA extraction. As a result of cDNA-AFLP, TDFs (transcript derived fragments) that were specific to OK91G117 were sequenced. GO functions of each sequenced TDF were annotated by Blast2GO using standard parameter with cut-off level 3 and mapped to the GO term (i.e. biological process; BP, molecular function; MF, cellular component; CC). The term with “organic substance metabolic process”, “primary metabolic process”, and “cellular metabolic process” account for almost 50 % of BP. The most represented terms among probes classified to MF were “transferase activity” and most of TDF were annotated in “cell part” of CC. In addition, rye-chromatin specific markers were developed by BLAST comparing sequence of TDF with wheat and rye genome data. RT-PCR was conducted to validate expression patterns of selected TDF. Further studies will be needed to elucidate functions of the highly expressed genes under drought stress.
