Searching for DNA introgressed from wheat and for wheat-like grain proteins in a rice x wheat hybridization derivative

The DNA of a putative rice x wheat hybridization derivative (X Oryticum oryzoides) from China, the DNA of its parental rice cultivar and the DNA of a wheat line were digested with ten different restriction endonucleases, resolved by agarose electrophoresis, Southern blotted and hybridized using genomic wheat DNA as a probe. Phenol extracted, ethanol and cetyltrimethylammonium bromide precipitated DNA of the putative hybrid showed a restriction fragment length polymorphism (RFLP) different from that of the parental rice. When the DNA was further purified by Qiagen chromatography, the RFLP differences were not detected. Hence the apparent RFLP differences were probably due partial digestion of the less pure DNA preparations by the restriction endonucleases. No real introgressed fragments from wheat genome could be shown. The HpaII/MspI sites were more frequently digested with MspI than with HpaII in rice and hybridization derivative DNA, but the sites were evidently more frequently methylated in wheat DNA. Thus, in terms of methylation of the DNA, the hybridization derivative was much more like the rice parent than the wheat parent. The hybridization derivative showed a single endospermal protein (mass 19 kg mol^-1) not detected in the parental rice cultivar. This minor protein was soluble in buffered 50% isopropanol and precipitable with methanol. The results indicate that there are no or only short introgressed sequences from wheat in the rice/wheat derivative, a result which might be considered in breeding efforts with the hybrid derivative.