Is Nor region variability in wheat invariably caused by tissue culture?

In a previous study we observed extensive Nor region variability in tissue-culture derived plants of only one out of three tested wheat cultivars. This finding prompted us to further question whether or not this variability was invariably caused by in vitro culture. In the present study, the upper halves of spikes from four source plants of the inbred cultivar 'ND7532' were removed 12 days after anthesis. The immature embryos from these halves were cultured and regenerated into plants. The lower halves of the same spikes were retained on the plants to obtain mature caryopses. DNA was extracted from seedlings, cut with TaqI endonuclease, run on agarose gels, and the respective Southern blots were probed with the plasmid pTA71 to reveal the Nor region patterns. The sexual progeny of regenerants from three out of four source plants derived from the immature embryos provided Nor region patterns which were exactly identical to the patterns obtained from seedlings which germinated from the caryopses matured on the respective source spikes. The regenerants from the fourth source plant provided variable Nor region patterns. Analyses of the Nor region patterns of 21 individual seedlings germinated from caryopses of this source plant showed that 18 had a three-fragment pattern (consisting of 3.0, 2.7 and 1.9 kb fragments) while three seedlings lacked one (2.7 of 1.9 kb) fragment. Furthermore, the next sexual progeny of the regenerants which had a three-fragment pattern further segregated into three- and two-fragment patterns.These results, in conjunction with previous reports on Nor region variability among tissue-culture derived plants, suggest that this variability is not invariably related to in vitro culture.