Author: Zhang Geng yun, Guo Yan, Liu Feng-hua, Chen Shou-yi and Chen Shao-lin
J Integr Plant Biol 1994, 36 (5): 0-.
The NaCl-tolerant calli of "77–170" were selected from 1800 anthers, treated with ethylmethanesulphonate (EMS) or untreated, on N6 agar medium containing 0. 5%, 0. 8 %, and 1.0 % NaC1 respectively. Twenty-two seed-setting plants were obtained from R1 pollen-plants among the NaCl-tolerant green plantlets. Through selection in soil cotaining 0. 5% NaCI for 10 successive generations, five NaCl-tolerant lines named as mutant-15, 16, 17, 19, 20 were obtained at length. In the soil containing 0.5 % NaC1, tolerant R10 plants of those five selected lines were much better than the control in growth and yield. Other NaC1-tolerant lines named as mutant-P2, R2, R3, YCR were selected from young inflorescence of "98" and "77–170" (P2 only). The procedure of selection and regeneration was nearly the same as the above-mentioned. As the six probes used, three of them (Rabl6, 21, 10E6) were specific cDNA clones isolated from rice callus, which transcription was induced by ABA or salt treatment, other two probes (RG4, RG711) were discovered in our laboratory and located on chromosome 7, and salt was a specific cDNA clone which only expressed in roots and sheaths under salt treatment. The RFLP survey showed that eight tolerant mutants were different from their respective controls at the six detectable chromosome loci. The number of mutants showing polymorphism for different probes was as follows, 6 for RG4, 6 for RG711, 5 for Rabl6, 2 for Rabl0E6, 2 for salt and 1 for Rab21 respectively. This result suggested that the loci RG4, RG711 and Rabl6 might be correlated with salt tolerance in rice. The weak salt tolerant one did not show any polymorphism with control by using six DNA probes. 70. 8% (17/24) of all polymorphic autoradiograms showed polymorphism on patterns which were obtained by digestion of at least two restriction enzymes. It seemed that the majority of mutations were caused by insertions or deletions. Comparing the RFLP results of R3 resistant callus and single regenerative R3 stalk, the existance of mosaic in R3 resistant callus was obviously observed. This suggested that protoplast screening before regeneration is an efficient way to obtain stable resistant line. When Rabl6 was used as probe, the autoradiogram was complicated by the appearance of many weak bands which indicated that Rabl6 could be a gene family.