Regeneration of Doubled Haploid Plants from in vitro Selected Microspores to improve Aluminium Tolerance in Wheat

The present study was designed to investigate the possibility of selection for aluminium tolerant doubled haploid lines from wheat anther culture. To carry out the expt?riment anthers from Mv16 were cultured on a solidified W14 induction media supplemented with 50 and lOOμmol/L aluminium sulfate, at low pH (pH 4.5). After 5 weeks of induction period the obtained embryoids were regenerated on 190-2 regeneration medium supplemented with AI in the concentrations as mentioned above. The offspring generation was tested in hematoxilin staining test to define the AI tolerance of the obtained DH lines. According to the results obtained AI added to the induction media had no negative effect on the androgenic responses (anther response, induction frequency of embryo like structures and their regeneration ability) of the genotype examined. The application of in vitro Al selection significantly increased the probability of obtaining DH lines with significantly higher tolerance, compared to the original genotype. Seeds of 10 selected DH lines were multiplied under field conditions for physiological studies. Seeds of the R4 generation were germinated in 10, 50 and lOOμmol/L AI containing media, and the root and shoot growth characteristics were measured after 10 days ofgermination. The chlorophyll and AI contents were also determined. In the physiological tests the shoot and root elongation ofthe AI tolerant lines was also inhibited but the degree ofinhibition was significantly less pronounced up to 50 μmol/L Al. Similar concentration dependent responses were obtained by comparing the fresh and dry weight of sensitive and tolerant lines. The toxic effect of AI was also reflected in chlorophyll content ofthe leaves. The increasing AI concentration decreased the chlorophyll content ofthe sensitive lines, but not in the AI tolerant ones.