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Selection for frost tolerance in anther culture-derived embryos and regeneration of frost-tolerant fertile DH plants in winter wheat



Selection for frost tolerance in anther culture-derived embryos and regeneration of frost-tolerant fertile DH plants in winter wheat



Acta Agronomica Hungarica 45(3): 285-293



The use of anther culture to screen for frost tolerance and to select mutants at haploid level is still limited in cereals. However, the in vitro haploid and dihaploid induction of wheat may provide a useful tool for the selection of frost resistance, as the existing or induced tolerance in the microspore or haploid embryo population can easily be screened and fixed in homozygotic form. The possibility of developing frost resistant lines by exposing somatic tissue cultures to subzero temperatures after hardening had been already reported by several research groups, but up till now no data on the use of haploid cultures is available. In the present experiment the possibility of selecting frost-tolerant haploid embryoids and the regeneration of fertile DH lines from these selected embryos was studied. In the course of the experiment anthers from the F1 generation of Mv15 (frost-tolerant) X GK Kincso (sensitive) reciprocal hybrids and their parents were cultured on a solidified W14 induction medium. After 5 weeks of induction in the dark the haploid embryos were harvested and cold-treated under controlled phytotronic conditions. The hardened embryos were frozen at -15degreeC. After freezing, regeneration was carried out using 190-2 regeneration medium and the regenerants were grown till maturity in phytotronic growth chambers. The fertile regenerants were self-pollinated. The progeny generation was tested using the usual frost resistance testing method and the results of the freezing test suggested that about 80% of the DH lines obtained were frost tolerant. According to the data obtained in the experiment, it can concluded that, using the haploid selection method, there is a strong possibility of producing frost-tolerant DH lines from crosses between frost-resistant and sensitive genotypes in a relatively short time. Some of the DH lines regenerated from the frozen embryos of the sensitive parent showed relatively high viability in the freezing test. These results suggest the possibility of selecting for frost-tolerant mutants from a haploid culture.

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