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Amplified fragment length polymorphism of DNA has been used to analyse the equality of plants obtained from isolated microspores. Although the control parental material was regarded as being highly homozygous, the analysis of the banding patterns of single plants showed a certain level of polymorphism. The analysis of regenerants with a doubled chromosome number did not show any diversity within the progeny of a single line. The differences in banding patterns coming from single plants were only observed in microspore donor lines. These results have proven the high purity of homozygous lines obtained via androgenesis from isolated microspores.
Conventional breeding methods are now supplemented by modern in vitro techniques. However, long term maintenance of cultures has many disadvantages. It incurs risk of loss through microbial contamination, somaclonal variation or human error, but above all the regeneration capacity can decrease gradually during extended maintenance. Cryopreservation as a method of long term storage of biological material without genetic alteration was adapted for embryogenic triticale calli preservation. Callus of both winter and spring genotypes were successfully cryopreserved. The best viability rates (80-85 %) were achieved with 6 weeks old winter genotypes treated with cryoprotective solution containing DMSO. This simple and efficient method of cryopreservation requires no special devices for controlled freezing and can be easily adapted for other cereals.
The androgenetic response of several selected male sterilitymaintainer genotypes of triticale was investigated. Androgenesis induction was obtained in all cultivars, but a large genotypic variation in green plant regeneration was observed. The number of regenerated triticale plants varied from 0.1 to 4.7 green plants per spike, depending on genotype. Spontaneous doubling of chromosomes varied from 14 to 60 % for particular genotypes and, on average, reached the value of 34 % for all genotypes. Fertile DH lines obtained in this study will find practical application in the development of triticale male sterile lines that are desirable in hybrid breeding.
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