Acta Univ Latv (2004) 676: 153–158

Embryogenesis, callogenesis and plant regeneration from anther cultures of spring rape (Brassica napus L.)

Abstract

Anther culture is a very important and useful tool in plant breeding for haploid production. The investigation was carried out with three spring rape cultivars, 'Trend', 'Landmark' and 'Auksiai'. Rape anthers were cultivated in modified Nitch and Nitch induction medium supplemented with 13 % sucrose and 0.4 % agarose. Embryo regeneration took place in a modified B5 Gamborg nutrient medium supplemented with 0.1 mg l-1 GA3 and 0.8 % agar. Temperature pretreatment of flower buds was at 35 °C for 72 h. After 72 h thermal shock pretreatment, the callus formation frequency differed for the tested genotypes. Thermal shock pretreatment on the callogenesis process appeared to be genotype dependent, since this treatment failed to significantly increase for 'Auksiai' and significantly decrease callus formation for the 'Landmark' and 'Trend' cultivars. Further embryo development was noted only in genotype 'Trend'. The frequency of embryo formation from anthers treated with high temperature was higher than the control. It was observed that in haploids the stomata length was reduced by 1.4 and their number in the vision field increased by approximately 1.1 in comparison with the diploid parent plants. The height of the haploid plants was 1.4 times less than parent plants. Plant regenerants were 100 % haploids.