Haploidisasi melalui androgenesis dan ginogenesis pada anyelir (Dianthus sp.)

Abstract

Haploidization technology ensures important advantages in obtaining pure lines by rapid fixation of homozygosity. Anther culture, ovule culture, ovary culture and irradiated pollen technique were used in this study. The objective of the research was to develop appropriate haploid technology in order to obtain haploid plants through androgenesis, gynogenesis by ovule culture and ovary culture and pseudofertilization on Dianthus chienesis. Androgenic callus was induced in four WT basic medium inductions supplemented with 2.4D, NAA, TDZ and BA. Four explants originated from ovule and ovary cultures of six genotypes of D. chinensis (Dchi-11, Dchi-12, Dchi-13, Dchi-14, Dchi-15 and Dchi-16) were applied in five media. For the parthenogenesis induction, 100-200 Gray gamma irradiations were applied to the Dchi-14 pollens and pollinated to the Dchi-11. Pollinations were conducted on the day after irradiation. Eight media were selected to obtain regenerated plants. Results showed that androgenic callus formation needed high auxin and sitokinin ratio, while regenerated callus required lower auxin and sitokinin ratio. 2,4-D was better than NAA in callus induction. Callus originated from anther culture, multi ovule slice culture, ovule culture and ovary slice culture regenerated producing early flowering plants and expressed of abnormal flowering mutant. Regeneran from anther culture was diploid, but two putative double haploids came up from multi ovule slice culture and ovary culture. Cytology and flow cytometry observations showed that seven haploid plants were obtained from pseudofertilization. Spontaneous chromosome doubling was inferred to have occurred during the callus culture period. In conclusion, gynogenesis through multiovule culture, ovary slice culture and pseudofertilization was more effective in inducing haploid and double haploid plants than androgenesis.