PLANT REGENERATION FROM ANTHER CULTURE OF ANCHOTE [Coccinia abyssinica (Lam.) Cong.]

Anchote is one of the most important but underutilized indigenous root tuber crops in Ethiopia. Lack of improved varieties is among the most important factors limiting production of the crop. Considering the significant contribution of in vitro haploid production in the development of improved variety, an experiment was conducted to develop in vitro protocol for haploid plant regeneration from anther culture of KUWE and 223098 genotypes of anchote. Determination of the responsive anthers by flower bud size and optimization of flower bud surface sterilization were done as a preliminary experiment. Callus induction was investigated using 2, 4 -D and BAP. Shoot regeneration was assessed using BAP on a medium supplied with 1 g/L activated charcoal. Root induction was conducted on a medium supplied with 0.5 mg/L IBA. Anthers obtained from flower buds of 0.8 to 1 cm in length were responsive. Flower buds sterilized with 1% NaOCl for 5 minutes resulted in 84.94% clean anthers. The sole application of 2, 4-D and BAP did not induce callus. The highest percent (84.37%) of callus induction frequency was recorded in genotype KUWE with a combination of 1 mg/L of each 2, 4-D and BAP. Maximum callus induction frequency (62.50%) in genotype 223098 was obtained at 1 mg/L 2, 4-D + 2 mg/L BAP. Lower concentrations of 2, 4-D induced massive greenish-white, loose to friable callus in earlier days of culture while higher concentration of 2, 4-D produced compact greenish callus. The highest (4.25) shoot number was recorded in genotype KUWE at 2 mg/L BAP while maximum (3.75) shoot number in genotype 223098 was recorded at 4 mg/L BAP. The lower concentration of BAP resulted in small number but longer shoots. The highest number of shoots recorded in genotype KUWE produced the highest leaf (4.5) and node (3.6) numbers. Whereas, maximum leaf (3.8) and node (2.7) numbers in genotype 223098 were recorded at 1 mg/L BAP. Out of 10 sample plants of each genotype tested for ploidy level, five in genotype KUWE and three in genotype 223098 were haploids. About 32% and 28% of in vitro rooted plantlets in KUWE and 223098 genotypes respectively survived after a month of acclimatization. In general, an in vitro protocol for haploid plant regeneration through anther culture has been successfully developed for both genotypes of anchote investigated in this study.