Androgenesis in rice (Oryza Sativa L.) breeding

Abstract

An investigation was carried out in rice, at the College of Horticulture, Vellanikkara to produce doubled haploid lines through anther culture technique. The study included production of a hybrid between IR 36 (ovule parent) and PTB 45 (pollen parent), evaluation of parents and hybrid, culturing the anthers of F1 to produce doubled haploids and analyse the effect of growth regulators (2,4-D, Kn, IAA and IBA) and carbon source (sucrose and maltose) on callus induction.

There was significant difference among the parents and hybrid. The hybrid recorded significantly different values for plant height, panicle length, grains / panicle, grain weight and grain yield / plant.

Among the 24 treatments used for anther culture, callus induction was observed in 11 treatments. Callus induction percentage ranged from 0 to 4.18. Maximum callus induction percentage was recorded in N6 medium supplemented with maltose 60 mg/l, 2, 4-D 2mg/l and Kn 0.5 mg/l (4.18%) followed by N6 medium supplemented with maltose 60 mg/l, 2, 4-D 2 mg/l and Kn 1mg/l (3.32%).

A significant increase in anther culture efficiency was observed when sucrose was replaced by maltose in the presence of growth regulators 2,4-D/Kn but not in the presence of IAA/Kn. Callus induction frequency was high in 2,4-D/Kn combination followed by IAA/Kn combination irrespective of carbon source. IBA/Kn combination was totally non-responsive.

Main effects of 2, 4-D and Kn were significant, but there was no interaction between 2, 4-D and Kn when sucrose was used as carbon source. Among the two levels of 2, 4-D, 3 mg/l (0.6%) was superior to 2 mg/l (0.24%) and among the two levels of kinetin 0.5 mg/l (0.71%) was superior to 1 mg/l (0.12%). Main effects of 2, 4-D only was significant when maltose was used as carbon source. Main effect of 2, 4-D indicated that there was significant reduction in callus induction response when level of 2, 4-D increased from 2mg/l (3.75%) to 3mg/l (2.31%). Sixty six calli were obtained through anther culture of which 57 were embryogenic calli and the rest non-embryogenic calli. The percentage of embryogenic calli (86.45%) was higher than non-embryogenic calli (13.6%). 2, 4-D/Kn combination produced more number of embryogenic calli (88.3%) than IAA/Kn combination (66.7%).

Thirty green plants and five albinos were obtained from regeneration medium. The frequency of green plant and the albinos were 85.71 per cent and 14.29 per cent respectively. Regeneration frequency of embryogenic and non-embyogenic calli were 27.19 per cent and 22.22 per cent respectively. Among 30 green plants obtained , 21 were homozygous diploids and the rest haploids. Frequency of spontaneous doubling was 70 per cent. Of the 30 regenerated plants taken for hardening 24 survived and all were observed for grains per panicle and seed setting percentage. The values of grains per panicle and seed setting percentage ranged from 87 to 106 and 89.79 to 95.33 respectively.