Use of mannitol for cold pretreatment of isolated spikes and as a component of nutrient medium for spring barley anther culture in vitro

Abstract

Aim. The investigation was aimed to compare the efficiency of mannitol as a component of spike cold pretreatment solution and of medium for spring barley haploid production via anther culture in vitro. Methods. Field-grown plants of two genotypes with a contrast androgenic capacity were used as donors. Cut tillers were pretreated in water at 4⁰C for 5 days, while aseptically isolated spikes were kept in 0.3 M mannitol for 10 days at the same temperature. Isolated anthers were inoculated on inductive media containing N6 macro-, MS micronutrients, organic supplements, maltose (9.0%); the media with and without 0.1M mannitol differed in solidifying agents. Results. In highly-responsive line DH00-126, there was an almost two-fold increase in mmgreen plant regeneration frequency (from 48 to 58 plants per 100 anthers) in all experimental variants in comparison to the control. In recalcitrant variety Mebere, the combination of tiller pretreatment in water with anther cultivation on inductive starch-gelling media supplemented with 0.1 M mannitol resulted in the highest green plant yield. Conclusions. Both methodical approaches were proved to be applicable to increase the efficiency of haploid production in spring barley anther culture in vitro.