Organogenesis and plant re-generation from callus culture of Acacia hybrid clones BV350 and BV376

Abstract

This study determines the appropriate media in the tissue culture, and perfected the technical process of propagating some acacia hybrid species by the method of shoot regeneration through callus tissue and somatic embryogenesis. The results showed that stem segments (without dormant buds) and leaf fragments of two lines BV350 and BV376 cultured on MS medium supplemented with 1 mg/l 2,4-D and 0.5 mg/l BAP gave callus formation rates of 81.11% and 84.44% (stem segments), 85.56% and 87.33% (leaf fragments), respectively. Callus induction to form somatic embryos on MS medium supplemented with 1 mg/l TDZ and 0.25 mg/l IAA gave somatic embryogenesis rates of BV350 and BV376 lines of 36.67% and 58.34% in stem segments, 50.67% and 60.56% in leaf fragments, respectively. Shoot regeneration from somatic embryos on MS medium supplemented with 1 mg/l TDZ and 0.25 mg/l IAA gave shoot regeneration rates of 36.24% in BV350 line and 50.62% in BV376 line after 6-9 weeks of culture. Shoot multiplication on MS medium supplemented with 1.5 mg/l BAP and 0.5 mg/l NAA gave an average number of shoots per cluster of 5.81 (BV350) and 6.92 (BV376). Rooting stimulation to create complete plants on 1/2MS medium supplemented with 1.5 mg/l IBA, rooting rate reached 87.62% (BV350) - 91.34% (BV376). The system of regenerating hybrid Acacia lines through callus and somatic embryogenesis was built and can be applied in gene transfer/gene editing to create Acacia varieties with desired traits.