Increasing In Vitro Shoot Elongation and Proliferation of ‘G.30’ and ‘G.41’ Apple by Chilling Explants and Plant Growth Regulators

Abstract

These studies were conducted to determine the most effective methods for increasing shoot elongation during the initial proliferation stage of micropropagation in two dwarfing apple, Malus ×domestica (Borkh.), rootstock cultivars. Several experiments were conducted to compare explant collection date, exposure to chilling (5 ± 1 °C) temperatures, and varying concentrations of plant growth regulators in Murashige and Skoog (MS) media. Microshoot growth of ‘Geneva 41’ (‘G.41’) was very low and unaffected by chilling duration from 0 to 8 weeks or by gibberellic acid (GA3) concentration from 0 to 1.0 mg·L−1, but was improved by an additional subculture which increased shoot length from 1 to 15 mm. In ‘Geneva 30’ (‘G.30’), shoot elongation was most affected by date, chilling explants, and by optimizing cytokinin concentration and type. Explant collection date in April increased shoot growth compared with August or November. Microshoot growth of ‘G.30’ was increased by chilling nodal explants for 4 and 6 weeks when explants were collected in August and November, but not in April. Eight weeks chilling was detrimental for explants collected in April, and generally had little or no effect with August and November. The cytokinin 6-benzylaminopurine (BA) increased shoot number to a greater extent than thidiazuron (TDZ) or zeatin (ZT), and was also more effective for increasing shoot elongation with concentrations of 0 to 2.0 mg·L−1. In ‘G.30’, GA3 increased shoot growth at the optimum concentration of BA, but not with lower concentrations. ‘G.30’ microshoots were fewer and shorter with 24-epi-brassinolide (EBR) at concentrations of 0.1 and 1.0 mg·L−1. Chemical names: N-phenyl-N’-(1,2,3-thiadiazol-5-yl)urea (TDZ), 6-(4-hydroxy-3-methylbut-2-enylamino)purine (ZT).