issue culture protocol of limber pine of North America

The tender apical bud of limber pine was taken as explant to investigate the induction and multiplication of adventitious buds and the formation of adventitious roots. It was found that the induction frequency of adventitious buds was 86.67% when the tender apical bud was cultured on SH medium supplemented with 0.5 mg/L BA and 0.1 mg/L NAA. Four weeks later, the induced buds were subcultured on SH basal medium for elongation. Then the elongated shoots were transferred to SH medium containing 2.0 mg/L BA for multiplication for 4 weeks, and the number of average buds per shoot was 3.75. After cultured on the SH basal medium or supplemented with 0.05% AC, shoots with a height of 1--1.5 cm were transferred on 1/2 GD medium without growth regulator to induce roots. Six weeks later, 1--3 adventitious roots were formed at the base part of adventitious buds. Finally, the plantlets were transplanted in soils (peat∶vermiculite=1∶1) and the survival rate was 64.86%. 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