Optimization of High-Efficiency Tissue Culture Regeneration Systems in Gray Poplar

Abstract

A series of tissue culture regeneration protocols were conducted on gray poplar (P. tremula × P. alba) to select the most efficient callus induction medium, adventitious shoot differentiation medium, shoot elongation medium and rooting medium, which laid the foundation for the optimization of genetic transformation technology for gray poplar. The results showed that the Woody Plant Medium (WPM) supplemented with 0.10 mg L−1 kinetin (KT) and 1.00 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) was the most suitable medium for callus induction. The callus induction rates of different tissues were greater than 85.7%. The optimal adventitious shoot differentiation medium was the WPM supplemented with 0.02 mg L−1 thidiazuron (TDZ), and the adventitious shoot differentiation rates of young tissues were 22.2–41.9%. The optimal direct differentiation medium was the Murashige and Skoog (MS) medium supplemented with 0.20 mg L−1 6-benzylaminopurine (6-BA), 0.10 mg L−1 indole butyric acid (IBA) and 0.001 mg L−1 TDZ, and the differentiation rate of adventitious shoots was greater than 94%. The best shoot elongation medium for adventitious shoots was the MS medium with 0.10 mg L−1 naphthylacetic acid (NAA). After 45 days of cultivation in the MS medium with 0.10 mg L−1 NAA, the average plant height was 1.8 cm, and the average number of elongated adventitious shoots was 11 per explant. The 1/2 MS medium with 0.10 mg L−1 NAA showed the best performance for rooting, and later, shoot growth. The direct shoot induction pathway can induce adventitious shoots much faster than the indirect adventitious shoot induction pathway can, and the time cost via the direct adventitious shoot induction pathway can be shortened by 2–6 weeks compared to that of the indirect shoot induction pathway.