‘Jincuilei’ is a mutant selected from Lonicera macranthoides Hand.-Mazz. It produces abundant flowers that never open with a chlorogenic acid (CGA) content up to 6.0%. Propagation through rooting or grafting has only a 30% survival rate. This study was undertaken to establish an efficient protocol for rapidly regenerating this mutant. Leaf explants were inoculated on Gamborg's B5 medium supplemented with different concentrations of 6-benzyladenine (BA) and 2,4-dichlorophenozyacetic acid (2,4-D). The optimal combination for callus induction was 4.4 μm BA with 2.26 μm 2,4-D, which resulted in 86.7% of leaf explants producing calluses in 4 weeks. Calluses produced from this optimal medium were cultured on B5 medium containing different concentrations of kinetin (KT) and α-naphthalene acetic acid (NAA). The best formulation for shoot induction was B5 medium containing 0.9 μm KT and 5.4 μm NAA in which 73.4% of cultured calluses produced shoots in 8 weeks, and shoot numbers ranged from three to six per callus piece (1 cm3). Adventitious shoots were cut and rooted in half-strength Murashige and Skoog medium supplemented with 14.8 μm 3-indolebutyric acid. Roots initiated 10 d after culture, and rooting percentages ranged from 98% to 100%. Plantlets grown in a container substrate in a shaded greenhouse had over a 95% survival rate. During the last 6 years, over four million plantlets were regenerated using this established procedure, and there was no somaclonal variation. Fresh and dry weights of 1000 flowers, CGA contents, and dry flower yields of the regenerated plants were not significantly different from those of the stock ‘Jincuilei’ propagated by cutting, indicating that plants regenerated from this established procedure were stable. This established in vitro culture method has led to rapid commercial production of this medicinal plant on more than 1500 ha of production field.
Xiaoming Wang, Jianjun Chen, Yongxin Li, Qiying Nie and Junbin Li
Huijie Zeng, Yun Li, Jianjun Chen, Xiaoming Wang, Zhongquan Qiao, Yongxin Li, Neng Cai and Sisi Liu
Xiaoming Wang, Yongxin Li, Huijie Zeng, Neng Cai, Zhongquan Qiao, Xiangying Wang and Jianjun Chen
Weigela florida (Bunge) A. DC. is a popular flowering shrub adapted to a wide range of environmental conditions. Efficient methods for micropropagation of this species have not been well developed. The present study established a protocol for in vitro shoot culture of W. florida ‘Tango’ after a systematic evaluation of different culture media, cytokinins, and auxins on axillary shoot induction. Single-node stems were cultured on Driver and Kuniyuki Walnut (DKW) medium for initial production of axillary shoots. The shoots were used as explants and cultured on DKW medium supplemented with 8.88 μm 6-benzylaminopurine (BA) and 0.27 μm naphthaleneacetic acid (NAA), resulting in the production of more than six axillary shoots per explant. The axillary shoots could either be used as explants for additional shoot production or be cultured on ½ DKW medium supplemented with 0.25 μm indole-3-butyric acid (IBA) for rooting. Plantlets were transplanted into a substrate with 99% survival rate in a shaded greenhouse. This established method could be used for rapid propagation of W. florida to speed the introduction of new hybrids or cultivars for commercial production.