Micropropagation of Morus latifolia Poilet using axillary buds from mature trees
Introduction
Mulberry is well known for its economic importance in the sericulture industry as its foliage is used as food for silkworms. About 1000 species of mulberry are cultivated worldwide (Hotta, 1958) and could be classified into foliage, fruit, timber or ornamental use. Conventionally, techniques for the propagation of mulberries involve cutting, grafting and sowing seeds, depending on the cultivar. Certain limitations exist for cutting and grafting, such as low survival rate, difficulty rooting and low multiplication rate due to the physiological state or environmental factors (Bapat et al., 1987, Narayan et al., 1989). Morus latifolia Poilet is famous for its z-bending shoots that can be shaped into different styles for ornamental use. Rooting ability of its cuttings is poor and time-consuming using conventional vegetative propagation methods (personal communication with J.T. Lin). The rapid development of tissue culture technology provides an alternative method to mass produce certain plants with desirable traits. For mulberries, in vitro propagation via nodal segments (Jain et al., 1990, Sharma and Thorpe, 1990, Yadav et al., 1990, Hossain et al., 1992, Katase, 1993, Pattnaik et al., 1996, Pattnaik and Cland, 1997, Vijaya Chitra and Padmaja, 1999), adventitious buds (Kim et al., 1985, Saito, 1992), and encapsulated shoot buds (Bapat et al., 1987) have been successfully reported. Comparing micropropagated plants with cutting-derived mulberries revealed that the former plants grew significantly more than the latter in the field, although their foliar nutrition contents were similar (Zaman et al., 1997).
Here I demonstrated the factors influencing the micropropagation efficiency of M. latifolia Poilet in order to establish a protocol for rapid multiplication. The role of light intensity, salt concentration, carbon source, plant growth regulators, and charcoal on propagation and rooting were evaluated in this study.
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Materials and methods
Six-month-old shoots were excised from 15-year-old M. latifolia Poilet trees grown in a field at the Miaoli District Agriculture Improvement Station. The shoots were rinsed thoroughly in running water for 10 min, disinfested in 2% (w/v) HClO with sonication for 15 min, and then rinsed 4–5 times in sterile distilled water. Each nodal segment was cut to 1.5 cm long, the scale leaves were removed from axillary buds and then buds were placed on induction medium.
All explants were cultured on a medium
Axillary bud induction in M. latifolia Poilet
Initially, in vitro propagated plantlets were obtained by inoculating axillary buds from mature M. latifolia Poilet trees on MS medium supplemented with 1 mg/l of BA and 2% glucose. The shoots as well as inflorescence developments were observed during bud induction. The shoots formed in vitro were isolated and sectioned into single-node pieces after removing the inflorescences and leaves. The axillary bud to shoot development could be classified into six stages as follows. First the axillary bud
Discussion
The nodal segment, which had a pre-existing meristem, is suitable for clonal propagation because it is easily manipulated, has a high proliferation rate and maintains clonal fidelity (Jain et al., 1990, Sharma and Thorpe, 1990, Yadav et al., 1990, Hossain et al., 1992, Katase, 1993, Pattnaik et al., 1996, Pattnaik and Cland, 1997, Vijaya Chitra and Padmaja, 1999). Six-month-old shoots with axillary buds from 15-year-old trees were used in this study. During shoot elongation of initial
Conclusion
A mulberry in vitro propagation system was established with high frequency using axillary buds. The multiplication rate using this method was about sixfold per month. This high multiplication rate could only be achieved using micropropagation rather than traditional vegetative propagation methods. Starting from a single axillary bud, estimated that 612 plantlets can be produced in 1 year. The clones were uniform and vigorous in the field. This result provided an efficient and rapid method for
Acknowledgements
This work was supported by Council of Agriculture, Executive Yuan, Republic of China (89-AST-1.1-FAD-08(38)). Thanks are given to S.M. Lin and Y.B. Wang for their technical assistance and Dr. Y.W. Liao for useful comments on the manuscript.
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