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Bulletin of Botanical Research ›› 2020, Vol. 40 ›› Issue (3): 368-375.doi: 10.7525/j.issn.1673-5102.2020.03.007

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Shoot Regeneration from Transverse Thin Cell Layers of Pseudobulbs and Micropropagation of Bletilla striata

FAN Li, WANG Ren-Rui, YU Ma, LIU Jin-Huan, HOU Da-Bin, TANG Zhi-Kang   

  1. School of Life Science and Engineering, Southwest University of Science and Technology, Mianyang 621010
  • Received:2019-08-20 Online:2020-05-05 Published:2020-05-29
  • Supported by:
    Research on key technique of seedling establishment with virus-free,rapid propagation and efficient cultivation of Bletilla striata,Key research and development plan of Sichuan province(2018SZ0095); Research on seedling cultivation of Bletilla striata "wf-1" based on virus-free with ultra-low temperature and rapid propagation,MiaoZi Project of Scientific and Technological Innovation in Sichuan Province(2019-06);Study on Rapid Propagation of Bletilla striata and cryopreservation of Germplasm Resources,Training Program of Innovation and Entrepreneurship for College students in Sichuan Province(S201910619049)

Abstract: Pseudobulbs of Bletilla striata were taken as initial explants and cut averagely for three transverse thin cell layers-upper position, middle position and lower position to culture on different media. Different parts of pseudobulbs, plant growth regulators and their concentrations were used to study the effects on shoots regeneration from pseudobulb transverse thin cell layers. The thickness of pseudobulb transverse thin cell layers were also used for explants for comparing the browning rate and the number of shoots. Orthogonal design was used to study the effects of BA and NAA for shoots micropropagation. The micropropagated shoots were cultured on a medium for being strong and rooting prior to transplanting to soil. The results showed that:Position of pseudobulb transverse thin cell layers significantly affected shoot induction. Lower part of pseudobulbs were significant easier to induce shoots than upper part and middle part. TDZ and BA also had obvious effect on shoot induction. NAA had no obvious effect on shoot induction. Lower parts of pseudobulb transverse thin cell layers cultured on basic medium+2.0 mg·L-1 BA+1.0 mg·L-1 TDZ for 4 weeks was the optimal treatment to induce shoots. The induction rate was 93.3% and the number of shoots was 15. Pseudobulb transverse thin cell layers(1.6-2.0 mm in thickness) were scarcely browning. Basic medium+1.5 mg·L-1 BA+0.3 mg·L-1 NAA was the optimal medium for micropropagation. The multiplication factor was 4.3, and the length of shoots was 7.8 cm. Shoots regenerated directly from pseudobulb transverse thin cell layers and micropropagation system of B.striata was established successfully, which laid a foundation for innovation of germplasm resources.

Key words: Bletilla striata, pseudobulb transverse thin cell layer, micropropagation

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