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Organogenesis and high-frequency plant regeneration in Caryopteris terniflora Maxim. using thidiazuron

  • Medicinal Plants
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Abstract

In vitro culture is employed as a means of producing plant materials from medicinal plants to meet the increasing demands of the market. Herein, we focused on an efficient protocol for organogenesis through two pathways in six explant types from Caryopteris terniflora Maxim (C. terniflora), a medicinal and ornamental plant. For direct organogenesis, the maximum number of shoots per internode explant (62.5) was achieved on Murashige and Skoog (MS) medium containing 3.0 mg L−1 thidiazuron (TDZ) and the maximum number of shoot tips per explant (45.5) was obtained by 1.5 mg L−1 TDZ, with 100% response. A protocol was also developed for indirect organogenesis from leaf, leaf-petiole, petiole, and root explants on MS basal medium containing a range of TDZ concentrations. Leaf-petiole explants induced more adventitious shoots (57.6 per explant) and leaves (31.4 per explant) than petioles (37.5 per explant) or roots (25.0 per explant), and the differentiation rate ranged from 87.2 to 100%. The regenerated shoots were elongated on MS medium containing 0.3 mg L−1α-naphthaleneacetic acid (NAA) and 1.0 mg L−1 gibberellic acid (GA3). The elongated shoots developed roots on ½ MS medium containing 0.3 mg L−1 NAA. Successful acclimatization of rooted plantlets in a greenhouse was achieved with 100% survival. Furthermore, the embryogenic-like structure was observed and the morphological characteristics of C. terniflora assessed at different stages of in vitro development. Our protocol offers, for the first time, an efficient, scalable propagation procedure for C. terniflora useful for large-scale cultivation for the purposes of conservation and genetic manipulation.

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Funding

The work was supported by Applied Basic Project of Sichuan Science and Technology Department (2018JY0192, 2018JY0244), Scientific research initiation project of Mianyang normal university (QD2019A12), the National Natural Science Foundation of China (31972258), the Open Project from the Ecological Security and Protection Key Laboratory of Sichuan Province (ESP1602), and the Open Program of Beijing Key Laoratory of Flavor Chemistry (SPFW2019YB05).

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Authors and Affiliations

  1. Ecological Security and Protection Key Laboratory of Sichuan Province, Mianyang Normal University, Mianyang, 621000, China

    Qinggui Wu, Honglin Yang, Yuxi Sun, Jinyao Hu & Lijuan Zou

  2. Beijing Key Laboratory of Flavor Chemistry, Beijing Technology and Business University (BTBU), Beijing, 100048, People’s Republic of China

    Yuxi Sun

Authors
  1. Qinggui Wu
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  2. Honglin Yang
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  3. Yuxi Sun
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  4. Jinyao Hu
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  5. Lijuan Zou
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Corresponding author

Correspondence to Lijuan Zou.

Additional information

Editor: Ted Klein

Electronic supplementary material

Figure S1

A Caryopteris terniflora Maxim. plants with purple petals. B Germinated three-wk-old seedlings. bar = 1 cm (A), 3 cm (B) (PNG 1139 kb)

High Resolution Image (TIF 2141 kb)

Figure S2

In vitro shoot regeneration from leaf explants of Caryopteris terniflora Maxim. on medium with the same concentration (1.0 mg L−1) of 6-BA (6-Benzylaminopurine), TDZ (thidiazuron) and ZT (zeatin). A Adventitious bud induction with 1.0 mg L−1 6-BA after 20 d. B Cluster of adventitious buds on medium with 1.0 mg L−1 TDZ after 15 d. C Brown leaves and a few adventitious buds were observed only near the leaf vein with 1.0 mg L−1 ZT after 27 d. bar = 5 mm (A,B), 1 cm (C) (PNG 2183 kb)

High Resolution Image (TIF 11792 kb)

Figure S3

Shoot organogenesis via a somatic embryogenesis-like process in Caryopteris terniflora Maxim. A Green global callus (arrow 1) and shoot bud induction from pale green compact callus (arrow 2). B, b, and C (red arrow) are globe-shaped structures. D Somatic embryo-like structure at heart stage (black arrow). E Somatic embryo-like structure at the cotyledon stage (red arrow). F Development of the shoot cluster. Bar = 5 mm (A, B, C, E, F), 2 mm (D) (PNG 4503 kb)

High Resolution Image (TIF 7717 kb)

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Wu, Q., Yang, H., Sun, Y. et al. Organogenesis and high-frequency plant regeneration in Caryopteris terniflora Maxim. using thidiazuron. In Vitro Cell.Dev.Biol.-Plant 57, 39–47 (2021). https://doi.org/10.1007/s11627-020-10114-8

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  • DOI: https://doi.org/10.1007/s11627-020-10114-8

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