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Transverse thin cell layer (tTCL) technology: a promising tool for micropropagation of Centratherum punctatum Cass.

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Abstract

The present study describes an efficient protocol for the micropropagation of Centratherum punctatum Cass. through transverse thin cell layers (tTCLs). The effect of plant growth regulators (PGRs), thickness of tTCLs, and source of tTCLs were evaluated. The tTCLs of varying thickness (0.5 to 5.0 mm) were excised from 10-d-old leaf and 45-d-old node and cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of PGRs (BAP, TDZ, KN; 0.1 to 2.0 mg L−1), alone or in combination with NAA (0.1 to 1.0 mg L−1) for shoot induction. The leaf and node tTCLs responded with direct shoot regeneration. A significant effect of thickness or width on shoot induction was observed. For leaf, 1.0-mm-wide and for node 2.0-mm-thick tTCLs showed a maximum response. MS medium supplemented with 1.5 mg L−1 TDZ and 1.0 mg L−1 BAP in combination with 0.2 mg L−1 NAA was found to be optimum for shoot induction from tTCLs of leaf (96% response with 41.5 shoots per explant) and node (87% response with 11.9 shoots per explant), respectively. Rhizogenesis was obtained when micropropagated shoots were transferred to half strength MS medium supplemented with various concentrations (0.25 to 2.0 mg L−1) of IBA. The rooted plantlets were eventually acclimatized and transferred to soil. The clonal fidelity assessment by SCoT markers of the mother plant and micropropagated plants revealed their genetic similarity. Chlorophyll content estimation indicated lower levels in in vitro plants, which increased to a comparable level to that of field-grown plants after 3 mo of acclimatization, showing absence of stress during acclimatization. The procedure described here is a promising tool for micropropagation of C. punctatum as it produces high-frequency healthy shoots from a minimum explant source.

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Funding

Aswathi N. V. thankfully acknowledges the financial support from CSIR, Govt. of India, in the form of JRF and SRF (Award No. 09/1108(0019)/2017-EMR-I).

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  1. Department of Plant Science, Central University of Kerala, Tejaswini Hills, Periye (P.O.), Kasaragod, Kerala, 671320, India

    N. V. Aswathi & T. Dennis Thomas

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ANV collected materials and performed the experiments, evaluated data, and wrote the manuscript. TDT designed the experiments, contributed to writing, and corrected the manuscript.

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Correspondence to T. Dennis Thomas.

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Aswathi, N.V., Thomas, T.D. Transverse thin cell layer (tTCL) technology: a promising tool for micropropagation of Centratherum punctatum Cass.. In Vitro Cell.Dev.Biol.-Plant 59, 340–353 (2023). https://doi.org/10.1007/s11627-023-10348-2

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