Mitigating in vitro induced macro- and micro-morphological anomalies using seismic stress in Vitex negundo L.
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ORIGINAL ARTICLE
Mitigating in vitro induced macro- and micro-morphological anomalies using seismic stress in Vitex negundo L. M. Manokari1 · S. Priyadharshini2 · N. Arumugam3 · Mahipal S. Shekhawat2 Received: 5 August 2020 / Revised: 8 September 2020 / Accepted: 9 September 2020 © Springer Nature B.V. 2020
Abstract Present study explores the potential of seismo-mechanical stress in alleviating the macro- and micro-morphological aberrations observed in the in vitro regenerated plantlets of Vitex negundo L. (Chaste tree). Untreated in vitro plantlets showed nonfunctional stomata, trichomes, and photosynthetic pigments. This study demonstrates that these structural and physiological anomalies are altered to near natural normalcy by application of a deliberate seismic stress (mechanical agitation) before ex vitro transplantation. The stress could be applied to the rooted in vitro plantlets by placing them on an incubator shaker set at 40, 60, 80 and 100 revolutions per minute (rpm) for 2–16 minutes twice a day during 4 weeks of experiment. The treated plantlets on evaluation revealed that a mechanical shaking of plantlets at 80 rpm for 12 min has induced optimal increase in shoot number (48.0 ± 0.32), leaves per shoot (12.8 ± 0.26) and fresh weight (4.8 g) as compared to the control (non-stressed). The treatment also improved the photosynthetic pigment ratio (Chl a, b and total carotenoids) thus, making them resemble normal plantlets (4.00 ± 0.30 µg g−1 FW). The foliar micro-morphological analysis revealed that the seismic stress helps in the development of structurally organized functional stomata, epidermal cells and trichomes in the in vitro derived plants. This is the first report on the use of seismic stress for alleviating the in vitro induced abnormalities and acclimatization of V. negundo plantlets. The study paves the way for rapid production of healthy plants by tissue culture and their acclimatization in a short time period for conservation and sustainable utilization of medicinal plants. Key message This is the first report on the application of in vitro seismic stress in the micropropagation of Vitex negundo. Seismic stress has potential role in alleviating in vitro induced macro- and micro-morphological aberrations. Stress enhanced the level of photosynthetic pigments and fresh weight of the plantlets. It promotes the survival rate of tissue cultured plantlets. Keywords Abiotic stress · Photosynthetic pigments · Seismo-morphogenesis · Stomata · Vitex negundo Abbreviations BAP 6-Benzylaminopurine IAA Indole-3-acetic acid Communicated by Francisco de Assis Alves Mourão Filho. * Mahipal S. Shekhawat [email protected] 1
Department of Botany, Siddha Clinical Research Unit, (CCRS, M/o AYUSH), Palayamkottai, Tirunelveli, Tamil Nadu, India
2
Biotechnology Unit, Kanchi Mamunivar Government Institute for Postgraduate Studies and Research, Puducherry, India
3
Department of Biotechnology, Pondicherry University, Puducherry, India
IBA Indole-3-butyric acid NAA α-Naphthalene acetic acid NaOC
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