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Genetic homogeneity assessment of in vitro-regenerated plantlets of Nyctanthes arbor-tristis L. and comparative evaluation of bioactive metabolites and antioxidant activity


In vitro propagation of Nyactanthes arbor-tristis L. was achieved by culturing N-phenyl-N′-benzothiazol-6-yl-urea (PBU)-pretreated nodal explants in Murashige and Skoog (MS) medium without any phytohormones. Pretreatment of nodal explants in liquid MS medium with 100 μM N-phenyl-N′-benzothiazol-6-yl-urea for 4 d showed the highest shoot proliferation by producing maximum number of shoots (17.40 ± 1.02) per explant, with average shoot length of 5.96 ± 0.08 cm at the end of 8 wk. Effective rooting was accomplished by preincubating the cut-end of shoots with half-strength MS medium containing 6 μM indole-3-butyric acid for 1 wk, followed by implantation into half-strength MS medium; an average of 6.20 ± 0.049 roots per shoot were produced. Seventy-eight percent of the plantlets regenerated in vitro were successfully acclimatized and transferred to soil. These plantlets appeared to be morphologically similar to the donor plants. The genetic fidelity of these in vitro-regenerated plantlets was confirmed by start codon targeted polymorphism (SCoT) marker analysis, followed by comparative evaluations of the bioactive metabolites (ursolic acid, rengyolone, arbortristoside-A, and nyctanthoside), antioxidant-rich phytochemicals, and radical scavenging activities. This optimized in vitro propagation protocol should be an aid for the conservation of N. arbor-tristis germplasm, as well as cater to the needs of herbal industries for the production of therapeutic molecules.

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This study is supported by the UGC, Govt. of India, and the Vice Chancellor, Sambalpur University, Odisha, India, through financial support and the facilities to carry out this work.

Author information

JP conceived the project, designed the experiment, interpreted the results, and contributed to the writing of the manuscript; SCR performed the experiments, analyzed and interpreted the data, and contributed to the writing of the manuscript; SS, SKM, and PKY contributed towards the analysis of antioxidant activity and estimation of biomolecule content. AKG contributed towards critical evaluation of the findings and writing of the manuscript. All the authors have contributed towards the final version of the manuscript by writing and editing.

Correspondence to Jogeswar Panigrahi.

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The authors declare that the study was carried out following scientific ethics and conduct. However, this study did not involve any use of animals; hence, no ethical approval has been obtained from the concerned committee.

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Editor: Pamela Weathers

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Rath, S.C., Seth, S., Mishra, S.K. et al. Genetic homogeneity assessment of in vitro-regenerated plantlets of Nyctanthes arbor-tristis L. and comparative evaluation of bioactive metabolites and antioxidant activity. In Vitro Cell.Dev.Biol.-Plant 56, 72–87 (2020).

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  • Antioxidant activity
  • Bioactive metabolites (arbortristoside-A, nyctanthoside, rengyolone, and ursolic acid)
  • N-Phenyl-N′-benzothiazol-6-yl-urea (PBU)
  • Start codon targeted polymorphism (SCoT) marker analysis