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Impact of silver nanoparticles on the micropropagation of Hybanthus enneaspermus and assessment of genetic fidelity using RAPD and SCoT markers

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Abstract

A prompt and dynamic clonal propagation system was developed for Hybanthus enneaspermus a medicinally significant plant using shoot tip as explants. Multiple shoots were induced on Murashige and Skoog (MS) medium invigorated with different levels of 6-benzylaminopurine (BAP), kinetin (Kin) and thidiazuron (TDZ) and AgNPs. The optimum concentration of BAP (1.5 mg L─1 ) with AgNPs (3.0 mg L─1) exhibited a determined response of 90.66% and created a maximum of 77.23 shoots with a maximum shoot length of 3.72 cm after 4 weeks. The vigorous shoots were embedded on an MS medium modified with optimum concentration of IBA 1.0 mg L─1 and AgNPs 1.0 mg L─1, which engendered the highest number of roots (9.65) with an average root length of 4.77 cm, with the response of 87%. Further, rooted plantlets were efficiently acclimatized in the soil. In addition, genetic homogeneity analysis was performed using RAPD and SCoT molecular marker, which revealed that the regenerated plants were genetically uniform and no detectable genetic difference was observed when compared to the mother plant. This efficient micropropagation protocol provides a new way for targeted gene editing for functional analysis of genes in Hybanthus enneaspermus.

Key message

The green synthesized AgNPs increased the regeneration frequency and there is no genetic variation among the invitro regenerated plants.

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Fig. 1
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Abbreviations

AgNPs:

Silver nanoparticles

PCR:

Polymerase chain reaction

SCoT:

Start codon targeted polymorphism

RAPD:

Randomly amplified polymorphic DNA

FTIR:

Fourier transform infrared

SEM:

Scanning electron microscope

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Acknowledgements

The authors thank to “RUSA, 2.0 - Biological Sciences, Bharathidasan University (TN RUSA: 311/RUSA (2.0)/2018 dt.02/12/2020)” for providing financial assistance.

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Authors and Affiliations

  1. Department of Biotechnology, Bharathidasan University, Tiruchirappalli, Tamil Nadu, 620 024, India

    Selvam Sathish, Venkatachalam Vasudevan, Sivabalan Karthik, Chandrasekaran Ajithan, Shanthi Pandurengan Parthasarathy & Markandan Manickavasagam

  2. Department of Biotechnology, A.V.C. College (Autonomous), Mayiladuthurai, Tamil Nadu, 609 305, India

    Venkatachalam Vasudevan

  3. Department of Biological Sciences, Kangwon National University, Chuncheon, 24341, South Korea

    Sivabalan Karthik

  4. School of Bio Sciences and Technology, Vellore Institute of Technology, Vellore, Tamil Nadu, 632014, India

    Ramamoorthy Siva

Authors
  1. Selvam Sathish
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  2. Venkatachalam Vasudevan
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  3. Sivabalan Karthik
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  4. Chandrasekaran Ajithan
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  5. Ramamoorthy Siva
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  6. Shanthi Pandurengan Parthasarathy
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  7. Markandan Manickavasagam
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Contributions

SS performed experiments, analyzed, compiled data and prepared manuscript. VV helped to carry out AgNPs synthesis. SK, CA and SPP contributed to results analysis. RS evaluated the data. MM supervised the findings of this work and evaluated the manuscript. 

Corresponding author

Correspondence to Markandan Manickavasagam.

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Conflict of interest

 The authors declared that they have no conflicts of interest to this work.

Additional information

Communicated by Amita Bhattacharya.

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Supplementary Information

Below is the link to the electronic supplementary material.

11240_2022_2350_MOESM1_ESM.docx

Supplementary material 1 (DOCX 15.6 kb) Supplementary Table 1 Effect of cytokinins on multiple shoot induction from shoot tip explants of H.enneaspermus on MS medium after 6 weeks of culture.

11240_2022_2350_MOESM2_ESM.docx

Supplementary material 2 (DOCX 14.8 kb) Supplementary Table 2 Effect of different concentrations of AgNPs along with 1.5 mgL─1BAP on multiple shoot induction from shoot tip explants of H.enneaspermus on MS medium after 6 weeks of culture.

11240_2022_2350_MOESM3_ESM.docx

Supplementary material 3 (DOCX 14.9 kb) Supplementary Table 3 Effects of different concentrations of IBA and NAA on root induction of H.enneaspermus after 4 weeks of culture.

11240_2022_2350_MOESM4_ESM.docx

Supplementary material 4 (DOCX 14.8 kb) Supplementary Table 4 Effect of different concentrations of AgNPs along with IBA 1.0 mg L─1on root induction of H.enneaspermus on MS medium after 4 weeks of culture.

11240_2022_2350_MOESM5_ESM.docx

Supplementary material 5 (DOCX 13 kb) Supplementary Table 5 List of primers, their sequences, number and size of the amplified fragments generated by RAPD primers in H.enneaspermus.

11240_2022_2350_MOESM6_ESM.docx

Supplementary material 6 (DOCX 13 kb) Supplementary Table 6 List of primers, their sequences, number and size of the amplified fragments generated by SCoT primers in H.enneaspermus.

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Sathish, S., Vasudevan, V., Karthik, S. et al. Impact of silver nanoparticles on the micropropagation of Hybanthus enneaspermus and assessment of genetic fidelity using RAPD and SCoT markers. Plant Cell Tiss Organ Cult 151, 443–449 (2022). https://doi.org/10.1007/s11240-022-02350-0

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  • DOI: https://doi.org/10.1007/s11240-022-02350-0

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