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Indirect organogenesis-mediated high frequency conversion of non-embryonic synthetic seeds, essential oil profiling and antibacterial activity in genetically stable plants of Patchouli

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Abstract

Patchouli is a prized tropical medicinal herb with broad-spectrum therapeutic importance. The present research work describes development of an efficient callus-mediated plant regeneration protocol along with associated germplasm portability system (via alginate-encapsulation). Using 1.5 mg/l α-naphthalene acetic acid (NAA) and 1.0 mg/l 2, 4-dichlorophenoxy acetic acid (2, 4-D), highly proliferative friable calli were produced that subsequently underwent organogenesis in combinatorial cytokinin treatment to yield multiple shoot clusters. The highest frequency of shoot formation was achieved using 1.5 mg/l NAA with 1.5 mg/l 6-benzylaminopurine (BAP) in Murashige and Skoog (MS) medium. In vitro-derived shoot tips were encapsulated with 3% sodium alginate and 100 mM CaCl2 solution. The encapsulated beads were germinated in MS media with various concentrations of polyamines, where the highest regeneration frequency was observed with 1.5 mg/l spermidine. The regenerated shoots were rooted in basal MS medium and were successfully acclimatized with 96% survival rate. Genetic homogeneity amongst the regenerated plantlets was validated using Start Codon Targeted polymorphism (SCoT) and CAAT box-derived polymorphism (CBDP) ascertaining a high degree of clonal fidelity. The essential oil (EO) profiling of the donor plant and the in vitro-derived plantlets revealed identical composition. Furthermore, the antibacterial activities of various tissue extracts and extracted EOs were evaluated against the opportunistic pathogens viz. Klebsiella pneumoniae (MTCC 109), Salmonella typhii (MTCC 733), Micrococcus luteus (MTCC 2470) and Staphylococcus aureus (MTCC 96). The minimum inhibitory concentration (MIC) ranged from 0.31 to 5.0 mg/ml and 2.5 to 5.0 mg/ml against Gram-positive and Gram-negative bacteria, respectively. Eventually, the present research provides a holistic insight into the rapid regeneration of quality planting material as well as pharmacological bioprospection of patchouli along with the scope of further qualitative improvement via genetic transformation.

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Acknowledgements

All the authors sincerely thank Dr. G. Narahari Sastry Director, CSIR-NEIST, Jorhat and CSIR for providing the research platform to carry out the present work. PB is also thankful to Mr.Anil Kumar, Dr. Dinesh Kumar of CSIR-IHBT and Dr. Mamta Arora, ASBASJSM College, Bela Ropar, Punjab for assisting in profiling/analysis GC-MS data. CSIR-NEIST Publication No: NMN-202138.

Funding

Authors sincerely acknowledges Ministry of Tribal affairs, Government of India for providing financial support in the form of National fellowship for SC/ST viding file number (No:201718-NFST-MIZ-00504), Academy of Scientific and Innovative Research (AcSIR), Ghaziabad – 201002 and CSIR Scientist Pool Scheme (Pool Number 9057A).

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  1. Brijmohan Singh Bhau

    Present address: Department of Botany, Central University of Jammu, Rahya-Suchani (Bagla), District - Samba, Jammu, Jammu & Kashmir, 181143, India

Authors and Affiliations

  1. Agrotechnology and Rural Development Division (ARDD), CSIR-North East Institute of Science and Technology, Jorhat, Assam, 785006, India

    Lucy Lalthafamkimi, Brijmohan Singh Bhau & Dipanwita Banik

  2. Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, 201002, India

    Lucy Lalthafamkimi & Dipanwita Banik

  3. Biotechnology Division, Council of Scientific and Industrial Research-Institute of Himalayan Bioresource Technology, P.O. Box 6, Palampur, Himachal Pradesh, 176061, India

    Sanjay Kumar, Srijana Mukhia, Rakshak Kumar & Paromik Bhattacharyya

  4. Department of Microbiology, Guru Nanak Dev University (GNDU), Amritsar, Punjab, 143005, India

    Srijana Mukhia

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Lalthafamkimi, L., Bhau, B.S., Kumar, S. et al. Indirect organogenesis-mediated high frequency conversion of non-embryonic synthetic seeds, essential oil profiling and antibacterial activity in genetically stable plants of Patchouli. 3 Biotech 12, 349 (2022). https://doi.org/10.1007/s13205-022-03302-3

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