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<p>The in vitro propagation protocol for <i>Anoectochilus elatus</i> was developed using rhizome explants. The explants were cultured on Mitra medium supplemented with various plant growth regulators for shoot bud induction, multiplication and in vitro rooting. After 12 weeks of culture, a maximum of 25.5 multiple shoots per explant, with a shoot length of 2.9&#xa0;cm were obtained from Mitra medium supplemented with 9.6 µM jasmonic acid and 4.2 µM adenine sulphate. The elongated micro-shoots were treated with 7.9 µM phloroglucinol for in vitro rooting. A maximum of 3.0 roots per shoot were produced within 17 days of culture, with a root length of 2.1&#xa0;cm and a 66% rooting rate. Subsequently, endophytic mycobionts, including <i>Rhizoctonia solani</i>, <i>Fusarium oxysporum</i> and <i>Fusarium proliferatum</i> were isolated from root segments and identified using morphological traits and internal transcribed spacer molecular markers. These mycobionts were co-cultured with in vitro propagated plantlets, and host root recolonization was confirmed. Furthermore, the antioxidant activity was assessed in leaves collected from in vitro propagated plants acclimatized with and without mycobionts using the Ferric Ion Reducing Antioxidant Power Assay. In these mycobionts, <i>R. solani</i> and <i>F. proliferatum</i> significantly increased the biomass content, antioxidant levels and promoted the <i>ex vitro</i> survival rate of in vitro propagated plants.</p>

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Isolation, characterization of endophytic mycobionts and their utilization for the acclimatization of in vitro propagated plantlets of Anoectochilus elatus – a near threatened terrestrial jewel orchid

  • 期刊论文

  • 发表日期：

    2026年4月28日

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  • N. Ahamed Sherif,
  • J. H. Franklin Benjamin,
  • M. N. Abubacker,
  • T. Senthil Kumar,
  • M. V. Rao

摘要

The in vitro propagation protocol for Anoectochilus elatus was developed using rhizome explants. The explants were cultured on Mitra medium supplemented with various plant growth regulators for shoot bud induction, multiplication and in vitro rooting. After 12 weeks of culture, a maximum of 25.5 multiple shoots per explant, with a shoot length of 2.9 cm were obtained from Mitra medium supplemented with 9.6 µM jasmonic acid and 4.2 µM adenine sulphate. The elongated micro-shoots were treated with 7.9 µM phloroglucinol for in vitro rooting. A maximum of 3.0 roots per shoot were produced within 17 days of culture, with a root length of 2.1 cm and a 66% rooting rate. Subsequently, endophytic mycobionts, including Rhizoctonia solani, Fusarium oxysporum and Fusarium proliferatum were isolated from root segments and identified using morphological traits and internal transcribed spacer molecular markers. These mycobionts were co-cultured with in vitro propagated plantlets, and host root recolonization was confirmed. Furthermore, the antioxidant activity was assessed in leaves collected from in vitro propagated plants acclimatized with and without mycobionts using the Ferric Ion Reducing Antioxidant Power Assay. In these mycobionts, R. solani and F. proliferatum significantly increased the biomass content, antioxidant levels and promoted the ex vitro survival rate of in vitro propagated plants.