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<p><i>Mucuna pruriens</i> is a leguminous plant known for its neuroprotective and antimicrobial properties. However, its large-scale utilization is limited due to poor seed germination, sensitivity to environmental stress, and inefficient propagation methods. Therefore, developing an efficient in vitro clonal propagation system, along with conservation strategies and bioactivity validation, is essential for its sustainable use. This study presents an enhanced protocol for clonal propagation of <i>Mucuna pruriens</i> using nodal segments integrated with synseed (encapsulation) technology for short-term conservation. While encapsulation approaches have been reported in several medicinal plants and micropropagation studies exist for <i>M. pruriens</i>, the present work uniquely combines synseed production with low-temperature storage, regeneration efficiency, physiological and biochemical assessment during acclimatization, genetic fidelity validation, and antibiofilm activity against MDR-ESKAPE pathogens. A 3% sodium alginate gelling matrix combined with calcium chloride (CaCl<sub>2</sub>; 100 mM) produced optimal beads, facilitating the induction of encapsulated nodal segments convert into plantlets. The highest shooting (91.60 ± 0.81%) was recorded on Murashige and Skoog (MS) basal medium with meta-Topolin (mT; 6.5 µM) and Putrescine (Put; 10.0 µM). Micro-shoots from synseeds rooted highest in ½ MS nutrient with combination of indole-3-butyric acid (IBA; 0.2 µM). Low-temperature storage (4&#xa0;°C) was evaluated for up to 70 days to assess the regeneration potential of encapsulated and non-encapsulated nodal segments. During acclimatization, photosynthetic pigments, oxidative stress markers, and antioxidant enzyme activities were monitored over 54 days. Encapsulated plantlets were successfully hardened and transferred to field conditions. Genetic fidelity analysis using 10 ISSR primers revealed a monomorphic banding pattern, confirming true-to-type regeneration.Tissue-cultured plant parts (stem, leaves, and seeds) of <i>M. pruriens</i> showed significant antibiofilm activity against multi-drug resistant ESKAPE bacteria (<i>Enterococcus faecium</i>,<i> Staphylococcus aureus</i>,<i> Klebsiella pneumoniae</i>,<i> Acinetobacter baumannii</i>,<i> Pseudomonas aeruginosa</i>, and <i>E. aerogenes</i>). However, the observed differences in bioactivity between tissue culture–raised and wild plant materials are based on biological assays. Therefore, these variations may be associated with differential accumulation of secondary metabolites under in vitro conditions, which requires further validation through advanced analytical techniques. This research supports the high-rate multiplication and conservation of new biologically active phyto-compounds.</p>

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Artificial seed mediated micropropagation of Mucuna pruriens L. (DC) and its novel antibiofilm potential against MDR ESKAPE pathogens

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年7月10日

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  • Naushad Alam,
  • Firoz Ahmad Ansari,
  • Fohad Mabood Husain,
  • Amit Kumar,
  • Mohd Hashim Khan,
  • Mohammad Anis

摘要

Mucuna pruriens is a leguminous plant known for its neuroprotective and antimicrobial properties. However, its large-scale utilization is limited due to poor seed germination, sensitivity to environmental stress, and inefficient propagation methods. Therefore, developing an efficient in vitro clonal propagation system, along with conservation strategies and bioactivity validation, is essential for its sustainable use. This study presents an enhanced protocol for clonal propagation of Mucuna pruriens using nodal segments integrated with synseed (encapsulation) technology for short-term conservation. While encapsulation approaches have been reported in several medicinal plants and micropropagation studies exist for M. pruriens, the present work uniquely combines synseed production with low-temperature storage, regeneration efficiency, physiological and biochemical assessment during acclimatization, genetic fidelity validation, and antibiofilm activity against MDR-ESKAPE pathogens. A 3% sodium alginate gelling matrix combined with calcium chloride (CaCl2; 100 mM) produced optimal beads, facilitating the induction of encapsulated nodal segments convert into plantlets. The highest shooting (91.60 ± 0.81%) was recorded on Murashige and Skoog (MS) basal medium with meta-Topolin (mT; 6.5 µM) and Putrescine (Put; 10.0 µM). Micro-shoots from synseeds rooted highest in ½ MS nutrient with combination of indole-3-butyric acid (IBA; 0.2 µM). Low-temperature storage (4 °C) was evaluated for up to 70 days to assess the regeneration potential of encapsulated and non-encapsulated nodal segments. During acclimatization, photosynthetic pigments, oxidative stress markers, and antioxidant enzyme activities were monitored over 54 days. Encapsulated plantlets were successfully hardened and transferred to field conditions. Genetic fidelity analysis using 10 ISSR primers revealed a monomorphic banding pattern, confirming true-to-type regeneration.Tissue-cultured plant parts (stem, leaves, and seeds) of M. pruriens showed significant antibiofilm activity against multi-drug resistant ESKAPE bacteria (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and E. aerogenes). However, the observed differences in bioactivity between tissue culture–raised and wild plant materials are based on biological assays. Therefore, these variations may be associated with differential accumulation of secondary metabolites under in vitro conditions, which requires further validation through advanced analytical techniques. This research supports the high-rate multiplication and conservation of new biologically active phyto-compounds.