The study revealed, for the first time, accumulation of spilanthol, an antiseptic alkylamide, in in vitro
cultures of Spilanthes acmella Murr., a medicinal plant of immense commercial value. To achieve this,
in vitro shoots were regenerated via direct organogenesis from leaf-disc explants of Spilanthes. Shoots
were induced in the presence of N6-benzylaminopurine (BAP) alone or in combination with either -
naphthalene acetic acid (NAA) or Indole-3-acetic acid (IAA) in Murashige and Skoog medium. The best
treatment for shoot regeneration was MS+ BAP (5.0M)+ IAA (5.0M), which promoted adventitious
shoot proliferation in >82% cultures with an average of 5.3 shoots per explant. Regenerated shoots
rooted spontaneously with a frequency of 100% on half strength MS medium (major salts reduced to
half strength) containing 50 g l−1 sucrose. The plantlets were acclimatized successfully with 90% survival
rate. Additionally, ploidy stability of the regenerated plants was assessed by flow cytometry which
showed that all investigated plants had the similar ploidy as that of the mother plant. For spilanthol
identification, peaks eluted from HPLC were analyzed by mass spectrometry with its characteristic fragmentation
pattern. For quantification studies, calibration curve was generated, which revealed a higher
amount of spilanthol content (3294.36±12.4g/gDW) in the leaves of in vitro plants compare to those of
in vivo plants (2703.66±9.6g/gDWof spilanthol). An efficient multiplication frequency, ploidy stability
and enhanced spilanthol accumulation ensure the efficacy of the protocol developed for this industrially
important medicinal plant.
The study revealed, for the first time, accumulation of spilanthol, an antiseptic alkylamide, in in vitro
cultures of Spilanthes acmella Murr., a medicinal plant of immense commercial value. To achieve this,
in vitro shoots were regenerated via direct organogenesis from leaf-disc explants of Spilanthes. Shoots
ถูกชักนำในการปรากฏตัวของ n6 benzylaminopurine ( BAP ) คนเดียว หรือในการรวมกันกับทั้ง -
แนพทาลีน acetic acid ( NAA ) หรือกรดกระเพาะ ( IAA ) ในอาหารสูตร Murashige and Skoog ) การรักษาที่ดีที่สุดสำหรับการยิง คือ นางสาว BAP
( 5.0 M ) IAA ( 5.0 เมตร ) ซึ่งเลื่อนการยิงปากดี
ใน > 82 % วัฒนธรรม เฉลี่ย 5.3 ยอดต่อชิ้นส่วนพืช . ได้ยิง
rooted spontaneously with a frequency of 100% on half strength MS medium (major salts reduced to
half strength) containing 50 g l−1 sucrose. The plantlets were acclimatized successfully with 90% survival
rate. Additionally, ploidy stability of the regenerated plants was assessed by flow cytometry which
showed that all investigated plants had the similar ploidy as that of the mother plant. For spilanthol
identification, peaks eluted from HPLC were analyzed by mass spectrometry with its characteristic fragmentation
pattern. For quantification studies, calibration curve was generated, which revealed a higher
amount of spilanthol content (3294.36±12.4g/gDW) in the leaves of in vitro plants compare to those of
in vivo plants (2703.66±9.6g/gDWof spilanthol). An efficient multiplication frequency, ploidy stability
and enhanced spilanthol accumulation ensure the efficacy of the protocol developed for this industrially
important medicinal plant.
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