In Vitro Regeneration and ISSR-Based Genetic Fidelity Analysis of Orthosiphon stamineus Benth

Abstract

Orthosiphon stamineus has been widely used as traditional remedy for various illnesses and diseases, such as cardiovascular diseases and epileptic seizures. In this study, direct regeneration through nodal segment of this species was attempted using Kinetin (6-Furfurylaminopurine) and IAA (indole-3-acetic acid). Optimum regeneration media was identified as MS media supplemented with 2.0 mg L−1 Kin plus 0.5 mg L−1 IAA. This yielded the highest number of shoots (5.57 ± 0.42) and leaves (20.53 ± 1.91) per explant. Acclimatization of the resulting in vitro regenerants was successful in all potting mixtures tested. However, potting mixture PF (1:1:1 ratio of black soil/red soil/compost) was identified as the best medium for acclimatization of this species, as it yielded 100% survival percentage after 90 days of acclimatization. Ten in vitro regenerants of O. stamineus were randomly collected after the third subculture and subjected to genetic variation analysis using inter-simple sequence repeat (ISSR) markers. Out of 20 ISSR markers tested, 10 working primers were observed to produce satisfactory amplification of bands, with an average of 7.11 bands per primer. A total of 610 bands were produced by the 10 primers. The percentage of polymorphism was observed to be very low, yielding only 7.32% polymorphism among all samples. Jaccard dissimilarity analysis was also conducted and very low genetic distance (about 0.1) was found among the in vitro regenerants and between the regenerants with the mother plant, thus ascertaining the clonal nature of the plantlets produced in this study.

Micropropagation, Genetic Fidelity and Phenolic Compound Production of Rheum rhabarbarum L.

“…In this regard, in vitro propagation could be a very effective alternative, since even a very small plant part, a few millimeters or even less, can be used as starting material for in vitro propagation which results in numerous clones in a much shorter time. Therefore, in vitro propagation of important medicinal plants and agricultural crops would become a reliable and necessary technique for high amount plant material production identical to mother plant [25,26].…”

Somaclonal Variation—Advantage or Disadvantage in Micropropagation of the Medicinal Plants

“…Govinden-Soulange et al (2010) [69] tested 30 different RAPD primers in order to assess the variation in micropropagated plantlets of Hibiscus sabdariffa L., and only 3 of them produced polymorphic bands, while Haque et al (2017) [28], working on Hibiscus cannabinus L., screened 25 decametric primers and selected only 3 for the analysis of the whole sample set. However, the ISSR markers, applied comparatively between the plantlets and the mother plant revealed, in general, the clonal nature of the analyzed regenerants, highlighting the genetic fidelity of in vitro propagation, even after several years [70][71][72].…”

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“…Change in the levels of biochemical markers such as AST, ALT, ALP and lipid peroxides were assayed in both paracetamol treated and control (untreated) groups. Treatment with the methanolic extract of Orthosiphon stamineus leaves (200 mg/kg) has accelerated thereturn of the altered levels of biochemical markers to the near normal profile in the dose dependant manner 34 .…”