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.
Highlights
Orthosiphon stamineus, commonly known as Misai Kucing, has been widely used as a traditional remedy for various illnesses
Emergence of new shoots could be observed after 1 week of culture
Media supplemented with combinations of Kin and indole-3-acetic acid (IAA) hormones showed better production of shoots and leaves than media supplemented with either Kin or IAA only
Summary
Orthosiphon stamineus, commonly known as Misai Kucing, has been widely used as a traditional remedy for various illnesses. This species can be found throughout Southeast Asia and tropical. The leaves have been introduced to Europe and Japan as a health tea, known as Java tea. The flowers resemble cat whiskers, with long wispy-shaped stamen [1]. This species has been reported to improve general health effectively, as well as to be a known remedy for kidney diseases, bladder inflammation, gout, and diabetes [2]. The leaves were found to possess diuretic [6,7]
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