PHYTOCHEMICAL ANALYSIS OF CALLUS TWO VARIETIES ORTHOSIPHON ARISTATUS (BLUME) MIQ ON MURASHIGE AND SKOOG MEDIA: A STRATEGIC STEP OF SECONDARY METABOLITE PRODUCTION

Objective: The research aimed to provide new information regarding the secondary metabolites content of purple and white-purple Orthosiphon aristatus (Blume) Miq. callus, which can then be used as a basis for developing towards cell suspension and ultimately producing secondary metabolites using bioreactors. Methods: Callus induction of two varieties of O. aristatus were performed by inoculating sterile leaf explants grown on Murashige and Skoog basal media supplemented with 2,4-dichlorophenoxyacetis acid 0.4 ppm. The secondary metabolites were analysed and quantified using high-performance liquid chromatography with gradient elution. Results: The results showed the growth of callus two varieties of O. aristatus in growth media MS with 2,4-D 0.4 ppm. Rosmarinic acid content in the acetone extract of the purple variety callus was 1.28% w/w, and the white-purple variety was 2.22% w/w. Conclusion: This study could form the basis for the development of rosmarinic acid production by In vitro culture modification

Effect of Media Variation on the Induction and Phytochemical Profile of Callus in Two Varieties of Cat's Whiskers (Orthosiphon aristatus Blume Miq)

The levels of rosmarinic acid and sinensetin in purple and white-purple varieties of Orthosiphon aristatus, cat's whiskers, can be increased using modified in vitro culture. This work focused on callus induction of the purple and white-purple variety of cat's whiskers grown on Gamborg (B5) and CHU (N6) with the addition of growth regulators 2,4-dichlorophenoxyacetis acid. Our observation suggested that the callus could grow within three weeks and produce rosmarinic acid and sinensetin. The level of sinensetin from various extraction methods is relatively low; in contrast, the rosmarinic acid from purple callus was detected at about 5% w/w, while the white-purple variety was around 2% w/w. The results of this study also provided new information on the basic media other than MS that can grow cat's whiskers callus while producing active compounds

A COMPARATIVE PHARMACOGNOSTIC STUDY OF THE TWO Orthoshipon aristatus (BLUME) MIQ. VARIETIES

The use of a plant as an ingredient in traditional medicine requires scientific evidence to determine its properties. Cat's whiskers (Orthosiphon aristatus) is one of the widely used traditional medicinal plants in various Asian and European countries. This study aimed to determine the pharmacognostic properties of purple and white varieties of O. aristatus. Aqueous and ethanolic extract of the stem and leaves of purple and white varieties of O. aristatus was prepared and investigated for the presence of active ingredients. The AlCl3 colorimetric method was used for the estimation of flavones and flavonols. The level of flavonoid was reported 13.06 ± 0.13 mg QE / 1g and  6.17 ± 0.049 mg QE / 1g for the leaves and stem extracts of purple varieties respectively while this value was reported 9.76 ± 0.15 mg QE / 1g and 3.79 ± 0.03 mg QE / 1g for the white variety. From the results of the study, it can be concluded that the purple variety has a significantly higher amount of flavonoid then the white variety

Performance of Transgenic Chrysanthemum Harbouring Wasabi Defensin Gene for White Rust Disease Resistance

This study was intended to obtain white rust (Puccinia horiana) disease resistance Chrysanthemum transformed with wasabi defensin gene through mediation of Agrobacterium tumefaciens from three explant sources, i.e., leaf, lateral shoot bud, and internode. Observations were made on transformation efficiency, PCR analysis, in vitro and ex vitro disease resistance tests. Results showed that efficiency of transgenic callus and shoot regeneration was found both highest from lateral shoot buds (57.5% and 50.0%, respectively). PCR analysis showed that three putative transgenic plantlets from lateral shoot buds and one from leaf explant were putative transgenic carrying the wasabi, hpt, and nptII genes. Rooting test showed that the highest number of rooted plants was found in treatment of hygromycin (Hg) 25 mg L-1 (81%) and lowest was in treatment combination of kanamycin (Km) 50 mg L-1 + Hg 25 mg L-1 (25%). In vitro disease resistance test with sorus inoculation of P. horiana, directly on the leaves, resulted in 20 resistant plants out of 30 putative transgenic plants (66.67%). Ex vitro testing on adult plants of the same samples in a confined closed greenhouse (CGH) resulted in average of 80% transgenic Chrysanthemum plants were resistant, whereas in control plants caused white rust disease symptom

Changes in Surface Structure during Direct Somatic Embryogenesis in Rice Scutellum Observed by Scanning Electron Microscopy

Direct somatic embryogenesis in rice scutella was studied by scanning electron microscopy paying attention to the morphological change of surface structure. Somatic embryos were formed when the scutella of immature embryos of cv. Nipponbare were cultured on MS media containing 2 mg L−1 2,4-D. Embryos were then transferred to the media containing 1 mg L−1 2,4-D and germinated on hormone-free media. The developmental stages of typical monocotyledonous somatic embryo were classified into proembryo, globular, scutellar and coleoptilar stages by referring to the terminology used for zygotic embryos. Each stage had a characteristic surface structure. The surface of proembryo was smooth. Fibrillar material was observed during the transition from proembryo to globular stage. Fibrillar material was then gradually replaced by a mesh-like structure at the globular stage. With subsequent development of globular embryo, the apical scutellum region became flattened and the ridge of mesh-like structure became lower. Development of scutellum followed by the emergence of coleoptile from the scutellar notch was observed at the scutellar stage. The coleoptile surface was covered with elongating cells. The coleoptilar stage was characterized by the growth of coleoptile and the development of root. However, aberrant somatic embryos which had leafy scutellum and multiple shoot meristem, were also observed

Improvement of Direct Somatic Embryogenesis in Rice by Selecting the Optimal Developmental Stage of Explant and Applying Desiccation Treatment

By selecting the optimal developmental stage of zygotic embryos used as expiants and applying desiccation treatment, we improved direct somatic embryogenesis in rice scutellum from two cultivars, Nipponbare and Sasanishiki. Zygotic embryos isolated 14-17, 21, 28-30 and 35-40 d after anthesis (DAA) from Nipponbare and 14-17, 18-21, 28-30 and 40-42 DAA from Sasanishiki were cultured on the embryo induction medium (EIM). Then they were transferred to embryo maturation medium (EMM) and germinated on the embryo germination medium (EGM). Only immature zygotic embryo isolated 14-17 DAA from Nipponbare and Sasanishiki could develop somatic embryos that germinated. Explants from embryos at other developmental stages could develop somatic embryos only until the elongating or scutellar stage. They enlarged and formed callus without further development. The EIM and EMM consisted of N6 macronutrients, B5 micronutrients, and B5 vitamins, supplemented with 0.1 g L-1 casein hydrolysate, 1.5 g L-1 proline and 1 g L-1 MES buffer. EGM consisted of MS macro- and micronutrients and MS vitamins without organic supplement. In addition, 2 mg L-1 2,4-D was added to EIM, 1 mg L-1 2,4-D to EMM and 0.01 mg L-1 zeatin to EGM. Developmental processes of somatic embryos derived from the expiants were observed by scanning electron microscopy. Desiccation treatment of maturing somatic embryo was proved to produce fully mature somatic embryos capable of germinating vigorously

Review: Flavonoid pada Tanaman Kumis Kucing (Orthosiphon stamineus Benth.): Review: Flavonoid Compounds in Orthosiphon stamineus

Orthosiphon stamineus has long been used in traditional medicine in East India, Indo China, Southeast Asia, and tropical Australia, where this plant is usually found. Based on the color of the flowers and petals, Orthosiphon stamineus is classified into two varieties: white flowers (white varieties) and purple flowers (purple varieties). Orthosiphon stamineus has traditionally been used to treat hypertension, diabetes, bladder and kidney disorders, gallstones, gout, and rheumatism. The leaves of Orthosiphon stamineus were introduced to Europe and Japan as tea for health. The main compounds possessed by Orthosiphon stamineus are rosmarinic acid, eupatorium, and sinensetin. In addition, several studies have isolated the plant of Orthosiphon stamineus. This journal review aims to review studies related to the content of secondary metabolites, traditional uses, pharmacological activities, and levels of flavonoids contained in plants