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Direct Somatic Embryogenesis in Rice (Oryza sativa L.) : Structural and Developmental Patterns Totik Sri Mariani; Hiroshi Miyake; Yoji Takeoka
Jurnal Matematika & Sains Vol 7, No 2 (2002)
Publisher : Institut Teknologi Bandung

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In this study, we used light microscopy to observe the structural and developmental pattern during direct somatic embryogenesis in rice (Oryza sativa L). Initiation of somatic embryogenesis revealed the origin of somatic embryo. It was unicellular origin from the epidermis of scutellum zygotic embryo. The polarity of somatic embryo established when the elongated embryo flattened at the apical end. On the onset of scutellum development, the scutellar notch was formed. In germinated somatic embryo, the shoot apex was connected with the root by the vascular bundle strands. This is an important characteristic of somatic embryo. The result led us to the conclusion that rice somatic embryos underwent the same structural and developmental pattern as that of rice zygotic embryo.

Effect of 2,4-D on Indirect Somatic Embryogenesis and Surface Structural Changes in Garlic (Allium sativum L.) cv. Lumbu Hijau Totik Sri Mariani; Hiroshi Miyake; Rizkita Rachmi Esyanti; Ida Nurwendah
Jurnal Matematika & Sains Vol 8, No 4 (2003)
Publisher : Institut Teknologi Bandung

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A study concerning the effect of 2,4-D on indirect somatic embryogenesis and surface structural changes in garlic (Allium sativum L.) cv. Lumbu Hijau was conducted. The purposes of this study were to evaluate the effect of 2,4-D on the induction of somatic embryogenesis and to observe the developmental stage of somatic embryo as well as the surface structural changes of somatic embryo in garlic. Root tips explants were cultured on embryogenic callus induction medium (ECIM). Somatic embryos were then transferred to the embryo maturation medium (EMM), desiccated and subsequently transferred to the repetitive somatic embryogenesis medium (RSEM). Observation by dissecting microscope showed that embryogenic callus and somatic embryo was formed at ECIM containing 0.1 Î¼M 2,4-D. At EMM containing 0.01 Î¼M 2,4-D somatic embryo developed into mature somatic embryo. Somatic embryo underwent repetitive somatic embryogenesis that consisted of globular and mature somatic embryo at RSEM, without plant growth regulator. Observation by scanning electron microscopy showed that the development al stages of garlic somatic embryo consisted of proembryo, transition phase from proembryo to globular, globular, mature embryo and single cotyledon stages. Effect of 2,4-D on the cell surface structure could be seen on each somatic embryo developmental stage. In ECIM containing 0,1 Î¼M 2,4-D, proembryo, transition embryo and globular embryo were formed. The surface of proembryo was smooth whereas fibrillar material as cellulose microfibril, was observed during the transition from proembryo to globular stage. The surface of globular stage was entirely covered by a new cell wall. Subsequently, in 0.01 Î¼M 2,4-D, mature embryo and single cotyledon were formed, which their cell wall surface was stable. From this study, it could be concluded that the optimum concentration of 2,4-D for induction of embryogenic callus and initiation of somatic embryo was 0.1 Î¼M. Decreasing of 2,4-D concentration to 0.01 Î¼M resulted in the development of globular embryo to mature and single cotyledon stage. Repetitive somatic embryogenesis occurred in RSEM medium without plant growth regulator. Based on this result it can be concluded that each somatic embryo developmental stage had a typical characteristic of surface structure.

Development of Somatic Embryo in Lithospermum erythrorhizon Siebb. et Zucc and The Study on The Effect of Methyl Jasmonate on its Maturation Mariani, Totik Sri; Ramayanti, Octavia; Yazaki, Kazufumi; Miyake, Hiroshi
ANNALES BOGORIENSES Vol 9, No 2 (2004): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

A research on the somatic embryogenesis in Lithospermum erythrorhizol1 has been conducted. Embryogenic callus was inoculated in EIM9 liquid medium (modification ofLS medium), i.e. L2PVP (initiation medium) and the development of somatic embryo was observed. Ten J.1M and 100).lM methyl jasmonate was added into L3PVP medium (differentiation medium) for somatic emb yo maLUration. The purposes of this research were to observe tlle development of somatic embryo and to observe the effect of methyl jasmonate on maturation of the somatic embryo in L erylhrorhizol/ . The results showed that somatic embryogenesis in L erythrorhizon derived from single cells differentiated further forming proembryo, globular. heart, torpedo and cotyledon stage. Treatment with 10 f.1M and 100).lM methyl jasmonate induced maturation of somatic embryos (cotyledon stage) after transferring them to embryo development medium (L4PVP).

Development of Somatic Embryo in Lithospermum erythrorhizon Siebb. et Zucc and The Study on The Effect of Methyl Jasmonate on its Maturation Totik Sri Mariani; Octavia Ramayanti; Kazufumi Yazaki; Hiroshi Miyake
ANNALES BOGORIENSES Vol 9, No 2 (2004): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

A research on the somatic embryogenesis in Lithospermum erythrorhizol1 has been conducted. Embryogenic callus was inoculated in EIM9 liquid medium (modification ofLS medium), i.e. L2PVP (initiation medium) and the development of somatic embryo was observed. Ten J.1M and 100).lM methyl jasmonate was added into L3PVP medium (differentiation medium) for somatic emb yo maLUration. The purposes of this research were to observe tlle development of somatic embryo and to observe the effect of methyl jasmonate on maturation of the somatic embryo in L erylhrorhizol/ . The results showed that somatic embryogenesis in L erythrorhizon derived from single cells differentiated further forming proembryo, globular. heart, torpedo and cotyledon stage. Treatment with 10 f.1M and 100).lM methyl jasmonate induced maturation of somatic embryos (cotyledon stage) after transferring them to embryo development medium (L4PVP).

Review: Flavonoid pada Tanaman Kumis Kucing (Orthosiphon stamineus Benth.): Review: Flavonoid Compounds in Orthosiphon stamineus Fahrauk Faramayuda; Silvy Julian; Ari Sr Windyaswari; Totik Sri Mariani; Elfahmi Elfahmi; Sukrasno Sukrasno
Proceeding of Mulawarman Pharmaceuticals Conferences Vol. 13 (2021): Proc. Mul. Pharm. Conf.
Publisher : Fakultas Farmasi, Universitas Mulawarman, Samarinda, Indonesia

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.

Comparison of Polyphenol Levels of Callus and Wild Type of Cat's Whiskers Plant ( Orthosiphon aristatus (Blume) Miq.) Purple Varieties Fahrauk Faramayuda; Soraya Riyanti; Ari Sri Widyaswari; Zaini Alfahmi; Sultan Salahudin Jamal; Totik Sri Mariani; Elfahmi Elfahmi; Sukrasno Sukrasno
Journal of Applied Agricultural Science and Technology Vol. 6 No. 2 (2022): Journal of Applied Agricultural Science and Technology
Publisher : Green Engineering Society

The cat's whiskers plant ( Orthosiphon aristatus (Blume) Miq) that grows in Indonesia has white flowers, purple flowers, and white flowers with purple patterns. The quality of the secondary metabolite content of each cat's whiskers plant is different because it depends on environmental influences such as rainfall or soil mineral content. Plant tissue culture methods such as callus culture or cell culture are used to overcome these obstacles because these methods do not depend on the environment and can produce secondary metabolites such as those produced by the original plant. Cat's whiskers contain the main secondary metabolite compounds: rosmarinic acid, eupatorin, and sinensetin. Rosmarinic acid belongs to the phenolic group, while sinensetin and eupatorin are flavonoids. This study aimed to determine the total levels of polyphenols and flavonoids in purple cat whiskers and determine the ratio of secondary metabolites of cat whiskers callus with wild-type plants—determination of polyphenol content using Folin Ciocalteau reagent. Determining flavonoid levels is based on colorimetry using AlCl 3 reagent, specific for flavone and flavonol compounds. The levels of polyphenols contained in the ethanolic extract of callus and cat whiskers of the purple variety were 16.056 ± 0.204 mgQE/g and 30.780 ± 0.425 mgQE/g, while the polyphenol content of the callus ethyl acetate extract and the purple variety of cat whiskers leaves was 15,489 ± 0.425 mgQE/g and 13,262 ± 0.044 mgQE/g. The levels of flavonoids contained in the ethanol extract of the cat whiskers leaf were 1.39±0.03 mgQE/g and callus 0.16±0.02 mgQE/g in the ethyl acetate extract of the leaves of the cat whiskers 2.16±0.012 mgQE/g and callus 2.24±0.010 mgQE/g. The polyphenols and flavonoids in the callus ethyl acetate extract were more significant than the ethyl acetate extract of the purple variety ( wild type ) cat whiskers leaf.

Effect of Media Variation on the Induction and Phytochemical Profile of Callus in Two Varieties of Cat's Whiskers (Orthosiphon aristatus Blume Miq) Fahrauk Faramayuda; Akhirul Kahfi Syam; Totik Sri Mariani; Elfahmi; Sukrasno
HAYATI Journal of Biosciences Vol. 30 No. 1 (2023): January 2023
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.30.1.159-170

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.

ANALYSIS OF NON-SPECIFIC STANDARDIZED PARAMETERS ON KUMIS KUCING LEAVES (Orthosiphon aristatus (Blume) Miq.) PURPLE VARIETY Fahrauk Faramayuda; Soraya Riyanti; Totik Sri Mariani; Rizka Khoirunnisa Guntina; Winda Nur Halimah
Medical Sains : Jurnal Ilmiah Kefarmasian Vol 9 No 1 (2024)
Publisher : Sekolah Tinggi Farmasi Muhammadiyah Cirebon

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37874/ms.v9i1.977

One of the plants in Indonesia that can be used as a medicine is the purple variety of cat's whiskers (Orthosiphon aristatus (Blume) Miq.). This study aims to examine the quality and safety of the purple variety of Cat’s Whiskers as a raw material for traditional medicine so that it meets the requirements of being free from heavy metal, microbial, and aflatoxin contamination. The results of setting the standardization parameters on the dry material of cat’s whiskers leaves obtained drying losses, water content, total ash content, and acid insoluble ash content, respectively 8.45 ± 0.68%; 6.67 ± 1.15 %; 8.70 ± 0.09 %; 1.31 ± 0.16 %. The results of testing for the heavy metals lead (Pb) and mercury (Hg) on ??dry material and extracts were £ 0.0001 mg/kg. The results of testing the levels of microbial contamination in dry material and extracts obtained total bacteria 1.45x101 and 2.35x101 Cfu/gr, respectively. Test results for aflatoxin contamination on the dry material of Cat’s Whiskers leaves B1, B2, G1, G2 and total aflatoxin £ 0.50 ?g/kg and total aflatoxin £ 2 ?g/kg, respectively. The dried material and leaf extract of the purple variety of cat’s whiskers that were tested met the requirements stated in the regulation of the Food and Drug Supervisory Agency number 32 of 2019 about traditional medicines' safety and quality requirements.

Development of Somatic Embryo in Lithospermum erythrorhizon Siebb. et Zucc and The Study on The Effect of Methyl Jasmonate on its Maturation Mariani, Totik Sri; Ramayanti, Octavia; Yazaki, Kazufumi; Miyake, Hiroshi
Annales Bogorienses Vol. 9 No. 2 (2004): Annales Bogorienses
Publisher : BRIN

Show Abstract | Download Original | Original Source | Check in Google Scholar

A research on the somatic embryogenesis in Lithospermum erythrorhizon has been conducted. Embryogenic callus was inoculated in EIM9 liquid medium (modification of LS medium), i.e. L2PVP (initiation medium) and the development of somatic embryo was observed. Ten uM and 100 uM methyl jasmonate was added into L3PVP medium (differentiation medium) for somatic embryo maturation. The purposes of this research were to observe the development of somatic embryo and to observe the effect of methyl jasmonate on maturation of the somatic embryo in L. erythrorhizon. The results showed that somatic embryogenesis in L. erythrorhizon derived from single cells differentiated further forming proembryo, globular heart, torpedo and cotyledon stage. Treatment with 10 uM and 100 uM methyl jasmonate induced maturation of somatic embryos (cotyledon stage) after transferring them to embryo development medium (L4PVP).

Short Communication: Callus induction in purple and white-purple varieties of Orthosiphon aristatus (Blume) Miq. Fahrauk Faramayuda; TOTIK SRI MARIANI; ELFAHMI ELFAHMI; SUKRASNO SUKRASNO
Biodiversitas Journal of Biological Diversity Vol. 21 No. 10 (2020)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d211063

Abstract. Faramayuda F, Mariani TS, Elfahmi, Sukrasno. 2020. Short Communication: Callus induction in purple and white-purple varieties of Orthosiphon aristatus (Blume) Miq. Biodiversitas 21: 4967-4972. Orthosiphon aristatus (Blume) Miq. are known to have many benefits, including stimulating urine expenditure (diuretics) and dissolving kidney stones. O. aristatus widely planted in Indonesia are purple and white-purple. The main secondary metabolite components of O. aristatus are sinensetin, rosmarinic acid, and eupatorin. One of the initial steps to increase secondary metabolites in O. aristatus is by induction of callus using plant tissue, which later can be developed into a culture suspension for secondary metabolites. The materials used are the leaf of two varieties of O. aristatus that have been sterilized and grown on Murashige and Skoog media with growth regulatory 2,4-dichlorophenoxyacetic acid at a concentration of 0.4;1.0; 2.0 mg/L. The identification of secondary metabolites of callus was carried out by thin-layer chromatography. The best growth regulating agent for callus induction on the leaves of purple and white-purple varieties of O. aristatus is 2,4-D 0.4 mg/L on Murashige and Skoog media. These media can grow callus at a faster time, friable, and slightly white-yellow color. The identification of secondary metabolites in callus acetone extract showed the presence of sinensetin and rosmarinic acid.  

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