Biosafety and Toxicological Evaluation of Tissue-Cultured Echinacea purpurea Adventitious Roots

Murthy, Hosakatte Niranjana,Park, So-Young,Lee, Eun Jeong,Paek, Kee Yoeup Korean Society of Horticultural Science 2015 원예과학기술지 Vol.33 No.1

Echinacea purpurea (L.) Moench (purple cone flower) is an important medicinal plant; it can enhance immunity, relieve pain, and reduce inflammation, and also has hormonal, antiviral, and antioxidant effects. Adventitious root biomass of Echinacea purpurea was produced in commercial-scale bioreactors for use as a dietary supplement in the food industry and in traditional medicine. Biosafety and toxicological evaluations of tissue-cultured Echinacea purpurea adventitious roots (TCEPARs) were performed. Reverse mutation and chromosomal aberration tests showed no significant mutagenicity. Furthermore, repeated four-week oral dose tests performed in Sprague-Dawley rats did not show any notable changes in the general behavior of the rats, in the gross appearance of their internal organs, or in their mortality rate. There were no differences between the control group and the treatment group in parameters such as absolute body weight, hematology, blood chemistry, and absolute and relative organ weights. These findings indicate that TCEPARs are safe and nontoxic when consumed at an average dietary level and can be used as raw material for traditional medicine and the food industry.

Ginsenosides: prospective for sustainable biotechnological production

Murthy, Hosakatte Niranjana,Georgiev, Milen I.,Kim, Yun-Soo,Jeong, Cheol-Seung,Kim, Sun-Ja,Park, So-Young,Paek, Kee-Yoeup Springer-Verlag 2014 Applied microbiology and biotechnology Vol.98 No.14

<P>Panax ginseng C.A. Meyer (ginseng) is a well-known medicinal plant that has been traditionally used in the oriental countries for centuries. Wild ginseng is a scarce and rare commodity. Field cultivation of the ginseng plant is a time-consuming and labor-intensive process. Ginsenosides, a group of glycosylated triterpenes, also known as saponins, are the principal bioactive constituents of ginseng. The use of cell and organ culture processes has been sought as a potential alternative for the efficient mass production of ginseng raw material. Various bioprocessing strategies have been developed to date. Cells and adventitious roots have been cultured in large-scale bioreactors and various strategies have been developed accordingly for the enhancement of biomass and ginsenoside accumulation. This review highlights the recent progress in the cultivation of ginseng cell and organ cultures for the production of ginsenosides from bioreactor cultures. In addition, the metabolism and biochemistry of ginsenoside biosynthesis, genomic and proteomic studies in ginseng, metabolic engineering, biosafety, toxicological evaluation, and efficacy assessment of ginseng raw material are also summarized and thoroughly discussed.</P>

Biosafety and Toxicological Evaluation of Tissue-Cultured Echinacea purpurea Adventitious Roots

Hosakatte Niranjana Murthy,So-Young Park,Eun Jeong Lee,Kee Yoeup Paek 한국원예학회 2015 원예과학기술지 Vol.33 No.1

Echinacea purpurea (L.) Moench (purple cone flower) is an important medicinal plant; it can enhance immunity, relieve pain, and reduce inflammation, and also has hormonal, antiviral, and antioxidant effects. Adventitious root biomass of Echinacea purpurea was produced in commercial-scale bioreactors for use as a dietary supplement in the food industry and in traditional medicine. Biosafety and toxicological evaluations of tissue-cultured Echinacea purpurea adventitious roots (TCEPARs) were performed. Reverse mutation and chromosomal aberration tests showed no significant mutagenicity. Furthermore, repeated four-week oral dose tests performed in Sprague-Dawley rats did not show any notable changes in the general behavior of the rats, in the gross appearance of their internal organs, or in their mortality rate. There were no differences between the control group and the treatment group in parameters such as absolute body weight, hematology, blood chemistry, and absolute and relative organ weights. These findings indicate that TCEPARs are safe and nontoxic when consumed at an average dietary level and can be used as raw material for traditional medicine and the food industry.

Production of secondary metabolites from cell and organ cultures: strategies and approaches for biomass improvement and metabolite accumulation

Murthy, Hosakatte Niranjana,Lee, Eun-Jung,Paek, Kee-Yoeup Springer-Verlag 2014 Plant cell, tissue and organ culture Vol.118 No.1

Establishment of bioreactor cultures for the production of chlorogenic acid and ferulic acid from adventitious roots by optimization of culture conditions in Angelica acutiloba (Siebold & Zucc.) Kitag

황호동,한종은,Murthy Hosakatte Niranjana,권혁준,이건명,신재혁,박소영 한국식물생명공학회 2022 Plant biotechnology reports Vol.16 No.2

Adventitious roots were induced from root explants of Angelica acutilobaand suspension cultures were established in shake flasks. The effect of different concentrations of indole acetic acid (IBA, 0–38.4 µM), the strength of the Murashige and Skoog (MS) medium (0.5–2 ×), the concentration of sucrose (10–70 g·L−1), inoculum density (2.5–10 g·L−1) were determined for accumulation biomass. The optimized conditions for adventitious root biomass accumulation were found to be full-strength MS medium supplemented with 29.5 µM IBA, 30 g·L−1 of sucrose, and 5 g·L−1 of inoculum density. With optimized con- dition, adventitious root suspension cultures were established in 3 L capacity airlift bioreactors using 2 L of MS medium. Chlorogenic acid and ferulic acid were increased 4.9 (117.7 mg g·g−1 DW) and 1.9 times (47.2 mg g·g−1 DW), respectively, compared to control (0 week) after 4 weeks of culture in the optimized conditions. The results suggested that the harvested adventitious root biomass could able accumulate higher concentrations of chlorogenic acid and ferulic acid which could be used industrial purpose as a biomaterials.

Effect of nutritional factors on the accretion of secondary metabolites in Malaysian ginseng adventitious root cultures

Cui Xi-Hua,Hosakatte Niranjana Murthy,Zhang Ji-De,Song Hang-Lin,Jiang Yin-Ji,Qi Wen-Wen,Li Yong Yi,백기엽,박소영 한국식물생명공학회 2020 Plant biotechnology reports Vol.14 No.3

In this study, we aimed to verify the effect of nutritional factors on the accretion of secondary metabolites in the adventitious root (AR) cultures of Malaysian ginseng (Eurycoma longifolia Jack) grown in small-scale bioreactors. AR were induced from leaf explants and cultured in different types of media including Murashige and Skoog (MS) medium, Driver Kuniyuki Walnut (DKW) medium, Gamborg’s B5 medium, Woody Plant Medium (WPM), and ¾ MS medium. Among these media, the MS and Gamborg’s B5 media induced lateral root development from initial inoculum, which accounted for the increase in AR biomass accretion. By contrast, the DKW and WPM media did not induce lateral root formation from the cultured explants. The ¾ MS medium was optimal for the growth of AR and accretion of secondary metabolites, after 7 weeks of culture, the biomass of AR increased by 8.6-fold in ¾ MS medium, and the total phenolic and flavonoid contents reached 5.23 and 2 mg g−1 of tissue dry weight, respectively. Analysis of mineral elements in the spent medium revealed that ¾ MS medium was most suitable for nutrient supply to developing AR. LC–MS analysis showed the accretion of eurycomanone, a therapeutically useful metabolite, in the AR of Malaysian ginseng.

Role of Phenolic Compounds in Root Induction of Virus-free Apple Rootstocks and Scions

한종은,Bo-Min Kwon,Hosakatte Niranjana Murthy,Cheol-Seung Jeong,So-Young Park1 한국원예학회 2022 원예과학기술지 Vol.40 No.5

The objective of the current study was to identify and quantify endogenous phenolic compounds in easy-to-root M.9 and M.26 rootstocks and difficult-to-root cultivars ‘Hongro’ (HR) and ‘Fuji’ (FJ) (scions) of apple (Malus domestica Borkh.) during in vitro rooting of shoots for 40 days in culture. Phloretic acid, coumaric acid, salicylic acid, chlorogenic acid, catechin hydrate, hesperetin, rutin, quercetin, and biochanin were the major endogenous phenolic compounds detected in the apple shoots by high-performance liquid chromatography (HPLC). Among the various phenolic compounds, chlorogenic acid was most abundant in shoots, favoring the induction and growth of adventitious roots from in vitro shoots. In contrast, salicylic acid was another phenolic acid that was reduced during the culture of shoots in the easy-to-root rootstocks (M.9 and M.26). However, salicylic acid levels remained constant or increased throughout the culture in the difficult-to-root cultivars (HR and FJ). This study indicates the role of endogenous phenolic compounds on the rooting of shoots and lays the foundation for further studies investigating the mechanism of rooting in these apple rootstocks and cultivars.

Improved Production of Caffeic Acid Derivatives in Suspension Cultures of Echinacea purpurea by Medium Replenishment Strategy

Chun-Hua Wu,Hosakatte Niranjana Murthy,Eun-Joo Hahn,백기엽 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.8

The aim of this study was to produce caffeic acid derivatives from adventitious root cultures of Echinacea purpurea, which are of high pharmaceutical value. The effects of both media optimization and replenishment strategies were adopted to achieve improved production of E. purpurea adventitious roots and caffeic acid derivatives. Of the different media strengths (0.25 MS, 0.5 MS, 0.75 MS and 1 MS) tested for the culturing of adventitious roots in 5 L bioreactors, 0.5 MS medium was found to be most suitable for the growth of adventitious roots. The optima accumulation of biomass (73.6 g L-1 FW and 10.03 g L-1 DW), phenolics (61.14 mg g-1 DW) and flavonoids (38.30 mg g-1 DW) were achieved in this medium. Furthermore, fed batch cultivations (media replenishment with 0.25 MS, 0.5 MS, 0.75 MS and 1 MS at the end of 2nd and 3rd weeks) to further enhance the production of adventitious root biomass and metabolites were also attempted. High adventitious root biomasses (83.1 g L-1 FW and 15.30 g L-1 DW) were achieved with feeding of the 0.5 MS medium at the end of 2nd week. This led to slight decreases in the total production of phenolics and flavonoids; however, this feeding was responsible for increases in the accumulation of caftaric acid (5.76 mg g-1 DW) and cichoric acid (26.12 mg g-1 DW).

CO₂-Enrichment and Photosynthetic Photon Flux Affect the Growth of In Vitro-Cultured Apple Plantlets

Li, Ru-Yu,Murthy, Hosakatte Niranjana,Kim, Seon-Kyu,Paek, Kee-Yeoup 충북대학교 첨단원예기술개발연구센터 2000 연구보고서 Vol.5 No.-

Micro-cutting (shoots with two small leaves) of cultivar M9 apple were cultured in-vitro for 40 d under CO2-enriched and non-enriched (i.e., ambient air) conditions, and at a PPF of 40 or 100 μmol m-2 s-1. Afterward, shoot length, number of leaves, leaf area, chlorophyll content, shoot and root fresh weights, and % survival were recorded. Those plantlets grown under CO2 -and PPF-enriched treatments were healthy and vigorous, and showed higher values for their growth parameters. In contrast, those grown without supplemental CO2 or PPF often showed hyperhydricity. We also demonstrated that CO2 enrichment and a relatively high PPF during in-vitro culture promoted normal photosynthesis and growth after ex-vitro transplantation.

Production of Biomass and Bioactive Compounds from Cell Suspension Cultures of Eurycoma longifolia in Balloon Type Bubble Bioreactors

Kyu-Man Shim,Hosakatte Niranjana Murthy,So-Young Park,Ibrahim Rusli,Kee-Yoeup Paek 한국원예학회 2015 원예과학기술지 Vol.33 No.2

Eurycoma longifolia is an important rare medicinal plant that contains valuable bioactive compounds. In the present study, cell suspension culture of E. longifolia was established for the production of biomass and phenolic compounds. Various medium parameters, such as concentration of auxin, salt strength of the medium, and sucrose and nitrogen concentrations, were optimized for the production of biomass at the flask-scale level. Full strength Murashige and Skoog (MS) medium supplemented with 3.0 ㎎·L<SUP>-1</SUP> naphthaleneacetic acid (NAA), 3% (w/v) sucrose, 0:60 NH₄⁺:NO₃⁻ was found suitable for biomass accumulation. Based on the optimized flask-scale parameters, cell suspension cultures were established in balloon-type bubble bioreactors, and bioprocess parameters such as inoculum density and aeration rate were optimized. Inoculum density of 50 g·L<SUP>-1</SUP> and increasing aeration rate from 0.05 to 0.3 vvm, with increases every 7 days, were suitable for the accumulation of both biomass and phenolic compounds. With the optimized conditions, 14.70 g·L<SUP>-1</SUP> dry biomass, 10.33 ㎎·g<SUP>-1</SUP> DW of phenolics and 3.89 ㎎·g<SUP>-1</SUP> DW of flavonoids could be achieved. Phenolics isolated from the cell biomass showed optimal free radical scavenging activity.