Production Of Active Compounds Research Articles

Effect of biotic and abiotic elicitors on Salvadora persica callus in vitro

أجري البحث لغرض دراسة تأثير تراكيز مختلفة من الجسيمات النانوية (الكيتوزان وثاني أكسيد التيتانيوم) و Fusarium oxysporum كمحفزات لزيادة إنتاج المركبات الفعالة من كالس نبات المسواك. اظهرت نتائج المحتوى الكلي مركبات الفلافونويد والقلويدات التي ازاد انتاجها عند اضافة الكيتوسان بتركيز 5 ملغم / لتر، وثاني أكسيد التيتانيوم بتركيز 1 ملغم / لتر، و Fusarium oxysporum بتركيز 5 ملغم / لتر حيث اعطت 93.10 ملغم / 100 ملغم، 128.7 ملغم / 100 ملغم، و 107.61 ملغم / 100 ملغم على التوالي للفلافينودات بينما ساعدت المحفزات على انتاج القلويدات بمعدل 2.39 %، 3.91%، و2.20% على التوالي. وأوضحت النتائج التفوق في وجود مركبات الفلافونويد في العينات مقارنة مع القلويدات. بين تحليل HPLC( تحليل كروماتوغرافي سائل عالي الأداء) اختلافات كبيرة في زيادة إنتاج الفلافونويد (Rutin, Kaempferol, Quercetin, Catechin, Luteolin, and Apigenin) أدت إضافة الكيتوسان عند التركيز 5 ملغم / لتر إلى زيادة إنتاج Rutin ، بمقدار 39.89 ملغم/لتر. أما الكالس المستحث المعالج بثاني أكسيد التيتانيوم بتركيز 1 ملغم/لتر فقد أدى إلى زيادة Rutin بمعدل 35.89 ملغم/لتر. بينما ارتفع إنتاج Rutin إلى 30.12 ملغم/لتر عند معاملته بـ Fusarium oxysporum بتركيز 5 ملغم/لتر.

Producción de beauveria bassiana para la formulación de bioplaguicidas

The harmful effects of chemicals in conventional agriculture and the growing demand for food free of toxic residues has developed environmentally sustainable strategies. An effective alternative for integrated pest management in agricultural crops are biopesticides formulated with microorganic structures or from the production of active compounds. This paper describes the production processes of Beauveria bassiana for formulating biopesticides for agricultural use. The information was collected through a systematic search in Research Gate, Google Scholar, Science Direct and PubMed, using keywords such as production, Beauveria bassiana, solid fermentation, liquid fermentation and metabolites. The results affirm that B. bassiana is one of the microorganisms with great potential to produce biopesticides, due to the entomopathogenic mechanism of action and secondary metabolites, which can be used for the biological control of phytophagous insects. Likewise, for the formulation of B. bassiana it should be considered a profitable culture medium for large-scale production, also the control of environmental variables such as temperature at 25 oC, relative humidity 65-70 %, pH of 5.4, propagation time between 4 to 8 days, and for the liquid fermentation process, a constant agitation between 200 to 400 rpm must be maintained. Biological products represent an alternative to minimize the use of synthetic pesticides, reduce environmental pollution and ensure food safety and security.

Phosphatidylcholine and ceramide derivatives from white rot fungus Microporus xanthropus PP17-20

The undescribed phosphatidylcholine (1), along with twelve known compounds, was isolated from the cultures of white rot fungus Microporus xanthropus PP17-20. In this work the fungus was cultivated in Yeast-Malt extract medium to explore active compound production. The chemical structures were elucidated on the basis of spectroscopic and HRESIMS data. Several isolated compounds were evaluated for anti-proliferative activity against A549 and MCF-7 cancer cell lines.

Identification of Growth-Promoting Bacterial Resources by Investigating the Microbial Community Composition of Polyporus umbellatus Sclerotia.

The sclerotium of the edible mushroom Polyporus umbellatus (Zhuling) exhibits various medicinal properties. However, given its long growth cycle and overexploitation, wild resources are facing depletion. Macrofungal growth depends on diverse microbial communities; however, the impact of soil bacteria on P. umbellatus development is unknown. Here, we combined high-throughput sequencing and pure culturing to characterize the diversity and potential function of bacteria and fungi inhabiting the P. umbellatus sclerotium and tested the bioactivities of their isolates. Fungal operational taxonomic units (OTUs) were clustered and classified, revealing 1275 genera. Bacterial OTUs yielded 891 genera. Additionally, 81 bacterial and 15 fungal strains were isolated from P. umbellatus sclerotia. Antagonism assays revealed three bacterial strains (FN2, FL19, and CL15) promoting mycelial growth by producing indole-3-acetic acid, solubilizing phosphate, and producing siderophores, suggesting their role in regulating growth, development, and production of active compounds in P. umbellatus. FN2-CL15 combined with bacterial liquid promoted growth and increased the polysaccharide content of P. umbellatus mycelia. This study reports new bioactive microbial resources for fertilizers or pesticides to enhance the growth and polysaccharide accumulation of P. umbellatus mycelia and offers guidance for exploring the correlation between medicinal macrofungi and associated microbial communities.

INCREASING SOME FLAVONOIDS COMPOUNDS FOR ECHINACEA PURPUREA L. USING COPPER OXIDE NANOPARTICLES IN VITRO.

This study was aimed to increase some flavonoids active compounds production in Echinacea purpurea seedlings through plant seeds treatment with different concentrations of copper oxide nanoparticles that daignosed and characterized using AFM technique. This research was implemented at plant tissue culture labrotary of College of biotechnology - Al Nahrain University, during the period of 2021 and 2023. The experiment designed factorial within CRD using three factors and ten replicates (3X3). Sodium hypochlorite concentration (S1, S2, S3, S4) (0.0, 1, 2, 3%) represented the first factor, treatment duartion time (T1, T2, T3) (5, 10, 15min) represented the second factor and copper oxide nanoparticles concentrations (C1, C2, C3, C4) (0.0, 25, 50, 75mg/ml) represented the third factor. Results showed that the full reduction in the contamination rate of the selected E. purpurea explant recorded in 3% sodium hypochlorite at 10 and 15min. The results also showed that there were a significant increase in the shoots numbers in 50mg/ml CuNPs that recording the highest shoots numbers, the shoot length increased significantly within the 25mg/ml recording 13.5cm then decreased in 50 and 75 mg/ml and the seedlings dry weight increased significantly up to 50mg/ml CuNPs that recording the highest seddlings dry weight, then the seedlings dry weight also decreased significantly in 75mg/ml CuNPs. Also, all the analyzed flavonoids compounds using HPLC device as Echinolone, Humulene, Coumarin, Myricetin, Heperidin and Naringin concentrations were significantly increased in 50 and 75 mg/L-1 Cu ONPs, except the Humulene and Coumarin compound that significantly decreased in 75 mg/L-1 Cu ONPs in comparsion to the control.

Withania somnifera (L.) Dunal: Enhanced production of withanolides and phenolic acids from hairy root culture after application of elicitors

Withania somnifera (L.) Dunal is an important indigenous medicinal plant with extensive pharmaceutical potential. The root is the main source of major bioactive compounds of this plant species including withanolides, withanine, phenolic acids, etc. Hairy root culture (HRC) is a crucial method for low-cost production of active compounds on a large scale. Four different Agrobacterium rhizogenes strains have been used for the hairy root induction. Maximum transformation efficiency (87.34 ± 2.13%) was achieved with A4 bacterial strain-mediated transformed culture. The genetic transformation was confirmed by using specific primers of seven different genes. Seven HR (Hairy root) lines were selected after screening 29 HR lines based on their fast growth rate and high accumulation of withanolides and phenolic acids content. Two biotic and three abiotic elicitors were applied to the elite root line to trigger more accumulation of withanolides and phenolic acids. While all the elicitors effectively increased withanolides and phenolic acids production, among the five different elicitors, salicylic acid (4.14 mg l–1) induced 11.49 -fold increase in withanolides (89.07 ± 2.75 mg g–1 DW) and 5.34- fold increase in phenolic acids (83.69 ± 3.11 mg g− 1 DW) after 5 days of elicitation compared to the non-elicited culture (7.75 ± 0.63 mg g–1 DW of withanolides and 15.66 ± 0.92 mg g–1 DW of phenolic acids). These results suggest that elicitors can tremendously increase the biosynthesis of active compounds in this system; thus, the HRC of W. somnifera is cost-effective and can be efficiently used for the industrial production of withanolides and phenolic acids.

Multi-drops and Cross-sections, Efficient Methods for Establishing Cell Suspension Culture of Cuminum cyminum L. and Plant Regeneration

The current study aims of the current study is to establish cell suspension cultures of the medicinal plant, cumin Cuminum cyminum L. by the application of multiple drops and cross-section techniques. These methods were used in cultivating cell suspensions and in vitro plant regeneration. Leaf, stem, hypocotyl and root explants were cultured in Murashige and Skoog (MS) medium containing different concentrations (0.05, 0.5 and 1.0 mg.l-1) of naphthalene acetic acid (NAA) and (0.05, 0.1, 1.0 and 2.0 mg.l-1) benzyl adenine BA for callus production. The results indicated the high response of cumin, as the percentage of callus initiation was 100%.The plant regeneration percentage reached 91.6%. Moreover, a friable callus of hypocotyls was appropriate for initiation of cell suspension culture in MS medium with 0.5mg.l-1 NAA and 1.0 mg.l-1benzyl adenine (BA). The best density for primordial callus formation was 51.0 ×105 cells ml-1 in both multiple-drops and cross-sections embedding methods. Callus that had been produced from multi drops and sectors had the ability for shoot regeneration. This study clarified the efficiency of these techniques in establishing cell suspension culture and shoot regeneration, which could be promising sources for high production of active compounds in cumin plant.

DNA methylation regulates biosynthesis of tanshinones and phenolic acids during growth of Salvia miltiorrhiza.

DNA methylation plays a crucial role in the regulation of plant growth and the biosynthesis of secondary metabolites. Danshen (Salvia miltiorrhiza) is a valuable Chinese herbal medicine commonly used to treat cardiovascular diseases; its active ingredients are tanshinones and phenolic acids, which primarily accumulate in roots. Here, we conducted a targeted metabolic analysis of S. miltiorrhiza roots at 3 distinct growth stages: 40 d old (r40), 60 d old (r60), and 90 d old (r90). The contents of tanshinones (cryptotanshinone, tanshinone I, tanshinone IIA, and rosmariquinone) and phenolic acids (rosmarinic acid and salvianolic acid B) gradually increased during plant development. Whole-genome bisulfite sequencing and transcriptome sequencing of roots at the 3 growth stages revealed an increased level of DNA methylation in the CHH context (H represents A, T, or C) context at r90 compared with r40 and r60. Increased DNA methylation levels were associated with elevated expression of various genes linked to epigenetic regulations, including CHROMOMETHYLASE2 (SmCMT2), Decrease in DNA Methylation 1 (SmDDM1), Argonaute 4 (SmAGO4), and DOMAINS REARRANGED METHYLTRANSFERASE 1 (SmDRM1). Moreover, expression levels of many genes involved in tanshinone and salvianolic acid biosynthesis, such as copalyldiphosphate synthase 5 (SmCPS5), cytochrome P450-related enzyme (SmCYP71D464), geranylgeranyl diphosphate synthase (SmGGPPS1), geranyl diphosphate synthase (SmGPPS), hydroxyphenylpyruvate reductase (SmHPPR), and hydroxyphenylpyruvate dioxygenase (SmHPPD), were altered owing to hyper-methylation, indicating that DNA methylation plays an important role in regulating tanshinone and phenolic acid accumulation. Our data shed light on the epigenetic regulation of root growth and the biosynthesis of active ingredients in S. miltiorrhiza, providing crucial clues for further improvement of active compound production via molecular breeding in S. miltiorrhiza.

Titanium dioxide nanoparticles improve element uptake, antioxidant properties, and essential oil productivity of Melissa officinalis L. seedlings under in vitro drought stress.

In vitro drought stress has a considerable impact on the mass production of active compounds in medicinal plants. Nevertheless, photosynthesis, nutrient uptake, and protein synthesis may be negatively affected by drought, which results in poor growth. Titanium dioxide nanoparticles (TiO2 NPs) have recently been shown to play an important role in increasing nutrient uptake, resistance to various environmental stresses, and better plant growth. Regarding the importance of pharmaceutical metabolites of Melissa officinalis L., this experiment aimed to assess the role of TiO2 NPs in improving physiological responses and phytochemical properties in M. officinalis under in vitro drought stress. For this, two-week-old seedlings were cultured on Murashige and Skoog (MS) medium supplemented with 0, 50, and 100mg L-1 TiO2 NPs and 0, 3, and 6% (w/v) polyethylene glycol (PEG). Two weeks after treatments, a reduction of chlorophyll, protein content, essential elements, and enhancement of H2O2 and malondialdehyde (MDA) levels were seen as a result of drought stress. It was observed that M. officinalis partially responded to the drought by increasing non-enzymatic antioxidants, including phenolics, flavonoids, and anthocyanin and ascorbate peroxidase activity. Moreover, PEG-induced drought stress increased some important essential oil content such as limonene, alpha-pinene, myrcene, γ-3-carene, citral, and carvacrol; however, the results showed that TiO2 NPs not only increased the quantity of essential oils but also led to tolerance to the drought stress by increasing photosynthetic pigments, antioxidant systems, absorption of essential nutrients, and decreasing H2O2 and MDA levels.

Improving propagation of Echinacea purpurea and its content of some active compounds by using Tyrosine and Salicylic acid in vitro

The seed propagation is the predominant method of Echinacea propagation, which has been criticized for its time-consuming control over the separation factor and the uncertainty of pathogen-free plants produced by this method. The technology of tissue culture has provided multiple opportunities for the production of secondary metabolites continuously without being restricted to a specific season, due to the possibility of controlling the environmental conditions and the components of the nutrient medium needed by the plant. This study was conducted to investigate the effects of salicylic acid as elicitor and tyrosine as precursor on propagation and some secondary compounds production in coneflower in vitro. The result showed the superiority of 2 mg / liter SA over the other treatments, which gave the highest average number of branches formed reaching 33.6 branches / plant part, number of leaves 33.6 leaves/vegetable part, fresh 1.067g and dry 0.058g weights as well as echinacoside concentration of 291.3427 μg / g DW. However, the treatment 100 mg/l tyrosine gave a maximum of average number of branches 11.80 branches / plant part, number of leaves 11.80 leaves/vegetable part, fresh and dry weight 0.152 and 0.023 g respectively. Concerning echinacoside value, its maximum 318.5203 μg / g DW was recorded with 150 mg/l tyrosine. From this study, we concluded that caffeic acid derivatives such as echinacosides are one of the main phytochemical components of Echinacea extracts. The system of cell culture is a tool for the large-scale, year-round culture of plant cells, so it is a continuous source for the production of active compounds

Methods for Long-Term Storage of Pure Cultures of Macrofungi

Basidiomycetous macrofungi have significant biotechnological potential and are promising objects for use in va-rious industrial sectors, such as food production, pharmaceuticals, the production of active compounds and polysaccharides. The industrial use of macrofungi implies the presence of large collections of cultures using sto-rage protocols that ensure the preservation of viability, reproduction, genetic stability and the ability to produce active compounds. With the expansion of the list of industrially used species, it is advisable to develop new protocols for the storage of strains and optimize the existing ones for new, promising types of macrofungi. It seems necessary to study in detail the effect of long periods of storage on morphological and cultural characteristics, genetic stability, enzymatic activity, and the ability to form sexual structures.

Characterization of Aroma Active Compound Production during Kombucha Fermentation: Towards the Control of Sensory Profiles.

Since the sensorial profile is the cornerstone for the development of kombucha as a beverage with mass market appeal, advanced analytical tools are needed to gain a better understanding of the kinetics of aromatic compounds during the fermentation process to control the sensory profiles of the drink. The kinetics of volatile organic compounds (VOCs) was determined using stir bar sorptive extraction-gas chromatography-mass spectrometry, and odor-active compounds were considered to estimate consumer perception. A total of 87 VOCs were detected in kombucha during the fermentation stages. The synthesis of mainly phenethyl alcohol and isoamyl alcohol probably by Saccharomyces genus led to ester formation. Moreover, the terpene synthesis occurring at the beginning of fermentation (Δ-3-carene, α-phellandrene, γ-terpinene, m- and p-cymene) could be related to yeast activity as well. Principal component analysis identified classes that allowed the major variability explanation, which are carboxylic acids, alcohols, and terpenes. The aromatic analysis accounted for 17 aroma-active compounds. These changes in the evolution of VOCs led to flavor variations: from citrus-floral-sweet notes (geraniol and linalool domination), and fermentation brought intense citrus-herbal-lavender-bergamot notes (α-farnesene). Finally, sweet-floral-bready-honey notes dominated the kombucha flavor (2-phenylethanol). As this study allowed to estimate kombucha sensory profiles, an insight for the development of new drinks by controlling the fermentation process was suggested. Such a methodology should allow a better control and optimization of their sensory profile, which could in turn lead to greater consumer acceptance.

Production of Taxol by Endophytic Fungi Isolated from Roots of Himalayan Yew (Taxus wallichiana Zucc.)

Taxol® (generic name – Paclitaxel), the most promising chemotherapeuticagent was isolated from bark of different Taxus sp. As Taxus species arethreatened with extinction (endangered in Himalaya), thus it is imperativeto develop alternate and sustainable method for commercialization and scaleup production of paclitaxel. In this respect, physical and chemical parametersare effective and important key points for active compound production par-ticularly by using endophytic microbes. In the present study, five endophyticfungi isolated from the roots of Taxus wallichiana, are tested for paclitaxelproduction using biochemical and molecular methods. Subsequently, effectof physico-chemical parameters like temperature, pH, incubation time, andmedium constituents i.e., salt concentration, carbon and nitrogen sources onpaclitaxel production were also analyzed. Among isolates, two of the fungiviz. GBPI TWR F1 (Penicillium sp.) and GBPI TWR F5 (Aspergillus sp.)were found to be paclitaxel producing. The genomic DNA samples were sequenced to confirm the presence of two genes viz. 10-deacetylbaccatinIII-10-O-acetyl transferase (DBAT) and C-13 phenylpropanoid side chain-CoA acyltransferase (BAPT), implicated in paclitaxel biosynthesis. Boththe endophytes showed the amplicons of DBAT and BAPT genes. Resultsrevealed that after optimization of medium components and physical condi-tion, paclitaxel production was increased in both the endophytes, maximumpaclitaxel production i.e., 5.45 ± 0.98 mg/L was obtained by GBPI TWR F5(Aspergillus sp.) following 10 days of incubation at 15◦C in optimized S7liquid medium composition

Atmospheric Reactive Oxygen Species and Some Aspects of the Antiviral Protection at the Respiratory Epithelium.

—The review summarizes literature data on molecular and biochemical mechanisms of nonspecific protection of respiratory epithelium. The special attention is paid to comprehensive analysis of up-to-date data on the activity of the lactoperoxidase system expressed on the surface of the respiratory epithelium which provides the generation of hypothiocyanate and hypoiodite in the presence of locally produced or inhaled hydrogen peroxide. Molecular mechanisms of production of active compounds with antiviral and antibacterial effects, expression profiles of enzymes, transporters and ion channels involved in the generation of hypothiocyanite and hypoiodite in the mucous membrane of the respiratory system in physiological and pathological conditions (inflammation) are discussed. A hypothesis about the effect of atmospheric air composition on the efficiency of hypothiocyanate and hypoiodite generation in the respiratory epithelium in the context of its antibacterial and antiviral protection is presented. The causes and consequences of insufficiency of the lactoperoxidase system caused by the action of atmospheric factors are discussed in the context of controlling the sensitivity of the epithelium to the action of bacterial agents and viruses. Good evidence exists that restoration of the lactoperoxidase system activity can be achieved by application of pharmacological agents aimed to compensate for the deficit of halides in tissues, and by the control of chemical composition of the inhaled air.

Foliar Spraying of Salicylic Acid Enhances Growth, Yield, and Curcuminoid Biosynthesis Gene Expression as Well as Curcuminoid Accumulation in Curcuma longa

The application of exogenously applied salicylic acid plays important roles in improving the growth, yield, and bioactive compound compositions of different plant species. Curcuma longa is a medicinal plant that is commonly used as a spice and food additive, and has antioxidant potential. In this study, an innovative strategy for enhancing active compound production was investigated by applying a natural plant growth enhancer—namely, salicylic acid (SA)—to C. longa plants. The experiment was conducted using a complete randomized block design. The effects of SA on the growth, yield, and chemical compound contents of C. longa were recorded. Our findings demonstrated that SA significantly improved C. longa growth, yield, and curcuminoid content when compared to control treatment, with SA at 10−3 M having the greatest effect. The study also indicated that the increase in the curcuminoid content was accompanied by the overexpression of the curcumin synthase 1 (CURS1), 2 (CURS2), and 3 (CURS3) genes, as well as the diketide-CoA synthase (DCS) gene, which have been implicated in the synthesis of curcuminoids.

Plant tissue culture of cat whiskers (Orthosiphon aristatus Blume Miq): A review of secondary metabolite production and micropropagation

Background: The cat whiskers plant (Orthosiphon aristatus Blume Miq) is widely used as a raw material in traditional medicine for one of its many properties, i.e. antiviral activity. Three varieties of cat whiskers grow in Indonesia, classified according to the colour of their flower: white, white-purple, and purple. The purple variety of cat whiskers is endangered, so it is necessary to propagate this plant. Objectives: This research aimed to obtain appropriate protocols for plant propagation and production of active compounds of cat whiskers by in vitro culture. Methods: Data were collected from various online journal sites such as PubMed, ResearchGate, and Scopus. This review discusses several aspects, including callus induction efforts, modification of cell suspension cultures, and efforts to propagate the cat whiskers plant by in vitro culture. Results: Callus induction of white cat whiskers with purple and purple hues could take place on Murashige and Skoog (MS) media added with growth regulator 2,4-dichlorophenoxyacetic acid (2,4-D) 0.4 mg/L. Suspension culture medium (MS+ 2,4-D 1 mg/L + NAA 1 mg/L) can increase cell biomass and rosmarinic acid levels. Media MS + 6-benzyl amino purine (BAP) 3 mg/L + 1-naphthaleneacetic acid (NAA) 2 mg/L can induce the growth of shoots of white cat whiskers with purple and purple patterns. Root induction of the two varieties of cat whiskers could take place on MS + Indole-3-butyric acid (IBA) 0.75 mg/L medium. Conclusion: Efforts to produce secondary metabolites and plant propagation of cat whiskers by in vitro culture have been successful. It is necessary to enhance the production and propagation using bioreactors to yield more active compounds and raw materials of the cat whiskers plant.

ANTITUBERCULOSIS EFFECTIVENESS OF ENDOPHYTIC ANDALAS MUTANT BACTERIA PRODUCS INDUCED BY UV LIGHT WILL BE CONDUCTED

Indonesia is the 3rd largest country in the world in cases of tuberculosis (TB) infection. Infections caused by the bacteria Mycobacterium Tuberculosis have claimed 100,000 lives per year. This death rate is expected to continue to increase due to resistance to Antituberculosis Drugs (OAT). One strategy to overcome the increasing number of resistance of M. tuberculosis bacteria to OAT is to find new and better antituberculosis active compounds. This study aims to determine the potential of Andalas Endophytic Bacteria mutated by UV light on antituberculosis activity. This research is descriptive research. The research was conducted at the Research Laboratory of the Faculty of Mathematics and Natural Sciences, Padang State University. The mutation process is carried out as a form of increasing the production of active compounds. Natural anti microbial compounds can be obtained through the fermentation process. The active compounds from the fermentation products were extracted using methanol and aquadest as solvents. Testing against M. tuberculosis using the Mycobacterium Growth Indicator Tube (MGIT) 960 System as a medium for sensitivity testing. The fermented supernatant with absolute concentration and extract of each solvent with a concentration of 75% were mixed into the MGIT 960 tube. The results showed that the length of time UV irradiation could reduce the number of endophytic bacterial cells of Andalas isolate ATB 10-6. UV light also causes changes in the morphology of mutant bacterial colonies. UV irradiation can produce mutant isolates that have better antifungal activity, where of the 8 mutant bacteria produced, 7 mutants have better antifungal activity than the wild type, and the growth of mutant bacteria is faster than the wild type.

Advances in Pilot-Scale Stirred Bioreactors in Solid-State and Submerged Cultivations of Medicinal Mushrooms.

Great interest for large-scale production of medicinal mushroom biomass and various pharmaceutically active compounds production dictates the development of comprehensive technologies. Solid state and submerged cultivations in bioreactors represent the most promising technologies for fast and large amount production of medicinal fungi biomass and pharmaceutically active products for human and veterinary need. There are many stages from shaking culture studies to large-scale industrial production. Pilot-scale studies represent the bridge and the balance between the gap of laboratory and industrial scale. Therefore it is not a surprise that most of pilot-scale results and experiences remain uncovered industrial secrets. This chapter is an overview of available engineering achievements in submerged and solid-state cultivation experiences in pilot-scale bioreactors.

Production of ginsenoside by Chaetomium sp. and its effect on enhancing the contents of ginsenosides in Panax ginseng adventitious roots

Using endophytic fungi elicitors to enhance the yield of valuable secondary metabolites in plant tissue culture is an effective biotechnology strategy. In this study, we isolated an endophytic fungus Chaetomium sp. from cultivated Panax ginseng root and detected a high concentration of Rg3 in its mycelium. By comparing the effects of different treatments of Chaetomium sp. on ginsenoside production in Panax ginseng adventitious roots (PGARs), the immobilized spores of Chaetomium sp. (ISC) were proved to be the best elicitor. An initial pH of 6.5 and cocultivation time of 24 h yielded the highest ginsenosides content (56.29 ± 0.62 mg/g) in ISC-treated PGARs, increasing ginsenosides production by 3.2-fold compared with the blank group (17.56 ± 0.49 mg/g). Moreover, we show that ISC could stimulate the temporary accumulation of defense-related signal molecules, intensify the expression of defense genes in PGARs, and upregulate biosynthetic genes involved in the biosynthetic pathway of ginsenosides. Besides, the expression of the biosynthetic gene FPS (farnesyl diphosphate synthase) was deemed the most relevant factor for ginsenosides production during the elicitation period, and the L-aspartic acid secreted by Chaetomium sp. was proven to play a key role in the elicitation process. In conclusion, this study provides an accessible way of improving pharmacologically active compounds production in medicinal plants.

Effect of 2,4-D and BAP on Morphological Characters and Genetic Stability of Kaffir Lime (Citrus hystrix DC.) Callus Cultures Among Generations

Kaffir lime has medicinal properties for the treatment of many diseases. We used in vitro culture techniques to maintain quality and increase the production of active compounds in kaffir lime. The objective of this study is to determine the phenotype and genotype of kaffir lime callus cultures grown on media with the addition of 2,4-Dichlorophenoxyacetic (2,4-D) and Benzylaminopurine (BAP) on three generations (G0, G1, and G2). The callus was induced and subcultured on Murashige and Skoog medium added with 2,4-D: BAP at concentrations of 1:0.5; 2:0 and 5:0. Results showed no significant difference in callus initiation time in all treatment groups. Morphological characteristics, including color, texture, and biomass, varied among growth regulator concentrations and generation level. A high level of callus generation corresponds to a more friable texture and a more yellowish callus. Callus grown with the addition of 2,4-D and BAP 2:0 and 5:0 showed a more friable structure and yellowish color than 1:0.5. Growth regulator concentrations in each generation did not affect the callus growth curves, but the length of each phase between generations was different. The exponential phases of G1 and G2 were faster than that of G0. Despite slight differences in phenotype, the DNA profiles of callus suggest the same pattern between treatment groups, thereby indicating that our kaffir lime callus has genetic stability and that the callus can be used as raw material for medicinal purposes. Keywords: ISSR markers, Kaffir lime callus, Generations, Genotype, Growth factor, Phenotype