SH Medium Research Articles

Influence of medium and gelling agents concentration on in vitro rooting of Polygonum tinctorium

Polygonum tinctorium has long been employed in the medical and dye industries in many nations, particularly in Korea and Japan, for traditional fabric staining as a source of blue color. The plant tissue culture method has proven to be a rapid and sustainable approach for the regeneration of various plant species, particularly those with medicinal or ornamental value. In this study, we explored the in vitro root regeneration and growth of P. tinctorium in response to different growth media and gelling agents. Among the tested media, Schenk and Hildebrandt medium (SH medium) demonstrated superior performance, resulting in the highest number and length of roots per explant. SH media produced about 53 and 18% higher root per explant and a 40% and 14% increase in root length compared to B5 and MS media, respectively. Half strength of SH medium proved to be the optimal condition for both root number (7.46 roots per explant) and root length (3.81 mm). Among the gelling agents, 2 g/L of Gelrite medium was most effective in promoting the highest and tallest roots. These findings have the potential to enhance rooting abilities in various crops, particularly in the case of medicinal and ornamental plants, and may offer valuable insights for future industrial-scale root production of P. tinctorium.

Influence of cytokinins and yeast extract on growth and flavone production in hairy root cultures of Scutellaria baicalensis

Hairy roots produce various bioactive chemical compounds than wild-type roots which offer a promising in vitro approach for synthesizing important nutraceutical compounds. The purpose of this study is to increase the production of flavone compounds in hairy root cultures of Scutellaria baicalensis by the addition of elicitors such as cytokinins and yeast extract. Cytokinins such as kinetin (KIN), 6-benzylaminopurine (BAP), and Thidiazuron (TDZ) were utilized at 0.1, 0.5, and 1.0 mg/L, whereas for yeast extract treatment 50, 100, and 150 mg/L concentrations were added to the ½ SH medium. Effects of elicitors were measured in terms of dry biomass and flavone contents (baicalin, baicalein, and wogonin) using high-performance liquid chromatography (HPLC). The highest dry weight was achieved in the control hairy root than that of all cytokinins-treated hairy root cultures. In all the cytokinin-treated hairy root cultures, as the concentration increased the dry weight of the hairy root decreased. In contrast, in all the yeast extract-treated hairy root cultures as the concentration increases the dry weight of the hairy root increased, whereas the highest dry weight was achieved in 150 mg/L of yeast extract. Moving to the flavone content, baicalin was detected highest content in all the hairy root cultures supplied with cytokinin and yeast extract. The highest total flavone content was achieved in the hairy root culture treated with 1.0 mg/L of TDZ and 50 mg/L of yeast extract. This result might help the commercial agronomic sector by facilitating the in vitro mass production of nutraceuticals using S. baicalensis hairy root cultures.

Production of Phenolic Compounds and Antioxidant Activity in Hairy Root Cultures of Salvia plebeia.

Salvia plebeia (Lamiaceae) is a medicinal plant containing diverse bioactive constituents that have biological properties. In this study, we determined the optimal conditions (media and auxin) for the hairy root culture of S. plebeia for the growth and accumulation of phenolic compounds and evaluated its antioxidant activities. Rosmarinic acid and five phenylpropanoids were detected using high-performance liquid chromatography. The hairy roots grown in 1/2 SH medium with 1 mg/L NAA had a high level of rosmarinic acid content. Hairy roots cultured in 1 mg/L NAA had the highest total content of five phenylpropanoids. Compared to wild-type roots grown in the field, hairy roots (NAA 1) expressed similar levels of rosmarinic acid but significantly enhanced phenylpropanoid accumulation. Furthermore, the total phenolic content and total flavonoid content of hairy roots (NAA 1) were 2.22 and 1.73 times higher than those of wild-type roots. In the results of DPPH, ABTS, and reducing power assays, the hairy roots (NAA 1) showed higher free radical scavenging effects and reduction potential than the wild-type roots. These results suggest that S. plebeia hairy roots cultured under optimal conditions, which exhibit enhanced phenolic compound accumulation and antioxidant activity, can potentially be used as sources of antioxidants.

Intracellular trafficking and glycosylation of hydroxyproline-O-glycosylation module in tobacco BY-2 cells is dependent on medium composition and transcriptome analysis

Expression of recombinant proteins in plant cells with a “designer” hydroxyproline (Hyp)-O-glycosylated peptide (HypGP), such as tandem repeats of a “Ser-Pro” motif, has been shown to boost the secreted protein yields. However, dramatic secretion and Hyp-O-glycosylation of HypGP-tagged proteins can only be achieved when the plant cells were grown in nitrogen-deficient SH medium. Only trace amounts of secreted fusion protein were detected in MS medium. This study aims to gain a deeper understanding of the possible mechanism underlying these results by examining the intracellular trafficking and Hyp-O-glycosylation of enhanced green fluorescent protein (EGFP) fused with a (SP)32 tag, consisting of 32 repeats of a "Ser-Pro" motif, in tobacco BY-2 cells. When cells were grown in MS medium, the (SP)32-EGFP formed protein body-like aggregate and was retained in the ER, without undergoing Hyp-O-glycosylation. In contrast, the fusion protein becomes fully Hyp-O-glycosylated, and then secreted in SH medium. Transcriptome analysis of the BY-2 cells grown in SH medium vs. MS medium revealed over 16,000 DEGs, with many upregulated DEGs associated with the microtubule-based movement, movement of subcellular component, and microtubule binding. These DEGs are presumably responsible for the enhanced ER-Golgi transport of HypGP-tagged proteins, enabling their glycosylation and secretion in SH medium.

Rindera graeca (A. DC.) Boiss. & Heldr. (Boraginaceae) In Vitro Cultures Targeting Lithospermic Acid B and Rosmarinic Acid Production.

The in vitro cultures of Rindera graeca, a rare endemic plant, were developed as a sustainable source of phenolic acids. Various shoot and root cultures were established and scaled up in a sprinkle bioreactor. A multiplication rate of 7.2 shoots per explant was achieved. HPLC-PDA-ESI-HRMS analysis revealed the presence of rosmarinic acid (RA) and lithospermic acid B (LAB) as the main secondary metabolites in both the shoot and root cultures. The maximum RA (30.0 ± 3.2 mg/g DW) and LAB (49.3 ± 15.5 mg/g DW) yields were determined in root-regenerated shoots. The strongest free radical scavenging activity (87.4 ± 1.1%), according to 2,2-diphenyl-1-picrylhydrazyl-hydrate assay, was noted for roots cultivated in a DCR medium. The highest reducing power (2.3 µM ± 0.4 TE/g DW), determined by the ferric-reducing antioxidant power assay, was noted for shoots cultivated on an SH medium containing 0.5 mg/L 6-benzylaminopurine. A genetic analysis performed using random amplified polymorphic DNA and start codon targeted markers revealed genetic variation of 62.8% to 96.5% among the investigated shoots and roots. This variability reflects the capacity of cultivated shoots and roots to produce phenolic compounds.

Effects of culture medium composition on the <italic>in vitro</italic> growth of stem cuttings of <italic>Kirengeshoma koreana</italic> Nakai, a rare species in Korea

Background and objective: <i>Kirengeshoma koreana</i> Nakai is an endemic and endangered species in South Korea. We conducted <i>in vitro</i> propagation and regeneration of <i>K. koreana</i> from stem cuttings to investigate the effects of nine different basal culture media and five different carbohydrate sources on its growth.Methods: Apical segments (at least 1 cm long) collected randomly from a six-week-old <i>K. koreana</i> plantlet grown <i>in vitro</i> were used as explants. In the first experiment, the explants were transferred into square vessels containing 50 mL of nine different basal culture media supplemented with 30 g⋅L<sup>-1</sup> sucrose and 3 g⋅L<sup>-1</sup> Phytagel. In the second experiment, the explants were transferred into square vessels containing 50 mL half-strength SH medium supplemented with five different carbohydrate sources at 30 g⋅L<sup>-1</sup>. Each medium was solidified with 3 g⋅L<sup>-1</sup> Phytagel. All experiments contained 4 cultures, and the experiments were repeated four times to enhance reproducibility. Data on stem length, shoot fresh weight, leaf width, leaf length, root count, and root length were collected at the end of 8 weeks of culture.Results: ANOVA showed that the basal culture medium had a significant effect on <i>K. koreana</i> growth (<i>p</i> < .001). The half-strength SH medium was the best condition for stem length, shoot fresh weight, leaf width and root length (3.76 ± 0.12 cm, 0.60 ± 0.06 g, 1.19 ± 0.05 cm, and 2.83 ± 0.13 cm, respectively). However, the highest percentage increase in root count (13.00 ± 0.90) was found to occur with half-strength WPM. The effect of different carbohydrate sources on <i>K. koreana</i> growth was significantly different (<i>p</i> < .001), with the exception of stem length and leaf width (<i>p</i> = .26 and <i>p</i> = .09, respectively). Maltose was the best condition for shoot fresh weight (0.90 ± 0.09 g). Although there was no significant difference, sucrose was found to be best for leaf width, leaf length and root length (1.34 ± 0.07 cm, 2.34 ± 0.10 cm, and 3.86 ± 0.19 cm, respectively).Conclusion: This <i>in vitro</i> propagation and regeneration system for <i>K. koreana</i> shows promise in terms of scalability and could help greatly with germplasm conservation and restoration efforts for the species.

Bully Anak Artis Lesti Kejora dalam Analisis Wacana Kritis Norman Fairclough

This article describes Fairclough’s normative critical discourse on physical insults to children with Lesti Kejora using Norman theory. The method used in this research is descriptive qualitative method, listening method and literature study. The observation method was also applied in this study. This study also uses a note-taking approach. The survey consists of three phases: a data acquisition phase, a classification phase, and a data analysis phase. This study uses two news media, Suara.com and Herstory, based on the analysis of Fairclough's theory, bullying the child of artist Lesti Kejora. The data analysis technique used is reducing data, presenting data, and drawing conclusions. The purpose of the study was to find out how the two SH media (Suara.com and Herstory.ac.id) obtained information about the physical insults of Lesti Kejora's children. The theory used in this study is a three-dimensional model from Norman Fairclough, namely the micro, mesostructural, and macrostructural dimensions. Microstructural analysis that describes the negative and positive sides of physical abuse of children with Lesti Kejora, mesostructurally how the media obtains information related to physical abuse of children. Social situations depicting children's physical appearance as monkeys are associated with children's physical humiliation.

Cellulose Production by Rhodococcus sp. MI 2 in a Low-Cost Medium Supplemented with Vegetable Extracts

At present the industrial production of bacterial cellulose is limited due to the high cost of carbon substrates used. This study aimed to screen a number of vegetable extracts that could be used instead of SH medium as substrates for cellulose production by Rhodococcus sp. MI 2. There were 3 (Cha-poo, Pak-Lheang, and sweet potato) out of 26 different vegetable preparations chosen as medium supplements. Rhodococcus sp. MI 2 gave the highest cellulose yield in a medium supplemented with Cha-poo (Piper sarmentosum Roxb.) extract. It was increased by about 3 folds compared with the initial conditions, and cost about one quarter of the optimum SH medium. In addition, the structure of cellulose produced by Rhodococcus sp. MI 2 in medium supplemented with Cha-poo extract observed by SEM was different from those produced in the medium supplemented with sweet potato extract.

Establishment of an efficient in vitro propagation system for Holostemma annulare (Roxb.) K. Schum. using SH medium

Holostemma annulare (Roxb.) K. Schum. (Asclepiadaceae) is an important medicinal herb and its root tubers are an important ingredient of the drug ‘jivanti’. Here, a more reliable procedure of in vitro propagation of the plant is discussed using SH nutrient medium formulation that facilitates the in vitro multiplication and establishment without callusing from the basal region of shoots that was prominent in the earlier protocols. Surface sterilization of the nodal explants with 10% (v/v) NaOCl for 12 minutes followed by 0.1% (w/v) HgCl2 for 8 minutes exhibited 90% shooting response. The in vitro initiated shoots subcultured to SH medium fortified with 2.0 mgl-1 BA in combination with 0.5 mgl-1 Kinetin and 1.0 mgl-1 IBA produced maximum 7.16±0.15 shoots. Better root induction of the in vitro raised shoots was noticed in SH medium augmented with 0.01 mgl-1 NAA that induced a mean number of 5.65±0.23 thin, long, white coloured roots. The plantlets were transferred to polythene bags containing river sand:garden soil (1:1) for acclimatization, further transplanted in garden soil and river sand mixture (3:1) in earthern pots in the greenhouse and subsequently to the field got established at 80% efficiency. They produced tuberous roots after 4 months of field establishment. Thus, the study standardized an improved system of in vitro propagation of H. annulare using SH medium and this protocol can be extended for long-term conservation methods such as cryoconservation and also for the in vitro production of bioactive metabolites.

In Vitro Propagation, Genetic Assessment, and Medium-Term Conservation of the Coastal Endangered Species Tetraclinis articulata (Vahl) Masters (Cupressaceae) from Adult Trees.

Tetraclinis articulata (Vahl) Masters is an endangered tree growing in coastal and arid environments that is widely exploited by the timber and resin industry, among other applications. In this context, the use of in vitro techniques is highly encouraged for its propagation. We present a protocol for micropropagation using twigs from adult trees as a source of explants. The Schenk and Hildebrandt basal medium (SH) supplemented with 30 g L−1 sucrose, 6.5 g L−1 plant agar, 4.0 mg L−1 6-benzyladenine (BA), and 0.05 mg L−1 1-naphthaleneacetic acid (NAA) provided the optimum multiplication rate (90.48 ± 9.52 explants with basal shoots and 2.58 ± 0.29 basal shoots per explant). Application of activated charcoal (AC) or ½ Knop solution in a liquid overlay produced significantly longer shoots. Supplementation of solid media with indole-3-butyric acid (IBA) or NAA gave low rooting percentages (<17%). Addition of 0.9 g L−1 AC improved rooting (40%) but rooting performance was optimal (66.7%) after a pulse treatment consisting of 4 h immersion in liquid SH medium without growth regulators, followed by 8 weeks of cultivation. Rooted microplants were successfully acclimatized (93.33%) in a peat moss and vermiculite mixture (1:1 v/v ratio). The genetic stability of the in vitro regenerated plantlets was confirmed using the randomly amplified polymorphic DNA (RAPD) technique. Explant survival and growth remained higher than 90% after 28 weeks of cold storage at both 4 °C and 10 °C. The protocol presented here allows for largescale T. articulata production and could be applied for both ex situ conservation strategies and industrial purposes.

Highly Stretchable Bacterial Cellulose Produced by Komagataeibacter hansenii SI1.

A new strain of bacteria producing cellulose was isolated from Kombucha and identified as Komagataeibacter hansenii, named SI1. In static conditions, the strain synthesises bacterial nanocellulose with an improved ability to stretch. In this study, utilisation of various carbon and nitrogen sources and the impact of initial pH was assessed in terms of bacterial nanocellulose yield and properties. K. hansenii SI1 produces cellulose efficiently in glycerol medium at pH 5.0–6.0 with a yield of 3.20–3.60 g/L. Glucose medium led to the synthesis of membrane characterised by a strain of 77%, which is a higher value than in the case of another Komagataeibacter species. Supplementation of medium with vitamin C results in an enhanced porosity and improves the ability of bacterial nanocellulose to stretch (up to 123%). The properties of modified membranes were studied by scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction and mechanical tests. The results show that bacterial nanocellulose produced in SH medium and vitamin C-supplemented medium has unique properties (porosity, tensile strength and strain) without changing the chemical composition of cellulose. The method of production BNC with altered properties was the issue of Polish patent application no. P.431265.

Effect of factors on growth and development of Dendrobium officinale Kimura et Migo cultured in vitro and ex vitro

Dendrobium officinate Kimura et Migo, a species of orchid for beautiful flowers, is used in traditional medicine in many Asian countries because there are many important pharmaceuticals (chrysotoxene, erianin, confusarin, polysaccharide, alkaloid ...) in anti-cancer, anti-aging, boosting immunity and vasodilation, etc. In recent years, more than 4,000 hectares of Dendrobium officinate Kimura et Migo artificial planting has been available in China and the price of dry product was around ¥ 80,000/kg. Currently, plantlet quality is an important factor influencing the acclimatization stage, growth, and development of plants in the greenhouse. In this study, the effect of a number of factors medium, plant regulator, ventilation, and substrate to improve plantlet quality as well as further growth and development in the greenhouse conditions were investigated. The results showed that SH medium was suitable for in vitro shoot growth in terms of monitoring parameters after 90 days of culture. In shoot multiplication stage, the shoots culture on SH medium supplemented with 2 mg/L BA, 30 g/L sucrose, 9 g/L agar, 1g /L activated charcoal (AC) gave the best results with 4.53 shoots/per shoot. The SH medium supplemented with 1.0 mg/L NAA, 30 g/L sucrose, 9 g/L agar, 1.0 g/L AC combined with ventilation conditions was suitable for rooting stage with plant height (5.73 cm), number of roots (4.77), root length (5.00 cm), fresh weight (3.36 g), dry weight (0,31 g), and total chlorophyll (SPAD) (45.76 nmol/cm2). Plantlets derived from culture ventilation conditions cultivated on the mixture of pine bark and fern fiber (50:50) was the highest survival rate (100%) and growth after 12 months in the greenhouse.

Zygotic embryo culture is an efficient way to optimize in vitro growth in Panax ginseng

Despite many studies on tissue culture of Panax ginseng, the propagation of cultivars is still dependent on seeds due to low survival rate of plants produced in vitro. Healthy in vitro grown plants guarantee high survival rates after they are transferred to soil, but limited information exists on the optimum medium for the in vitro growth of P. ginseng. In this study, zygotic embryo culture was used to develop an optimum medium suitable for in vitro cultivation of P. ginseng. After 4–6 months of culture, the aged aerial parts turned yellow, and the in vitro grown roots (IGRs) had thickened taproots with dormant rhizomes. The seedling growth was evaluated using various concentrations of sucrose and basal media at various strengths. Of the tested media, 2% sucrose and 1/3 strength SH medium showed superior results. Subsequently, IGRs were transferred to soil and evaluated for their morpho-anatomical characteristics. Furthermore, their plants were analyzed for genetic fidelity using molecular markers. IGRs showed outstanding morphological differences compared with conventionally grown 1-year-old ginseng roots; however, there was no difference in the sprouting and survival rate after soil transfer. 2-year-old IGRs deprived from the IGRs have intermediate properties between 1-year-old and 2-year-old root. Unlike the 2-year-old roots, the morphology of 2-year-old IGRs had shorter primary roots with several lateral roots. Despite the morpho-anatomical differences, molecular marker analysis also confirmed that the IGRs had genetic fidelity.

Establishment of Codonopsis pilosula (Franch.) Nannf. transformed roots, influence of the culture conditions on root growth and production of essential oil

Codonopsis pilosula (Franch.) Nannf. is a species whose radix has long been used in traditional folk medicine in China, Japan and Korea. Codonopsis Radix is obtained from cultivations mainly in Gansu, Shanxi, Sichuan, and Guizhou Provinces of China. This study describes an efficient protocol for genetic transformation of C. pilosula by Rhizobium rhizogenes and the obtainment of transformed roots of this species with valuable medicinal properties. It was found that both the bacterial strain (A4 and ATCC 15834) and acetosyringone (AcS) addition influenced the efficiency of transformation. The highest frequency of root induction (69 %) was observed on the Murashige and Skoog (MS) agar medium supplemented with AcS after transformation with A4 strain. The transformation was confirmed by PCR analysis. Of the tested media (MS, Schenk and Hildebrandt - SH, Gamborg - B5, and Woody Plant - WPM) those with full macro- and microsalt concentrations elicited significantly greater C. pilosula transformed root (CP2) biomass accumulation than the half-strength media, apart from MS medium. The greatest CP2 root biomass was observed in SH medium (1.65 g flask−1 DW). Among the tested sucrose concentrations (10−70 g L−1), the best result was obtained by using 30 g L−1 sucrose. Essential oils isolated by hydrodistillation from CP2 transformed roots and commercially available raw material, C. pilosula roots, had very similar qualitative composition. The most important group of common constituents of both oils were sesquiterpene hydrocarbons. The main among twenty two compounds in this group possessed skeletons of barbatane, thujopsane and bisabolane; the predominant ones being α- and β-barbatene, followed by β-chamigrene and thujopsene. This study presented that C. pilosula transformed roots can be a potential source for the industrial production of the raw material from species of the genus Codonopsis used in traditional Chinese medicine.

Effects of explant type, plant growth regulator and cultivation system on multiplication of Polygonum multiflora Thunb. cultured in vitro

Polygonum multiflora Thunb. (Belongs to Polygonaceae family) is a medicinal plant of economic value and should be protected, according to the Red Book of Vietnam. Many studies have shown that P. multiflora Thunb. was effective for many diseases such as hair loss, premature gray hair, lower back pain, knee weakness, muscle weakness, hemiplegia, nervousness, dizziness, insomnia, etc. Propagation and preservation of this herbal species are significantly important because of increased demand. Adventitious root biomass culture has been successful on a number of plant species, and that it has potential for P. multiflorum breeding. In this study, the effect of explants, plant growth regulators and different culture systems on adventitious root formation was investigated. The adventitious roots were induced from the explants and the optimum culture medium were transferred to four different culture systems (3-liter triangular flask, shaking triangle, bioreactor self-made and Korean bioreactor) containing 5 g adventitious roots per 1 liter medium. The results showed that the leaf cultured on SH medium supplemented with 1.5 mg L-1 IBA, 30 g L-1 sucrose, 8.5 g L-1 agar and pH 5.8 gave the higher of adventitious root regeneration rate (100%), root number/explant (25.33 roots), fresh weight (131.67 mg) and dry weight 13.35 mg) compared to those cultured on other condition after 6 weeks of culture. The adventitious root multiplication in the Korea bioreactor system is more effective than other culture systems shown in the fresh biomass (17.04 g), dry biomass (1.56 g) and proliferation rate (3.40 fold) after 6 weeks of culture.

Potential of Lemnoideae Species for Phytoremediation of Fresh Water with Elevated Manganese Concentration

Background . Wastewater treatment using physical, chemical, and biological methods is primary solution for the reduction of water pollution that reaching the critical thresholds. The members of subfamily Lemnoideae , commonly called duckweed, are considered the most efficient aquatic plants for wastewater remediation. Although properties of duckweed to survive in water with high concentration of heavy metal ions such as chromium, cobalt, lead, nickel and cuprum are well documented, the growth of duckweed in water with high concentrations of manganese and the efficiency of retention of manganese from water by these species has not been estimated. Objective . Four duckweed species ( Spirodela polyrhiza , Landoltia punctata , Lemna aequinoctialis , and L. turionifera ) were used for establishment of influence Mn on their vitality and growth and for studying their potential for phytoremediation of fresh water with elevated manganese concentration. Methods . Duckweed collected in Eastern China was introduced in tissue culture in vitro by surface sterilization. The identification of the collected duckweed species was determined by DNA barcoding using primers specific for chloroplast intergenic spacers atpF - atpH (ATP) and psbK - psbL (PSB). The experiments for establishment of influence Mn on duckweed growth carried out in aseptic condition. To determinate concentration of Mn, the samples of different water type (Hongze Lake, ponds around Hongze Lake, Huaian local municipal sewage plant and industrial sewage plant) were analyzed by the Inductively Coupled Plasma Optical Emission Spectrometry. Results . The most sensitive duckweed to Mn was S. polyzhiza , the first characteristic symptoms of toxicity like brown spots have appeared when concentration of Mn was 40 mg/L, the concentration 200 mg/L Mn resulted in chlorosis and death of fronds. L . aequinoctialis and L. turionifera had similar effects in SH medium supplemented with 650 mg/L and 975 mg/L Mn, respectively. L . punctata was the most tolerant duckweed to Mn plants continued to grow even at concentration 975 mg/L. Response of duckweed on Mn was dependent on availability of nitrogen in nutrient medium. Using four duckweed species for treatment of water containing 4.12 mg/L Mn allowed to reduce concentration until safe level of standard (0.1 mg/L Mn). Conclusions . All investigated duckweed species ( S. polyrhiza, L. punctata , L. aequinoctialis , and L. turionifera ) were characterized by a high level of resistance to manganese, especially L. punctata. Response of duckweed on Mn was dependent on availability of nitrogen in nutrient medium. The tested species of subfamily Lemnoideae were high effective for phytoremediation of water with elevated manganese concentration.

The in vitro shoot regeneration of Ehretia asperula Zol. and Mor.

Xa den young branches in the orchard were sterilized and used as explants for shoot initiation and growth experiments. The shoot induction was carried out with BA (benzyl adenine) or TDZ (thidiazuron). New shoots sprouted after one week of culture and the highest shoots (2.02 cm) were on MS medium (Murashige and Skoog medium) with BA 0.6 mg/L after 3 weeks. Furthermore, the number of leaves per shoot was also higher than other treatments (8.31 leaves per shoot). In vitro shoots were used in other experiments to investigate the effects of explants, biotin concentrations, minerals, type and concentration of cytokinins on the formation and elongation of shoots and clusters. When 5 mg/L biotin was added to MS medium, shoots grew better. The MS medium appeared to be most suitable for the initiation and elongating of shoots, followed by WPM (woody plant medium) and SH medium (Schenk and Hildebrandt medium), while B5 medium (Gamborg B5 medium) was the least effective. The spouting from three-week-old explants was earlier than others (4 and 5 weeks of age), which in turn affected on the shoot elongating. BA 1.5 mg/L was suitable to induce shoot clusters (3.91 shoots per explant) after 4 weeks.

Enhancement of Phenolic Compounds and Antioxidative Activities by the Combination of Culture Medium and Methyl Jasmonate Elicitation in Hairy Root Cultures of Lactuca indica L.

Lactuca indica L. has been traditionally used as a wild vegetable and as a medicinal plant for centuries. The various compounds present in it and their biological activities have been extensively reported. Hairy-root culture combined with agrobacterium-meditated metabolic engineering is a useful technique to achieve stable production of biologically active plant compounds. Here, we evaluated the enhancement of secondary metabolites in L. indica L. and their bioactivities by testing culture media composition and the use of an elicitor. Hairy roots were induced and cultured in MS or SH liquid media for 2 weeks prior to treatment with various concentrations of MeJa, for different periods. The resulting phenolic contents and physiological activities were analyzed. Higher total phenolic, flavonoid, and hydroxycinnamic acids contents were attained by elicitation with MeJa. Metabolite accumulation, especially in SH media and in the presence of MeJa, was time dependent. Particularly, accumulation of chicoric acid increased markedly with time. Similarly, we observed time dependent positive and negative responses of antioxidant activity in DPPH and ABTS assays, respectively. As in previous studies, the highest correlation was found between total phenolic content and total flavonoid content. Further, 3,5-DCQA showed the highest correlation with total phenolic content, total flavonoid content, and antioxidant activities in hydroxycinnamic acids. Our data effectively identified optimal culture conditions to increase the accumulation of secondary metabolites and antioxidant activity in hairy roots cultures of L. indica L.

Ứng dụng đèn led trong vi nhân giống cây gừng (Zingiber officinale Rosc.)

Currently, fluorescent lamps are often used as lighting systems in the tissue culture. These lamps consumed a lot of power. Recently, the use of LED lamp systems for planting was being noticed because LEDs were more conducive to tissue culture and less power consumption. In this study, the effects of mineral content, sugar concentration and monochromatic light on ginger’s growth were investigated. The results showed that ginger plant cultured in SH medium (Schenk and Hildebrandt, 1972) had higher height and diameter than in other media. The experiment using different sugar concentration, explants get the best growth when they were cultured in medium supplemented with 30 g/l sucrose. The next experiment, explants were placed in different types of light sources including, red LED, blue LED, or combination of red with blue light in 9: 1; 8: 2; 7: 3; 6: 4; 5: 5 ratios, fluorescent lamps, red fluorescent lamps, blue fluorescent lamps. The results showed that the red LED was suitable for the growth of ginger. The plant height (4.55 cm), leaf area (2.66 mm2) and root diameter (3.83 mm) was significantly higher than that of the other treatments. In the nursery, plants under cultivation of red LED were good growth and a survival plantlet ratio obtained over 96%.

Studies on micropropagation of Rosa setigera michx. through multiple shoot induction

The present paper describes the studies carried out to determine the effect of nutrition on micropropagation of Rosa setigera. The nodal explants harvested from 4th to 8th node of juvenile shoots were inoculated over the media composed of salts and vitamins of MS, WPM, SH and B5 medium. These media contained 2–4% of sucrose or glucose or maltose as the carbon source. The explants harvested in rainy season were inoculated over the medium fortified with 2.2μM BAP, while those harvested in winter season were inoculated over the medium containing 0.5μM NAA and 8.8μM BAP. It was observed that the regeneration efficiency of explants harvested in rainy season was best with SH salts and vitamins, 3% glucose and 2.2μM BAP. Similarly, the explants harvested in winter season displayed their regeneration potential to maximum over the medium composed of WPM salts and vitamins, 3% glucose, 0.5μM NAA and 8.8μM BAP. The maximum elongation of microshoots was obtained with the medium containing 0.6% agar and rooting over the ½ strength MS medium containing 9.8μM IBA.