Influence of inoculum density and aeration volume on biomass and bioactive compound production in bulb-type bubble bioreactor cultures of Eleutherococcus koreanum Nakai

This study deals with the effects of initial inoculum density and aeration volume on biomass and bioactive compound production in adventitious roots of Eleutherococcus koreanum Nakai in bulb-type bubble bioreactors (3-L capacity). While the fresh and dry weights of the roots increased with increasing inoculum density, the highest percentage dry weight and accumulation of total target compounds (eleutheroside B and E, chlorogenic acid, total phenolics, and flavonoids) were noted at an inoculum density of 5.0 g L⁻¹. Poor aeration volume (0.05 vvm) stunted root growth, and high aeration volume (0.4 vvm) caused physiological disorders. Moreover, an inoculum density of 5.0 g L⁻¹ and an aeration volume of 0.1 vvm resulted in the highest concentration of total target compounds and least root death. Such optimization of culture conditions will be beneficial for the large-scale production of E. koreanum biomass and bioactive compounds.     

Induction and cultures of mountain ginseng adventitious roots and AFLP analysis for identifying mountain ginseng

Among the various plant growth regulators (PGR), 9.0 μM 2,4-D(2,4-dichlorophenoxyacetic acid) turned out to be the best for callogenesis and 4.5 μM 2,4-D was revealed to be more suitable for callus proliferation. Of the four basal media, the highest number of 8.2 adventitious roots was induced in Woody plant medium (WPM) containing 14.8 μM IBA(indole 3-butyic acid). F tests on each auxin treatment in the ANOVA revealed highly significant difference at α=0.01. The number of multiple adventitious roots (MARs) formed in each adventitious roots highly depended on the types and levels of auxin tested. At both 24.6 μM and 34.4 μM IBA, more than 25 MARs were induced after 4 weeks of culture. When different medium were applied, immersion type showed the highest fresh weight of MARs at the time of harvest. With 160 g fresh weight of inoculum, about 14.2 times of biomass was obtained after 6 weeks of culture. Identical fractionation pattern of the amplified fragment length polymorphism (AFLP) analysis was observed between the adventitious root and donor of a mountain ginseng. This observation suggested that any rearrangements of the genomic DNA had not occurred in the adventitious roots after 1 year of cultures. The cultivated ginseng and the mountain ginseng groups could be distinguished by 2 AFLP markers.     

Psoralen production in hairy roots and adventitious roots cultures of Psoralea coryfolia

Psoralea corylifolia is an endangered plant producing various compounds of medical importance. Adventitious roots and hairy roots were induced in cultures prepared from hypocotyl explants. Psoralen content was evaluated in both root types grown either in suspension cultures or on agar solidified medium. Psoralen content was ~3 mg g⁻¹ DW in suspension grown hairy roots being higher than in solid grown hairy roots and in solid and suspension-grown adventitious roots.     

Influence of medium salt strength and nitrogen source on biomass and metabolite accumulation in adventitious root cultures of Pseudostellaria heterophylla

We investigated the effects of medium salt strength and ammonia/nitrate ratio on biomass production and metabolites accumulation of adventitious root. The medium with full-strength Murashige and Skoog (MS) reached the highest growth rate (16.77), and the contents of saponin and polysaccharide reached the peak (i.e., 0.65 and 24.85 %) at 3/4 MS and 1 MS, respectively. In case of ammonia/nitrate ratio, a NH₄ ⁺/NO₃ ^? ratio of 20:40 was optimal for the production of biomass and polysaccharide (23.27 %). In contrast, the content of saponin achieved the optimum (0.74 %) at a NH₄ ⁺/NO₃ ^? ratio of 30:30. In 5-L balloon-type bubble bioreactor (BTBB) cultures, an approximately 23-fold increase in biomass was recorded. The fresh weight (FW) and dry weight (DW) were 72.78 and 6.79 g per bioreactor with the contents of saponin (0.62 %) and polysaccharide (17.32 %), respectively. It indicated potential application to produce adventitious roots of pseudostellaria heterophylla with bioreactors on large scale in commercial.     

诱导子对雷公藤不定根生长和次生代谢产物含量的影响

为探讨真菌诱导子以及硝酸银等非生物诱导子对雷公藤不定根生长及次生代谢产物含量的影响。以雷公藤不定根为材料,通过向培养基中添加不同种类的真菌诱导子以及硝酸银等非生物诱导子,采用高效液相色谱检测不定根中雷公藤甲素和生物碱含量。结果表明:各种真菌诱导子不影响不定根的生长,但对次生代谢产物含量有显著影响,其中,苹果炭疽和柿子炭疽诱导子的加入不仅使不定根中雷公藤甲素的含量分别提高了2.24和1.93倍,生物碱的含量也各提高了2.02和2.07倍。苹果炭疽诱导子浓度为50μg/m L时比较适合雷公藤不定根生长及雷公藤甲素和生物碱的积累。硝酸银抑制不定根的生长和生物碱的积累,但促进雷公藤甲素的积累。硝酸银浓度为25μmol/L时雷公藤甲素含量为对照的1.71倍。茉莉酸甲酯浓度为50μmol/L时,不定根增长量为对照的1.04倍,雷公藤甲素和生物碱含量分别为对照的1.64倍和2.12倍。酵母提取物浓度为2 g/L时,雷公藤甲素含量为对照的1.48倍。表明培养基中添加硝酸银和酵母提取物对不定根中雷公藤甲素的合成具有明显的促进作用,苹果炭疽和茉莉酸甲酯的协同作用既能促进雷公藤甲素的合成又能促进雷公藤生物碱的合成。     

Combined effects of phytohormone, indole-3-butyric acid, and methyl jasmonate on root growth and ginsenoside production in adventitious root cultures of Panax ginseng C.A. Meyer

Indole-3-butyric acid at 25 μM with methyl jasmonate (MJ) at 100 μM in Panax ginseng synergistically stimulated both root growth and ginsenoside accumulation compared with 100 μM MJ alone. Productivity of ginsenoside was 10 mg l⁻¹ d⁻¹ compared to 7.3 mg l⁻¹ d⁻¹ with MJ elicitation alone.     

Cortical Aerenchyma formation in hypocotyl and adventitious roots of Luffa cylindrica subjected to soil flooding

BACKGROUND AND AIMS: Aerenchyma formation is thought to be one of the important morphological adaptations to hypoxic stress. Although sponge gourd is an annual vegetable upland crop, in response to flooding the hypocotyl and newly formed adventitious roots create aerenchyma that is neither schizogenous nor lysigenous, but is produced by radial elongation of cortical cells. The aim of this study is to characterize the morphological changes in flooded tissues and the pattern of cortical aerenchyma formation, and to analyse the relative amount of aerenchyma formed. METHODS: Plants were harvested at 16 d after the flooding treatment was initiated. The root system was observed, and sections of fresh materials (hypocotyl, tap root and adventitious root) were viewed with a light or fluorescence microscope. Distributions of porosity along adventitious roots were estimated by a pycnometer method. KEY RESULTS: Under flooded conditions, a considerable part of the root system consisted of new adventitious roots which soon emerged and grew quickly over the soil surface. The outer cortical cells of these roots and those of the hypocotyl elongated radially and contributed to the development of large intercellular spaces. The elongated cortical cells of adventitious roots were clearly T-shaped, and occurred regularly in mesh-like lacunate structures. In these positions, slits were formed in the epidermis. In the roots, the enlargement of the gas space system began close to the apex in the cortical cell layers immediately beneath the epidermis. The porosity along these roots was 11-45 %. In non-flooded plants, adventitious roots were not formed and no aerenchyma developed in the hypocotyl or tap root. CONCLUSIONS: Sponge gourd aerenchyma is produced by the unique radial elongation of cells that make the expansigeny. These morphological changes seem to enhance flooding tolerance by promoting tissue gas exchange, and sponge gourd might thereby adapt to flooding stress.     

GL生根剂对扶桑插条不定根皮层细胞超微结构的影响

研究了GL生根剂对扶桑插条不定根皮层细胞超微结构的影响。GL生根剂使皮层细胞中线粒体和内质网增加,液胞、线粒体和内质网膜有局部损害．经GL生根剂处理的皮层细胞含有的淀粉粒和脂质体数目较对照皮层细胞多．该实验结果为了解GL生根剂促进不定根形成的原因提供了细胞学基础．     

Elicitation enhanced the production of bioactive compound and biomass accumulation in callus cultures of Glycyrrhiza glabra L.

In Vitro Cellular & Developmental Biology - Plant - Licorice (Glycyrrhiza glabra L.), Fabaceae, has a wide range of medicinal properties due to the metabolites found in the plant tissues....     

Effect of polyploidy induction on biomass and ginsenoside accumulations in adventitious roots of ginseng

Adventitious roots ofPanax ginseng C.A. Meyer (a natural tetraploid) were treated with 50 or 100 mg L^-1 colchicine for 12, 24,36, 48, or 60 h to induce polyploid (octoploid) roots. The largest number of octoploid roots was obtained with a 100 mg L^-1 colchicine treatment over 60 h. To verify that ginsenoside was being accumulated in the developing tissues, the tetraploid (control) and octoploid roots were cultured for 40 d in Murashige and Skoog media that lacked NH₄NO₃ but was supplemented with 2 mg L^-1 naphthaleneacetic acid and 50 g L_-1 sucrose. Levels of fresh and dry biomass were greater in the octoploid roots. Although total ginsenoside and Rb-group ginsenoside contents were less in the octoploid roots than in the tetraploids, the former had a higher amount of Rg-group ginsenosides (especially Rg1). These results demonstrate the benefit that polyploid adventitious roots provide in enhancing the production of secondary metabolites in ginseng.     

Synergistic effects of plant growth retardants and IBA on the formation of adventitious roots in hypocotyl cuttings of mung bean

The synergistic effect of plant growth retardants, such as daminozide, paclobutrazol and triadimefon, and of indole-3-butyric acid (IBA) on the formation of adventitious roots in hypocotyl cuttings of mung bean was studied. The three retardants and IBA all stimulated adventitious root growth, but IBA was the most effective. However, mixtures of the retardants with IBA have proven generally more effective than IBA alone in promoting adventitious root formation. When IBA was applied to the hypocotyls one day after cutting preparation followed by the growth retardant on the second day, there were even more adventitious roots produced than if applied in the reverse order. The effectiveness of the treatments were in the order, IBA followed by growth retardant, IBA + growth retardant together, and IBA alone.Abbreviations IBA indole-3-butyric acid - GA gibberellin     

Influence of sucrose concentration and phosphate source on biomass and metabolite accumulation in adventitious roots of Pseudostellaria heterophylla

In this study, we investigated the induction of Pseudostellaria heterophylla adventitious root and the effects of sucrose concentration and phosphate source on biomass increase and metabolites accumulation. These roots were initially cultured in Murashige and Skoog medium for 4 weeks. IBA 3.0 mg L^?1 proved to be the best auxin for inducing adventitious roots and the frequency of adventitious roots induced from roots (100 %) was higher than that from leaves (78 %) and stems (27 %). The medium with 4 % sucrose resulted in the optimum biomass i.e. 1.04 g/flask DW, and the content of saponin and polysaccharides reached the peak i.e. 0.676 and 24.4 %, respectively. With regards to phosphate source, 1.25 mM phosphate concentration was more favorable for biomass of roots (0.87 g/flask of DW), whereas the optimum saponin (0.74 %) and polysaccharides (22.09 %) were achieved with 2.5 mM phosphate. However, the saponin content at 2.5 mM phosphate did not show significant difference from the saponin content at 0.625 mM (0.69 %) or 3.75 mM phosphate (0.69 %).     

Influence of carbon source on shoot multiplication and adventitious bud regeneration in in vitro beech cultures

Experiments were performed to determine the effects ofcarbon source and concentration on shootmultiplication in shoot cultures of Fagussylvatica (one clone) and F. orientalis (twoclones) and on the induction of adventitious shootbuds from leaf and internode explants of F.orientalis. In general, glucose was the best carbonsource for both axillary branching and adventitiousshoot regeneration. Shoot-tip explants grown on 3–4%glucose medium produced more shoots than those onsucrose or fructose. For maximum shoot length, glucosemedium was best for two of the three clones, and 4%sucrose for the other. The number of shoots was theparameter most influenced by glucose concentration inthe adventitious shoot regeneration experiments, thenumber increasing with sugar concentration. The lowesthyperhydricity rate occurred in the presence ofsucrose in both species. Shoot growth and quality wasnegatively affected by fructose supplied media. Theuse of filter-sterilized rather than autoclavedfructose neither stimulated shoot growth nor reducedthe incidence of hyperhydricity in all three clones.The response of shoot cultures to differentcarbohydrate treatments appears to some extent to begenotype dependent.     

Effect of various bud induction treatments on elongation and rooting of adventitious shoots of Canary Island pine (Pinus canariensis)

Three-day-old cotyledonary explants of Pinus canariensis were subjected to 30 induction treatments using half-strength Bornman's medium containing various combinations of N⁶- benzyladenine, zeatin, kinetin and 2-isopentenyl-adenine. The highest numbers of buds were obtained with 10 M 6-benzyladenine, but both kinetin and zeatin influenced shoot elongation. Shoots were maintained on half-strength Schenk and Hildebrandt medium with 2% sucrose and 0.05% activated charcoal. For rooting, shoots were pulsed for 4 h in a 100 M indole-3-butyric acid aqueous solution (pH 4.2–4.5), and planted in peat:vermiculite:perlite (1:1:1). After 8 weeks, the numbers of rooted shoots were similar for most treatments. Therefore, the bud induction treatments did not significantly influence rooting of adventitious shoots of Canary Island pine.     

Effect of explant type and genotype on the accumulation of bioactive compounds in adventitious root cultures of Polygonum multiflorum

Plant Cell, Tissue and Organ Culture (PCTOC) - The correlation between growth and accumulation of bioactive compounds was investigated in six genotypes of Polygonum multiflorum, grown in vitro,...     

Hairy roots induced by Agrobacterium rhizogenes and production of regenerative plants in hairy root cultures in maize

Hairy roots of maize were induced by infecting 15-d calli with Agrobacterium rhizogenes. The hairy roots cultured in hormone-free media showed the vigorous growth and typical hairy root features. The regenerated plants were produced from hairy roots in MS media supplemented with 1.6 mg/L ZT and 0.4 mg/L NAA. The PCR-Southern hybridization demonstrated that T-DNA had been integrated into the chromosome of regenerated plants. These authors contributed equally to this work.     

Effects of earthworms on biomass production,nitrogen allocation and nitrogen transfer in wheat-clover intercropping model systems

The effects of earthworms (Lumbricidae) on plant biomass production and N allocation in model intercropping systems of winter wheat and white clover were evaluated in two pot experiments. Wheat and wheat-clover mixtures were grown in a low-organic loam soil, earthworms were added at densities comparable to field population densities and the experiments were terminated 48 and 17 d after earthworm introductions. In both experiments, earthworms significantly increased the biomass and N uptake of wheat while they had generally no effects on clover. As a result, earthworm activity increased the proportion of wheat biomass in the total plant biomass of the mixture. Nitrogen budgets of the experiment lasting 48 d indicated that additional N in the system made available by earthworm activity was primarily taken up by the wheat. Earthworms also affected intra-plant N allocation in wheat which had significantly higher shoot:root N ratios when earthworms were present. When clover was labelled with ¹⁵N in the experiment which lasted 17 d, endogeic earthworms significantly reduced the amounts of ¹⁵N excess transferred from living or decomposing clover roots to accompanying wheat plants. Earthworms assimilated small quantities of ¹⁵N tracer from decomposing clover roots but not from living clover roots. The results of these model experiments suggest that earthworms can affect the balance between intercropped cereals and legumes by altering intra- and inter-plant N allocation. This revised version was published online in June 2006 with corrections to the Cover Date.     

Hairy roots induced by Agrobacterium rhizogenes and production of regenerative plants in hairy root cultures in maize

Before the late 1980s, although the majority of Agrobacterium-mediated gene transfer experiments have been performed with A. tumefaciens[1―3], some work has also been done with its close relative, Agro-bacterium rhizogene. It has been considered that onl…     

Effect of oxygen supply on biomass and helvolic acid production in submerged fermentation of Cordyceps taii

The entomogenous fungus Cordyceps taii, a traditional Chinese medicinal mushroom, exhibits potent important pharmacological effects and it has great potential for health foods and medicine. In this work, the effects of oxygen supply on production of biomass and bioactive helvolic acid were studied in shake-flask fermentation of C. taii mycelia. The value of initial volumetric oxygen transfer coefficient (K_La) within 10.1–33.8 h⁻¹ affected the cell growth, helvolic acid production and expression levels of biosynthetic genes. The highest cell concentration of 17.2 g/L was obtained at 14.3 h⁻¹ of initial K_La. The highest helvolic acid production was 9.6 mg/L at 10.1 h⁻¹ of initial K_La. The expression levels of three genes encoding hydroxymethylglutaryl-CoA synthase, hydroxymethylglutaryl-CoA reductase and squalene synthase were down-regulated on day 2 and day 8 but up-regulated on day 14 at an initial K_La value of 10.1 h⁻¹ vs. 33.8 h⁻¹, which well corresponded to the helvolic acid biosynthesis in those conditions. The information obtained would be helpful for improving the biomass and helvolic acid production in large-scale fermentation of C. taii.