Induction of trans-resveratrol and extracellular pathogenesis-related proteins in elicited suspension cultured cells of Vitis vinifera cv Monastrell

Suspension-cultured cells of Vitis vinifera cv Monastrell were used to investigate the effects of methyljasmonate, ethylene and salicylic acid separately or in combination with cyclodextrins on both trans-resveratrol production and the induction of defense responses.     

Contents of <Emphasis Type="Italic">Trans</Emphasis>-resveratrol in Korean Grape Cultivars,including “Kyoho”

Concentrations of trans-resveratrol were analyzed in 36 Korean-grown grape cultivars, including cv. Kyoho. This cultivar has large irregular berries and a deep-black skin and accounts for >15% of all grapevines grown in Korea. The content of trans-resveratrol is an important quality parameter, and its pattern of variability increased gradually until the time of harvest. Its distribution also fluctuated significantly among leaves, seeds, and exocarps, with the highest amount (1,477 μg g⁻¹ dry weight) occurring in the leaves. Among all cultivars evaluated here, trans-resveratrol contents were significantly higher in “Cheongsoo” and “High Bailey,” their levels being 50% above the overall mean (i.e., 2.0-mg g⁻¹ dry weight), whereas 14 cultivars had contents that were 10% less than that mean value     

Pseudozyma sp. SPJ: an economic and eco-friendly approach for degumming of flax fibers

A cellulase free, alkaline, thermo-tolerant pectinase was produced by a novel yeast strain Pseudozyma sp. SPJ using citrus peel as inexpensive carbon source. The crude enzyme showed good prospects in degumming of flax fibers for textile industry. An optimum pectinase dose of 80 U g⁻¹ resulted in reduction of 15 ± 1.92% dry weight of the fibers, releasing maximum galacturonic acid (10825.5 ± 34.2 μg g⁻¹ dry fiber) after the incubation of 6 h. The yeast culture could grow on the flax fibers (as sole carbon source) without addition of any other nutrient and produce good enzyme yield (9235.5 ± 21.51 U g⁻¹ dry fiber). After 12 h incubation of the fibers with the isolated yeast strain, 4471 ± 19.5 μg g⁻¹ dry fiber galacturonic acid was achieved with maximum weight loss of 11 ± 1.2%. This process reduced the amount of chemicals and energy used in conventional methods. It also contributed to enhance fineness and overall quality of the fiber strands. This study is relevant to the textile industry as it provided a fast, economical and eco-friendly method for degumming of flax fibers.     

Statistical medium optimization for enhanced azadirachtin production in hairy root culture of Azadirachta indica

Azadirachtin, a well-known biopesticide, is a secondary metabolite extracted from the seeds of Azadirachta indica. In the present study, azadirachtin was produced in hairy roots of A. indica, generated by Agrobacterium rhizogenes-mediated transformation of leaf explants. Liquid cultures of A. indica hairy roots were developed with a liquid-to-flask volume ratio of 0.15. The kinetics of growth and azadirachtin production were established in a basal plant growth medium containing MS medium major and minor salts, Gamborg’s medium vitamins, and 30 g l⁻¹ sucrose. The highest azadirachtin accumulation in the hairy roots (up to 3.3 mg g⁻¹) and azadirachtin production (∼44 mg l⁻¹) was obtained on Day 25 of the growth cycle, with a biomass production of 13.3 g l⁻¹ dry weight. To enhance the production of azadirachtin, a Plackett–Burman experimental design protocol was used to identify key medium nutrients and concentrations to support high root biomass production and azadirachtin accumulation in hairy roots. The optimal nutrients and concentrations were as follows: 40 g l⁻¹ sucrose, 0.19 g l⁻¹ potassium dihydrogen phosphate, 3.1 g l⁻¹ potassium nitrate, and 0.41 g l⁻¹ magnesium sulfate. Concentrations were determined by a central composite design protocol and verified in shake-flask cultivation. The optimized medium composition yielded a root biomass production of 14.2 g l⁻¹ and azadirachtin accumulation of 5.2 mg g⁻¹, which was equivalent to an overall azadirachtin production of 73.84 mg l⁻¹, 68% more than that obtained under non-optimized conditions.     

Elicitation of guggulsterone production in cell cultures of <Emphasis Type="Italic">Commiphora wightii</Emphasis> by plant gums

Plant gum as an elicitor for guggulsterone production in cell cultures of Commiphora wightii is reported for the first time. Guggulsterone production increased 2.4 fold in the cell cultures by gum Arabic (100 mg l⁻¹), while mesquite gum elicited 2 fold. The cells treated with gum Arabic at 7th and 9th day accumulated enhanced guggulsterones within 24 h, which increased further up to 48 h and then declined. The cells treated at 9th day accumulated higher amount (218 μg l⁻¹) of guggulsterones after 48 h of elicitation as compared to cells treated at 7th day (164 μg l⁻¹). The optimized elicitation conditions were used in vessels of varying capacity where maximum yield of 285 μg l⁻¹ of guggulsterones was recorded in 3 l shake flasks. These experiments enabled highest guggulsterones yield in a short duration of 11 days in cell cultures of C. wightii.     

Efficient production of lactic acid from sucrose and corncob hydrolysate by a newly isolated <Emphasis Type="Italic">Rhizopus oryzae</Emphasis> GY18

The aim of this study is to investigate production of l-lactic acid from sucrose and corncob hydrolysate by the newly isolated R. oryzae GY18. R. oryzae GY18 was capable of utilizing sucrose as a sole source, producing 97.5 g l⁻¹ l-lactic acid from 120 g l⁻¹ sucrose. In addition, the strain was also efficiently able to utilize glucose and/or xylose to produce high yields of l-lactic acid. It was capable of producing up to 115 and 54.2 g l⁻¹ lactic acid with yields of up to 0.81 g g⁻¹ glucose and 0.90 g g⁻¹ xylose, respectively. Corncob hydrolysates obtained by dilute acid hydrolysis and enzymatic hydrolysis of the cellulose-enriched residue were used for lactic acid production by R. oryzae GY18. A yield of 355 g lactic acid per kg corncobs was obtained after 72 h incubation. Therefore, sucrose and corncobs could serve as potential sources of raw materials for efficient production of lactic acid by R. oryzae GY18.     

Drought,warming and soil fertilization effects on leaf volatile terpene concentrations in <Emphasis Type="Italic">Pinus halepensis</Emphasis> and <Emphasis Type="Italic">Quercus ilex</Emphasis>

The changes in foliar concentrations of volatile terpenes in response to water stress, fertilization and temperature were analyzed in Pinus halepensis and Quercus ilex. The most abundant terpenes found in both species were α-pinene and Δ³-carene. β-Pinene and myrcene were also abundant in both species. P. halepensis concentrations were much greater than those of Q. ilex in agreement with the lack of storage in the latter species (15205.60 ± 1140.04 vs. 0.54 ± 0.08 μg g⁻¹ [d.m.]). The drought treatment (reduction to 1/3 of full watering) significantly increased the total terpene concentrations in both species (54% in P. halepensis and 119% in Q. ilex). The fertilization treatment (addition of either 250 kg N ha⁻¹ or 250 kg P ha⁻¹ or both) had no significant effects on terpene foliar concentrations. The terpene concentrations increased from 0.25 μg g⁻¹ [d.m.] at 30°C to 0.70 μg g⁻¹ [d.m.] at 40°C in Q. ilex (the non-storing species) and from 2,240 μg g⁻¹ [d.m.] at 30°C to 15,621 μg g⁻¹ [d.m.] at 40°C in P. halepensis (the storing species). Both species presented negative relationship between terpene concentrations and relative water contents (RWC). The results of this study show that higher terpene concentrations can be expected in the warmer and drier conditions predicted for the next decades in the Mediterranean region.     

Enhanced fructooligosaccharides and inulinase production by a <Emphasis Type="Italic">Xanthomonas campestris</Emphasis> pv. <Emphasis Type="Italic">phaseoli</Emphasis> KM 24 mutant

Xanthomonas campestris pv phaseoli produced an extracellular endoinulinase (9.24 ± 0.03 U mL⁻¹) in an optimized medium comprising of 3% sucrose and 2.5% tryptone. X. campestris pv. phaseoli was further subjected to ethylmethanesulfonate mutagenesis and the resulting mutant, X. campestris pv. phaseoli KM 24 demonstrated inulinase production of 22.09 ± 0.03 U mL⁻¹ after 18 h, which was 2.4-fold higher than that of the wild type. Inulinase production by this mutant was scaled up using sucrose as a carbon source in a 5-L fermenter yielding maximum volumetric (21,865 U L⁻¹ h⁻¹) and specific (119,025 U g⁻¹ h⁻¹) productivities of inulinase after 18 h with an inulinase/invertase ratio of 2.6. A maximum FOS production of 11.9 g L⁻¹ h⁻¹ and specific productivity of 72 g g⁻¹ h⁻¹ FOS from inulin were observed in a fermenter, when the mutant was grown on medium containing 3% inulin and 2.5% tryptone. The detection of mono- and oligosaccharides in inulin hydrolysates by TLC analysis indicated the presence of an endoinulinase. This mutant has potential for large-scale production of inulinase and fructooligosaccharides.     

Effective biotic elicitation of <Emphasis Type="Italic">Ruta graveolens</Emphasis> L. shoot cultures by lysates from <Emphasis Type="Italic">Pectobacterium atrosepticum</Emphasis> and <Emphasis Type="Italic">Bacillus</Emphasis> sp.

Growth of Ruta graveolens shoots was induced when Bacillus sp. cell lysates were added to the culture medium. Elicitation of coumarin by this lysate was also very effective; the concentrations of isopimpinelin, xanthotoxin and bergapten increased to 610, 2120 and 1460 μg g⁻¹ dry wt, respectively. It also had a significant effect on the production of psoralen and rutamarin (680 and 380 μg g⁻¹ dry wt) and induced the biosynthesis of chalepin, which was not detected in the control sample, up to 47 μg g⁻¹ dry wt With lysates of the Pectobacterium atrosepticum, their effect on growth was not so significant and had no effect on the induction of coumarin accumulation. But elicitation with this lysate was much more effective for inducing the production of furoquinolone alkaloids; the concentrations of γ-fagarine, skimmianine, dictamnine and kokusaginine rose to 99, 680, 172 and 480 μg g⁻¹ dry wt, respectively.     

Seasonal changes in growth rate, carrageenan yield and lectin content in the red alga Kappaphycus alvarezii cultivated in Camranh Bay, Vietnam

The three color morphotypes of the red alga Kappaphycus alvarezii (brown, red and green) were cultured in Camranh Bay, Vietnam, using the fixed off-bottom monoline culture method to evaluate the growth rate, carrageenan yield, 3,6-anhydrogalactose, gel strength and lectin content. The brown morphotype was cultivated over a 12-month period; the red and green morphotypes were over a 6-month period. At the 60-day culture timepoint, the brown morphotype showed a higher growth rate (3.5–4.6% day⁻¹) from September to February, and lower growth rate (1.6–2.8% day⁻¹) from March to August. Significant (P < 0.05) differences in growth rate between culture months were found with the brown morphotype. High growth rates for the red (3.6–4.4% day⁻¹) and green (3.7–4.2% day⁻¹) morphotypes were obtained from September to February. The carrageenan yield, 3,6-anhydrogalactose and gel strength of the three morphotypes showed little variation, with the highest values obtained in November–December. At the 30-day sampling point, the brown morphotype had a higher lectin content (167–302 μg g⁻¹ dry alga) from August to March and a lower lectin content (23–104 μg g⁻¹ dry alga) from April to July. High lectin contents were recorded for the red (139–338 μg g⁻¹ dry alga) and green (124–259 μg g⁻¹ dry alga) morphotypes from September to February. This study shows that the different morphotypes of K. alvarezii can be grown in the tropical waters of the Camranh during the northeast monsoon, and part of the southwest monsoon, especially the brown morphotype, which can be grown during any season.     

Identification of QTL underlying isoflavone contents in soybean seeds among multiple environments

Soybean isoflavones are valued in certain medicines, cosmetics, foods and feeds. Selection for high-isoflavone content in seeds along with agronomic traits is a goal of many soybean breeders. The aim of the study was to identify the quantitative trait loci (QTL) underlying seed isoflavone content in soybean among seven environments in China. A cross was made between ‘Zhongdou 27’, a soybean cultivar with higher mean isoflavone content in the seven environments (daidzein, DZ, 1,865 μg g⁻¹; genistein, GT, 1,614 μg g⁻¹; glycitein, GC, 311 μg g⁻¹ and total isoflavone, TI, 3,791 μg g⁻¹) and ‘Jiunong 20’, a soybean cultivar with lower isoflavone content (DZ, 844 μg g⁻¹; GT, 1,046 μg g⁻¹; GC, 193 μg g⁻¹ and TI, 2,061 μg g⁻¹). Through single-seed-descent, 130 F₅-derived F₆ recombinant inbred lines were advanced. A total of 99 simple-sequence repeat markers were used to construct a genetic linkage map. Seed isoflavone contents were analyzed using high-performance liquid chromatography for multiple years and locations (Harbin in 2005, 2006 and 2007, Hulan in 2006 and 2007, and Suihua in 2006 and 2007). Three QTL were associated with DZ content, four with GT content, three with GC content, and five with TI content. For all QTL detected the beneficial allele was from Zhongdou 27. QTL were located on three (DZ), three (GC), four (GT) and five (TI) molecular linkage groups (LG). A novel QTL was detected with marker Satt144 on LG F that was associated with DZ (0.0014 > P > 0.0001, 5% < R ² < 11%; 254 < DZ < 552 μg g⁻¹), GT (0.0027 > P > 0.0001; 4% < R ² < 9%; 262 < GT < 391 μg g⁻¹), and TI (0.0011 > P > 0.0001; 4% < R ² < 15%; 195 < TI < 871 μg g⁻¹) across the various environments. A previously reported QTL on LG M detected by Satt540 was associated with TI across four environments and TI mean (0.0022 > P > 0.0001; 3% < R ² < 8%; 182 < TI < 334 μg g⁻¹) in China. Because both beneficial alleles were from Zhongdou 27, it was concluded that these two QTL would have the greatest potential value for marker-assisted selection for high-isoflavone content in soybean seed in China. G. Zeng, D. Li and Y. Han have equal contributions to the paper.     

Increased miroestrol, deoxymiroestrol and isoflavonoid accumulation in callus and cell suspension cultures of Pueraria candollei var. mirifica

Miroestrol and deoxymiroestrol are highly active phytoestrogens derived from the tuberous roots of Pueraria candollei var. mirifica. To date, there have been no reports regarding the production of miroestrol and deoxymiroestrol in in vitro cell culture. In this study, callus and cell suspension cultures were established for the purpose of investigating miroestrol and deoxymiroestrol content in P. candollei var. mirifica cells. Stem-derived callus cultured on Murashige and Skoog (MS) medium supplemented with 0.1 mg l⁻¹ thidiazuron (TDZ), 0.5 mg l⁻¹ naphthaleneacetic acid (NAA), and 1.0 mg l⁻¹ benzyladenine (BA) provided optimal conditions for the accumulation of deoxymiroestrol and total isoflavonoids. The calli produced 184.83 ± 20.09 μg g⁻¹ dry weight of total chromene and 20.72 ± 2.38 mg g⁻¹ dry weight of total isoflavonoid. This is the first report to suggest that callus culture is a suitable alternative method for producing miroestrol and deoxymiroestrol. Carbon sources were evaluated for the cell suspension cultures of P. candollei var. mirifica. Sucrose provided optimal conditions for biomass production, whereas fructose was the most suitable carbon source for deoxymiroestrol and isoflavonoid production. The information from our study can be employed for enhancing the production of miroestrol, deoxymiroestrol, and total isoflavonoids using in vitro cell culture of P. candollei var. mirifica.     

Rate of dehydration,state of subcellular organisation and nature of cryoprotection are critical factors contributing to the variable success of cryopreservation: studies on recalcitrant zygotic embryos of <Emphasis Type="Italic">Haemanthus montanus</Emphasis>

Effects of sequential procedures required for cryopreservation of embryos excised from the recalcitrant seeds of Haemanthus montanus were assessed ultrastructurally and in conjunction with respiratory activity and the rate of protein synthesis. Fresh material (water content, 5.05 ± 0.92 g g⁻¹ dry mass) afforded ultrastructural evidence of considerable metabolic activity, borne out by respiratory rates. Neither exposure to glycerol nor sucrose as penetrating and non-penetrating cryoprotectants, respectively, brought about degradative changes, although increased vacuolation and autophagy accompanied both, while respiratory and protein synthetic activity were not adversely affected. Glycerol-cryoprotected embryos flash dried to water contents >0.4 g g⁻¹ showed organised ultrastructural features and considerable autophagy consistent with metabolic activity, and although respiratory activity was lower, protein synthesis rate was enhanced relative to fresh material. However, at water contents <0.4 g g⁻¹, embryo tissue presented a mosaic of cells of variable density and ultrastructural status, but trends in rates of respiration and protein synthesis remained similar. Flash drying after sucrose exposure was accompanied by considerable ultrastructural abnormality particularly at water contents <0.4 g g⁻¹, lysis of individual and groups of cells and considerable depression of respiration, but not of protein synthesis. Success, assessed as ≥50% axes forming seedlings after cryogen exposure, was obtained only when glycerol-cryoprotected embryos at water contents >0.4 g g⁻¹—in which the degree of vacuolation remained moderate—were rapidly cooled. The outcomes of this study are considered particularly in terms of the stresses imposed by prolonged, relatively slow dehydration and ultimate water contents, on embryos showing considerable metabolic activity.     

Genetic variation of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> isolates forming fumonisin B<Subscript>1</Subscript> and moniliformin

Thirty single-spore isolates of a toxigenic fungus, Fusarium oxysporum, were isolated from asparagus spears and identified by species-specific polymerase chain reaction (PCR) and translation elongation factor 1-α (TEF) sequence analysis. In the examined sets of F. oxysporum isolates, the DNA sequences of mating type genes (MAT) were identified. The distribution of MAT idiomorph may suggest that MAT1-2 is a predominant mating type in the F. oxysporum population. F. oxysporum is mainly recognised as a producer of moniliformin—the highly toxic secondary metabolite. Moniliformin content was determined by high-performance liquid chromatography (HPLC) analysis in the range 0.05–1,007.47 μg g⁻¹ (mean 115.93 μg g⁻¹) but, also, fumonisin B₁ was detected, in the concentration range 0.01–0.91 μg g⁻¹ (mean 0.19 μg g⁻¹). There was no association between mating types and the mycotoxins biosynthesis level. Additionally, a significant intra-species genetic diversity was revealed and molecular markers associated with toxins biosynthesis were identified.     

Influence of the KDEL signal,DMSO and mannitol on the production of the recombinant antibody 14D9 by long-term <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> cell suspension culture

We have established two transgenic cell suspension culture lines of Nicotiana tabacum that express the catalytic antibody 14D9 as a secretory product (sec-Ab) or as a KDEL-tagged product in the endoplasmic reticulum (Ab-KDEL), respectively. After 3 years of culture, the performance improved to a production level of 0.15 ± 0.03 μg ml⁻¹ on the seventh day of culture for the sec-Ab line and 0.48 ± 0.05 μg ml⁻¹ on the third day for Ab-KDEL line. Analysis of the effect of osmotic stress using mannitol (90 g l⁻¹) as an osmolite revealed that there was a 12-fold increase in antibody yield (1.96 ± 0.20 μg ml⁻¹) on the seventh day of culture in line sec-Ab and a fivefold increase (2.31 ± 0.18 μg ml⁻¹) on the seventh day for line Ab-KDEL. The concentration of the antibody in the culture medium was not significant. Dimethyl sulfoxide used as a permeabilizing agent was not effective in increasing 14D9 yield, but it did cause distinctive cell damage at all concentrations tested.     

Establishment of anthocyanin-producing cell suspension cultures of Cleome rosea Vahl ex DC. (Capparaceae)

The effects of different levels of Murashige and Skoog (MS) basal medium, 2,4-dichlorophenoxyacetic acid (2,4-D), and sucrose on anthocyanin production and biomass accumulation of cell suspension cultures of Cleome rosea were investigated. Cultures were established in liquid MS medium containing 30 g l⁻¹ sucrose and supplemented with 0.90 μM 2,4-D. Proliferating cell suspension cultures achieved the highest growth capacity, a fourfold increase in biomass accumulation, following subculture at the exponential growth phase, 14–18 days of culture. Moreover, the presence of 2,4-D was essential for anthocyanin production and biomass accumulation. On the other hand, increasing levels of sucrose above 30 g l⁻¹ resulted in a drastic reduction in biomass accumulation. Anthocyanin production was highest in cell suspension cultures grown on half-strength MS medium (1/2 MS), 30 g l⁻¹ sucrose, and 0.45 μM 2,4-D. These cell suspension cultures were mainly composed of small aggregates of spherical cells with similar morphology observed in anthocyanin-producing and non-producing cultures. Moreover, microscopic analysis of anthocyanin-producing cultures showed the presence of mixtures of non-pigmented, low-pigmented, and high-pigmented cells.     

Increased lignan biosynthesis in the suspension cultures of Linum album by fungal extracts

Linum album accumulates anti-tumor podophyllotoxin (PTOX) and its related lignans, which were originally isolated from an endangered species Podophyllum. In the present study, we examined the effects of five fungal extracts on the production of lignans in L. album cell cultures. Fusarium graminearum extract induced the highest increase of PTOX [143 μg g⁻¹ dry weight (DW) of the L. album cell culture], while Rhizopus stolonifer extract enhanced the accumulation of lariciresinol up to 364 μg g⁻¹ DW, instead of PTOX. Typical elicitors, such as chitin, chitosan, or methyl jasmonate (MeJA), were shown to be less effective in lignan production in L. album cell cultures. These results verified the advantages of fungal extracts to increase lignan production in L. album cell culture, and suggested potential on-demand metabolic engineering of lignan biosynthesis using differential fungal extracts.     

Drift-ice and under-ice water communities in the Gulf of Bothnia (Baltic Sea)

The algal, protozoan and metazoan communities within different drift-ice types (newly formed, pancake and rafted ice) and in under-ice water were studied in the Gulf of Bothnia in March 2006. In ice, diatoms together with unidentified flagellates dominated the algal biomass (226 ± 154 μg ww l⁻¹) and rotifers the metazoan and protozoan biomass (32 ± 25 μg ww l⁻¹). The under-ice water communities were dominated by flagellates and ciliates, which resulted in lower biomasses (97 ± 25 and 21 ± 14 μg ww l⁻¹, respectively). The under-ice water and newly formed ice separated from all other samples to their own cluster in hierarchical cluster analysis. The most important discriminating factors, according to discriminant analysis, were chlorophyll-a, phosphate and silicate. The under-ice water/newly formed ice cluster was characterized by high nutrient and low chlorophyll-a values, while the opposite held true for the ice cluster. Increasing trends in chlorophyll-a concentration and biomass were observed with increasing ice thickness. Within the thick ice columns (>40 cm), the highest chlorophyll-a concentrations (6.6–22.2 μg l⁻¹) were in the bottom layers indicating photoacclimation of the sympagic community. The ice algal biomass showed additional peaks in the centric diatom-dominated surface layers coinciding with the highest photosynthetic efficiencies [0.019–0.032 μg C (μg Chl-a ⁻¹ h⁻¹) (μE m⁻² s⁻¹)⁻¹] and maximum photosynthetic capacities [0.43-1.29 μg C (μg Chl-a ⁻¹ h⁻¹)]. Rafting and snow-ice formation, determined from thin sections and stable oxygen isotopic composition, strongly influenced the physical, chemical and biological properties of the ice. Snow-ice formation provided the surface layers with nutrients and possibly habitable space, which seemed to have favored centric diatoms in our study.     

High stilbenes accumulation in root cultures of <Emphasis Type="Italic">Cayratia trifolia</Emphasis> (L.) Domin grown in shake flasks

The Root cultures of Cayratia trifolia (Vitaceae) a tropical lianas, were maintained in liquid Murashige and Skoog’s medium containing 0.5 mg l⁻¹ NAA, 0.1 mg l⁻¹ kinetin with 3% sucrose. These root cultures when grown with 6% sucrose accumulated stilbenes (piceid, resveratrol, viniferin, ampelopsin) in high amounts, which on elicitation by 500 mg l⁻¹ yeast extract, 50 μM salicylic acid (SA), 50 μM methyl jasmonate (MeJa), 500 μM ethrel added at 25th day, increased up to ninefolds (7.1 mg l⁻¹). Addition of alar or phenylalanine along with the elicitors further enhanced the stilbenes content. In the present study, stilbenes accumulation up to 12 folds (9.2 mg l⁻¹) was obtained with SA and alar. The SA was the most effective in increasing the stilbenes contents while less than control values were recorded in the cells treated with MeJa. The roots could be grown up to 2 l flasks. The present work demonstrates that presence of precursor and sucrose during elicitation at an appropriate time combined with growth retardation significantly increased the production of stilbenes in C. trifolia cell cultures.     

Succinic acid production by a newly isolated bacterium

A new bacterial strain producing succinic acid was enriched from bovine rumen content. It is facultatively anaerobic, belongs to the family Pasteurellaceae and has similarity to the genus Mannheimia. In batch cultivations with D-glucose or sucrose the strain produced up to 5.8 g succinic acid l⁻¹ with a productivity and a yield of up to 1.5 g l⁻¹ h⁻¹ and 0.6 g g⁻¹, respectively. With crude glycerol up to 8.4 g l⁻¹, 0.9 g l⁻¹ h⁻¹ and 1.2 g g⁻¹ were obtained. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.