Production of a Bioflocculant by Rhodococcus erythropolis S-1 Grown on Alcohols

Rhodococcus erythropolis strain S-1, which was isolated from soil, produces a bioflocculant. We have found that alcohols are useful carbon sources for its flocculant production. Ethanol was best for flocculant production and culture time. The bioflocculant produced on ethanol medium flocculated a wide range of suspended soils, alkaline and acid.     

Simultaneous production of hydrogen and bioflocculant by Enterobacter sp. BY-29

Hydrogen and a bioflocculant could be produced simultaneously by anaerobic culture of Enterobacter sp. BY-29. For production of hydrogen and the bioflocculant by cell culture of the bacterium in batch cultures, cultivation at 37 °C in a medium containing glucose as a carbon source and Polypepton as a nitrogen source was found to be suitable. In continuous production of hydrogen and the bioflocculant by cell culture or immobilized cells of the bacterium, the hydrogen production rate and hydrogen yield by the immobilized cells on porous glass beads in stirred and column reactors were higher than those by the cell culture in a stirred reactor. However, production of the bioflocculant by the cell culture was superior to that by the immobilized cells in continuous production.     

Novel bioflocculant produced by salt-tolerant,alkaliphilic strain Oceanobacillus polygoni HG6 and its application in tannery wastewater treatment

The optimized production of MBF-HG6, which is a novel salt-tolerant alkaliphilic bioflocculant produced by Oceanobacillus polygoni with its application in tannery wastewater treatment was investigated in this study. It was found the optimal carbon source, nitrogen source, cation, and initial pH of the medium for bioflocculant production were starch, urea, Fe²⁺, and pH 9.0, respectively. The best stability in the temperature range was from 0 to 80°C and the purified MBF-HG6 contained polysaccharides of 81.53% and proteins of 9.98%. The carboxyl, hydroxyl, and amino groups were determined in bioflocculants, while the optimized bioflocculating activity was observed as 90.25% for the dosages of 6.96mL MBF-HG6, 4.77mL CaCl₂ (1%, m/v), and 19.24g/L NaCl using response surface methodology. In addition, SS and turbidity removal rates of the tannery wastewater (4g/L MBF-HG6) could, respectively, reach 46.49% and 91.08%, indicating that the great potential was emerged in enhancement of tannery wastewater treatment by MBF-HG6.     

Production of an exopolysaccharide bioflocculant by Sorangium cellulosum

AIMS: To isolate a new exopolysaccharide bioflocculant produced by the myxobacterium Sorangium cellulosum NUST06, and to characterize its chemical composition and expolysaccharide production relative to carbon source. METHODS AND RESULTS: Exopolysaccharide levels and biomass production by S. cellulosum NUST06 were analysed relative to carbon source. Glucose in the medium at a level of 3 g l(-1) completely inhibited cell growth and exopolysaccharide production, but low concentrations of glucose (1-2 g l(-1)) could stimulate cell utilization of starch. The chemical composition and flocculating activity of the NUST06 exopolysaccharide was investigated. The flocculant comprised 38.3% proteins and 58.5% carbohydrates, of which glucose, mannose and glucuronic acid were present at 51.3%, 39.2% and 10.5%, respectively. The flocculating activity of the NUST06 flocculant depended strongly on cations. CONCLUSIONS: It is feasible to produce an exopolysaccharide bioflocculant by the strain NUST06 in a mineral salts medium using starch as a carbon source. SIGNIFICANCE AND IMPACT OF THE STUDY: This strain may be advantageous for commercial bioflocculant production and may enrich existing knowledge of myxobacteria.     

Production of a novel bioflocculant MNXY1 by Klebsiella pneumoniae strain NY1 and application in precipitation of cyanobacteria and municipal wastewater treatment

Aims: To isolate and characterize the novel bioflocculant‐producing bacteria, to optimize the bioflocculant production and to evaluate its potential applications. Methods and Results: Klebsiella pneumoniae strain NY1, a bacterium that produces a novel bioflocculant (MNXY1), was selected on the chemically defined media. It was classified according to the 16S rRNA gene sequence, morphological and microscopic characteristics. MNXY1 was characterized to contain 26% protein and 66% total sugar. The constituent sugar monomers of MNXY1, revealed by NMR analysis, are glucose, galactose and quinovose. Favourable culture conditions for MNXY1 production were determined. Strain NY1 produces a high level (14.9 g l^?1) of MNXY1. MNXY1 is thermostable and tolerant to the extreme pH. It precipitated 54% of cyanobacteria from laboratory culture and 72% of the total suspended solids from raw wastewater. Conclusions: Strain NY1 was identified to produce a novel bioflocculant MNXY1. The outstanding performance of MNXY1 in practical applications and its availability in copious amounts make it attractive for further investigation and development for industrial scale applications. Significance and Impact of the Study: This is first report for the identification of a quinovose‐containing bioflocculant and application of a protein–polysaccharide complex bioflocculant in precipitation of cyanobacteria. These findings suggest that MNXY1 holds great potential for use in management of harmful algae and city wastewater treatment.     

In situ measurements confirm the seasonal dominance of benthic algae over phytoplankton in nearshore primary production of a large lake

1. Despite the recognition of its importance, benthic primary production is seldom reported, especially for large lakes. We measured in situ benthic net primary production by monitoring flux in dissolved inorganic carbon (DIC) concentration in benthic incubation chambers, based on continuous measurements of CO_2(aq) flux, alkalinity, and the temperature‐dependent dissociation constants of carbonic acid (K₁ and K₂). This methodology has the advantages of monitoring net primary production directly as change in carbon, maintaining continuous water recirculation, and having sufficient precision to detect change in DIC over short (i.e. 15 min) incubations, even in alkaline waters. 2. Benthic primary production on Cladophora‐dominated rocky substrata in western Lake Ontario was measured biweekly. Maximum biomass‐specific net photosynthetic rates were highest in the spring (2.39 mgC g Dry Mass^?1 h^?1), decreased to negative rates by early summer (?0.76 mgC g DM^?1 h^?1), and exhibited a regrowth in late summer (1.98 mgC g DM^?1 h^?1). 3. A Cladophora growth model (CGM), previously validated to predict Cladophora biomass accrual in Lake Ontario, successfully simulated the seasonality and magnitude of biomass‐specific primary production during the first cohort of Cladophora growth. Averaged over this growing season (May–Aug), mean areal net benthic production at the estimated depth of peak biomass (2 m) was 405 mg C m^?2 d^?1. 4. We measured planktonic primary production in proximity to the benthic study and constructed a depth‐resolved model of planktonic production. Using the CGM, benthic primary production was compared with planktonic primary production for the period May–Aug. Net benthic production from the shoreline to the 12 m contour (1–2 km offshore) equalled planktonic production. Closer to shore, benthic primary production exceeded planktonic primary production. Failure to account for benthic primary production, at least during abundant Cladophora growth, will lead to large underestimates in carbon and nutrient flows in the nearshore zone of this Great Lake.     

Contribution of benthic microalgae to the temporal variation in phytoplankton assemblages in a macrotidal system

Suspended marine benthic microalgae in the water column reflect the close relationship between the benthic and pelagic components of coastal ecosystems. In this study, a 12‐year phytoplankton time‐series was used to investigate the contribution of benthic microalgae to the pelagic system at a site along the French‐Atlantic coast. Furthermore, all taxa identified were allocated into different growth forms in order to study their seasonal patterns. The highest contribution of benthic microalgae was observed during the winter period, reaching up to 60% of the carbon biomass in the water column. The haptobenthic growth form showed the highest contribution in terms of biomass, dominant in the fall‐winter period when the turbidity and the river flow were high. The epipelic growth form did not follow any seasonal pattern. The epiphytic diatom Licmophora was most commonly found during summer. As benthic microalgae were found in the water column throughout the year, the temporal variation detected in the structure of pelagic assemblages in a macrotidal ecosystem was partly derived from the differentiated contribution of several benthic growth forms.     

Characterization of diatom assemblages in mid-altitude streams of NW Italy

Following the European Water Framework Directive, this study aims to be the first step to (i) identify diatom type assemblages in unpolluted streams in NW Italy, and (ii) find which ecological factors explain most of the variation. To achieve this, we collected physical, chemical and benthic community data from four streams in NW Italy from 2001 to 2004, for a total of 72 samples. All sampling sites were between 200 m a.s.l. and 800 m a.s.l., but differed in the dominant lithological substrate, i.e. alluvial or siliceous. Relationships between diatom communities and environmental factors were examined using canonical correspondence analysis (CCA), while Indicator Species Analysis was used to define characterizing species and accompanying species of three environmental groups identified by CCA: (1) high water quality and medium saline content, (2) high water quality and low saline content, (3) poor water quality. The diatom assemblages of the three groups of sites differed significantly, as shown by the Multi-Response Permutation Procedure. There were strong correlations between diatoms and environmental factors, especially chlorides (also highly correlated with sulphates and carbonate hardness), nitrate concentration and conductivity. The group 1 assemblage was typical of the alluvial Alpine streams with medium saline content and was characterized by mostly oligosaprobic/β-mesosaprobic taxa such as Cymbella affinis, Diatoma ehrenbergii and Staurosira pinnata. The species assemblage found in the siliceous Alpine rivers with good water quality make them suitable reference sites for a benthic diatom community. Electronic Supplementary Material Supplementary material is available for this article at and accessible for authorised users Handling editor: K. Martens     

一种富集高温栅藻Desmodesmus sp. F51的新型生物絮凝剂

[背景]海科贝特氏菌(Cobetia marina)可产生大量具有絮凝活性的胞外产物,可视为一种新型的生物絮凝剂。高温栅藻(Desmodesmus sp.F51)是一种具有较高叶黄素含量的微藻,被认为是一种新兴的叶黄素来源,但利用该生物絮凝剂高效富集高温栅藻的相关研究迄今尚未见报道。[目的]以高温栅藻为对象,研究该新型生物絮凝剂的絮凝效率,并对絮凝机理进行初步探讨。[方法]探索在不同生长阶段微藻培养液添加生物絮凝剂、添加量、絮凝时间、pH对絮凝效率的影响,分析生物絮凝剂的功能基团,并测定在不同pH条件下添加生物絮凝剂前后高温栅藻的Zeta电位变化,以及在显微镜下分析藻细胞在添加生物絮凝剂前后的形态。[结果]在高温栅藻生长至稳定期(pH 8.0)添加2 mL生物絮凝剂,絮凝15 min絮凝效果最佳,达82.1%。傅里叶红外光谱(fourier transform infrared spectroscopy,FTIR)显示了多糖及酰胺结构的特征吸收峰,由此推测生物絮凝剂主要是多糖的混合物,含有少量蛋白质。根据Bradford法测定絮凝剂中蛋白含量约为0.4%(质量比),通过苯酚-硫酸法测定总糖质量分数约为34.5%(质量比),与FTIR分析结果基本相符。生物絮凝剂在pH 4.0-11.0保持60%以上的絮凝效率,说明无论是酸性或是碱性条件下絮凝效率都较高,结合Zeta电位的分析表明,推测生物絮凝剂对高温栅藻的絮凝机理中占主导地位的可能是吸附架桥作用。[结论]该研究对微藻生物絮凝具有重要的理论和实践意义。     

NO EVIDENCE FOR AN ALLELOPATHIC ROLE OF OKADAIC ACID AMONG CIGUATERA-ASSOCIATED DINOFLAGELLATES

Serine/threonine protein phosphatases are regulatory enzymes critical to growth and replication in eukaryotes. Okadaic acid (OA), a potent serine/ threonine protein phosphatase inhibitor, has been well studied in vertebrates; however, little is known about the role this phycotoxin plays in the ecology of its producer, the benthic dinoflagellate Prorocentrum lima Ehrenberg. The frequency and diversity of toxins produced by benthic dinoflagellates suggests that toxin production might play a role in competition among algal species. To assess the role of OA in growth competition, dinoflagellate species that co-occur with P. lima were grown in medium preconditioned by P. lima. Exudates from P. lima inhibited the growth of three of the four co-occurring species. All four dinoflagellate species were found to possess protein phosphatases that are inhibited by low concentrations of OA (IC₅₀∼5 nM). Furthermore, when treated in culture for 24 h with either purified OA or medium preconditioned by P. lima, their protein phosphatase activity was substantially inhibited. In contrast, P. lima 's protein phosphatase activity was refractory to OA both in vitro and in culture. To determine whether its growth inhibitory action was dependent on its phosphatase inhibitory activity, the P. lima -preconditioned medium was fractionated by HPLC and fractions tested for growth-inhibitory and protein phosphatase–inhibitory activity. We found that the two activities reside in different fractions. These results suggest that although OA has growth-inhibitory potential against other microalgae, it does not represent the major growth-inhibitory activity found in P. lima 's medium.     

Global research trends on bioflocculant potentials in wastewater remediation from 1990 to 2019 using a bibliometric approach

The preference of biofloculants over chemical flocculants in water and wastewater remediation systems has gained wider attention due to their biodegradability, innocuousness, safety to human and environmental friendliness. The present study aimed to evaluate research outputs on bioflocculant potentials in wastewater remediation from 1990 to 2019 using bibliometric analyses. To the best of our knowledge, this is the first bibliometric report in bioflocculant research. The subject bibliometric dataset was extracted from the Web of Science Core Collection (WoSCC) and Scopus using the Boolean, ‘bioflocculant* and waste*’ and analysed for indicators such as a yearly trend, productivity (authors, articles, country, institution and journal source), conceptual framework and collaboration network. We found 119 documents with 347 authors from 78 journal sources on the subject, an annual growth rate of 12·1%, and average citations/document of 15·08. Guo J. and Wang Y. were the top researchers with 15 and 12 outputs respectively. China (42%) and South Africa (9·24%) ranked the top two dominant countries in the field. The top journals were Bioresource Technology (9 papers, 506 citations), Applied Microbiology and Biotechnology (5 papers, 268 citations), whereas, the top institution was Chengdu University of Information and Technology (n = 9 documents) followed by Sichuan Univ. Sci. & Engn, China (n = 8 documents). This study found that lack of intercountry collaboration and research funding adversely affects research participants in the field.     

Harvest of Scenedesmus sp. with bioflocculant and reuse of culture medium for subsequent high-density cultures

The optimal flocculating conditions for harvesting high-density cultures of Scenedesmus sp. were investigated using inorganic coagulants and the bioflocculant produced by Paenibacillus polymyxa AM49. The flocculated medium as nutrients for subsequent algal cultivation was also tested. Consecutive treatment with 8.5 mM CaCl(2) and 0.2 mM FeCl(3) as coagulants and 1% bioflocculant from the culture broth of P. polymyxa AM49 showed the highest flocculating activity of up to 95% for high density algal cultures. The medium flocculated with the coagulants and bioflocculant showed less than 8% decrease in the growth yield in the subsequent algal cultivation. Furthermore, a 20% or 50% fresh BG11 medium supplement allowed the flocculated medium to maintain a high growth yield in subsequent algal cultivation. These results suggest that the flocculation method presented here is efficient and bio-friendly, and allows the reuse of the flocculated medium, thereby contributing to the economic cultivation and harvest of microalgae.     

Benthic microbial production of oxygen supersaturates the bottom water of a stratified hypersaline lake

Lake Hayward is a hypersaline lake that stratifies seasonally and maintains oxygen supersaturation in its bottom water for about 6 months each year. This phenomenon was found to be the result of photosynthesis by the benthic microbial communities, composed primarily of the cyanobacteria Cyanothece spp., Spirulina sp., and Oscillatoria sp. When these communities were present and the lake was stratified, the bottom water was supersaturated with oxygen (up to 370%). During illumination, the benthic microbial communities rapidly developed very high concentrations (e.g., >500 M) of oxygen, which then diffused into the overlying water. However, while the overlying water became supersaturated, the concentration in the water was lower than in the benthic microbial communities because (1) transport across the sediment-water interface was limited by diffusion, and (2) turbulence rapidly mixed the oxygen throughout the much larger volume of the bottom water (approximately 1.5 m deep). Thus, import of oxygen by the benthic microbial communities at night proceeded more slowly than daytime export, allowing supersaturation of the bottom water to develop.     

Manipulation of MS and B5 components for enhancement of growth and solasodine production in hairy root cultures of <Emphasis Type="Italic">Solanum khasianum </Emphasis>Clarke

Hairy root cultures of Solanum khasianumClarke were studied for growth and solasodine production by manipulating the major and minor salts, Fe-EDTA and vitamins of MS and B5 medium. Tissue growth and solasodine production are strongly affected by the culture medium. The experiments demonstrated that in part the effect of each componenet of B5 and MS is dependent on the growth phase of the roots. Solasodine production increased with a decrease in total nitrogen. The ratio between ammonium nitrate and potassium nitrate was crucial. Phosphate concentration greatly influences growth, although it showed no significant affect on solasodine production. Half strength of magnesium sulphate and calcium chloride as in MS medium increased growth and solasodine production as compared to the control. This combination of the major salts with minor salts from the MS medium and chelated iron and vitamins from Gamborgs B5 medium was found to be most suitable. Hence it was possible to enhance both the specific and volumetric yield of solasodine by formulating a medium that maximized secondary metabolite production and growth in hairy root cultures. The novelty of this article lies in manipulating the components in MS medium, which supported growth, and Gamborgs B5 medium, which supported secondary product formation in hairy root cultures of S. khasianum and combining the two for maximizing both the parameters through out the growth phase. Thus it shall find immense practical applicability in hairy root scale up, where the secondary metabolite production is directly related to growth.     

RECRUITMENT OF BENTHIC MICROCYSTIS (CYANOPHYCEAE) TO THE WATER COLUMN: INTERNAL BUOYANCY CHANGES OR RESUSPENSION?1

In some lakes, large amounts of the potentially toxic cyanobacterium Microcystis overwinter in the sediment. This overwintering population might inoculate the water column in spring and promote the development of dense surface blooms of Microcystis during summer. In the Dutch Lake Volkerak, we found photochemically active Microcystis colonies in the sediment throughout the year. The most vital colonies originated from shallow sediments within the euphotic zone. We investigated whether recruitment of Microcystis colonies from the sediment to the water column was an active process, through production of gas vesicles or respiration of carbohydrate ballast. We calculated net buoyancy, as an indication of relative density, using the amounts and densities of the major cell constituents (carbohydrates, proteins, and gas vesicles). Carbohydrate content of benthic Microcystis cells was very low throughout the year. Buoyancy changes of benthic Microcystis were mostly a result of changes in gas vesicle volume. Before the summer bloom, net buoyancy and the amount of buoyant colonies in the sediment did not change. Therefore, recruitment of Microcystis from the sediment does not seem to be an active process regulated by internal buoyancy changes. Instead, our observations indicate that attachment of sediment particles to colonies plays an important part in the buoyancy state of benthic colonies. Therefore, we suggest that recruitment of Microcystis is more likely a passive process resulting from resuspension by wind‐induced mixing or bioturbation. Consequently, shallow areas of the lake probably play a more important role in recruitment of benthic Microcystis than deep areas.     

干旱胁迫条件下AMF促进小马鞍羊蹄甲幼苗生长的机理研究

采用温室水分控制试验,在干旱胁迫条件下,定量化研究优势丛枝菌根真菌(AMF)影响优势乡土植物小马鞍羊蹄甲(Bauhinia faberi var.microphylla)幼苗生长的机理,主要通过研究干旱胁迫条件下摩西球囊霉菌(Funneliformis mosseae)与小马鞍羊蹄甲的共生关系,阐明AMF在植物生长初期的作用。结果表明,干旱胁迫条件下,摩西球囊霉菌能够很好地侵染幼苗,侵染率高达89%—97%,并且不受水分条件影响。接种的幼苗最大光合速率、水分利用效率随着干旱胁迫程度从重度到轻度(水分从低到高)逐渐增大,相反地,叶片脯氨酸含量逐渐减小。接种显著地促进幼苗株高、叶片数、叶面积、根长、根面积等生长指标,提高幼苗各部分生物量、地上地下磷(P)含量。当含水量为60%田间持水量时,AMF促进小马鞍羊蹄甲幼苗吸收P的效果最好。接种还显著影响幼苗的生物量分配,在重度干旱胁迫时影响P分配,水分条件也显著影响幼苗的生物量分配。此外,接种和水分的交互作用对叶生物量、总生物量、生长指标以及地上部氮(N)总量影响显著。结果表明干旱胁迫条件下菌根效应显著,并在干旱条件下显著促进了小马鞍羊蹄甲幼苗的生长,这为进一步干旱河谷植被恢复提供了理论依据。     

Biological,chemical and physical factors influencing the seasonal change of polysaccharide concentration in irrigation-channel sediment

Variation of polysaccharide concentration in irrigation-channel sediment was determined concurrently with biological, chemical and physical factors influencing the benthic algal community. Phenol-sulphuric acid method was used to measure polysaccharide concentration. Polysaccharide concentration, biomass of benthic algae, and species composition changed spatially and temporally. Fluctuations of total suspended solid (TSS) concentration and exposure of channel bed to direct sunlight had major effects on algal growth and polysaccharide production. Polysaccharide concentration was correlated to chlorophyll a concentration (r=0.73, P<0.001) and algal biomass (r=0.57, P<0.001). Fragilaria construens and Aulacoseira (Melosira) italica were the most common diatoms in the benthic flora. Chlorophyll a concentration in the sediment showed a strong negative correlation (r=-0.99, P<0.001) with the seasonal variation of TSS concentration in channel water. The polysaccharides produced by benthic microorganisms play a major role in clogging channel bed and thereby reducing seepage from earthen irrigation channels. Correlations between polysaccharide concentration and chlorophyll a (and algal biomass) further indicate the importance of benthic algae for polysaccharide production. Since availability of light to the algal flora is critical for the production of polysaccharides, the effect of clogging can be maximized by exposing the channel bed to direct sunlight during non-irrigation period (winter).     

Purification and Characterization of Lipid Bioflocculant Produced by Rhodococcus erythropolis

The bioflocculant produced by Rhodococcus erythropolis S-1 was found to exist as huge assemblies, the molecular mass of which is over one million daltons, composed of many polypeptides and lipids in aqueous solution. We have isolated and purified this lipid bioflocculant by ultracentrifugation, extracting with 90% acetone, and two successive silica gel chromatographies from the culture broth. It was homogeneous on silica gel thin-layer chromatography. ¹H-NMR and HPLC studies showed that it was a kind of glycolipid that contained a C₁₆ methylene chain on the average and glucose in its chemical structure. The flocculating activity against kaolin clay suspension was dependent on the Ca²⁺ concentration.     

Medium optimization for the production of a novel bioflocculant from Halomonas sp. V3a′ using response surface methodology

The novel exopolysaccharide bioflocculant HBF-3 is produced by Halomonas sp. V3a′, which is a mutant strain of the deep-sea bacterium Halomonas sp. V3a. Response surface methodology (RSM) was employed to optimize the production medium for increasing HBF-3 production. Using a Plackett–Burman experimental design to aid in the first step of optimization, edible glucose, MgSO₄·7H₂O, and NH₄Cl were found to be significant factors affecting HBF-3 production. To determine the optimal concentration of each significant variable, a central composite design was employed. Based on response surface and canonical analysis, the optimum concentrations of the critical components were obtained as follows: edible glucose, 16.14 g/l; MgSO₄·7H₂O, 2.73 g/l; and NH₄Cl, 1.97 g/l. HBF-3 production obtained by using the optimized medium was 4.52 g/l, which was in close agreement with the predicted value of 4.55 g/l. By scaling up fermentation from flask to fermenter, HBF-3 production was further increased to 5.58 g/l.     

Effect of Cullural Conditions on the Growth of Agrostemma githago Cells in Suspension Culture and the Concomitant Production of an Anti-Plant Virus Substance

An extract of cultured Agroxieinma githago L. cells was found to show potent inhibitory activity against plans virus infection. The effects of cultural conditions on the growth of the cell suspension and on the production of the inhibitor were examined. Since the production of substance was dependent on growth. experiments were made to improve growth. The optimum temperature was 26 to 30°C and optimum pH of the medium before autoclaving was between 5 and 7. In a medium of higher osmotic pressure, the water content of the cultured cells was lowered markedly. The growth rate in a small volume of the medium was higher than that in a larger volume at an early stage of the cultivation, but it was not changed by different inoculum sizes. The cells required thiamine and 2,4-D for growth but no other vitamins or growth regulators. The optimum level of 2,4-D was 0.1 mg/l. Higher sucrose concentration in the medium gave higher production of cell mass and of the inhibitor. However, 3% of sucrose was selected as the most economical concentration. For normal cell growth, the presence of both NH₄NO₃ and KNO₃ as nitrogen sources was required. The use of a single nitrogen source caused a long lag period or inhibition of the cell growth. KH₂PO₄ stimulated the growth when in was used in the level of 2.5 to 5 mM. The cell adhesion on the surface of the fermentor sometimes causes trouble in a large-scale cultivation. It was found that reducing the Ca²⁺ level in the medium prevented the cell adhesion and foaming remarkably. Based on the results obtained, a modified medium was established which was excellent for shortening the culture period and for efficient production of the anti-plant virus inhibitor.