Liquid Culture Conditions Research Articles

A methyltransferase LaeA regulates ganoderic acid biosynthesis in Ganoderma lingzhi.

The methyltransferase LaeA is a global regulator involved in the biosynthesis of secondary metabolites by ascomycete fungi. However, little is known of its regulatory role in basidiomycete fungi. In this study, the laeA gene was identified in the basidiomycete Ganoderma lingzhi and its function in regulating the biosynthesis of anti-tumor ganoderic acids was evaluated. A laeA deletion (ΔlaeA) Ganoderma strain exhibited significantly reduced concentration of ganoderic acids. qRT-PCR analysis further revealed that the transcription levels of genes involved in the biosynthesis of ganoderic acids were drastically lower in the ΔlaeA strain. Moreover, deletion of laeA resulted in decreased accumulation of intermediates and abundances of asexual spores in liquid static culture of G. lingzhi. In contrast, constitutive overexpression of laeA resulted in increased concentration of ganoderic acids. These results demonstrate an essential role of LaeA in the regulation of ganoderic acid biosynthesis in Ganoderma.

Regeneration of tea [<em>Camellia sinensis</em> (L.) Kuntze] cv. TRI 5001 plants through somatic embryogenesis in a liquid dynamic medium

Somatic embryogenesis is an efficient micro-propagation technique, which can lead to mass multiplication of tea with a tap root. Hence, the development of a viable somatic embryogenesis and multiplication protocol through liquid culture conditions is needed to increase the efficiency of the protocol. Cotyledon-derived somatic embryos, leaf and stem callus of TRI 5001 cultivar, were tested with different liquid MS media. A significantly higher cotyledon-derived somatic embryo multiplication rate was observed in MS +3 mg/L Thidiazuron (TDZ) medium. Callus proliferation and compact and embryonic callus formation were comparatively higher in MS + 1.1 mg/L TDZ, 1.86 mg/L NAA medium, and subsequent somatic embryo formation was observed in MS + 0.1 mg/L NAA,1 mg/L BAP and 0.2 mg/L GA3 medium on callus produced on leaf explants. Higher nodal callus proliferation and compact callus formation were recorded in MS+0.11 mg/L TDZ 0.1 mg/L IBA and higher embryonic callus was observed in MS+0.1 mg/L TDZ, 1.86 mg/L NAA media. Plant regeneration was resulted in MS+3 mg/LBAP 0.1 mg/L NAA medium. Identified treatment combinations can be used as a foundation to establish somatic embryogenesis protocol through different explants of tea.

Acceleration of Polybutylene Succinate Biodegradation by Terribacillus sp. JY49 Isolated from a Marine Environment.

Polybutylene succinate (PBS) is a bioplastic substitute for synthetic plastics that are made from petroleum-based products such as polyethylene and polypropylene. However, the biodegradation rate of PBS is still low and similar to that of polylactic acid (PLA). Moreover, our knowledge about degrader species is limited to a few fungi and mixed consortia. Here, to identify a bacterial degrader to accelerate PBS degradation, we screened and isolated Terribacillus sp. JY49, which showed significant degradability. In order to optimize solid and liquid culture conditions, the effect of factors such as temperature, additional carbon sources, and salt concentrations on degradation was confirmed. We observed a degradation yield of 22.3% after 7 days when adding 1% of glucose. Additionally, NaCl was added to liquid media, and degradation yield was decreased but PBS films were broken into pieces. Comparing the degree of PBS degradation during 10 days, the degradation yield was 31.4% after 10 days at 30 °C. Alteration of physical properties of films was analyzed by using scanning electron microscopy (SEM), gel permeation chromatography (GPC), and Fourier transform infrared (FT-IR). In addition, Terribacillus sp. JY49 showed clear zones on poly(butylene adipate-co-terephthalate) (PBAT), polycaprolactone (PCL), and copolymers such as P(3HB-co-3HV) and P(3HV-co-4HB), exhibiting a broad spectrum of degradation activities on bioplastics. However, there was no significant difference in absorbance when esterase activity was examined for different types of bioplastics. Overall, Terribacillus sp. JY49 is a potential bacterial strain that can degrade PBS and other bioplastics, and this is the first report of Terribacillus sp. as a bioplastic degrader.

Enhancement of cordycepin production from Cordyceps militaris culture by epigenetic modification.

Cordycepin (3'-deoxyadenosine) is a nucleoside analogue and biosynthesised by Cordyceps militaris, an entomopathogenic fungus. In this study, an epigenetic modifier was applied to static liquid cultures to enhance cordycepin production. C. militaris was cultured in a static liquid culture, and valproic acid (VPA), a histone deacetylase (HDAC) inhibitor, was supplemented in order to modifying the epigenetic status. Gene regulatory network was explored to understand the molecular mechanisms underlying cordycepin production. 50 micromolar of VPA enhanced cordycepin production by 41.187% via the upregulation of 5'-nucleotidase, adenylate kinase, phosphorybosyltransferase, Cns1, Cns2, Cnsa3, and Cns4 of C. militaris for at least 2days after VPA treatment. The maximum production of cordycepin was 2,835.32 ± 34.35mg/L in 400mL-working volume. A scaled-up culture was established with a working volume of 10 L, which led to the slight decrease of cordycepin production. This might due to multifactorial effects, for instance limited aeration and an uneven dispersion of nutrients in the culture system. This scaled-up culture was still needed further optimization. The modification of epigenetic status by VPA significantly enhanced cordycepin production by altering key gene regulatory network of C. militaris. The strategy established in this study might be applicable to other microorganism culture in order to improving the production of bioactive compounds. This work aimed to enhance the production of cordycepin by modifying the epigenetic status of C. militaris, in which subsequently altered gene regulatory network of cordycepin biosynthesis pathway. The weekly supplementation of valproic acid (VPA), a histone deacetylase (HDAC) inhibitor, significantly improve cordycepin production over 40%, compared to the untreated control, and the gene regulatory network of C. militaris was also adapted.

Physiological responses of Aechmea setigera Mart. ex Schult. & Schult. f. (Bromeliaceae) in vitro cultured

In vitro propagation of bromeliads is widely used to produce plantlets intended for conservation. This study aims at evaluating physiological responses of Aechmea setigera in vitro cultured under different concentrations of 6-benzylaminopurine (BAP). The experiment was conducted in completely randomized design with five treatments (0, 1.0, 2.0, 4.0 and 8.0 mg L-1 of BAP). After 90 days of in vitro cultured in stationary liquid MS culture medium it were assessed morphological and biochemical parameters. The BAP increment increased stomatal density, reduced plant size and root number, but increased the number of shoots. There was significant effect of BAP dose on fresh and dry mass. High BAP doses (4.0 and 8.0 mg L-1) promoted the reduction of chlorophyll a and total chlorophyll content, but there was no change of chlorophyll b and carotenoids. The concentration of starch and total soluble carbohydrates decreased above 2.0 and 4.0 mg L-1 of BAP, respectively. The total phenolic content decreased with increasing BAP concentrations. Evaluated changes in metabolism elucidate the phenotypic plasticity for in vitro multiplication process in A. setigera.

High production of jasmonic acid by Lasiodiplodia iranensis using solid-state fermentation: Optimization and understanding.

Jasmonic acid (JA) is a plant hormone involved in regulating developmental and growth controls as well as photosynthesis. In addition, this hormone protects the plant against insects and has good applications in agriculture, the flavored industry and other fields. Filamentous fungus generally produces JA using liquid static culture. In the present study, a solid-state fermentation (SSF) method is developed for high production of JA using Lasiodiplodia iranensis. By selecting the solid substrate and optimizing the initial water content, inoculum volume, loading volume and other culture conditions, the maximum JA yield reached 5306.38mg kg-1 when fermented for 12 days in a petri dish containing a medium with crushed wheat as the solid substrate and 75% initial water content. The logistic and Luedeking-Piret models were used to characterize the relationship between microbial growth and product synthesis in the SSF process, and the maximum JA production is predicted to be 5263.23mg kg-1 , which is close to the experimental value. Liquid chromatography with tandem mass spectrometry (LC-MS/MS) is used to examine the metabolic changes that develop during fermentation. The results indicate that JA biosynthesis occurs in the α-linolenic acid metabolic pathway, of which 13(S)-HpOTrE is a key intermediate metabolite and both 13(S)-HOTrE and traumatic acid are byproducts of the branches of its synthesis. The results of this study provide a method for obtaining high JA yields by SSF, and offer new insights for understanding the production of JA by fungal fermentation.

Lindane removal in contaminated soil by defined microbial consortia and evaluation of its effectiveness by bioassays and cytotoxicity studies.

Lindane contamination in different environmental matrices has been a global concern for long. Bacterial consortia consisting of Paracoccus sp. NITDBR1, Rhodococcus rhodochrous NITDBS9, Ochrobactrum sp. NITDBR3, NITDBR4 and NITDBR5 were used for the bioremediation of soil artificially contaminated with lindane. The bacteria, Paracoccus sp. NITDBR1 and Rhodococcus rhodochrous NITDBS9, have been selected based on their lindane degrading capacity in liquid culture conditions (~80-90 %). The remaining three bacteria were chosen for their auxiliary properties for plant growth promotion, such as nitrogen fixation, phosphate solubilization, indole-3-acetic acid production and ammonia production under in vitro conditions. In this study, market wastes, mainly vegetable wastes, were added to the soil as a biostimulant to form a biomixture for assisting the degradation of lindane by bioaugmentation. Residual lindane was measured at regular intervals of 7 days to monitor the biodegradation process. It was observed that the consortium could degrade ~80% of 50 mgkg-1 lindane in soil which was further increased in the biomixture after six weeks of incubation. Bioassays performed on plant seeds and cytotoxicity studies performed on human skin fibroblast and HCT116 cell lines revealed that the groups contaminated with lindane and treated with the bacterial consortium showed lower toxicity than their respective controls without any bacteria. Hence, the use of both pesticide degrading and plant growth-promoting bacteria in a consortium can be a promising strategy for improved bioremediation against chemical pesticides, particularly in soil and agricultural fields, simultaneously enhancing crop productivity in those contaminated soil.

Optimization of mycelial growth of <i>Xylaria papulis</i> Lloyd (Xylariaceae) in indigenous liquid culture conditions, science city of Muñoz, Nueva Ecija, Philippines

The search for potential and inexhaustible sources of bioactive compounds with great functional activities is imperative for potential drug leads in treating various human diseases. In this regard, this study investigated the optimum liquid medium and physical culture conditions for the mycelial growth of <italic>X. papulis</italic> Lloyd for nutraceutical studies. The greatest mycelial biomass was achieved in PSB liquid medium among the indigenous media evaluated, such as potato sucrose broth (PSB), corn meal broth (CMB), rice bran broth (RBB), and coconut water (CW). In terms of pH, <italic>X. papulis</italic> Lloyd was observed to grow in a wide range of pH (5.0−8.5), but the best mycelial growth was observed at pH 6.5. Room temperature of 28 °C, lighted (137 lux), and static conditions were the other optimum physical culture conditions for mycelial growth of <italic>X. papulis</italic> Lloyd.

The Manganese Peroxidase Gene Family of Trametes trogii: Gene Identification and Expression Patterns Using Various Metal Ions under Different Culture Conditions.

Manganese peroxidases (MnPs), gene family members of white-rot fungi, are necessary extracellular enzymes that degrade lignocellulose and xenobiotic aromatic pollutants. However, very little is known about the diversity and expression patterns of the MnP gene family in white-rot fungi, especially in contrast to laccases. Here, the gene and protein sequences of eight unique MnP genes of T. trogii S0301 were characterized. Based on the characteristics of gene sequence, all TtMnPs here belong to short-type hybrid MnP (type I) with an average protein length of 363 amino acids, 5–6 introns, and the presence of conserved cysteine residues. Furthermore, analysis of MnP activity showed that metal ions (Mn2+ and Cu2+) and static liquid culture significantly influenced MnP activity. A maximum MnP activity (>14.0 U/mL) toward 2,6-DMP was observed in static liquid culture after the addition of Mn2+ (1 mM) or Cu2+ (0.2 or 2 mM). Moreover, qPCR analysis showed that Mn2+ obviously upregulated the Group I MnP subfamily (T_trogii_09901, 09904, 09903, and 09906), while Cu2+ and H2O2, along with changing temperatures, mainly induced the Group II MnP subfamily (T_trogii_11984, 11971, 11985, and 11983), suggesting diverse functions of fungal MnPs in growth and development, stress response, etc. Our studies here systematically analyzed the gene structure, expression, and regulation of the TtMnP gene family in T. trogii, one of the important lignocellulose-degrading fungi, and these results extended our understanding of the diversity of the MnP gene family and helped to improve MnP production and appilications of Trametes strains and other white-rot fungi.

Removal of lindane in liquid culture using soil bacteria and toxicity assessment in human skin fibroblast and HCT116 cell lines

ABSTRACT The development of effective measures for the remediation of lindane contaminated sites is the need of the hour. In this study, a potent lindane degrading bacteria, identified as Rhodococcus rhodochrous NITDBS9 was isolated from an agricultural field of Odisha that could utilize up to 87% of 100 mg L−1 lindane when grown under liquid culture conditions in mineral salt media in 10 days. The bacteria could produce biofilm in lindane-containing media. Rhodococcus rhodochrous NITDBS9 was further characterized for its plant growth-promoting properties and it was found that the bacteria showed abilities for phytohormone, ammonia and biosurfactant production, etc. This could be beneficial for the bioremediation and improvement of crop production in contaminated sites. Ecotoxicity studies carried out for lindane, and its degradation products in mung bean and mustard seeds showed a reduction in toxicity of lindane after treatment with NITDBS9. NITDBS9 was used with a previously isolated potent lindane degrading strain Paracoccus sp. NITDBR1 in a dual mixed culture for the enhanced removal of lindane in the liquid system i.e. up to 93% in 10 days. Cytotoxicity studies were conducted with lindane before and after treatment with the single and dual mixed cultures on human skin fibroblast and HCT116 cell lines. They revealed a significant reduction in toxicity of lindane after it was bioremediated with the single and dual mixed cultures. Therefore, our proposed strategy could be efficiently used for the detoxification of the lindane-contaminated system, and further work should be done to study the use of these cultures in the contaminated soil system.

Cellulase Production and Characterization from Thermomucor indicae-seudaticae

Cellulases are one of the important groups of industrial hydrolases which hydrolyze lignocellulosics into glucose, hence find numerous applications in the food, feed and beverage industry, textile, pulp and paper industry and biofuel production. With a view of industrial importance of cellulases, this study aimed to screen the cellulolytic potential of fungus Thermomucor indicae-seudaticae sp. nov. The zone of cellulose hydrolysis producing around the growth of fungus confirmed its cellulase-producing ability. Further, the enzyme production was carried out under liquid fermentation and various culture conditions were optimized. Among the physical conditions, maximum enzyme production was obtained at pH 7.0, incubation temperature of 50°C and incubation period of 7 days. Corn cob as carbon source followed by rice straw enhanced the enzyme production. Yeast extract was found to be the best nitrogen source. The crude enzyme exhibited highest enzyme activity at pH 7.0 and temperature 45oC. Among the various additives tested, some metal ions (Na+, K+, Ca+2 and Mg+2) stimulated the enzyme activity while Hg+2 strongly inhibited the enzyme. Among the detergents, SDS had stimulatory effect while Triton X-100 was found inhibitory for cellulase.

Development of high frequency cost-effective micropropagation protocol for Juncus rigidus using liquid culture medium and extraction of cellulose from their in vitro shoots - An important rush

Juncus rigidus is important halophyte with high cellulose content, but no studies has been made on micropropagation of the plant and cellulose extraction from in vitro shoots. To optimize an effective system for shoot multiplication, explants were grown in agar medium, liquid static culture, and bioreactor. Best shoot bud induction was obtained in liquid MS medium supplemented with2.27 μM thidiazuron (TDZ) over 0.46–4.65 μM Kinetin (KN) and 2.22–6.66 μM 6-benzyl aminopurine (BAP). Maximum number of shoot bud (36 ± 1.3) were achieved in static liquid MS medium supplemented with 2.22 μM BAP and 0.57 μM indole acetic acid (IAA). Upon subsequent subcultures >80 shoot buds were achieved in static liquid MS medium supplemented with 1.11 μM BAP, 0.57 μM IAA, and 0.46 μMKN within 30 days of culture. Being halophytic plant NaCl did not have any significant effect on shoot proliferation. Elongated shoots cultured in MS medium supplemented with 0.44 μM BAP, and 0.57 μM IAA developed 97% rooting in filter paper boat culture vessel. Sapling production cost in liquid culture medium was 90% less as compared to solid medium. Cellulose was extracted from the in vitro shoots and 17.64% cellulose achieved from liquid medium cultured shoots. For the first time micro-shoots are tested for in vitro cellulose production. • Maximum shoot bud (36 ± 1.2) induction was achieved in static liquid medium. • More than 80 shoot buds generated in static liquid MS medium supplemented with BAP, IAA, and KN. • 33.4 multiplication rate was recorded in static liquid medium. • 17.64 % cellulose extracted form in vitro shoots of J. rigidus. • In vitro plant based cellulose synthesis is important with future industrial application and bioengineering.

Determination of antifungal efficiency of some fungicides and secondary metabolites of Trichoderma species against Botrytis cinerea

Aim: The present study aimed to investigate the antifungal effects of fenhexamid, cyprodinil+fludioxanil, Bacillus subtilis QST 713, Trichoderma secondary metabolites (EGE-K-38 and EGE-K-71) on 6 isolates of necrotrophic fungi (Botrytis cinerea) under in- vitro conditions. Methodology: A dual culture technique was used to evaluate the effects of volatile compounds produced by Trichoderma in culture media against Botrytis cinerea isolates. For non-volatile and volatile metabolites, measurements for growth inhibition were made with the mixture of filter sterilized liquid medium of Trichoderma and potato dextrose agar. The fungicidal effects of these non-volatile metabolites and fungicides were first evaluated according to the effective concentration (EC50) data of growing mycelium inhibition of disease agents. Results: According to the results obtained, the drug/agent with the lowest fungicidal activity was determined as fenhexamide, which seems to have the lowest EC50 value of 0.05 ug m l-1. Both non-volatile metabolites and volatile metabolites of Trichoderma strains showed strong inhibition against B. cinerea isolates under experimental conditions. The highest growth inhibition percentage was determined with volatile metabolites of Trihoderma atroviride EGE-K-71 strain as 71.8%. Interpretation: The study suggest that filtrates that contain secondary metabolites and volatile compounds of Trichoderma strains produced in both liquid culture and dual culture conditions can be used as an effective fungal control agents against pre and postharvest contamination of grapes with Botrytis cinerea.

Tolerance to Abiotic Factors of Microsclerotia and Mycelial Pellets From Metarhizium robertsii, and Molecular and Ultrastructural Changes During Microsclerotial Differentiation.

Metarhizium species fungi are able to produce resistant structures termed microsclerotia, formed by compact and melanized threads of hyphae. These propagules are tolerant to desiccation and produce infective conidia; thus, they are promising candidates to use in biological control programs. In this study, we investigated the tolerance to both ultraviolet B (UV-B) radiation and heat of microsclerotia of Metarhizium robertsii strain ARSEF 2575. We also adapted the liquid medium and culture conditions to obtain mycelial pellets from the same isolate in order to compare these characteristics between both types of propagules. We followed the peroxisome biogenesis and studied the oxidative stress during differentiation from conidia to microsclerotia by transmission electron microscopy after staining with a peroxidase activity marker and by the expression pattern of genes potentially involved in these processes. We found that despite their twice smaller size, microsclerotia exhibited higher dry biomass, yield, and conidial productivity than mycelial pellets, both with and without UV-B and heat stresses. From the 16 genes measured, we found an induction after 96-h differentiation in the oxidative stress marker genes MrcatA, MrcatP, and Mrgpx; the peroxisome biogenesis factors Mrpex5 and Mrpex14/17; and the photoprotection genes Mrlac1 and Mrlac2; and Mrlac3. We concluded that an oxidative stress scenario is induced during microsclerotia differentiation in M. robertsii and confirmed that because of its tolerance to desiccation, heat, and UV-B, this fungal structure could be an excellent candidate for use in biological control of pests under tropical and subtropical climates where heat and UV radiation are detrimental to entomopathogenic fungi survival and persistence.

Production of Monacolin K in Monascus pilosus: Comparison between Industrial Strains and Analysis of Its Gene Clusters.

Monascus pilosus strains are widely applied to yield a cholesterol synthesis inhibitor monacolin K (MK), also called lovastatin (LOV). However, the mechanism of MK production by M. pilosus strains is still unclear. In this study, we firstly confirmed four Monascus strains, MS-1, YDJ-1, YDJ-2, and K104061, isolated from commercial MK products as M. pilosus and compared their abilities to produce MK in solid-state and liquid-state cultures. Then, we sequenced and analyzed their genomes and MK biosynthetic gene clusters (BGCs). The results revealed that the MK yields of MS-1, YDJ-1, YDJ-2, and K104061 in solid-state cultures at 14 days were 6.13, 2.03, 1.72, and 0.76 mg/g, respectively; the intracellular and extracellular MK contents of MS-1, YDJ-1, YDJ-2, and K104061 in liquid-state cultures at 14 days reached 0.9 and 1.8 mg/g, 0.38 and 0.43 mg/g, 0.30 and 0.42 mg/g, and 0.31 and 0.76 mg/g, respectively. The genome sizes of the four M. pilosus strains were about 26 Mb, containing about 7000–8000 coding genes and one MK gene cluster. The MK BGCs of MS-1, YDJ-2, and K104061 contained 11 genes, and the MK BGC of YDJ-1 contained 9 genes. According to the literature search, there are few comparisons of gene clusters and related genes responsible for the synthesis of LOV and MK. We also compared the LOV BGC in A. terreus with the MK BGCs in different species of Monascus spp., and the results revealed that although LOV and MK were the same substance, the genes responsible for the synthesis of MK were much less than those for LOV synthesis, and the gene functions were quite different. The current results laid a foundation to explore the mechanism of MK produced by Monascus spp. and compare the synthesis of LOV and MK.

Enhancement of ganoderic acid production by promoting sporulation in a liquid static culture of Ganoderma species

Ganoderic acids (GAs) produced by Ganoderma are a type of lanostane-type triterpenoids with anticancer and antimetastatic activities; however, low production of GAs limits its wide application. In this study, a novel strategy by promoting sporulation of Ganoderma was developed to increase GA production. First, a high-spore producing Ganoderma strain G. 260125 was obtained from dikaryotic strain CGMCC 5.0026, and the sporulation-specific gene of this strain exhibits a higher transcription level than CGMCC 5.0026. Then, the effect of promoting sporulation on GA content was investigated. The maximum ganoderic acid (GA)-T, GA-Mk, and GA-Me contents in G. 260125 in shake flasks were 358.97, 78.32, and 12.75 μg/100 mg dry weight, respectively, which were 3.42, 2.91, and 1.73 times higher than those obtained in CGMCC 5.0026. Moreover, total and individual GA contents in spores were significantly higher than those in liquid static culture. Both concentrations of intermediates and transcription levels of GA biosynthetic genes also improved in G. 260125 during fermentation compared with those in CGMCC 5.0026. For scaling-up experiments, GA-T, GA-Me, and GA-Mk production in G. 260125 improved by 2.2-, 2.6-, and 2.1-fold compared with those in CGMCC 5.0026. In addition, the effectiveness of the developed strategy was also confirmed in three different Ganoderma strains. This work illustrated that promoting sporulation efficiently improves GA production in liquid static cultures of Ganoderma.

Impact of different fermentation characteristics on the production of mycelial biomass, extra-cellular polysaccharides, intra-cellular polysaccharides, and on the antioxidant activities of Cordyceps militaris (L.) Fr. (strains AG-1, PSJ-1)

The mycelial biomass, antioxidant activity and production of extra- and intra-cellular polysaccharides production [EPS, IPS] of Cordyceps militaris strains AG-1, PSJ-1 were evaluated under different submerged liquid culture (SLC) conditions. At 24 0C mycelial biomass and polysaccharide production of AG-1, PSJ-1 was optimal using PVC media and static culture conditions; (AG-1: 21.85 ± 1.00; PSJ-1: 18.20 ± 1.84 g l-1 ), and oven drying at 40 oC (AG-1: 25.95 ± 0.84, PSJ-1: 23.55 ± 0.69 mg g-1) compared with hot water extraction (AG-1: 7.07 ± 0.15, PSJ-1: 7.39 ± 0.61 mg g-1). Maximum biomass, EPS and IPS production were observed when the initial pH was 6.7: AG-1: 12.92 ± 0.33, 209.70 ± 1.56, 32.62 ± 0.87; PSJ-1: 9.03 ± 0.24 g l-1 , 198.16 ± 0.85 mg g-1, 30.63 ± 1.96 mg g-1, respectively. The use 3.5 % coconut oil improved biomass, EPS, IPS production, which were 8.27 ± 0.09, 8.01 ± 0.01 g l-1 ; 1208.00 ± 8.60, 1110.40 ± 7.20 mg g-1; 32.43 ± 0.49, 29.74 ± 0.44, for AG-1 and PSJ-1, respectively. Both culture condition and oven drying methods had significant effects on H2O2 and radical scavenging activity, ABTS.+ radical activity, lipid peroxidation, and also had effects on total flavonoid and, total phenolic contents. The use of crude submerged liquid culture and oven drying on strains AG-1, PSJ-1 led to extracts with potent antioxidant activity, suggesting the therapeutic use of polysaccharides from strains AG-1, PSJ-1.

Valorization of low-cost, carbon-rich substrates by edible ascomycetes and basidiomycetes grown on liquid cultures.

Three ascomycetes (Morchella vulgaris AMRL 36, M. elata AMRL 63, Tuber aestivum AMRL 364) and four basidiomycetes strains (Lentinula edodes AMRL 124 and 126, Agaricus bisporus AMRL 208 and 209) were screened for their ability to grow on liquid static flask cultures of glucose, glycerol, molasses and waste flour-rich hydrolysates with C/N ratio of 20 and produce biomass, exopolysaccharides and lipids. The profile of lipid fatty acids was also assessed. Selected strains were furthermore cultivated in C/N=50. Results showed that substrate consumption, biomass formation and secondary metabolites production were strain, substrate and C/N ratio dependent. The maximum biomass (X), lipid (L) and exopolysaccharides (EPS) values noted were Xmax=25.2g/L (C/N=20; molasses) and Lmax=6.51g/L (C/N=50; rice cereal hydrolysates) by T. aestivum strain AMRL 364 and EPSmax=2.41g/L by M. elata strain AMRL 63 (C/N=50; molasses), respectively. When C/N ratio of 50 was applied, biomass, lipid production and substrate consumption seem to be negatively affected in most of the trials. The adaptation and capability of the mushroom strains to be cultivated on substrates based on agro-industrial waste streams and infant food of expired shelf date offers the opportunity to set a circular oriented bioprocess.

KIN10 promotes stomatal development through stabilization of the SPEECHLESS transcription factor

Stomata are epidermal structures that modulate gas exchanges between plants and the atmosphere. The formation of stomata is regulated by multiple developmental and environmental signals, but how these signals are coordinated to control this process remains unclear. Here, we showed that the conserved energy sensor kinase SnRK1 promotes stomatal development under short-day photoperiod or in liquid culture conditions. Mutation of KIN10, the catalytic α-subunit of SnRK1, results in the decreased stomatal index; while overexpression of KIN10 significantly induces stomatal development. KIN10 displays the cell-type-specific subcellular location pattern. The nuclear-localized KIN10 proteins are highly enriched in the stomatal lineage cells to phosphorylate and stabilize SPEECHLESS, a master regulator of stomatal formation, thereby promoting stomatal development. Our work identifies a module links connecting the energy signaling and stomatal development and reveals that multiple regulatory mechanisms are in place for SnRK1 to modulate stomatal development in response to changing environments.

Optimization of Liquid Culture Condition of a Novel Fungus Hygrophoropsis sp. and Antioxidant Activity of Extracts.

To evaluate pharmacological activities of a novel fungus Hygrophoropsis sp., the influence of aeration rate on the production of mycelial biomass, exopolysaccharides (EPS), and intrapolysaccharides (IPS) in the fungus Hygrophoropsis sp. was investigated. And the water extracts of cultured Hygrophoropsis sp. mycelia and the fermentation broth were analyzed for their antioxidation activity by using four different assay methods such as hydroxyl radical scavenging, superoxide radical scavenging, hydrogen peroxide scavenging, and reducing power. The Hygrophoropsis sp. was cultivated under various aeration rates in a 7 l bioreactor. The highest mycelial biomass (3.98 mg/mL) and IPS production (19.63 mg/g) were obtained at aeration rate 4.5 v.v.m. The results showed that Hygrophoropsis sp., in general, possesses a strong antioxidation activity in all assays tested. The deproteinized extracts had stronger antioxidation activity as compared to the un-deproteinized extracts by using superoxide radical scavenging, hydrogen peroxide scavenging, and reducing power. Besides, the un-deproteinized extracts had stronger antioxidation activity as compared to the deproteinized extracts by using hydroxyl radical scavenging. Thus, the polysaccharide extractions from the Hygrophoropsis sp. studied have antioxidant activities in vitro, which may be a good source of natural antioxidants or further investigation as potential natural antioxidants.