Multifunctional in vitro, in silico and DFT analyses on antimicrobial BagremycinA biosynthesized by Micromonospora chokoriensis CR3 from Hieracium canadense.

Among the actinomycetes in the rare genera, Micromonospora is of great interest since it has been shown to produce novel therapeutic compounds. Particular emphasis is now on its isolation from plants since its population from soil has been extensively explored. The strain CR3 was isolated as an endophyte from the roots of Hieracium canadense, and it was identified as Micromonospora chokoriensis through 16S gene sequencing and phylogenetic analysis. The in-vitro analysis of its extract revealed it to be active against the clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and Candida tropicalis (15 mm). No bioactivity was observed against Gram-negative bacteria, Escherichia coli ATCC 25922, and Klebsiella pneumoniae ATCC 706003. The Micromonospora chokoriensis CR3 extract was also analyzed through the HPLC-DAD-UV-VIS resident database, and it gave a maximum match factor of 997.334 with the specialized metabolite BagremycinA (BagA). The in-silico analysis indicated that BagA strongly interacted with the active site residues of the sterol 14-α demethylase and thymidylate kinase enzymes, with the lowest binding energies of - 9.7 and - 8.3 kcal/mol, respectively. Furthermore, the normal mode analysis indicated that the interaction between these proteins and BagA was stable. The DFT quantum chemical properties depicted BagA to be reasonably reactive with a HOMO-LUMO gap of (ΔE) of 4.390 eV. BagA also passed the drug-likeness test with a synthetic accessibility score of 2.06, whereas Protox-II classified it as a class V toxicity compound with high LD50 of 2644 mg/kg. The current study reports an endophytic actinomycete, M. chokoriensis, associated with H. canadense producing the bioactive metabolite BagA with promising antimicrobial activity, which can be further modified and developed into a safe antimicrobial drug.

Prevalence of mycorrhizae in host plants and rhizosphere soil: A biodiversity aspect.

Plants roots are colonized by soil inhabitants known as arbuscular mycorrhizal fungi (AMF), which increase plant productivity, and enhance carbon storage in the soil. We found mycorrhizal vesicles, arbuscles, and mycelium in the root of more than 89% of the selected plants of University of Rajshahi campus, Bangladesh. The rate of their presence differed in plant to plant of a family and different families. The highest root colonization (98±1.0%) was found to be present in Xanthium strumarium (Asteraceae). Mycorrhiza was not found in the root of Sphagneticola calendulacea (Asteraceae), Cestrun nocturnum (Solanaceae), Acacia nilotica and Acacia catechu (Mimosoidae), Rorippa nasturtium, Brassica oleracla var botrytis (Brasicaceae), Punica granatum (Lythraceae), Tecoma capensis (Bignoniacea), Spinacia oleracia (Chenopodiaceae), Chenopodium album (Goosefoot). Result of soil analysis reveals that the rhizospheric soils were deficient in nutrients which might be suitable for mycorrhizal symbiosis with plants. In the rhizospheric soils, 22 species of Glomus, Scutelospora, Gigaspora, Archaeospora, and Acullospora were found. We also found the genera 'Glomus' dominance in the plant root and rhizospheric soil. So, it can be concluded that the highly colonized roots as well as spores can be used to prepare mycorrhizal inoculum for future purposes.

OSMAC Strategy Integrated with Molecular Networking for Accessing Griseofulvin Derivatives from Endophytic Fungi of Moquiniastrum polymorphum (Asteraceae).

Three endophytic fungi isolated from Moquiniastrum polymorphum (Less.) G. Sancho (Asteraceae) were cultivated using the one strain many compounds (OSMAC) strategy to evaluate the production of griseofulvin derivatives. Extracts obtained were analyzed by HPLC-MS/MS and the chromatographic and spectrometric data used to elaborate a feature-based molecular network (FBMN) through the GNPS platform. This approach allowed the observation of differences such as medium-specific and strain-specific production of griseofulvin derivatives and variations of cytotoxic activity in most extracts. To evaluate the efficiency of the OSMAC approach allied with FBMN analysis in the prospection of compounds of biotechnological interest, griseofulvin and 7-dechlorogriseofulvin were isolated, and the relative concentrations were estimated in all culture media using HPLC-UV, allowing for the inference of the best strain-medium combinations to maximize its production. Malt extract-peptone broth and Wickerham broth media produced the highest concentrations of both secondary metabolites.

Anti-inflammatory activity of endophytic bacterial isolates from Emilia sonchifolia (Linn.) DC.

ETHNOPHARMACOLOGICAL RELEVANCE: In the traditional medicine system, plants have been utilized as a rich source of anti-microbial, anti-inflammatory, anti-cancer, anti-viral and anti-oxidant compounds. The biological properties of plant-based drugs depend on their interaction with endophytes which persist as an important provider of bioactive secondary metabolites. Bacterial endophytes secrete anti-inflammatory molecules whose activity can be the base for the anti-inflammatory property of the plant. AIM OF THE STUDY: During the screening of endophytes from Emilia sonchifolia, we isolated six different bacteria whose potential as the sources of anti-inflamamtory compounds have been aimed at in this study. MATERIALS AND METHODS: Anti-inflammatory activity of the ethyl acetate extract of endophytes was studied by both in vitro and in vivo analyses. In vitro study was done using protein denaturation, COX, LOX, iNOS, myeloperoxidase and nitric oxide assays and in vivo analysis was carried out by carrageenan-induced and formalin-induced paw oedema tests. The expression level of anti-inflammatory genes such as COX-2 and NfKb was confirmed by real time PCR. RESULTS: We confirmed anti-inflammatory activity of the ethyl acetate extract of bacterial endophytes of E sonchifolia by both in vitro and in vivo experiments. Carrageenan- and formalin-induced inflammations in mice were effectively reduced by the administration of the bacterial extract. Among the isolates, strain ES1effectively reduced inflammation. Gene expression studies confirmed reduction in the expression of COX-2 and NfKb genes in the presence of ES1 extract. CONCLUSION: The present investigation demonstrated the anti-inflammatory property of the isolated bacterial endophyte ES1 (Bacillus subtilis strain-MG 692780) and thus justifies the possible role of endophytes in contributing anti-inflammatory property to E sonchifolia which is ethno-botanically important as a source of anti-inflammatory drug.

Invasive Lactuca serriola seeds contain endophytic bacteria that contribute to drought tolerance.

The mutualistic relationship between alien plant species and microorganisms is proposed to facilitate or hinder invasive success, depending on whether plants can form novel associations with microorganisms in the introduced habitats. However, this hypothesis has not considered seed endophytes that would move together with plant propagules. Little information is available on the seed endophytic bacteria of invasive species and their effects on plant performance. We isolated the seed endophytic bacteria of a xerophytic invasive plant, Lactuca serriola, and examined their plant growth-promoting traits. In addition, we assessed whether these seed endophytes contributed to plant drought tolerance. Forty-two bacterial species were isolated from seeds, and all of them exhibited at least one plant growth-promoting trait. Kosakonia cowanii occurred in all four tested plant populations and produced a high concentration of exopolysaccharides in media with a highly negative water potential. Notably, applying K. cowanii GG1 to Arabidopsis thaliana stimulated plant growth under drought conditions. It also reduced soil water loss under drought conditions, suggesting bacterial production of exopolysaccharides might contribute to the maintenance of soil water content. These results imply that invasive plants can disperse along with beneficial bacterial symbionts, which potentially improve plant fitness and help to establish alien plant species.

Compact shoot architecture of Osteospermum fruticosum transformed with Rhizobium rhizogenes.

KEY MESSAGE: Improved compact shoot architecture of Osteospermum fruticosum Ri lines obtained through Rhizobium rhizogenes transformation reduces the need for chemical growth retardants. Compactness is for many ornamental crops an important commercial trait that is usually obtained through the application of growth retardants. Here, we have adopted a genetic strategy to introduce compactness in the perennial shrub Cape daisy (Osteospermum fruticosum Norl.). To this end, O. fruticosum was transformed using six different wild type Rhizobium rhizogenes strains. The most effective R. rhizogenes strains Arqua1 and ATCC15834 were used to create hairy root cultures from six Cape daisy genotypes. These root cultures were regenerated to produce transgenic Ri lines, which were analyzed for compactness. Ri lines displayed the characteristic Ri phenotype, i.e., reduced plant height, increased branching, shortened internodes, shortened peduncles, and smaller flowers. Evaluation of the Ri lines under commercial production conditions showed that similar compactness was obtained as the original Cape daisy genotypes treated with growth retardant. The results suggest that the use of chemical growth retardants may be omitted or reduced in commercial production systems of Cape daisy through implementation of Ri lines in future breeding programs.

Fusarium oxysporum as an Opportunistic Fungal Pathogen on Zinnia hybrida Plants Grown on board the International Space Station.

A plant production system called Veggie was launched to the International Space Station (ISS) in 2014. In late 2015, during the growth of Zinnia hybrida cv. 'Profusion' in the Veggie hardware, plants developed chlorosis, leaf curling, fungal growth that damaged leaves and stems, and eventually necrosis. The development of symptoms was correlated to reduced air flow leading to a significant buildup of water enveloping the leaves and stems in microgravity. Symptomatic tissues were returned to Earth on 18 May 2016 and were immediately processed to determine the primary causal agent of the disease. The presumptive pathogen was identified as Fusarium oxysporum by morphological features of microconidia and conidiophores on symptomatic tissues; that is, by epifluorescent microscopy (EFM), scanning electron microscopy (SEM), metabolic microarrays, and ITS sequencing. Both EFM and SEM imaging of infected tissues showed that germinating conidia were capable of stomatal penetration and thus acted as the primary method for infecting host tissues. A series of ground-based pathogenicity assays were conducted with healthy Z. hybrida plants that were exposed to reduced-airflow and high-water stress (i.e., encased in sealed bags) or were kept in an unstressed configuration. Koch's postulates were successfully completed with Z. hybrida plants in the lab, but symptoms only matched ISS-flown symptomatic tissues when the plants were stressed with high-water exposure. Unstressed plants grown under similar lab conditions failed to develop the symptoms observed with plants on board the ISS. The overall results of the pathogenicity tests imply that F. oxysporum acted as an opportunistic pathogen on severely high-water stressed plants. The source of the opportunistic pathogen is not known, but virulent strains of F. oxysporum were not recovered from unused materials in the Veggie plant pillow growth units assayed after the flight.

Secondary metabolites produced by endophytic Pantoea ananatis derived from roots of Baccharoides anthelmintica and their effect on melanin synthesis in murine B16 cells.

Five indole derivatives, 1H-indol-7-ol (1), tryptophol (2), 3-indolepropionic acid (3), tryptophan (4), 3,3-di(1H-indol-3-yl)propane-1,2-diol (5) and two diketopiperazines, cyclo(L-Pro-L-Tyr) (6), cyclo[L-(4-hydroxyprolinyl)-L-leucine (7) along with one dihydrocinnamic acid (8) were isolated from Pantoea ananatis VERA8, that endophytic bacteria derived from Baccharoides anthelmintica roots. This is a first report towards an isolation of endophytic strains (funji or bacteria) from the B. anthelmintica herb. The synergetic properties of the total extract compositions, as well as effects of the pure isolated secondary metabolites evaluated on their melanin synthesis in murine B16 cells towards for vitiligo treatment.

Sequencing and analysis of gerbera daisy leaf transcriptomes reveal disease resistance and susceptibility genes differentially expressed and associated with powdery mildew resistance.

BACKGROUND: RNA sequencing has been widely used to profile genome-wide gene expression and identify candidate genes controlling disease resistance and other important traits in plants. Gerbera daisy is one of the most important flowers in the global floricultural trade, and powdery mildew (PM) is the most important disease of gerbera. Genetic improvement of gerbera PM resistance has become a crucial goal in gerbera breeding. A better understanding of the genetic control of gerbera resistance to PM can expedite the development of PM-resistant cultivars. RESULTS: The objectives of this study were to identify gerbera genotypes with contrasting phenotypes in PM resistance and sequence and analyze their leaf transcriptomes to identify disease resistance and susceptibility genes differentially expressed and associated with PM resistance. An additional objective was to identify SNPs and SSRs for use in future genetic studies. We identified two gerbera genotypes, UFGE 4033 and 06-245-03, that were resistant and susceptible to PM, respectively. De novo assembly of their leaf transcriptomes using four complementary pipelines resulted in 145,348 transcripts with a N50 of 1124 bp, of which 67,312 transcripts contained open reading frames and 48,268 were expressed in both genotypes. A total of 494 transcripts were likely involved in disease resistance, and 17 and 24 transcripts were up- and down-regulated, respectively, in UFGE 4033 compared to 06-245-03. These gerbera disease resistance transcripts were most similar to the NBS-LRR class of plant resistance genes conferring resistance to various pathogens in plants. Four disease susceptibility transcripts (MLO-like) were expressed only or highly expressed in 06-245-03, offering excellent candidate targets for gene editing for PM resistance in gerbera. A total of 449,897 SNPs and 19,393 SSRs were revealed in the gerbera transcriptomes, which can be a valuable resource for developing new molecular markers. CONCLUSION: This study represents the first transcriptomic analysis of gerbera PM resistance, a highly important yet complex trait in a globally important floral crop. The differentially expressed disease resistance and susceptibility transcripts identified provide excellent targets for development of molecular markers and genetic maps, cloning of disease resistance genes, or targeted mutagenesis of disease susceptibility genes for PM resistance in gerbera.

[Antibacterial secondary metabolites of Clonostachys rosea, an endophytic fungus from Blumea balsamifera (L.) DC.]

Endophytic fungus is an important treasure trove for discovery of structurally unusual and biologically diverse compounds. A phytochemical investigation on a fungus Clonostachys rosea inhabits inner tissue of Blumea balsamifera (L.) DC. was initiatedrecently in our lab. Six pure compounds were isolated through silica gel column chromatography, sephadex LH-20, and semi-preparative HPLC techniques, with bio-guided strategy. Their structures were characterized as verticillin A (1), (S)-(+)-fusarinolic acid (2), 8-hydroxyfusaric acid (3), cerebroside C (4), 3-Maleimide-5-oxime (5), and bionectriol A (6) by analyses of NMR and MS data. All compounds were tested in vitro antibacterial activities against four strains of bacteria, Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Pseudomonas aeruginosa, and results revealed that 1, 4 and 6 display notableinhibition againstthree bacteria, with MIC values ranging from 2 to 16 µg/mL. Our findings provide references for mining novel antibiotics from endophytes originated from Li Minority medicinal plant B. balsamifera (L.) DC.

Cercospora beticola: The intoxicating lifestyle of the leaf spot pathogen of sugar beet.

Cercospora leaf spot, caused by the fungal pathogen Cercospora beticola, is the most destructive foliar disease of sugar beet worldwide. This review discusses C. beticola genetics, genomics, and biology and summarizes our current understanding of the molecular interactions that occur between C. beticola and its sugar beet host. We highlight the known virulence arsenal of C. beticola as well as its ability to overcome currently used disease management strategies. Finally, we discuss future prospects for the study and management of C. beticola infections in the context of newly employed molecular tools to uncover additional information regarding the biology of this pathogen. TAXONOMY: Cercospora beticola Sacc.; Kingdom Fungi, Phylum Ascomycota, Class Dothideomycetes, Order Capnodiales, Family Mycosphaerellaceae, Genus Cercospora. HOST RANGE: Well-known pathogen of sugar beet (Beta vulgaris subsp. vulgaris) and most species of the Beta genus. Reported as pathogenic on other members of the Chenopodiaceae (e.g., lamb's quarters, spinach) as well as members of the Acanthaceae (e.g., bear's breeches), Apiaceae (e.g., Apium), Asteraceae (e.g., chrysanthemum, lettuce, safflower), Brassicaceae (e.g., wild mustard), Malvaceae (e.g., Malva), Plumbaginaceae (e.g., Limonium), and Polygonaceae (e.g., broad-leaved dock) families. DISEASE SYMPTOMS: Leaves infected with C. beticola exhibit circular lesions that are coloured tan to grey in the centre and are often delimited by tan-brown to reddish-purple rings. As disease progresses, spots can coalesce to form larger necrotic areas, causing severely infected leaves to wither and die. At the centre of these spots are black spore-bearing structures (pseudostromata). Older leaves often show symptoms first and younger leaves become infected as the disease progresses. MANAGEMENT: Application of a mixture of fungicides with different modes of action is currently performed although elevated resistance has been documented in most employed fungicide classes. Breeding for high-yielding cultivars with improved host resistance is an ongoing effort and prudent cultural practices, such as crop rotation, weed host management, and cultivation to reduce infested residue levels, are widely used to manage disease. USEFUL WEBSITE: https://www.ncbi.nlm.nih.gov/genome/11237?genome_assembly_id=352037.

The effect of Funneliformis mosseae on the plant growth, Cd translocation and accumulation in the new Cd-hyperaccumulator Sphagneticola calendulacea.

The screening and identification of hyperaccumulators is the key to the phytoremediation of soils contaminated by heavy metal (HM). Arbuscular mycorrhizal fungus (AMF) can improve plant growth and tolerance to HM; therefore, AMF-assisted phytoextraction has been regarded as a potential technique for the remediation of HM-polluted soils. A greenhouse pot experiment was conducted to determine whether Sphagneticola calendulacea is a Cd-hyperaccumulator and to investigate the effect of the AMF-Funneliformis mosseae (FM) on plant growth and on the accumulation, subcellular distribution and chemical form of Cd in S. calendulacea grown in soils supplemented with different Cd levels. At 25, 50 and 100 mg Cd kg-1 level, S. calendulacea showed high Cd tolerance, the translocation factor and the bioconcentration factor exceeded 1, and accumulation of more than 100 mg Cd kg-1 was observed in the aboveground parts of the plant, meeting the requirements for a Cd-hyperaccumulator. Moreover, FM colonization significantly increased both biomasses and Cd concentration in S. calendulacea. After FM inoculation, the Cd concentrations and proportions increased in the cell walls, but exhibited no significant change in the organelles of the shoots. Meanwhile, FM symbiosis contributed to the conversion of Cd from highly toxic chemical forms (extracted by 80% ethanol and deionized water) to less toxic chemical forms (extracted by 1 M NaCl, 2% acetic acid, 0.6 M HCl) of Cd in the shoots. Overall, S. calendulacea is a typical Cd-hyperaccumulator, and FM symbiosis relieved the phytotoxicity of Cd and promoted plant growth and Cd accumulation, and thus greatly increasing the efficiency of phytoextraction for Cd-polluted soil. Our study provides a theoretical basis and application guidance for the remediation of Cd-contaminated soil by the symbiont of S. calendulacea with FM.

Biotechnological applications of the medicinal plant Pseudobrickellia brasiliensis and its isolated endophytic bacteria.

AIM: This study aimed to isolate Pseudobrickellia brasiliensis endophytic bacteria and evaluate the production of hydrolytic enzymes and antibiotics by these bacterial strains. The study also measured the antibacterial activity of P. brasiliensis. METHODS AND RESULTS: Thirteen endophytic bacteria strains were isolated from stem and leaf fragments of P. brasiliensis. Extracellular enzyme production by the isolated endophytic bacteria was evaluated in an agar plate-based assay. The highest protease production was achieved by Bacillus subtilis P4 in alkaline medium. Antimicrobial activity of endophytic bacteria and P. brasiliensis extracts was investigated using microbroth dilution. An MIC value of 1000 µg ml-1 against Pseudomonas aeruginosa was found for B. subtilis P3, B. subtilis P5, Pseudomonas sp. P8 and Pseudomonas sp. P12. Leaf extract of P. brasiliensis showed the highest antibacterial activity against P. aeruginosa, with an MIC value of 0·781 mg ml-1 . CONCLUSIONS: Pseudobrickellia brasiliensis is a source of bacterial endophytes, which can produce antibacterial compounds and enzymes. This work also demonstrated the antibacterial potential of P. brasiliensis. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study that revealed the antibacterial activity of P. brasiliensis and bioactive metabolite production by P. brasiliensis endophytic bacteria.

Biodegradation of diuron by endophytic Bacillus licheniformis strain SDS12 and its application in reducing diuron toxicity for green algae.

The endophytic bacteria live in close nuptial relationship with the host plant. The stress experienced by the plant is expected to be transferred to the endophytes. Thus, plants thriving at polluted sites are likely to harbor pollutant-degrading endophytes. The present study reports the isolation of phenylurea herbicides assimilating Bacillus sps. from Parthenium weed growing at diuron-contaminated site. The isolated endophytes exhibited plant growth-promoting (PGP) activities. Among five isolated diuron-degrading endophytes, the most efficient isolate Bacillus licheniformis strain SDS12 degraded 85.60 ± 1.36% of 50 ppm diuron to benign form via formation of degradation intermediate 3, 4-dichloroaniline (3,4-DCA). Cell-free supernatant (CFS) obtained after diuron degradation by strain SDS12 supported algal growth comparable with the pond water. The chlorophyll content and photosynthetic efficiency of green algae decreased significantly in the presence of diuron-contaminated water; however, no such change was observed in CFS of strain SDS12, thus, suggesting that strain SDS12 can be applied in aquatic bodies for degrading diuron and reducing diuron toxicity for primary producers. Further, the use of PGP and diuron-degrading bacteria in agriculture fields will not only help in remediating the soil but also support plant growth.

Arbuscular mycorrhizal fungal community composition determines the competitive response of two grassland forbs.

We performed a greenhouse experiment to assess how differences in AM fungal community composition affect competitive response of grassland plant species. We used a full factorial design to determine how inoculation with natural AM fungal communities from different habitats in Western Estonia affects the growth response of two grassland forbs (Leontodon hispidus L., Plantago lanceolata L.) to competition with a dominant grass (Festuca rubra L.). We used AM fungal inocula that were known to differ in AM fungal diversity and composition: more diverse AM fungal communities from open grasslands and less diverse AM fungal communities from former grassland densely overgrown by pines (young pine forest). The presence of AM fungi balanced competition between forb and grass species, by enhancing competitive response of the forbs. The magnitude of this effect was dependent on forb species identity and on the origin of the AM fungal inoculum in the soil. The grassland inoculum enhanced the competitive response of the forb species more effectively than the forest inoculum, but inoculum-specific competitive responses varied according to the habitat preference of the forb species. Our findings provide evidence that composition and diversity of natural AM fungal communities, as well as co-adaptation of plant hosts and AM-fungal communities to local habitat conditions, can determine plant-plant interactions and thus ultimately influence plant community structure in nature.

New antimicrobial compounds produced by Seltsamia galinsogisoli sp. nov., isolated from Galinsoga parviflora as potential inhibitors of FtsZ.

A total amount of 116 fungal strains, belonging to 30 genera, were acquired from the rhizosphere soil and plant of Galinsoga parviflora. A strain SYPF 7336, isolated from the rhizospheric soil, was identified as Seltsamia galinsogisoli sp. nov., by morphological and molecular analyses, which displayed high antibacterial activity. In order to study the secondary metabolites of Seltsamia galinsogisoli sp. nov., nine compounds were successfully seperated from the strain fermentation broth, including two new compounds and seven known compounds. Their structures were elucidated based on spectral analysis including 1D and 2D NMR. All the seperated compounds were evaluated for their antimicrobial activities. Compounds 2, 5 and 1 displayed antimicrobial activities against Staphylococcus aureus with MIC values of 25, 32 and 75 µg/mL, respectively. Moreover, morphological observation showed the coccoid cells of S. aureus to be swollen to a volume of 1.4 to 1.7-fold after treatment with compounds 1, 2 and 5, respectively. Molecular docking was carried out to investigate interactions of filamentous temperature-sensitive protein Z (FtsZ) with compounds 1, 2 and 5.

Aflatoxin B1 (AFB1) production by Aspergillus flavus and Aspergillus parasiticus on ground Nyjer seeds: The effect of water activity and temperature.

Nyjer oil seed cake supports high levels of aflatoxin B1 (AFB1) production. AFB1 is a secondary metabolite of Aspergillus flavus and A. parasiticus, classified as a Class 1A carcinogen. The aim of this study was to determine the effects of temperature (20, 27, and 35 °C) and water activity (0.82, 0.86, 0.90, 0.94, and 0.98 aw) on fungal growth and AFB1 production of A. flavus and A. parasiticus on ground Nyjer seeds over a 30-day incubation period. Linear regression models indicated that both fungal growth and AFB1 production were significantly influenced by water activity of Nyjer seeds and incubation temperature. The two fungi did not grow on Nyjer seeds at 0.82 aw at the three incubation temperatures. The most favorable growth conditions for both fungi were 0.90-0.98 aw at 27 °C or 0.90-0.94 aw at 35 °C. The optimum temperature for AFB1 production was 27 °C for both A. flavus and A. parasiticus (with regression coefficients of 6.01 and 9.11, respectively). Both fungi were likely to produce high levels of AFB1 at 0.90 aw (with regression coefficients of 3.56 for A. flavus and 7.17 for A. parasiticus). Aspergillus flavus only produced AFB1 on seeds with 0.90-0.98 aw at 27 °C (in the range of 203-282 µg/kg) and on seeds with 0.90 aw at 35 °C (212 µg/kg). No detectable AFB1 was produced by this fungus in any other culture conditions that were studied. Aspergillus parasiticus, in contrast, was able to produce AFB1 under all of the growth conditions. At 20 °C, this fungus produced the highest level of AFB1 (212 µg/kg) at high water activity (0.98 aw). At 27 °C, A. parasiticus produced high levels of AFB1 (in the range of 209-265 µg/kg) at a wide range of water activities (0.86-0.98 aw). In the entire study, the highest AFB1 concertation for A. parasiticus was detected on seeds incubated at high temperature (35 °C) and low water activity (0.86 aw). The findings of this study could help optimize the storage conditions of Nyjer oil seeds to reduce aflatoxin contamination.

Production of inulin- and neolevan-type fructooligosaccharides by Penicillium janczewskii Zaleski CCIBt 3352.

Fructooligosaccharides (FOS) are fructose-based oligosaccharides employed as additives to improve the nutritional and technological properties of foods. The rhizosphere of inulin-accumulating plants from the Cerrado (Brazilian savanna) harbor fungi capable of synthesizing FOS from sucrose through the transfructosylating activity of ß-fructosyltransferases and/or ß-fructofuranosidases. Here, we investigated the ability of Penicillium janczewskii Zaleski CCIBt 3352, a fungus isolated from the rhizosphere of Chrysolaena obovata (Asteraceae), to produce FOS in a medium supplemented with sucrose concentrations of 30, 100, or 150 g L-1 . Hydrolytic activity on sucrose was observed in culture filtrates; however, at 150 g L-1 sucrose, the accumulation of 8 g L-1 1-kestose (inulin-type FOS) and 7.3 g L-1 neokestose (neolevan-type FOS) was observed, the latter being a type of FOS not commonly produced by filamentous fungi. In addition, minor amounts of four unidentified oligosaccharides, with a high degree of polymerization, were detected. The production of FOS was also observed in enzymatic assays, indicating the presence of extracellular enzymes with transfructosylating activity in the culture filtrates. Our findings demonstrate the feasibility of isolating promising microorganisms, for the production of FOS-synthesizing enzymes, from the rhizosphere of fructan-producing plants of the Brazilian Cerrado.