Browsing by Author "Strobel, Gary A."

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3-Carbamoylquinoxalin-1-ium chloride
(2011-12) Harper, James K.; Strobel, Gary A.; Arif, Atta M.
The title compound, C9H8N3O+·Cl-, was isolated from a liquid culture of streptomyces sp. In the cation, the ring system makes a dihedral angle of 0.2 (2)° with the amide group. The protonation creating the cation occurs at ome of the N atoms in the quinoxaline ring system. In the crystal, the ions are linked through N-H...O and N-H...Cl hydrogen bonds, forming a two-dimensional network parallel to (10\overline{3}).
(6 S *)-6-[(1 S *,2 R *)-1,2-Dihydroxypentyl]-4-methoxy-5,6-dihydro-2 H -pyran-2-one
(2013-10) Valenti, Domenic J.; Arif, Atta M.; Strobel, Gary A.; Harper, James K.
The title compound, C11H18O5, was isolated from a liquid culture of Pestalotiopsis sp. In the molecule, the pyran-2-one ring assumes a half-chair conformation. The two terminal C atoms of the pentyl group were refined as disordered over two sets of sites, with refined occupancies of 0.881 (10) and 0.119 (10). In the crystal, molecules are linked via O-H...O hydrogen bonds forming a three-dimensional network.
Antifungal Activities of Volatile Secondary Metabolites of Four Diaporthe Strains Isolated from Catharanthus roseus
(2018-05) Yan, Dong-Hui; Song, Xiaoyu; Li, Hongchang; Luo, Tushou; Dou, Guiming; Strobel, Gary A.
Four endophytic fungi were isolated from the medicinal plant, Catharanthus roseus, and were identified as Diaporthe spp. with partial translation elongation factor 1-alpha (TEF1), beta-tubulin (TUB), histone H3 (HIS), calmodulin (CAL) genes, and rDNA internal transcribed spacer (ITS) region (TEF1-TUB-HIS--CAL-ITS) multigene phylogeny suggested for species delimitation in the Diaporthe genus. Each fungus produces a unique mixture of volatile organic compounds (VOCs) with an abundant mixture of terpenoids analyzed by headspace solid-phase microextraction (SPME) fiber-GC/MS. These tentatively-detected terpenes included Î±-muurolene, Î²-phellandrene, Î³-terpinene, and Î±-thujene, as well as other minor terpenoids, including caryophyllene, patchoulene, cedrene, 2-carene, and thujone. The volatile metabolites of each isolate showed antifungal properties against a wide range of plant pathogenic test fungi and oomycetes, including Alternaria alternata, Botrytis cinerea, Colletotrichum gloeosporioides, Fusarium graminearum, and Phytophthora cinnamomi. The growth inhibition of the pathogens varied between 10% and 60% within 72 h of exposure. To our knowledge, the endophytic Diaporthe-like strains are first reported from Catharanthus roseus. VOCs produced by each strain of the endophytic Diaporthe fungi were unique components with dominant monoterpenes comparing to known Diaporthe fungal VOCs. A discussion is presented on the inhibitive bioactivities of secondary metabolites among endophytic Diaporthe fungi and this medicinal plant.
Bioactive metabolites from an endophytic Cryptosporiopsis sp. inhabiting Clidemia hirta
(2013-11) Zilla, Mahesh; Qadri, Masroor; Pathania, Anup S.; Strobel, Gary A.; Nalli, Yedukondalu; Kumar, Sunil; Guru, Santosh K.; Bhushan, Shashi; Singh, Sanjay K.; Vishwakarma, Ram A.; Ul-Hassan, Syed Riyaz
An endophytic Cryptosporiopsis sp. was isolated from Clidemia hirta and analyzed for its secondary metabolites that lead to the isolation of three bioactive molecules. The compounds were purified from the culture broth of the fungus and their structures were determined by spectroscopic methods as (R)-5-hydroxy-2-methylchroman-4-one (1), 1-(2,6-dihydroxyphenyl)pentan-1-one (2) and (Z)-1-(2-(2-butyryl-3-hydroxyphenoxy)-6-hydroxyphenyl)-3-hydroxybut-2-en-1-one (3). Compound 1 exhibited significant cytotoxic activity against the human leukemia cell line, HL-60 with an IC50 of 4 μg/ml. This compound induced G2 arrest of the HL-60 cell cycle significantly. In addition, out of these compounds, 2 and 3 were active against several bacterial pathogens. Compound 2 was active against Bacillus cereus, Escherichia coli and Staphylococcus aureus with IC50 values varying from 18 to 30 μg/ml, and compound 3 displayed activity against Pseudomonas fluorescens with an IC50 value of 6 μg/ml. Compounds 2 and 3 are novel whereas compound 1 was reported earlier but the stereochemistry of its C-2 methyl is established for the first time.
Bioprospecting-fuels from fungi
(2015-02) Strobel, Gary A.
The world has a continuing demand and utility for liquid fuels to power its societies. The utilization of crude oil based fuels is leading to a dramatic increase in the CO2 content of the atmosphere which is being related to a dangerously warming earth. Having liquid fuels that are derived from biological sources is one solution to this growing problem since the carbon being utilized is only from recycled sources. Presently, the microbes, having the greatest impact on the worldâ€™s economies, producing liquid fuel are various yeasts producing ethanol. Other microbial sources need to be sought since ethanol is not the most desirable fuel and yeasts require simple sugars to carry out the fermentation processes. Recently, several endophytic fungi have been described that make hydrocarbons with fuel potential (Mycodiesel). Among others the compounds found in the volatile phases of these cultures include alkanes, branched alkanes, cyclohexanes, cyclopentanes, and alkyl alcohols/ketones, benzenes and polyaromatic hydrocarbons. Most importantly, generally these organisms make hydrocarbons while utilizing complex carbohydrates found in all plant-based agricultural wastes. Also discussed in this review is a rationale for finding hydrocarbon producing endophytes as well as examples of other promising hydrocarbon producers-Nodulisporium spp. which make 1,8-cineole and families of other hydrocarbons. Extremely favorable results of engine and fuel testing experiments recently completed on cineole and other products of Nodulisporium sp. are also presented. Finally, there is a brief discussion on the main limiting steps in the domestication of these fungi
Carbon chain length of biofuel- and flavor-relevant volatile organic compounds produced by lignocellulolytic fungal endophytes changes with culture temperature
(2017-09) Schoen, Heidi R.; Hunt, Kristopher A.; Strobel, Gary A.; Peyton, Brent M.; Carlson, Ross P.
Three fungal endophytes from the genus Nodulisporium were studied for volatile organic compound (VOC) production. All three fungi grew on a wide range of carbon substrates ranging from simple sugars to waste biomass sources. The fungi synthesized a number of long and short-chain VOCs, including eucalyptol; 1-butanol, 3-methyl; 1-octen-3-ol; and benzaldehyde, all with potential applications as biofuel or flavor compounds. As culture temperature decreased, average VOC carbon chain length increased, especially for VOCs associated with fatty acid metabolism. The results provide a template for controlling synthesis of desired VOCs through selection of species and culturing conditions.
Characterization of an Endophytic Gloeosporium sp. and Its Novel Bioactivity with “Synergistans”
(2014-12) Schaible, George A.; Strobel, Gary A.; Mends, Morgan Tess; Geary, Brad; Sears, Joe
Gloeosporium sp. (OR-10) was isolated as an endophyte of Tsuga heterophylla (Western hemlock). Both ITS and 18S sequence analyses indicated that the organism best fits either Hypocrea spp. or Trichoderma spp., but neither of these organisms possess conidiophores associated with acervuli, in which case the endophytic isolate OR-10 does. Therefore, the preferred taxonomic assignment was primarily based on the morphological features of the organism as one belonging to the genus Gloeosporium sp. These taxonomic observations clearly point out that limited ITS and 18S sequence information can be misleading when solely used in making taxonomic assignments. The volatile phase of this endophyte was active against a number of plant pathogenic fungi including Phytophthora palmivora, Rhizoctonia solani, Ceratocystis ulmi, Botrytis cinerea, and Verticillium dahliae. Among several terpenes and furans, the most abundantly produced compound in the volatile phase was 6-pentyl-2H-pyran-2-one, a compound possessing antimicrobial activities. When used in conjunction with microliter amounts of any in a series of esters or isobutyric acid, an enhanced inhibitory response occurred with each test fungus that was greater than that exhibited by Gloeosporium sp. or the compounds tested individually. Compounds behaving in this manner are hereby designated “synergistans.” An expression of the “median synergistic effect,” under prescribed conditions, has been termed the mSE50. This value describes the amount of a potential synergistan that is required to yield an additional median 50 % inhibition of a target organism. In this report, the mSE50s are reported for a series of esters and isobutyric acid. The results indicated that isoamyl acetate, allyl acetate, and isobutyric acid generally possessed the lowest mSE50 values. The value and potential importance of these microbial synergistic effects to the microbial environment are also discussed.
Characterization of Novel Endophytic Isolate Or10-4 and Analysis of Increased Bio-Activity by Addition of Esters
(2013-03) Schaible, George A.; Strobel, Gary A.
Today there is thought to be over a million species of fungi to exist and approximately 75,000 of those have been scientifically identified, leaving an enormous amount of species still unidentified and under-researched. The work done in Dr. Gary Strobel’s lab focuses on the collection of biologically diverse plant material for isolation and classification of fungal endophytic species. Research has shown that these species of endophytes contain novel characteristics that make them of high interest for further research and analysis. Or10-4 is an endophytic isolate from the Canadian hemlock that produces a valuable volatile organic compound (VOC), 6-Pentyl-2H-pyran-2-one, which can be used as an anti-pathogen for crop plants to be applied in large agricultural settings. Analysis of scanning electron microscope (SEM) pictures from spore structures have indicated that Or10-4 is a new species to be taxonomically classified calling for complete rDNA sequencing. Furthermore, tests done on Or10-4 with the addition of esters in bioactivity tests against pathogenic species has shown increased inhibition. The chemistry of the interaction between the esters and VOC’s is still being analyzed.
The diversity and antimicrobial activity of endophytic actinomycetes isolated from medicinal plants in Panxi Plateau, China
(2010-06) Zhao, Ke; Penttinen, Petri; Guan, Tongwei; Xiao, Jing; Chen, Qiang; Xu, Jun; Lindström, Kristina; Zhang, Lili; Zhang, Xiaoping; Strobel, Gary A.
Traditional Chinese medicinal plants are sources of biologically active compounds, providing raw material for pharmaceutical, cosmetic and fragrance industries. The endophytes of medicinal plants participate in biochemical pathways and produce analogous or novel bioactive compounds. Panxi plateau in South-west Sichuan in China with its unique geographical and climatological characteristics is a habitat of a great variety of medicinal plants. In this study, 560 endophytic actinomycetes were isolated from 26 medicinal plant species in Panxi plateau. 60 isolates were selected for 16S rDNA-RFLP analysis and 14 representative strains were chosen for 16S rDNA sequencing. According to the phylogenetic analysis, seven isolates were Streptomyces sp., while the remainder belonged to genera Micromonospora, Oerskovia, Nonomuraea, Promicromonospora and Rhodococcus. Antimicrobial activity analysis combined with the results of amplifying genes coding for polyketide synthetase (PKS-I, PKS-II) and nonribosomal peptide synthetase (NRPS) showed that endophytic actinomycetes isolated from medicinal plants in Panxi plateau had broad-spectrum antimicrobial activity and potential natural product diversity, which further proved that endophytic actinomycetes are valuable reservoirs of novel bioactive compounds.
The Emergence of Endophytic Microbes and Their Biological Promise
(2018-05) Strobel, Gary A.
As is true with animal species, plants also have an associated microflora including endophytes as well as microbes associated with the phyllosphere and rhizosphere (plant surfaces) and this is considered the plant microbiome. However, those organisms within virtually all tissues and organs of the plant are known as endophytes. Most often fungi are the most frequently recovered endophytes from plant tissues, but bacterial forms generally occur in greater numbers, but not in species varieties. The exact biological/biochemical role of the endophyte in the plant and how it interacts with the plant and other endophytes and plant associated organisms has not been intensely and carefully examined. However, this has not stopped investigators in exploring the direct utility of endophytes in boosting plant production, and discovering that endophytes can directly influence the plant to resist temperature extremes, drought, as well as the presence of disease causing organisms. Also, because of the relationships that endophytes seem to have with their host plants, they make a myriad of biologically active compounds some of which are classified as antibiotics, antioxidants, anticancer agents, volatile antimicrobial agents, immunosuppressive compounds, plant growth promoting agents, and insecticides. These endophytic compounds represent a wide range of organic molecules including terpenoids, peptides, carbohydrates, aromatics, hydrocarbons and others and it seems that these compounds may have a role in the host microbe relationship. Most recently and quite surprisingly, some endophytes have been discovered that make hydrocarbons of the types found in diesel and gasoline fuels. In addition, recently discovered are epigenetic factors relating to the biology and biochemistry of endophytes. Interestingly, only about 1–2% of the entire spectrum of 300,000 known plants have been studied for their endophyte composition. Additionally, only a few plants have ever been completely studied including all tissues for the microbes within them. Likewise, the vast majority of plants, including those in oceans and lower plant forms, have never been examined for their endophytes. Furthermore, endophytes representing the “microbiome” of world’s major food plants as they exist in their native “centers of origin” are largely unknown. This non-classical review is intended to provide background information on aspects of developments in endophyte biology and more importantly the identification of new questions in this field that need to be addressed. The review is primarily based on the author’s long held experience in this field.
An endophytic Coniochaeta velutina producing broad spectrum antimycotics
(2015-06) Xie, Jie; Strobel, Gary A.; Feng, Tao; Ren, Huishuang; Mends, Morgan Tess; Nhou, Zeyang; Geary, Brad
An endophyte (PC27-5) was isolated from stem tissue of Western hemlock (Tsuga heterophylla) in a Pacific Northwest temperate rainforest. Phylogenetic analyses, based on ITS-5.8S rDNA and 18S rDNA sequence data, combined with cultural and morphological analysis showed that endophyte PC27-5 exhibited all characteristics of a fungus identical to Coniochaeta velutina. Furthermore, wide spectrum antimycotics were produced by this endophyte that were active against such plant pathogens as Sclerotinia sclerotiorum, Pythium ultimum, and Verticillium dahliae and lethal to Phythophthora cinnamomi, Pythium ultimum, and Phytophthora palmivora in plate tests. The bioactive components were purified through organic solvent extraction, followed by silica column chromatography, and finally preparative HPLC. The minimum inhibitory concentration of the active fraction to Pythium ultimum, which was gained from preparative HPLC, was 11 Î¼g/ml. UPLC-HRMS analysis showed there were two similar components in the antimycotic fraction. Their molecular formulae were established as C30H22O11 (compound I) and C30H22O10 (compound II) respectively, and preliminary spectral results indicate that they are anthroquinone glycosides. Other nonâ€“biologically active compounds were identified in culture fluids of this fungus by spectral means as emodin and chrysophanol - anthroquinone derivatives. This is the first report that Coniochaeta velutina as an endophyte produces bioactive antifungal components.
An endophytic Myrothecium inundatum producing volatile organic compounds
(2010) Banerjee, Debdulal; Strobel, Gary A.; Booth, B.; Sears, Joe; Spakowicz, Daniel; Busse, S.
Myrothecium inunduatum was isolated as an endophyte from a euphorbeacean herb, Acalypha indica in NE India. This fungus when grown in shake culture produced an abundance of foam. Contained in the foam was a mixture of volatile organic compounds (VOCs) some of which were hydrocarbons and hydrocarbon derivatives. The most prevalent compounds were 3-octanone, 3-octanol, and 7-octen-4-ol. Numerous other volatile organic compounds were also produced including many terpenes, organic acids, ketones, and alcohols. The VOCs of this fungus demonstrated growth inhibitory activity against a number of plant pathogenic fungi including Pythium ultimum and Sclerotinia sclerotiorum. However, when grown in microaerophilic conditions, the organism produced a number of hydrocarbons of interest as fuel related hydrocarbons including octane and tentatively identified- 1,4- cyclohexadiene, 1-methyl- and cyclohexane, (1-ethylpropyl) and others. An NMR method was used to measure VOC production which peaked at day 15 in a time course experiment. Numerous substrates can serve to support the production of VOCs by this fungus including potato broth and beet pulp extracts.
An Endophytic Nodulisporium sp. from Central America Producing Volatile Organic Compounds with Both Biological and Fuel Potential
(2013) Hassan, Syed R.-U.; Strobel, Gary A.; Geary, Brad; Sears, Joe
A Nodulisporium sp. (Hypoxylon sp.) has been isolated as an endophyte of Thelypteris angustifolia (Broadleaf Leaf Maiden Fern) in a rainforest region of Central America. It has been identified both on the basis of its morphological characteristics and by scanning electron microscopy as well as ITS sequence analysis. The endophyte produces volatile organic compounds (VOCs) that have both fuel (mycodiesel) and use for biological control of plant disease. When grown on potato dextrose agar, the organism uniquely produces a series of ketones, including acetone; 2-pentanone; 3-hexanone, 4-methyl; 3-hexanone, 2,4- dimethyl; 2-hexanone, 4-methyl, and 5-hepten, 2-one and these account for about 25% of the total VOCs. The most abundant identified VOC was 1,8 cineole, which is commonly detected in this group of organisms. Other prominent VOCs produced by this endophyte include 1-butanol, 2- methyl, and phenylethanol alcohol. Moreover, of interest was the presence of cyclohexane, propyl, which is a common ingredient of diesel fuel. Furthermore, the VOCs of this isolate of Nodulisporium sp. were selectively active against a number of plant pathogens, and upon a 24 h exposure caused death to Phytophthora palmivora, Rhizoctonia solani, and Sclerotinia sclerotiorum and 100% inhibition to Phytophthora cinnamomi with only slight to no inhibition of the other pathogens that were tested. From this work, it is becoming increasingly apparent that each isolate of this endophytic Nodulisporium spp., including the Daldina sp. and Hypoxylon spp. teleomorphs, seems to produce its own unique set of VOCs.
An endophytic Nodulisporium sp. producing volatile organic compounds having bioactivity and fuel potential
(2012) Mends, Morgan Tess; Yu, Eizadora; Strobel, Gary A.; Hassan, S. R. U.; Booth, Eric; Geary, Brad; Sears, Joe; Taatjes, C. A.; Hadi, M.
Nodulisporium sp. has been isolated as an endophyte of Myroxylon balsamum found in the upper Napo region of the Ecuadorian Amazon. This organism produces volatile organic compounds (VOCs) that have both fuel and biological potential.Under microaerophilic growth environments, the organism produces 1, 4-cyclohexadiene, 1 methyl-,1-4 pentadiene and cyclohexene, 1-methyl-4-(1-methylethenyl)- along with some alcohols and terpenoids of interest as potential fuels. The fungus was scaled up in an aerated large fermentation flask, and the VOCs trapped by Carbotrap technology and analyzed by headspace solid –phase microextraction (SPME) fiber-GC/MS. Under these conditions, Nodulisporium sp. produces a series of alkyl alcohols starting with 1-butanol-3-methyl, 1- propanol-2-methyl, 1- pentanol, 1-hexanol, 1-heptanol, 1- octanol, 1-nonanol along with phenylethyl alcohol.The organism also produces secondary alkyl alcohols, esters, ketones, benzene derivatives, a few terpenoids, and some hydrocarbons. It appears that many of the products have fuel potential. Furthermore, the VOCs of Nodulisporium sp. were active against a number of pathogens causing death to both Aspergillus fumigatus and Rhizoctonia solani and severe growth inhibition produced in Phytophthora cinnamomi and Sclerotinia sclerotiorum within 48 hr of exposure. The Carbotrapped materials somewhat mimicked the bioactivities of the culture itself when certain test organisms were exposed to these VOCs. A brief discussion on the relationship of these fungal VOCs to those compounds found in transportation fuels is presented.
An endophytic/pathogenic Phoma sp. from creosote bush producing biologically active volatile compounds having fuel potential
(2011-05) Strobel, Gary A.; Singh, Sanjay K.; Ul-Hassan, Syed Riyaz; Mitchell, Angela M.; Geary, Brad; Sears, Joe
A Phoma sp. was isolated and characterized as endophytic and as a pathogen of Larrea tridentata (creosote bush) growing in the desert region of southern Utah, USA. This fungus produces a unique mixture of volatile organic compounds (VOCs), including a series of sesquiterpenoids, some alcohols and several reduced naphthalene derivatives. Trans-caryophyllene, a product in the fungal VOCs, was also noted in the VOCs of this pungent plant. The gases of Phoma sp. possess antifungal properties and is markedly similar to that of a methanolic extract of the host plant. Some of the test organisms with the greatest sensitivity to the Phoma sp. VOCs were Verticillium, Ceratocystis, Cercospora and Sclerotinia while those being the least sensitive were Trichoderma, Colletotrichum and Aspergillus. We discuss the possible involvement of VOC production by the fungus and its role in the biology/ecology of the fungus/plant/environmental relationship with implications for utilization as an energy source.
Evaluation of cellulose as a substrate for hydrocarbon fuel production by Ascocoryne sarcoides (NRRL 50072)
(2014-02) Mallette, Natasha D.; Pankrantz, E. M.; Busse, S.; Strobel, Gary A.; Carlson, Ross P.; Peyton, Brent M.
The fungal endophyte, Ascocoryne sarcoides, produced aviation, gasoline and diesel-relevant hydrocarbons when grown on multiple substrates including cellulose as the sole carbon source. Substrate, growth stage, culturing pH, temperature and medium composition were statistically significant factors for the type and quantity of hydrocarbons produced. Gasoline range (C5-C12), aviation range (C8-C16) and diesel range (C9-C36) organics were detected in all cultured media. Numerous non-oxygenated hydrocarbons were produced such as isopentane, 3,3-dimethyl hexane and d-limonene during exponential growth phase. Growth on cellulose at 23˚C and pH 5.8 produced the highest overall yield of fuel range organics (105 mg * g·biomass−1). A change in metabolism was seen in late stationary phase from catabolism of cellulose to potential oxidation of hydrocarbons resulting in the production of more oxygenated compounds with longer carbon chain length and fewer fuel-related compounds. The results outline rational strategies for controlling the composition of the fuel-like compounds by changing culturing parameters.
Functional Characterization of Endophytic Fungal Community Associated with Oryza sativa L. and Zea mays L
(2017-03) Potshangbam, Momota; Devi, S. Indira; Sahoo, Dinabandhu; Strobel, Gary A.
In a natural ecosystem, the plant is in a symbiotic relationship with beneficial endophytes contributing huge impact on its host plant. Therefore, exploring beneficial endophytes and understanding its interaction is a prospective area of research. The present work aims to characterize the fungal endophytic communities associated with healthy maize and rice plants and to study the deterministic factors influencing plant growth and biocontrol properties against phytopathogens, viz, Pythium ultimum, Sclerotium oryzae, Rhizoctonia solani, and Pyricularia oryzae. A total of 123 endophytic fungi was isolated using the culture-dependent approach from different tissue parts of the plant. Most dominating fungal endophyte associated with both the crops belong to genus Fusarium, Sarocladium, Aspergillus, and Penicillium and their occurrence was not tissue specific. The isolates were screened for in vitro plant growth promotion, stress tolerance, disease suppressive mechanisms and based on the results, each culture from both the cereal crops was selected for further study. Acremonium sp. (ENF 31) and Penicillium simplicisssum (ENF22), isolated from maize and rice respectively could potentially inhibit the growth of all the tested pathogens with 46.47 Â± 0.16 mm to 60.09 Â± 0.04 mm range zone of inhibition for ENF31 and 35.48 Â± 0.14 to 62.29 Â± 0.15 mm for ENF22. Both significantly produce the defensive enzymes, ENF31 could tolerate a wide range of pH from 2 to 12, very important criteria, for studying plant growth in different soil types, especially acidic as it is widely prevalent here, making more land unsuitable for cultivation. ENF22 grows in pH range 3-12, with 10% salt tolerating ability, another factor of consideration. Study of root colonization during 7th to 30th days of growth phase reveals that ENF31 could colonize pleasantly in rice, though a maize origin, ranging from 1.02 to 1.21 log10 CFU/g root and in maize, it steadily colonizes ranging from 0.95 to 1.18 log10 CFU, while ENF22 could colonize from 0.98 to 1.24 Log10CFU/g root in rice and 1.01 to 1.24Log10CFU/g root in maize, just the reverse observed in Acremonium sp. Therefore, both the organism has the potency of a promising Bio-resource agent, that we must definitely explore to fill the gap in the agriculture industry.
Functionalized para-substituted benzenes as 1,8-cineole production modulators in an endophytic Nodulisporium species
(2014-08) Nigg, Jered; Strobel, Gary A.; Knighton, W. Berk; Hilmer, Jonathan K.; Geary, Brad; Ul-Hassan, Syed Riyaz; Harper, James K.; Valenti, Domenic J.; Wang, Yuemin
A Nodulisporium species (designated Ti-13) was isolated as an endophyte from Cassia fistula. The fungus produces a spectrum of volatile organic compounds (VOCs) that includes ethanol, acetaldehyde and 1,8-cineole as major components. Initial observations of the fungal isolate suggested that reversible attenuation of the organism via removal from the host and successive transfers in pure culture resulted in a 50 % decrease in cineole production unrelated to an overall alteration in fungal growth. A compound (CPM1) was obtained from Betula pendula (silver birch) that increases the production of 1,8-cineole by an attenuated Ti-13 strain to its original level, as measured by a novel bioassay method employing a 1,8-cineole-sensitive fungus (Sclerotinia sclerotiorum). The host plant produces similar compounds possessing this activity. Bioactivity assays with structurally similar compounds such as ferulic acid and gallic acid suggested that the CPM1 does not act as a simple precursor to the biosynthesis of 1,8-cineole. NMR spectroscopy and HPLC-ES-MS indicated that the CPM1 is a para-substituted benzene with alkyl and carboxyl substituents. The VOCs of Ti-13, especially 1,8-cineole, have potential applications in the industrial, fuel and medical fields.
Fungi as Architects of the Rimstone Dams in Huanglong, NSD, Sichuan, China
(2017-01) Xie, Jie; Strobel, Gary A.; Wu, Wei-Fang; Chen, Jie; An, De-Jun; Geary, Brad
The Huanglong park area of the Sichuan Province of China is a unique scenic area of the world. It is known for its thousands of aquamarine-colored pools that are formed behind naturally formed rimstone dams of travertine (calcite) along a cold water stream. The travertine, based on its crystalline structural analysis, is of biological origin. This makes sense since the temperature of the waters of Huanglong varies from 5 to 7 A degrees C and thus geochemical crystallization does not occur as it does in other locations around the world possessing thermal pools whose structures are primarily formed through cooling processes. Fungi and bacteria were discovered associated with both leaves associated with the calcite dams as well as in the older parts of well-established dams. Several species of Phytium, a phycomycete and an endophyte, accounted for over 45 % of all of the fungi successfully isolated from the well-established dam samples and at least 85 % in the floating leaf samples. Saprolegnia spp. (Phycomycetes) along with Phoma spp. (Ascomycetes) were noted along with Mortierella sp. as other dam-associated fungi. The fungal hyphae observed on dead leaf material as well as in the calcite dams directly served as nucleation points for the formation of crystalline CaCO3. Eventually, these crystals grow large enough to fuse to make calcite plates which form the main structural feature of all of the travertine dams in this area. Interestingly, each of the individual crystals associated with the dams has an associated hole in its core where a fungal hypha used to reside as observed by scanning electron microscopy. While diatoms were present in the analysis, they too seem to contribute to the structure of the dams but in a minor way. The only bacteria isolated from the older dam of this aquatic environment were Pseudomonas spp. and their role in dam formation is uncertain. Huanglong is a unique and beautiful place, and the water features present in this area can definitely be attributed to those fungal architects that encourage calcite crystal formation.
Genomic and Functional Characterization of the Endophytic Bacillus subtilis 7PJ-16 Strain, a Potential Biocontrol Agent of Mulberry Fruit Sclerotiniose
(2018-09) Xu, Wei-Fang; Ren, Hui-Shuang; Ou, Ting; Lei, Ting; Wei, Jun-Hong; Huang, Chuan-Shu; Li, Tian; Strobel, Gary A.; Zhou, Ze-Yang; Xie, Jie
Bacillus sp. 7PJ-16, an endophytic bacterium isolated from a healthy mulberry stem and previously identified as Bacillus tequilensis 7PJ-16, exhibits strong antifungal activity and has the capacity to promote plant growth. This strain was studied for its effectiveness as a biocontrol agent to reduce mulberry fruit sclerotiniose in the field and as a growth-promoting agent for mulberry in the greenhouse. In field studies, the cell suspension and supernatant of strain 7PJ-16 exhibited biocontrol efficacy and the lowest disease incidence was reduced down to only 0.80%. In greenhouse experiments, the cell suspension (1.0â€‰Ã—â€‰106 and 1.0â€‰Ã—â€‰105 CFU/mL) and the cell-free supernatant (100-fold and 1000-fold dilution) stimulated mulberry seed germination and promoted mulberry seedling growth. In addition, to accurately identify the 7PJ-16 strain and further explore the mechanisms of its antifungal and growth-promoting properties, the complete genome of this strain was sequenced and annotated. The 7PJ-16 genome is comprised of two circular plasmids and a 4,209,045-bp circular chromosome, containing 4492 protein-coding genes and 116 RNA genes. This strain was ultimately designed as Bacillus subtilis based on core genome sequence analyses using a phylogenomic approach. In this genome, we identified a series of gene clusters that function in the synthesis of non-ribosomal peptides (surfactin, fengycin, bacillibactin, and bacilysin) as well as the ribosome-dependent synthesis of tasA and bacteriocins (subtilin, subtilosin A), which are responsible for the biosynthesis of numerous antimicrobial metabolites. Additionally, several genes with function that promote plant growth, such as indole-3-acetic acid biosynthesis, the production of volatile substances, and siderophores synthesis, were also identified. The information described in this study has established a good foundation for understanding the beneficial interactions between endophytes and host plants, and facilitates the further application of B. subtilis 7PJ-16 as an agricultural biofertilizer and biocontrol agent.