Classifications of Fungi

2,5-diamino-6-ribitylamino-4(3H)-pyrimidinone 5¢-phosphate synthases of fungi and archaea Werner Ro ¨ misch-Margl 1,2 , Wolfgang Eisenreich 1 , Ilka Haase 3 , Adelbert Bacher 1 and Markus Fischer 3 1 Lehrstuhl fu ¨ r Organische Chemie und Biochemie, Technische Universita ¨ tMu ¨ nchen, Garching, Germany 2 Institute of Bioinformatics and Systems Biology, Helmholtz Zentrum Mu ¨ nchen, Neuherberg, Germany 3 Institute of Food Chemistry, University of Hamburg, Germany The coenzymes FMN and FAD derived from vitamin B 2 are essential in all organisms. They are involved in a wide variety of redox processes, some of which are fundamental to central energy transduction functions. They are also involved in a variety of non-redox processes such as DNA photorepair, blue-light sensing in plants and a variety of enzyme reactions including certain dehydration and isomerisation reactions [1–3]. In view of the vital role of these coenzymes, it appears likely that biosynthesis of the parent compound, vita- min B 2 (riboflavin, compound 8 in Fig. 1), must already have been operative in the early phase of evolution. The pathway of riboflavin biosynthesis has been studied in considerable detail for more than five dec- ades (for review, see [4–7]). One of the driving forces for this research was the commercial requirement for bulk amounts (approximately 3000 tonnes per year) of the vitamin for use in human and animal nutrition and as a non-toxic food colorant [8]. However, fermenta- tion processes using yeasts and eubacteria have now completely replaced chemical synthesis of the trace nutrient [9]. The biosynthesis of the vitamin is summarised in Fig. 1. Although the final part of the pathway is universal in all organisms studied to date, the early section shows significant differences between taxo- nomic kingdoms. In eubacteria, fungi and plants, the first committed step, catalysed by the enzyme GTP cyclohydrolase II (reaction A in Fig. 1), consists of hydrolytic opening of the imidazole ring of GTP (compound 1 in Fig. 1) with concomitant removal of a pyrophosphate moiety; the reaction mechanism for this enzyme has been studied in considerable detail [10–13]. In archaea, the first committed step involves release of pyrophosphate and opening of the imidazole ring Keywords 2,5-diamino-6-ribitylamino-4(3H)-pyrimidinone 5¢-phosphate synthase; archaea; fungi; riboflavin biosynthesis; stereochemistry Correspondence M. Fischer, Institut fu ¨ r Lebensmittelchemie, Universita ¨ t Hamburg, Grindelallee 117, D-20146 Hamburg, Germany Fax: +49 40 428384342 Tel: +49 40 428384359 E-mail: markus.fischer@uni-hamburg.de (Received 18 April 2008, revised 21 June 2008, accepted 4 July 2008) doi:10.1111/j.1742-4658.2008.06586.x The pathway of riboflavin (vitamin B 2 ) biosynthesis is significantly different in archaea, eubacteria, fungi and plants. Specifically, the first committed intermediate, 2,5-diamino-6-ribosylamino-4(3H)-pyrimidinone 5¢-phosphate, can either undergo hydrolytic cleavage of the position 2 amino group by a deaminase (in plants and most eubacteria) or reduction of the ribose side chain by a reductase (in fungi and archaea). We compare 2,5-diamino-6- ribitylamino-4(3H)-pyrimidinone 5¢-phosphate synthases from the yeast Candida glabrata, the archaeaon Methanocaldococcus jannaschii and the eubacterium Aquifex aeolicus. All three enzymes convert 2,5-diamino-6- ribosylamino-4(3H)-pyrimidinone 5¢-phosphate into 2,5-diamino-6-ribitylami- no-4(3H)-pyrimidinone 5¢-phosphate, as shown by 13 C-NMR spectroscopy using [2,1¢,2¢,3¢,4¢,5¢- 13 C 6 ]2,5-diamino-6-ribosylamino-4(3H)-pyrimidinone 5¢-phosphate as substrate. The b anomer was found to be the authentic substrate, Classifications of Fungi Classifications of Fungi Bởi: OpenStaxCollege The kingdom Fungi contains five major phyla that were established according to their mode of sexual reproduction or using molecular data Polyphyletic, unrelated fungi that reproduce without a sexual cycle, are placed for convenience in a sixth group called a “form phylum” Not all mycologists agree with this scheme Rapid advances in molecular biology and the sequencing of 18S rRNA (a part of RNA) continue to show new and different relationships between the various categories of fungi The five true phyla of fungi are the Chytridiomycota (Chytrids), the Zygomycota (conjugated fungi), the Ascomycota (sac fungi), the Basidiomycota (club fungi) and the recently described Phylum Glomeromycota The Deuteromycota is an informal group of unrelated fungi that all share a common character – they use strictly asexual reproduction Note: “-mycota” is used to designate a phylum while “-mycetes” formally denotes a class or is used informally to refer to all members of the phylum Chytridiomycota: The Chytrids The only class in the Phylum Chytridiomycota is the Chytridiomycetes The chytrids are the simplest and most primitive Eumycota, or true fungi The evolutionary record shows that the first recognizable chytrids appeared during the late pre-Cambrian period, more than 500 million years ago Like all fungi, chytrids have chitin in their cell walls, but one group of chytrids has both cellulose and chitin in the cell wall Most chytrids are unicellular; a few form multicellular organisms and hyphae, which have no septa between cells (coenocytic) They produce gametes and diploid zoospores that swim with the help of a single flagellum The ecological habitat and cell structure of chytrids have much in common with protists Chytrids usually live in aquatic environments, although some species live on land Some species thrive as parasites on plants, insects, or amphibians ([link]), while others are saprobes The chytrid species Allomyces is well characterized as an experimental organism Its reproductive cycle includes both asexual and sexual phases Allomyces produces diploid or haploid flagellated zoospores in a sporangium 1/12 Classifications of Fungi The chytrid Batrachochytrium dendrobatidis is seen in these light micrographs as transparent spheres growing on (a) a freshwater arthropod and (b) algae This chytrid causes skin diseases in many species of amphibians, resulting in species decline and extinction (credit: modification of work by Johnson ML, Speare R., CDC) Zygomycota: The Conjugated Fungi The zygomycetes are a relatively small group of fungi belonging to the Phylum Zygomycota They include the familiar bread mold, Rhizopus stolonifer, which rapidly propagates on the surfaces of breads, fruits, and vegetables Most species are saprobes, living off decaying organic material; a few are parasites, particularly of insects Zygomycetes play a considerable commercial role The metabolic products of other species of Rhizopus are intermediates in the synthesis of semi-synthetic steroid hormones Zygomycetes have a thallus of coenocytic hyphae in which the nuclei are haploid when the organism is in the vegetative stage The fungi usually reproduce asexually by producing sporangiospores ([link]) The black tips of bread mold are the swollen sporangia packed with black spores ([link]) When spores land on a suitable substrate, they germinate and produce a new mycelium Sexual reproduction starts when conditions become unfavorable Two opposing mating strains (type + and type –) must 2/12 Classifications of Fungi be in close proximity for gametangia from the hyphae to be produced and fuse, leading to karyogamy The developing diploid zygospores have thick coats that protect them from desiccation and other hazards They may remain dormant until environmental conditions are favorable When the zygospore germinates, it undergoes meiosis and produces haploid spores, which will, in turn, grow into a new organism This form of sexual reproduction in fungi is called conjugation (although it differs markedly from conjugation in bacteria and protists), giving rise to the name “conjugated fungi” Zygomycetes have asexual and asexual life cycles In the sexual life cycle, plus and minus mating types conjugate to form a zygosporangium 3/12 Classifications of Fungi Sporangia grow at the end of stalks, which appear as (a) white fuzz seen on this bread mold, Rhizopus stolonifer The (b) tips of bread mold are the spore-containing sporangia (credit b: modification of work by "polandeze"/Flickr) Ascomycota: The Sac Fungi The majority of known fungi belong to the Phylum Ascomycota, which is characterized by the formation of an ascus (plural, asci), a sac-like structure that contains haploid ascospores Many ascomycetes are of commercial importance Some play a beneficial role, such as the yeasts used in baking, brewing, and wine fermentation, plus truffles and morels, which are held as gourmet delicacies ...Agriculture and Agri-Food Canada Agriculture et Agroalimentaire Canada Use of Fungi for Insect Control - Issues, Developments & Research Needs Mark Goettel Agriculture and Agri-Food Canada Research Centre, Lethbridge, AB Agriculture and Agri-Food Canada Agriculture et Agroalimentaire Canada Entomopathogenic Fungi • Classification • Pathogenesis • Why fungi as BCA’s? • Commercialization • Registration • Some examples • Research Agriculture and Agri-Food Canada Agriculture et Agroalimentaire Canada Classification • Kingdom Chromista (formerly Eumycota) • Division Oomycota – Water molds – – Pythiales Pythiales » » Lagenidium Lagenidium giganteum giganteum – – Reproduce asexually Reproduce asexually – – Motile zoospores Motile zoospores Agriculture and Agri-Food Canada Agriculture et Agroalimentaire Canada Classification • Kingdom Eumycota (the fungi) • Four Divisions – Chytridiomycota – Zygomycota – Ascomycota – Basidiomycota – (Deuteromycota) Agriculture and Agri-Food Canada Agriculture et Agroalimentaire Canada Division Chytridiomycota • Genera Coelomomyces • Pathogen of aquatic Diptera • Obligate pathogens • Require alternate crustacean hosts • Very host specific Agriculture and Agri-Food Canada Agriculture et Agroalimentaire Canada Division Zygomycota • Order Entomophthorales • Genera: Conidiobolus, Entomophaga, Entomophthora, Erynia, Pandora, Zoophthora • Mostly obligate pathogens • Sexual & asexual spores • Spores ejected forcibly • Mostly restricted host ranges Agriculture and Agri-Food Canada Agriculture et Agroalimentaire Canada Division Ascomycota • Order Clavicipitales • Genera: Cordyceps, Torrubiella – Teleomorphic (sexual) state of many spp within Deuteromycota • Order Ascosphaerales – Genera: Ascosphaera – – Mostly obligate pathogens Mostly obligate pathogens – – Sexual & asexual spores Sexual & asexual spores – – Mostly restricted host ranges Mostly restricted host ranges Agriculture and Agri-Food Canada Agriculture et Agroalimentaire Canada Division Deuteromycota • Genera: Aschersonia, Aspergillus, Beauveria, Hirsutella, Metarhizium, Nomuraea, Paecilomyces, Verticillium (Lecanicillium) • Most? anamorphic (asexual) state of Cordyceps • Mostly facultative pathogen • Assexual, immotile spores, conidia • Mostly wider host ranges Agriculture and Agri-Food Canada Agriculture et Agroalimentaire Canada Some examples Aschersonia aleyrodis Cordyceps Agriculture and Agri-Food Canada Agriculture et Agroalimentaire Canada Metarhizium flavoviride Hirsutella [...]... adapted to host • Great potential for epizootics Agriculture and Agriculture et Agri-Food Canada Agroalimentaire Canada Use of fungi • Classical (introduction) • Conservation • Augmentation (insecticidal) Agriculture and Agriculture et Agri-Food Canada Agroalimentaire Canada Commercialization • Insecticidal approach • Niche products • Regulations • Profit margins too low for the big conglomerates • Shelf... Agroalimentaire Canada Classical control • Worldwide, between 1888 and 1992 • > 5,500 programs with parasitoids & predators • < 50 programs with pathogens • Why? Agriculture and Agriculture et Agri-Food Canada Agroalimentaire Canada Pathophobia!! • Bacteria • Fungi • Protozoa • Viruses • Mutations Agriculture and Agriculture et Agri-Food Canada Agroalimentaire Canada Classical control • Fungi • Entomophaga maimaiga... Vertalec (Koppert) – Aphids • New formulations with improved efficacy • Registered in Europe and Japan • Applied on “1000’s of hectares” Agriculture and Agriculture et Agri-Food Canada Agroalimentaire Canada Innovative application: thinking outside of the box! Agriculture and Agriculture et Agri-Food Canada Agroalimentaire Canada Innovation Agriculture and Agriculture et Agri-Food Canada Agroalimentaire... Neem Institut für Nutzpflanzenwissenschaften und Ressourcenschutz der Rheinischen Friedrich-Wilhelms-Universität Bonn Characterisation of fitness parameters and population dynamics of Botrytis cinerea for the development of fungicide resistance management strategies in grapevine Inaugural-Dissertation zur Erlangung des Grades Doktor der Agrarwissenschaften (Dr agr.) der Landwirtschaftlichen Fakultät der Rheinischen Friedrich-Wilhelms-Universität Bonn vorgelegt am 08.08.2013 von Jürgen Derpmann aus Kalkar ABSTRACT Referent: Prof Dr H.-W Dehne Korreferent: Prof Dr H E Goldbach Tag der mündlichen Prüfung: 21.02.2014 Erscheinungsjahr: 2014 ABSTRACT Dedicated To My Parents ABSTRACT Jürgen Derpmann Characterization of Fitness Parameters and Population Dynamics of Botrytis cinerea for the Development of Fungicide Resistance Management Strategies in Grapevine Gray mold caused by the fungus Botrytis cinerea is an economically important disease in grapevine The pathogen has a high tendency to become resistant to frequently applied systemic fungicides Only a few years after introduction of the fungicide class of benzimidazoles (MBC), resistant strains appeared frequently in European vineyards Since the discontinuation of the use of benzimidazoles to control B cinerea in 1975, the frequency of MBC-resistant strains decreased significantly In the present study, the influence of fungicide resistance management strategies on the population dynamics of B cinerea isolates resistant to fungicides was investigated in a three year field trial at three sites near Bordeaux The tested strategies were mixture, alternation and annual alternation of thiophanate-methyl (TM) and mepanipyrim (MP) Strategies were compared to the solo application of TM and conventional fungicide treatments, where no TM was applied Frequencies of fungicide-resistant isolates were determined in monitoring procedures conducted prior and subsequent to fungicide applications In all three years, spray programs including TM resulted in significantly higher frequencies of TM-resistant isolates (BenR1 phenotype) compared to those detected in conventionally treated plots In the first year, all strategies tested led to similar BenR1 isolate frequencies compared to the solo application of TM (23%) In the second year, solo application of MP as part of the annual alternation resulted in significantly lower BenR1 isolate frequencies (16%) compared to spray programs including TM (39%) However, at the end of the study no significant differences in BenR1 isolate frequencies were detected between the strategies tested and the solo application of TM (47%) Different single nucleotide polymorphisms (SNP) in the β-tubulin gene confer resistance to MBC fungicides Allele-specific polymerase chain reactions (as-PCR) as well as EvaGreen® real-time as-qPCR showed a high correlation between the BenR1 isolate and E198A allele frequency Over the winter period 2009/10, a decrease of BenR1 isolate frequency was detected (-12%), which points to difference in fitness of MBC-sensitive (BenS) and BenR1 isolates Therefore, various fitness parameters were tested comparing ten BenS with ten BenR1 isolates At favourable conditions, no significant differences were detected between the two sensitivity groups At unfavourable conditions, mycelium growth, lesion size and spore production of BenS isolates were significantly higher than those of BenR1 isolates In a competitive assay on leaf discs as well as on grapevine plants a decrease in BenR1 conidia frequency of % per generation was observed Fitness costs associated with resistance could have reduced the frequency of BenR1 isolates within the primary inoculum, when the fungus was confronted with unfavourable development conditions If no MBC fungicides are applied during the season, then the short-distance dispersal of BenS conidia from the infected flowers and other sources leads to a decrease of the resistant fraction in the consecutive berry-associated population, as Tetrahedron Letters 51 (2010) 4344–4346 Contents lists available at ScienceDirect Tetrahedron Letters journal homepage: www.elsevier.com/locate/tetlet A facile construction of the tricyclic 5-7-6 scaffold of fungi-derived diterpenoids The first total synthesis of (±)-heptemerone G and a new approach to Danishefsky’s intermediate for a guanacastepene A synthesis Karol Michalak, Michał Michalak, Jerzy Wicha * Institute of Organic Chemistry, Polish Academy of Sciences, ul Kasprzaka 44/52, 01-224 Warsaw 42, Poland a r t i c l e i n f o Article history: Received 27 April 2010 Revised 27 May 2010 Accepted 11 June 2010 Available online 17 June 2010 a b s t r a c t The first total synthesis of (±)-heptemerone G, a diterpenoid metabolite of a submerged culture Coprinus heptemerus, and a new approach to an advanced intermediate for a synthesis of guanacastepene A are reported Ó 2010 Elsevier Ltd All rights reserved Keywords: Annulation reactions Medium-ring compounds Quaternary stereocenters Terpenoids Total synthesis Fungi-derived and microbial terpenoids, distinctive by the presence of medium rings in their structures, are important synthetic targets.1 Recently, the attention of several groups has focused on guanacastepenes, a family of diterpenoids isolated from endophilic fungi growing on the branches of the Daphnopsis americana tree (Guanacaste Conservation Area, Costa Rica).2 The first identified representative of this family, guanacastepene A (1, Fig 1), has a tricyclic structure with linearly fused five-, seven- and six-membered rings The ‘northern’ region of this molecule is highly polar while the opposite side is hydrophobic and bears two quaternary carbon atoms, and an isopropyl group More recently, structurally closely related terpenoids named heptemerones, including heptemerone G (2) were isolated from a broth of a submerged culture of Coprinus heptemerus.3 Interest in guanacastepene and heptemerone synthesis has been stimulated by their fascinating structures and biological activity The crude fermentation extracts of fungi from Daphnopsis as well as isolated guanacastepene A were found to be highly active against certain malicious antibiotic-resistant bacteria.2b Although the biological activity profile of guanacastepene A is encumbered with a detrimental side effect (lysis of human red blood cells), a new class of structures has been revealed for chemical and pharmacological exploration The first total synthesis of guanacastepene A (1) was reported by Danishefsky and co-workers.4 The synthesis of has also been accomplished by Shipe and Sorensen5 and formal total syntheses were reported by Hanna,6 Snider,7 and Mehta et al.8 Guanacastepene C was synthesized by Mehta et al.,8 guanacastepene N by Overman and co-workers,9 and guanacastepene E by Trauner and co-workers.10 To date, only one representative of the heptemerone family, heptemerone B, has been synthesized.10 Several approaches to advanced intermediates for guanacastepene synthesis have also been developed.11,12 15 H O O OH 14 AcO A C B 12 11 16 guanacastepene A 18 17 0040-4039/$ - see front matter Ó 2010 Elsevier Ltd All rights reserved doi:10.1016/j.tetlet.2010.06.064 O H OAc heptemerone G O O O * Corresponding author Tel.: +48 22 632 8117; fax: +48 22 6326681 E-mail address: jwicha@icho.edu.pl (J Wicha) O O O O Ot-Bu O Figure Structures of guanacastepene A, heptemerone G and the key synthetic intermediates 4345 K Michalak et al / Tetrahedron Letters 51 (2010) 4344–4346 O O O1 O H O OH 14 O O H a) O O O O O O O 15, X = H 16, X = SePh 14 t-BuO2C b) 13 14 X O O H OTMS c) O O O d) O O O e) O 8 17 18 Scheme Highlights of the proposed scheme for the synthesis of We now report the first total synthesis of heptemerone G (2) and, en route, a new synthetic approach to compound (which is a guanacastepene A precursor in the Danishefsky synthesis), via the versatile tricyclic intermediate The main features of the proposed synthetic route to are shown in Scheme The bicyclic MIT Center for Real Estate Week 12: Real Estate and Regional Economic Growth • Exports, transfers, investments and the determinants of regional growth: demand. • Population growth and migration: supply • 3-Q model of regional response. Factor supply elasticity and the role of real estate. • Wages, productivity and real estate costs – across MSAs. MIT Center for Real Estate Income and Product Accounts in States Summary of Output and Income Accounts for Florida and Pennsylvania, 1991 Florida ($ billions) Pennsylvania ($ billions) Income Accounts* Income (Y) 262 242 Wages (w) 126 127 Other Income (y + G) 136 115 Consumption (C) 260 193 Private 214 161 Government 46 32 Federal Taxes (T) 38 41 Savings (S) -36 8 Output Accounts** Output (Q) 219 211 Wages (w) 126 127 Profits and Rents (π) 93 84 Consumption (C) 260 193 Investment (I) 44 27 Imports (M) 175 153 Exports (X) 92 144 INCOME (Y) - OUTPUT (Q) 43 31 MIT Center for Real Estate Regional Accounts: Flow of Funds • Regions do not have to have individually balanced accounts. Surpluses in goods can be balanced by deficits in capital or government flows: the following cross border flows however must sum to zero. 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MIT Center for Real Estate Sources of Regional Demand . • Some variables are determined directly by the size of a state’s economy (Income or Output) : imports (M), Federal Taxes (T), consumption or savings (S) and profits earned in the state (π). • Other variables are determined by forces largely outside of the region and serve to bring money into the state, generating growth and ultimately determining state size (level of income or output): - Exports (X) - Investment (I) - Federal spending (G) - Unearned income: SS, retirement…(y) MIT Center for Real Estate Characterizing Export growth and Investment? ∑ e i n i = ∑ e i N+ ∑ e i (N i -N) + ∑ e i (n i -N i ) i i i i Share | Mix Competitive | Shift (i): industry n,e: regional growth in activity, level of activity N: national growth of activity • Share: a matter of timing • Mix: Historic industrial structure • Competitive: “our” companies versus “theirs” [innovation –vs- production costs: “product cycle] MIT Center for Real Estate Study of impact of each Demand factor on the Boston Area Economy over time (Coulson) Mix effect Share effect Competitive effect Impact on Region 0 1 2 3 4 years since start 8 9 10 11 MIT Center for Real Estate Regional Supply shifts are as important • Migration into a region that results from factors in the origin and not destination. [US history 1820-1920]. • Birth rates in the state – 20 years earlier! (Mass –vs- California Net Reproduction Rates). • Recent immigration from Mexico and Asia. MIT Center for Real Estate P Output Market Simultaneous Equilibrium in a region’s product, labor and structures markets. 1. Product Demand=production costs. 2. Costs = average of wages and rents. 3. Wages equilibrate labor supply with labor demand (proportional to output). 4. Rents do the same in structures market. Q D Q C=α K R + α L W W/P Labor Market L D =α L Q L S R Real Estate Market L K K D =α K Q W K S R MIT Center for Real Estate Changes in Regional output, prices, wages and rents in reaction to shift in product demand Qd to Qd’. 1). Prices (and costs) must rise. Ditto output. 2). Wages and employment rise. 3). Likewise for rents and stock of structures. 4). Reverse for downward demand shifts 5). Supply Elasticity and the Magnitude of price versus quantity changes? P Output Market Q Q D C=α Ecology of Fungi Ecology of Fungi Bởi: OpenStaxCollege Fungi play a crucial role in the ... act as agents of dispersal The benefit to the fungus outweighs the cost of producing fleshy fruiting bodies 11/12 Classifications of Fungi For each of the four groups of perfect fungi (Chytridiomycota,... formation of mycelia A basidiocarp is the fruiting body of a mushroom-producing fungus [link] D Review Questions The most primitive phylum of fungi is the 10/12 Classifications of Fungi Chytridiomycota... Classifications of Fungi The lifecycle of an ascomycete is characterized by the production of asci during the sexual phase The haploid phase is the predominant phase of the life cycle Which of