In Vitro Culture of Mycorrhizas PDF
Preview In Vitro Culture of Mycorrhizas
Soil Biology 4 Series Editor: Ajit Varma Stéphane Declerck Désiré-Georges Strullu J.-André Fortin (Eds.) In Vitro Culture of Mycorrhizas With84Figures,13inColor 123 Prof.Dr.StéphaneDeclerck Prof.Dr.Désiré-GeorgesStrullu UniversitécatholiquedeLouvain(UCL) Universitéd’Angers Mycothèquedel’Universitécatholique LaboratoiredePhytonique deLouvain(MUCL) 2Bd.Lavoisier Unitédemicrobiologie 49045AngersCedex 3PlaceCroixduSud France 1348Louvain-la-Neuve e-mail:[email protected] Belgium e-mail:[email protected] Prof.J.AndréFortin UniversitéLaval DépartementdeSciencesduBois etdelaForêt CentredeRechercheenBiologieForestière Sainte-Foy,QuébecG1K7P4 Canada e-mail:[email protected] LibraryofCongressControlNumber:2004116751 ISSN1613-3382 ISBN-103-540-24027-6SpringerBerlinHeidelbergNewYork ISBN-13978-3-540-24027-3SpringerBerlinHeidelbergNewYork This work is subject to copyright. All rights reserved, whether the whole or part of the materialisconcerned,specificallytherightsoftranslation,reprinting,reuseofillustrations, recitation,broadcasting,reproductiononmicrofilmorinanyotherway,andstorageindata banks.Duplicationofthispublicationorpartsthereofispermittedonlyundertheprovisions oftheGermanCopyrightLawofSeptember9,1965,initscurrentversion,andpermission forusemustalwaysbeobtainedfromSpringer.Violationsareliableforprosecutionunder theGermanCopyrightLaw. SpringerisapartofSpringerScience+BusinessMedia springeronline.com ©Springer-VerlagBerlinHeidelberg2005 PrintedinGermany Theuseofgeneraldescriptivenames,registerednames,trademarks,etc.inthispublication doesnotimply,evenintheabsenceofaspecificstatement,thatsuchnamesareexempt fromtherelevantprotectivelawsandregulationsandthereforefreeforgeneraluse. Coverdesign:design&production,Heidelberg,Germany Typesettingandproduction:LE-TEXJelonek,Schmidt&VöcklerGbR,Leipzig,Germany 31/3150-YL-543210-Printedonacid-freepaper Foreword Thefirst30cmoftheearth’ssurfacerepresentsafragileandvaluableecosys- tem,thankstowhichterrestrialplants,andindirectlyanimalsandhumans, canlive.Themicrobialactivityoccurringinsoilislargelyresponsiblefor itsphysicalandnutritionalquality.Amongthemicro-organismslivingin soil, the arbuscular mycorrhizal (AM) fungi play a major role. They are present in all types of soil, everywhere on the planet, living in symbiotic associationwiththerootsofmostplantspecies.Theyhaveco-evolvedwith plantsfor400millionyears,improvingtheirnutritionandresistancetovar- ious types of stress. Present practices in conventional agriculture, which introducegreatamountsofchemicals,haveeliminatedorunderexploited the AM symbiosis. The rational exploitation of AM fungi in sustainable agriculture,tohelpminimizetheuseofchemicalfertilizersandpesticides, has been hampered by several biological characteristics of these micro- organisms:theycannotbegrownintheabsenceofaplanthostandtheir geneticstructureisverycomplex. Despite these limitations, biologists have made important progress in understanding better the functioning of AM fungi. An in vitrotechnique has been developed using mycorrhizal root organ cultures, which made it possible to investigate the genetics, cell biology and physiology of AM fungi. We can now be objective enough to critically evaluate the impacts theinvitrotechniquehashadto improveourknowledgeonmycorrhizal symbiosis. Moreover, more experiences in using the technique allows us to appreciate its limits, as well as its yet unexploited scientific potential. AreviewonthesubjecthasbeenrecentlypublishedbyFortinetal(2002). Alongthesamelines,butinamuchmorecomprehensiveway,thisbook, throughcontributionsfromexperiencedspecialistsinthefield,offersvalu- ableinsightsintothemostrecentusesofthetechnique.Itillustrateshowim- portantquestionsregardinggermplasmcollection,taxonomy,physiology andmetabolismofarbuscularmycorrhizalfungicanbecleverlyaddressed bytakingadvantageoftheinvitrosystem.Italsoreportshowthetechnique hasbeenextendedtothecultureofothersymbioticfungi.Inauniqueway, aroot/fungussymbiosisnormallyoccurringinsoilismadeaccessiblefor various investigations: e.g. non-destructive microscope observations, re- liable cell physiology studies, clean biochemical and molecular analyses, andhighlycontrolledinteractionstudieswithothermicro-organisms.Be- causethesystemprovidesawaytocultivateinvitroanobligatebiotrophic micro-organism, it can even be used to produce aseptically, for the first time,AMfungalinoculaonanindustrialscale. Young scientists interested in mycorrhizal symbiosis will find in this book, not only valuable technical information, but also a rich source of inspiration for their research and for the further exploitation of the po- tential of mycorrhizal in vitro cultures. Like microscopy for cell biology, andthepolymerasechainreactionformolecularbiology,themycorrhizal rootorganculturesystemcanbeconsideredacriticalstepinthescientific history of mycorrhiza R&D. This book will certainly provide convincing evidencetosupportthisassertion. GuillaumeBécard Acknowledgements Wewouldliketothanktheauthorsfortheircontributionsandforalltheir efforts to produce and up-date a literature overview of so many different topics. The success of this book lies in their remarkable contributions. We owe special thanks to Professor Ajit Varma, who invited us to edit thisbookintheSeries,SoilBiology.Hisconstantencouragementishighly appreciated. We address special thanks to Dr. Jutta Lindenborn for her patience,dedicatededitorialassistanceandprofessionalismduringallsteps ofthisbookproductionandtoDr.DieterCzeschlik,whofoundourresearch avaluabletopicforthisSpringerseries. S.D.gratefullyacknowledgethefinancialsupportfromtheBelgianFed- eralSciencePolicyOffice(contractBCCMC3/10/003). S.Declerck,D.G.Strullu,J.A.Fortin November2004 Contents Part I State of the Art 1 InVitroCultureofMycorrhizas 3 J.AndréFortin,StéphaneDeclerck,Désiré-GeorgesStrullu 1 Introduction.................................................................. 3 2 AToolforGermplasmCollection...................................... 4 3 AToolforSystematicsandBiodiversity.............................. 5 4 LifeCycleofGlomusspp................................................... 6 5 LifeHistoryofGigasporaceae........................................... 6 6 EffectsofEnvironmentalFactors onHyphalGrowthandBranching..................................... 6 7 QuestioningtheValueofMonoxenicCultures..................... 7 8 AMFungi;HostandNon-Host ......................................... 7 9 CarbonandLipidMetabolism.......................................... 8 10 MonoxenicCultureandPhysiology ofinVitroGrownPlants.................................................. 9 11 NutrientDynamicsinAMMonoxenicCultures................... 9 12 AMFungiandRhizosphereMicro-Organisms .................... 10 13 CistusIncanusRootOrganstoStudyEctomycorrhizalFungi. 10 14 MonoxenicCultureofEdibleEctomycorrhizalFungi............ 11 15 TheUniqueGeosiphonSymbiosis...................................... 11 16 ShouldWeConsiderRoot-InhabitingSebacinaceae asMycorrhizalFungi?...................................................... 12 17 Industrial-ScaleMonoxenicAMFungusProduction............ 12 18 PreciseTechniquesforSuccessfulDevelopment ofAMMonoxenicCulture................................................ 13 19 Conclusion .................................................................... 13 References............................................................................. 14 X Contents Part II Systematics 2 TheMonoxenicCultureofArbuscularMycorrhizalFungi asaToolforGermplasmCollections 17 StéphaneDeclerck,SylvieSéguin,YolandeDalpé 1 Introduction.................................................................. 17 2 HistoricalPerspectiveofAMFungiCultureCollections........ 17 3 PrerequisitetoIncludeAMFungi inMonoxenicCultureCollections ..................................... 19 4 CultureProperties: Viability–Identity–Purity–Stability(VIPS)..................... 20 5 Long-TermConservation................................................. 22 6 StrengthsandWeaknessesofMonoxenicCultureCollections 25 7 Conclusion .................................................................... 28 References............................................................................. 29 3 TheMonoxenicCultureofArbuscularMycorrhizalFungi asaToolforSystematicsandBiodiversity 31 YolandeDalpé,SylvieCranenbrouck,SylvieSéguin, StéphaneDeclerck 1 Introduction.................................................................. 31 2 Systematics.................................................................... 32 2.1 SpeciesandStrainAvailability................................. 32 2.2 FungalMyceliaandSpores...................................... 33 2.3 BiochemicalStudies............................................... 36 2.4 MolecularStudies.................................................. 37 3 Biodiversity ................................................................... 39 3.1 TrappingofIsolates................................................ 39 3.2 Micro-Morphology................................................ 40 3.3 FunctionalDiversity .............................................. 41 4 Conclusion .................................................................... 43 References............................................................................. 43 4 LifeCycleofGlomusSpeciesinMonoxenicCulture 49 YolandeDalpé,FranciscoAdrianodeSouza,StéphaneDeclerck 1 Introduction.................................................................. 49 2 LifeCycle....................................................................... 49 3 AMFungiPropaguleGerminationStage ............................ 50 4 Pre-SymbioticMyceliumStage.......................................... 53 5 HostRootConnectingStage............................................. 54 6 SymbioticStage.............................................................. 55 6.1 IntraradicalMycelium............................................ 56 Contents XI 6.2 ExtraradicalMycelium........................................... 58 7 Spores........................................................................... 61 8 Conclusion .................................................................... 65 References............................................................................. 65 5 LifeHistoryStrategiesinGigasporaceae: InsightfromMonoxenicCulture 73 FranciscoAdrianodeSouza,YolandeDalpé,StéphaneDeclerck, IvanEnriquedelaProvidencia,Nathalie Séjalon-Delmas 1 Introduction.................................................................. 73 2 TheFamilyGigasporaceaeandItsOccurrence .................... 73 3 LifeCycle....................................................................... 74 3.1 Pre-SymbioticPhase.............................................. 74 3.2 SymbioticVegetative andReproductiveGrowingPhases........................... 77 4 GeneticDiversityandPhenotypicVariation ....................... 80 4.1 VegetativeCompatibilityTest(VCT)......................... 81 5 LifeHistoryStrategy(LHS)ofGigasporaceae, asRevealedUsingMonoxenicCultures .............................. 82 5.1 Co-ExistenceandCompetitionExperiments UnderDixenicCulture ........................................... 85 5.2 EcologicalImplications oftheGigasporaceaeLifeHistoryStrategy................. 85 6 Conclusions................................................................... 87 References............................................................................. 88 Part III In Vitro Development and Physiology of Glomeromycetes 6 EnvironmentalFactors ThatAffectPresymbioticHyphalGrowthandBranching ofArbuscularMycorrhizalFungi 95 GeraldNagahashi,DavidD.DoudsJr. 1 Introduction.................................................................. 95 2 InVitroTechniques......................................................... 96 2.1 SporeProductionandGermination.......................... 96 2.2 RootOrganCulturesandRootExudates................... 96 2.3 IncubationConditions withCO andExudateTreatments............................ 97 2 2.4 LightExperiments................................................. 97 2.5 SynergisticEffectsBetweenChemicalCompounds..... 98 XII Contents 3 ChemicalComponentsofExudatesorCompounds FoundintheSoilEnvironment ThatInfluencePresymbioticAMFungalGrowth................. 98 4 EffectsofVolatileCompounds onPresymbioticAMFungalGrowth.................................. 100 4.1 EffectsofCO ontheHyphalGrowth 2 ofGerminatedGigasporaGiganteaSpores................. 101 5 TheEffectofLightonthePresymbioticGrowth ofAMFungi................................................................... 103 6 Conclusions................................................................... 106 References............................................................................. 108 7 BreakingMythsonArbuscularMycorrhizasinVitroBiology 111 BertBago,CustodiaCano 1 Introduction.................................................................. 111 2 QuestioningAMMonoxenicCultures................................ 113 2.1 AreAMMonoxenicCultures DevicesTooArtificialtoTrust?................................ 113 2.2 AreTransformedRootOrgansaGoodHostMaterial toStudyAMFungalBiology?................................... 116 2.3 TheDownfallofTwoColonizationMyths.................. 118 2.4 AreBranchedAbsorbingStructures(BAS)Commonly FormedbyallGlomaleanFungi?AreTheyArtifacts FormedOnlyUnderinVitroConditions?.................. 119 2.5 AreThereAnyDifferences inAMFungalDevelopment inMonoxenicsVersusSoil? ..................................... 124 2.6 AreAMMonoxenicLiquidCulturesAccurate?........... 126 2.7 TheDangerofContamination inAMMonoxenicCultures ..................................... 128 2.8 WhatElseHaveMonoxenicCultures toOfferontheStudyofAMFungalBiology?.............. 129 3 Conclusion .................................................................... 133 References............................................................................. 133 8 HostandNon-HostImpactonthePhysiology oftheAMSymbiosis 139 HorstVierheilig,BertBago 1 Introduction.................................................................. 139 2 AsymbioticAMFungalGrowth......................................... 140 2.1 pH....................................................................... 141 2.2 Temperature......................................................... 141
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