AnumberofbacteriawereisolatedfromseawaterinSkagerrak,Denmark,at30mdepth.Twooftheisolates,strainsD28andD30^T,belongedtotheFlavobacteriaceaewithintheCytophaga–Flavobacterium–Bacteroidesgroup.Sequencingof16SrRNAgenesofthetwostrainsindicatedstronglythattheybelongedtothegenusTenacibaculumandthattheyshowedgreatestsimilaritytothespeciesTenacibaculumamylolyticumandTenacibaculummesophilum.DNA–DNAhybridizationvalues,DNAbasecompositionandphenotypiccharacteristicsseparatedtheSkagerrakstrainsfromtheotherspecieswithinTenacibaculum.Thus,itisconcludedthatthestrainsbelongtoanovelspecieswithinthegenusTenacibaculum,forwhichthenameTenacibaculumskagerrakensesp.nov.isproposed,withstrainD30^T(=ATCCBAA-458^T=DSM14836^T)asthetypestrain.

ThediversityofculturablebacteriaassociatedwithseaicefromfourpermanentlycoldfjordsofSpitzbergen,ArcticOcean,wasinvestigated.Atotalof116psychrophilicandpsychrotolerantstrainswereisolatedunderaerobicconditionsat4°C.TheisolatesweregroupedusingamplifiedrDNArestrictionanalysisfingerprintingandidentifiedbypartialsequencingof16SrRNAgene.Thebacterialisolatesfellinfivephylogeneticgroups:subclassesαandγofProteobacteria,theBacillus–Clostridiumgroup,theorderActinomycetales,andtheCytophaga–Flexibacter–Bacteroides(CFB)phylum.Over70%oftheisolateswereaffiliatedwiththeProteobacteriaγsubclass.Basedonphylogeneticanalysis(<98% sequence="" similarity),="" over="" 40%="" of="" arctic="" isolates="" represent="" potentially="" novel="" species="" or="" genera.="" most="" of="" the="" isolates="" were="" psychrotolerant="" and="" grew="" optimally="" between="" 20="" and="" 25°c.="" only="" a="" few="" strains="" were="" psychrophilic,="" with="" an="" optimal="" growth="" at="" 10–15°c.="" the="" majority="" of="" the="" bacterial="" strains="" were="" able="" to="" secrete="" a="" broad="" range="" of="" cold-active="" hydrolytic="" enzymes="" into="" the="" medium="" at="" a="" cultivation="" temperature="" of="" 4°c.="" the="" isolates="" that="" are="" able="" to="" degrade="" proteins="" (skim="" milk,="" casein),="" lipids="" (olive="" oil),="" and="" polysaccharides="" (starch,="" pectin)="" account="" for,="" respectively,="" 56,="" 31,="" and="" 21%="" of="" sea-ice="" and="" seawater="" strains.="" the="" temperature="" dependences="" for="" enzyme="" production="" during="" growth="" and="" enzymatic="" activity="" were="" determined="" for="" two="" selected="" enzymes,="" α-amylase="" and="" β-galactosidase.="" interestingly,="" high="" levels="" of="" enzyme="" productions="" were="" measured="" at="" growth="" temperatures="" between="" 4="" and="" 10°c,="" and="" almost="" no="" production="" was="" detected="" at="" higher="" temperatures="" (20–30°c).="" catalytic="" activity="" was="" detected="" even="" below="" the="" freezing="" point="" of="" water="" (at="" −5°c),="" demonstrating="" the="" unique="" properties="" of="" these="" enzymes.="">

TheGram-positive,alkali-andsalt-tolerantmarinebacteriumstrainP203^Tisdescribedtogetherwithitsclosestphylogeneticneighbour,terrestrialisolateLMG21005^T.StrainP203^TwasisolatedfrommaterialfromthespongePlakortissimplexthatwasobtainedfromtheSula-Ridge,NorwegianSea.StrainLMG21005^TwasanundescribedstrainthatwasisolatedfromachurchwallmuralinGermany.StrainsP203^TandLMG21005^TwereidentifiedasnovelalkalitolerantmembersoftheBacillusrRNAgroup6witha16SrRNAgenesequencesimilarityof99.5 %.Theclosestdescribedneighbour,BacillusgibsoniiDSM8722^T,showed99.0 %genesequencesimilaritywithP203^Tand98.8 %similaritywithstrainLMG21005^T.Despitethehigh16SrRNAgenesequencesimilarity,DNA–DNAcross-hybridizationrevealedonly25.8–34.1 %similarityamongstthethreestrains.TheDNAG+Ccontentswere41.1 mol%forstrainP203^Tand39.6 mol%forstrainLMG21005^T.BothstrainsgrewwellbetweenpH7andpH11.StrainP203Tshowedgrowthatmoderatetemperatures(from4to30°C)andinthepresenceofupto12 %(w/v)NaClatpH9.7,whereasstrainLMG21005^Twasnotsalttolerant(upto4 %NaCl)andnogrowthwasobservedat4°C.ThemajorfattyacidsofstrainsP203^T,LMG21005^TandthetypestrainofB.gibsoniiwerethesaturatedterminallymethyl-branchedcompoundsiso-C15 : 0(19.8,15.6and28.0 %,respectively)andanteiso-C15 : 0(57.1,48.6and45.2 %,respectively).PhysiologicalandbiochemicaltestsallowedgenotypicandphenotypicdifferentiationofstrainsP203^TandLMG21005^TfromthesixrelatedBacillusspecieswithvalidlypublishednamesandsupportedtheproposaloftwonovelspecies,Bacillusplakortidis[typestrainP203^T(=DSM19153^T=NCIMB14288^T)]andBacillusmurimartini[typestrainLMG21005^T(=NCIMB14102^T)].

Reclassificationof‘Pseudomonasfluorescenssubsp.cellulosa’NCIMB10462(Uedaetal.1952)asCellvibriojaponicussp.nov.andrevivalofCellvibriovulgarissp.nov.,nom.rev.andCellvibriofulvussp.nov.,nom.rev.
Pseudomonasfluorescenssubsp.cellulosa’NCIMB10462hasbeendemonstratedbyapolyphasictaxonomicapproachtobeamemberofthegenusCellvibrio.16SrDNAsequenceanalysissuggeststhatthisistheonlygenusthatcouldacceptthisspecimen.Thesequenceis95·5 %similartothatofCellvibriomixtussubsp.mixtusACM2601^T(thetypestrainofthetypespeciesofthegenus),whichisitsclosestrelation.ThegenomicDNAG+Ccontentwasdeterminedtobe53·3 mol%,whichissimilartothevaluesobtainedforthevalidlydescribedCellvibriospecies.DNA–DNAhybridizationexperimentshaveshownthatstrainNCIMB10462^T(=NCDO2697^T)representsanovelspecies;therefore,itisproposedthatitbedesignatedasthetypestrainofthenovelspeciesCellvibriojaponicassp.nov.Thisstudyalsoused16SrDNAanalysis,DNA–DNAhybridizationexperimentsandphenotypictestingtorevivethespeciesCellvibriovulgarissp.nov.,nom.rev.andCellvibriofulvussp.nov.,nom.rev.C.vulgarisNCIMB8633^T(=LMG2848^T)andC.fulvusNCIMB8634^T(=LMG2847^T)aretheproposedtypestrains.

Anovelaerobic,psychrotolerantmarinebacteriumwasisolatedat4°CfromseawatersamplescollectedfromSpitzbergenintheArctic.Thestrainwasapolar-flagellated,Gram-negativebacteriumthatgrewoptimallyat10–15°CandpH7–8inmediacontaining2–3 %NaCl(w/v),usingvariouscarbohydratesandorganicacidsassubstrates.Themainfattyacidcomponentsincluded16 : 0(12.7 %oftotalfattyacids),straight-chainsaturatedfattyacidmethylester(FAME)and16 : 1ω7c(40.2 %)monounsaturatedFAME.Phylogeneticanalysisrevealedacloserelationship(99 %16SrRNAgenesequencesimilarity)betweenthenovelisolateandPseudoalteromonaselyakoviiKMM162^TandsomeotherspeciesofthegenusPseudoalteromonas.TheDNAG+Ccontentofthenovelstrainwas39mol%.DNA–DNAhybridizationshowedonly47.6 %DNA–DNArelatednesswithP.elyakoviiKMM162^T,44.2 %withPseudoalteromonasdistinctaKMM638^Tand22.6 %withPseudoalteromonasnigrifaciensNCIMB8614^TBasedonphylogeneticandphenotypiccharacteristics,thisisolaterepresentsanovelspeciesofthegenusPseudoalteromonasforwhichthenamePseudoalteromonasarcticaisproposed;thetypestrainisA37-1-2^T(=LMG23753^T=DSM18437^T).

Introduction:Inwood-dwellingfungus-farmingweevils,theso-calledambrosiabeetles(Curculionidae:ScolytinaeandPlatypodinae),woodintheexcavatedtunnelsisusedasamediumforcultivatingfungibythecombinedactionofdigginglarvae(whichcreatemorespaceforthefungitogrow)andofadultssowingandpruningthefungus.Thebeetlesareobligatelydependentonthefungusthatprovidesessentialvitamins,aminoacidsandsterols.However,towhatextentmicrobialenzymessupportfungusfarminginambrosiabeetlesisunknown.Herewemeasure(i)13plantcell-walldegrADIngenzymesinthefungusgardenmicrobialconsortiumoftheambrosiabeetleXyleborinussaxesenii,includingitsprimaryfungalsymbionts,inthreecompartmentsoflaboratorymaintainednests,atdifferenttimepointsaftergalleryfoundationand(ii)fourspecificenzymesthatmaybeeitherinsectormicrobiallyderivedinX.saxeseniiadultandlarvalindividuals.Results:Wediscoveredthattheactivityofcellulasesinambrosiafungusgardensisrelativelysmallcomparedtotheactivitiesofothercellulolyticenzymes.Enzymeactivityinallcompartmentsofthegardenwasmainlydirectedtowardshemicellulosecarbohydratessuchasxylan,glucomannanandcallose.Hemicellulolyticenzymeactivitywithinthebroodchamberincreasedwithgalleryage,whereasirrespectiveoftheageofthegallery,thehighestoverallenzymeactivityweredetectedinthegallerydumpmaterialexpelledbythebeetles.Interestinglyendo-β-1,3(4)-glucanaseactivitycapableofcallosedegradationwasidentifiedinwhole-bodyextractsofbothlarvaeandadultX.saxesenii,whereasendo-β-1,4-xylanaseactivitywasexclusivelydetectedinlarvae.Conclusion:Similartocloselyrelatedfungiassociatedwithbarkbeetlesinphloem,themicrobialsymbiontsofambrosiabeetleshardlydegradecellulose.Instead,theirenzymeactivityisdirectedmainlytowardscomparativelymoreeasilyaccessIBLehemicellulosecomponentsoftheray-parenchymacellsinthewoodxylem.FurThermore,thedetectionofxylanolyticenzymesexclusivelyinlarvae(whichfeedonfunguscolonizedwood)andnotinadults(whichfeedonlyinfungi)indicatesthatonlylarvae(pre-)digestplantcellwallstructures.ThisimpliesthatinX.saxeseniiandlikelyalsoinmanyotherambrosiabeetles,adultsandlarvaedonotcompeteforthesamefoodwithintheirnests-incontrast,larvaeincreasecolonyfitnessbyfacilitatingenzymaticwooddegradationandfunguscultivation.

Fungus-growing(attine)antsandtheirfungalsymbiontspassedthroughseveralevolutionarytransitionsduringtheir50millionyearoldevolutionaryhistory.Thebasalattinelineagesoftenshiftedbetweentwomaincultivarclades,whereasthederivedhigher-attinelineagesmaintainedanassociationwithamonophyleticcladeofspecializedsymbionts.Inconjunctionwiththetransitiontospecializedsymbionts,theantsadvancedincolonysizeandsocialcomplexity.Hereweprovideacomparativestudyofthefunctionalspecializationinextracellularenzymeactivitiesinfungusgardensacrosstheattinephylogeny.Weshowthat,relativetosisterclades,gardensofhigher-attineantshaveenhancedactivityofprotein-digestingenzymes,whereasgardensofleaf-cuttingantsalsohaveincreasedactivityofstarch-digestingenzymes.However,theenzymeactivitiesoflower-attinefungusgardensaretargetedprimarilytowardpartialdegradationofplantcellwalls,reflectingaplesiomorphicstateofnondomesticatedfungi.Theenzymeprofilesofthehigher-attineandleaf-cuttinggardensappearparticularlysuitedtodigestfreshplantmaterialsandtoaccessnutrientsfromlivecellswithoutmajorbreakdownofcellwalls.Theadaptivesignificanceofthelower-attinesymbiontshiftsremainsunclear.Oneoftheseshiftswasobligate,butdigestiveadvantagesremainedambiguous,whereastheotherremainedfacultativedespiteprovidinggreaterdigestiveefficiency.

PenicilliumochrochloronBiourge,whichwasisolatedfromrottensweetpotato,canproduceplantcellwalldegradingenzymes(PCWDEs)withhighviscosityreducingcapABIlityforethanolproductionusingfreshsweetpotatotubersasfeedstock.Theenzymepreparationwascharacterizedbyabroadenzymespectrumincluding13kindsofenzymeswiththeactivitytohydrolyzecellulose,hemicellulose,pectin,starchandprotein.Themaximumviscosityreducingcapabilitywasobservedwhentheenzymepreparationwasobtainedafterfivedaysfermentationusing20g/Lcorncobassolecarbonsource,4.5g/LNH₄NO₃assolenitrogensource,andaninitialmediumpHof6.5.Thesweetpotatomashtreatedwiththeenzymepreparationexhibitedmuchhigherfermentationefficiency(92.58%)comparedwithcommercialcellulase(88.06%)andcontrol(83.5%).Theenzymeproductionwasthenscaledupto0.5,5,and100L,andtheviscosityreducingrateswerefoundtobe85%,90%,and91%,respectively.Thus,P.ochrochloronBiourgedisplayspotentialviscosityreducingcapabilityforethanolproduction.

Chineseliquorisoneoftheworld"sbest-knowndistilledspiritsandisthelargestspiritcategorybysales.Theuniqueandtraditionalsolid-statefermentationtechnologyusedtoproduceChineseliquorhasbeenincontinuoususeforseveralthousandyears.Thediverseanddynamicmicrobialcommunityinaliquorstarteristhemaincontributortoliquorbrewing.However,littleisknownabouttheecologicaldistributionandfunctionalimportanceofthesecommunitymembers.Inthisstudy,metatranscriptomicswasusedtocomprehensivelyexploretheactivemicrobialcommunitymembersandkeytranscriptswithsignificantfunctionsintheliquorstarterproductionprocess.Fungiwerefoundtobethemostabundantandactivecommunitymembers.Atotalof932carbohydrate-activeenzymes,includinghighlyexpressedauxiliaryactivityfamily9and10proteins,wereidentifiedat62°Cunderaerobicconditions.Somepotentialthermostableenzymeswereidentifiedat50,62,and25°C(maturestage).Increasedcontentandoverexpressedkeyenzymesinvolvedinglycolysisandstarch,pyruvateandethanolmetabolismweredetectedat50and62°C.Thekeyenzymesofthecitratecyclewereup-regulatedat62°C,andtheirabundantderivativesarecrucialforflavorgeneration.Here,themetabolismandfunctionalenzymesoftheactivemicrobialcommunitiesinNFliquorstarterwerestudied,whichcouldpavethewaytoinitiateimprovementsinliquorqualityandtodiscovermicrobesthatproducenovelenzymesorhigh-valueaddedproducts.