commiss decis septemb detail interim test scheme diagnosi detect identif pseudomona solanacearum smith smith potato ec commiss european commun regard treati establish european commun regard council direct eec decemb protect measur introduct commun organ harm plant plant product spread commun amend direct ec articl thereof commiss decis ec novemb author member state temporarili addit measur dissemin pseudomona solanacearum smith smith kingdom netherland amend decis ec articl paragraph bb thereof member state carri offici offici supervis test potato requir quarantin procedur pseudomona solanacearum smith smith establish european mediterranean plant protect organ eppo procedur approv accord procedur laid articl direct eec ad hoc expert committe bacteri diseas plant set servic commiss european commun auspic stand committe plant health determin detail interim test scheme take account improv detect test procedur develop public eppo quarantin procedur test scheme interim research anticip sensit specif individu test aim select standard optimum test inclus updat test method interim test scheme provid decis accord opinion stand committe plant health adopt decis articl purpos implement provis articl paragraph bb commiss decis ec amend quarantin procedur pseudomona solanacearum smith smith establish european mediterranean plant protect organ eppo replac interim test scheme diagnosi detect identif pseudomona solanacearum smith smith laid annex decis articl decis address member state brussel septemb commiss franz fischler member commiss oj oj oj oj bulletin eppo oepp annex interim test scheme diagnosi detect identif pseudomona solanacearum smith smith scope test scheme present scheme describ procedur involv diagnosi brown rot potato plant tuber ii detect pseudomona solanacearum sampl potato tuber iii identif pseudomona solanacearum appendic detail provid prepar test materi growth media buffer solut reagent content section applic test scheme diagnosi brown rot potato plant tuber detect identif pseudomona solanacearum sampl potato tuber section ii diagnosi brown rot potato plant tuber symptom brown rot rapid screen test isol procedur confirm test section iii detect identif pseudomona solanacearum sampl potato tuber prepar sampl test test elisa test pcr test select plate test bioassai test enrich test pathogen test appendix nutrient media isol cultur pseudomona solanacearum appendix materi sampl prepar appendix materi test appendix determin level contamin test appendix materi elisa test appendix materi pcr test appendix materi select plate enrich test refer section applic test scheme diagnosi brown rot potato plant tuber test procedur intend plant tuber symptom typic suspect brown rot involv rapid screen test isol pathogen infect vascular tissu diagnost media case posit result identif cultur pseudomona solanacearum flow chart diagram present gt refer graphic gt flowchart refer gt start graphic gt descript symptom provid section ii rapid screen test facilit presumpt diagnosi test stream test vascular stem tissu section ii test poli hydroxybutyr granul section ii test section iii elisa test section iii pcr test section iii isol pathogen plant materi typic symptom dilut plate straightforward cultur fail advanc stage infect saprophyt bacteria grow diseas tissu outgrow inhibit pathogen isol medium isol test neg diseas symptom typic isol repeat prefer select plate test reliabl identif pure cultur pseudomona solanacearum achiev test list section ii combin pathogen test section ii strain character option recommend case gt end graphic gt detect identif pseudomona solanacearum sampl potato tuber procedur intend detect latent infect potato tuber prefer screen test posit complement isol pathogen case isol typic coloni identif pure cultur pseudomona solanacearum flow chart diagram present gt refer graphic gt flowchart refer gt start graphic gt sampl size standard size tuber procedur appli conveni sampl tuber screen test singl test suffici sensit reliabl detect pseudomona solanacearum sampl test recommend test prefer base biolog principl immunofluoresc test test establish screen test advantag test fulli develop valid test statutori bacteria clavibact michiganensi subsp sepedonicu read paramet method sensit test sensit threshold cell ml potato extract pellet critic factor reliabl test result qualiti antiserum antiserum high titr minimum crude antiserum accept test carri antiserum titr dilut titr indirect method prefer direct method appli test level sensit specif equival indirect method test advantag subject interpret cell stain morpholog fluoresc intens provid inform reaction specif cross reaction serolog relat bacteria soil potato tissu cell morpholog pseudomona solanacearum common test sole screen test case cross reaction suspect addit screen test base biolog principl case select plate test select plate modifi smsa medium test methodolog method sensit select test pseudomona solanacearum result dai sampl prepar pathogen obtain directli cultur readili identifi full exploit potenti test requir care prepar heel end avoid secondari bacteria potato tuber competitor pseudomona solanacearum medium affect develop pathogen strain grow poorli compon medium affect target organ care requir differenti pseudomona solanacearum bacteria develop medium select plate sole screen test provid case inhibit pseudomona solanacearum bacteria plate suspect neg test result obtain sampl retest test confirm refut diagnosi case test test elisa test elisa test gener sensit test sensit threshold cell ml potato extract pellet test cheap fast gener vulner fals posit cross reaction fals neg result inhibit phenol molecul potato extract requir antiserum specif extrem high elisa test singl screen test pcr test pcr potenti sensit detect test readili inhibit plant tuber extract compon result fals neg potato cultivar inhibitor remov inhibitor inhibit reduc dilut popul pseudomona solanacearum dilut great care step sampl test prepar prevent contamin result fals posit test fals posit aris sequenc homolog organ reason direct pcr sole screen test enrich test incub potato extract pellet sampl semi select broth modifi smsa broth multipl pseudomona solanacearum import dilut potenti inhibitor elisa pcr test pseudomona solanacearum enrich broth detect elisa pcr recommend direct plate made enrich broth enrich method test includ good potenti rel lack experi sole detect method bioassai test bioassai test isol pseudomona solanacearum potato extract pellet select enrich host plant tomato plant eggplant test requir optim incub condit method bacteria inhibitori pseudomona solanacearum smsa medium interfer test confirm test reliabl identif pure cultur pseudomona solanacearum achiev test list section ii combin pathogen test section ii strain character option recommend case gt end graphic gt section ii diagnosi brown rot potato plant tuber symptom brown rot potato plant earli stage infect wilt leav top plant high temperatur dai recoveri night wilt rapidli irrevers result death plant vascular tissu transvers cut stem wilt plant brown milki ooz exud cut surfac easili express squeez cut stem vertic water thread slime stream vascular bundl potato tuber potato tuber cut transvers close heel stolon end earli stage infect glassi yellow light brown discolor vascular ring pale cream ooz emerg spontan minut gentl pressur appli thumb skin cut surfac vascular discolor distinct brown necrosi extend parenchymat tissu advanc stage infect break outward heel end ey result reddish brown slightli sunken lesion skin bacteria ooz caus soil particl adher rapid screen test rapid screen test facilit presumpt diagnosi test stem stream test presenc pseudomona solanacearum wilt potato stem assess simpl presumpt test cut stem soil level place cut surfac beaker water shortli thread bacteri slime stream spontan vascular bundl bacteria caus vascular infect potato plant show phenomenon detect poli hydroxybutyr phb granul phb granul cell pseudomona solanacearum visual stain nile blue sudan black prepar smear ooz suspend tissu microscop slide prepar smear hour cultur ypga spa appendix prepar posit control smear biovar race strain consid neg control smear heterolog strain dry pass lower surfac slide time rapidli flame smear fix nile blue test flood fix smear aqueou solut nile blue incub minut drain stain solut wash briefli gentli run tap water remov excess water tissu paper flood smear aqueou acet acid incub minut ambient temperatur wash gentli run tap water blot dry tissu paper remoisten drop water appli coverslip examin stain smear epifluoresc microscop nm oil immers magnif observ bright orang fluoresc phb granul observ normal light ensur granul intracellular cell morpholog typic pseudomona solanacearum sudan black test flood fix smear sudan black solut ethanol incub minut ambient temperatur drain stain solut wash briefli tap water remov excess water remov excess water tissu paper dip smear briefli xylol blot dry tissu paper caution xylol harm product work fume cabinet flood smear aqueou safranin leav second ambient temperatur caution safranin harm product work fume cabinet wash gentli run tap water blot dry tissu paper appli coverslip examin stain smear light microscop transmit light oil immers magnif phb granul cell pseudomona solanacearum stain blue black cell wall stain pink test screen test test section iii elisa test section iii pcr test section iii isol procedur remov ooz section discolour tissu vascular ring tuber vascular strand stem suspend small volum steril distil water mm phosphat buffer leav minut bench prepar seri decim dilut suspens gt num gt gt den gt gt num gt gt den gt consid transfer standard volum suspens dilut gener nutrient medium na ypga spa appendix kelman tetrazolium medium appendix smsa select medium appendix spread streak dilut plate techniqu consid prepar separ plate medium dilut cell suspens cultur virul biovar race strain pseudomona solanacearum posit control incub plate dai incub prolong dai growth slow coloni smsa plate atyp die gener nutrient media virul isol pseudomona solanacearum develop pearli white flat irregular fluidal coloni characterist whorl kelman tetrazolium medium typic coloni virul isol pseudomona solanacearum cream flat irregular fluidal blood red colour whorl centr avirul form pseudomona solanacearum develop butyr deep red coloni smsa medium typic coloni virul isol pseudomona solanacearum milki white flat irregular fluidal blood red colour centr avirul form pseudomona solanacearum develop fluidal coloni complet pink red smsa medium purifi coloni characterist morpholog subcultur gener nutrient medium avoid regular subcultur induc loss virul confirm test identif pseudomona solanacearum identifi pure cultur pseudomona solanacearum procedur nutrit enzymat test note includ control strain test phenotyp properti pseudomona solanacearum univers present absent gt tabl gt media method provid lelliott amp stead test prepar suspens cell ml cultur control strain prepar seri twofold dilut antiserum appli procedur section iii titr cultur equival posit control elisa test prepar suspens amp gt cell ml cultur control strain appli elisa procedur section iii elisa cultur equival posit control pcr test prepar suspens cell ml cultur control strain appli pcr procedur section iii pcr product cultur size restrict enzym analysi rea pattern posit control fluoresc situ hybridis fish prepar suspens cell ml cultur control strain appli fish procedur van beuningen al oli pcr primer seal al cultur show reaction posit control protein profil denatur cell protein separ polyacrylamid gel electrophoresi page stead fatti acid profil fap grow cultur posit control strain hour trypticas soi agar appli fap procedur jans stead stead profil cultur ident posit control condit characterist fatti acid und strain character strain character option recommend case biovar determin pseudomona solanacearum separ biovar basi abil produc acid hexos alcohol sugar hayward amp gt tabl gt addit test differenti biovar subphenotyp hayward gt tabl gt race determin race buddenhagen al determin basi pathogen test tomato plant eggplant tobacco plant hypersensit reaction hr test tobacco leav lozano sequeira gt tabl gt race character pathogen test tobacco hypersensit test highli reliabl deduc biovar natur host origin cultur character genom fingerprint molecular differenti strain pseudomona solanacearum complex rflp analysi cook al repetit sequenc pcr rep eric amp box pcr louw al smith al pathogen test test confirm diagnosi pseudomona solanacearum assess virul cultur identifi pseudomona solanacearum prepar inoculum cell ml cultur posit control strain inocul tomato plant eggplant prefer true leaf stage older section iii incub week high rel humid daili water observ wilt epinasti chlorosi stunt isol symptomat plant remov section tissu stem cm inocul point comminut suspend small volum steril distil water mm phosphat buffer plate incub check typic coloni pseudomona solanacearum section iii detect identif pseudomona solanacearum sampl potato tuber note standard sampl size tuber procedur appli conveni sampl tuber prepar sampl test note potato extract pellet obtain procedur detect clavibact michiganensi subsp sepedonicu pre test option consid incub sampl week encourag multipl low pseudomona solanacearum popul ii wash tuber run water disinfect deterg air dry tuber remov clean disinfect scalpel veget knife skin heel end tuber vascular tissu visibl carefulli cut small conic core mm diamet vascular tissu heel end amount vascular tissu minimum process tuber sampl note visual examin tuber section ii stage set tuber symptom sever rot test separ section ii collect heel end close contain prefer heel end process immedi store hour longer hour process heel end procedur transfer heel end contain add suffici volum macer buffer appendix cover heel end comminut heel end ware blender ultra thurrax complet homogen achiev avoid excess homogen macer soak minut ii transfer heel end contain add suffici volum macer buffer cover heel end place contain rotari shaker incub rpm hour hour iii transfer heel end strong dispos macer bag stomach bag dimens mm mm radiat steril crush heel end carefulli tool hammer complet homogen achiev add suffici volum macer buffer cover heel end macer settl minut extract bacteria process heel end procedur decant macer gentli centrifug tube leav debri contain bag decant macer cloudi centrifug minut temperatur centrifug decant macer supernat centrifug step minut minut temperatur discard supernat disturb pellet ii filter macer filtrat system pore size µm enhanc filtrat vacuum pump collect filtrat centrifug tube wash filter macer buffer centrifug filtrat minut minut temperatur discard supernat disturb pellet resuspend pellet ml pelletbuff appendix divid equal part transfer part microvi microvi test conserv remaind extract test add steril glycerol microvi vortex store week month test antiserum pseudomona solanacearum prefer race biovar determin titr suspens cell ml homolog strain pseudomona solanacearum dilut fluorescein isothiocyan fitc conjug manufactur recommend crude antiserum titr multiwel microscop slide prefer window mm diamet includ fitc conjug control test slide test repeat pb control includ posit cell observ fitc control prepar separ posit control slide suspens cell ml strain race biovar pseudomona solanacearum slide set test prepar test slide procedur pellet rel starch pipett measur standard volum µl mm window diamet scale volum larger window resuspend pellet row window remain row duplic sampl present figur ii pellet prepar decim dilut gt num gt gt den gt gt num gt gt den gt gt num gt gt den gt resuspend pellet pelletbuff pipett measur standard volum µl mm window diamet scale volum larger window resuspend pellet dilut row window remain row duplic sampl present figur droplet dry fix bacteri cell slide heat flame ethanol procedur test slide prepar prepar set twofold dilut antiserum buffer appendix titr gt num gt gt den gt titr gt num gt gt den gt titr titr ii test slide prepar ii prepar work dilut wd antiserum buffer work dilut dilut antiserum optimum specif half titr gt tabl gt gt tabl gt arrang slide damp tissu paper cover test window antiserum dilut appli pb fitc window volum antiserum appli window equival volum extract appli incub cover minut ambient temperatur shake droplet antiserum slide rins slide carefulli buffer wash minut buffer tween subsequ minut buffer appendix carefulli remov excess moistur arrang slide damp tissu paper cover test window fitc window dilut fitc conjug determin titr volum conjug appli window ident volum antiserum appli incub cover minut ambient temperatur shake droplet conjug slide rins wash carefulli remov excess moistur pipett µl phosphat buffer glycerol appendix similar mountant window appli coverslip read test examin test slide epifluoresc microscop filter suitabl excit fitc oil immers magnif scan window diamet angl perimet check posit control slide cell bright fluoresc complet stain note test repeat stain aberr read test slide observ absenc fluoresc cell fitc control window fluoresc cell fitc control specif bind conjug autofluoresc contamin note repeat test observ observ bright fluoresc cell characterist morpholog pseudomona solanacearum test window fluoresc intens equival posit control strain antiserum dilut cell incomplet stain weak fluoresc disregard cell interpret test result interpret test result bright fluoresc cell characterist morpholog found test neg ii bright fluoresc cell characterist morpholog found determin number cell microscop field calcul number cell ml resuspend pellet appendix popul approxim cell ml resuspend pellet consid limit detect test sampl amp gt cell ml resuspend pellet test consid posit sampl amp gt cell ml resuspend pellet test consid posit iii larg number amp gt cell ml incomplet weakli fluoresc cell titr antiserum test carri test base biolog principl repeat test antiserum ten fold dilut pellet elisa test base robinson smith al antiserum pseudomona solanacearum prefer race biovar determin titr suspens cell ml homolog strain pseudomona solanacearum nunc polysorb microtitr plate recommend includ neg potato extract control phosphat buffer salin pb control suspens amp gt cell ml strain race biovar pseudomona solanacearum posit control test ident manner sampl separ sampl microtitr plate pipett µl resuspend pellet microvi heat minut remov microvi ic add equal volum doubl strength carbon coat buffer appendix vortex appli µl aliquot well microtitr plate incub hour overnight flick extract well wash well time pb tween appendix leav wash solut well minut prepar dilut pseudomona solanacearum antiserum block buffer appendix appli µl antiserum dilut well incub hour flick antiserum well wash well prepar dilut alkalin phosphatas conjug block buffer appli µl conjug dilut well incub hour flick conjug well wash well prepar alkalin phosphatas substrat solut appendix appli µl well incub minut hour dark ambient temperatur read absorb nm interpret elisa test elisa test neg optic densiti od sampl amp lt od neg control elisa test posit optic densiti od sampl amp gt od neg control pcr test base seal al note filterplug pipett tip stage sampl prepar manipul involv pcr prepar suspens cell ml race biovar strain pseudomona solanacearum posit control test ident manner sampl pipett µl resuspend pellet microvi altern transfer µl resuspend pellet microvi µl naoh mix repeatedli invert microvi heat minut remov microvi immedi ic prepar decim dilut gt num gt gt den gt gt num gt gt den gt consid steril distil ultra pure water upw prepar pcr reaction mix appendix steril vial ad compon order gt tabl gt reaction calcul quantiti compon requir number reaction mix compon transfer µl µl mix steril pcr vial vial pcr reaction mix ic µl reaction volum scale compon pcr amplif option puls centrifug vial boil sampl posit control add order µl sampl water control posit control vial pcr reaction mix place vial heat block dna thermal cycler run programm cycl minut denatur templat cycl ii second denatur iii second anneal primer iv second extens copi cycl minut extens cycl vi hold note paramet perkin elmer thermal cycl requir miner oil overlai pcr reaction vial modif durat step ii iii iv amplif profil remov vial thermal cycler analys pcr product immedi store vial dai longer analysi pcr product pcr fragment detect agaros gelelectrophoresi stain ethidium bromid prepar agaros gel gentli bring boil agaros tri acet electrophoresi tae buffer cool molten agaros pour mould electrophoresi unit insert comb solut solidifi remov comb submerg gel tae cover mm buffer place µl droplet load buffer parafilm add µl pcr product sampl posit control water control mix gentl aspir pipett tip load volum modifi fit capac well agaros gel carefulli load well gel includ refer dna marker connect wire power suppli electrophoresi equip run gel cm front track indic cm end gel switch power suppli disconnect wire electrophoresi unit carefulli remov gel soak ethidium bromid solut minut note wear dispos glove time handl ethidium bromid power mutagen destain distil water minut visualis amplifi dna fragment uv transillumin pcr product pseudomona solanacearum primer set oli bp length check dna marker posit control note water control neg case posit repeat test photograph gel perman record requir confirm authent amplifi fragment restrict enzym analysi rea restrict enzym analysi rea transfer µl pcr product microvi add µl enzym buffer µl restrict enzym ava ii mix gentl aspir pipett tip drop remain wall vial puls spin microcentrifug incub hour analys digest pcr fragment agaros gelelectrophoresi interpret pcr test result pcr test neg characterist bp fragment detect fragment detect posit control strain pseudomona solanacearum pcr test posit bp fragment detect rea analysi amplifi fragment ident posit control strain pseudomona solanacearum select plate test base elphinston al perform test dilut plate techniqu prepar decim dilut gt num gt gt den gt gt num gt gt den gt consid resuspend pellet pelletbuff pipett measur standard volum µl resuspend pellet dilut modifi smsa select medium appendix spread glass rod surfac medium consid perform dilut streak µl loop resuspend pellet flame loop streak ii transfer measur standard volum µl resuspend pellet modifi smsa select medium spread glass rod surfac medium streak rod flame modifi smsa plate appli dilut plate techniqu suspens cell ml virul race biovar strain pseudomona solanacearum posit control set separ modifi smsa plate incub plate start read plate dai neg incub dai coloni virul isol pseudomona solanacearum milki white flat irregular fluidal blood red colour centr show intern streak whorl purifi coloni characterist morpholog subcultur gener nutrient medium appendix identifi pure cultur section ii confirm pseudomona solanacearum cultur pathogen test section ii interpret select plate test result select plate test neg bacteri coloni isol dai coloni characterist pseudomona solanacearum isol provid inhibit suspect coloni bacteria coloni characterist pseudomona solanacearum found posit control select plate test posit coloni characterist pseudomona solanacearum isol bioassai test base jans test plant suscept tomato eggplant seedl true leaf stage sampl water testplant hour inocul distribut µl resuspend pellet testplant inocul stem cotyledon place inocul techniqu seedl suspens cell ml virul biovar race strain pseudomona solanacearum posit control pelletbuff neg control separ posit control plant plant avoid cross contamin grow test plant week high rel humid daili water observ wilt epinasti chlorosi stunt isol infect plant section ii identifi pure cultur characterist morpholog section ii confirm pseudomona solanacearum cultur pathogen test section ii consid check absenc infect batch test plant show indic infect remov test plant cm section stem cm inocul point homogen tissu macer buffer perform dilut plate section iii posit identifi pure cultur characterist morpholog section ii confirm pseudomona solanacearum cultur pathogen test section ii interpret bioassai test result bioassai test neg test plant infect pseudomona solanacearum provid pseudomona solanacearum detect posit control bioassai test posit test plant infect pseudomona solanacearum enrich test base elphinston al transfer µl resuspend pellet ml modifi smsa broth appendix incub hour case longer hour cap tube loos fit aerat tighten cap vortex aliquot test section elisa test section pcr test section pathogen test refer section ii appendix nutrient media isol cultur pseudomona solanacearum nutrient agar na gt tabl gt prepar litr volum medium litr flask dissolv ingredi sterilis autoclav minut cool pour plate yeast pepton glucos agar ypga gt tabl gt prepar litr volum medium litr flask dissolv ingredi sterilis autoclav minut cool pour plate sucros pepton agar spa gt tabl gt prepar litr volum medium litr flask dissolv ingredi adjust ph sterilis autoclav minut cool pour plate kelman tetrazolium medium gt tabl gt prepar litr volum medium litr flask dissolv ingredi sterilis autoclav minut cool add filter steril aqueou solut triphenyl tetrazolium chlorid sigma obtain final concentr mg litr pour plate appendix materi sampl prepar macer buffer mm phosphat buffer ph buffer tissu macer gt tabl gt dissolv ingredi check ph aliquot consid sterilis autoclav minut addit polyvinylpyrrolidon mwt pvp recommend perform direct pcr test reduc incid amplif inhibit aromat molecul extract addit defloccul antifoam agent anti oxid recommend ware blender ultra turrax homogen procedur macer potato tissu core gt tabl gt autoclav separ add desir concentr pelletbuff mm phosphat buffer ph buffer resuspens dilut potato heel pellet gt tabl gt dissolv ingredi check ph aliquot consid sterilis autoclav minut appendix materi test buffer mm phosphat buffer salin pb ph buffer dilut antisera gt tabl gt dissolv ingredi check ph aliquot consid sterilis autoclav minut buffer tween buffer wash slide add tween buffer phosphat buffer glycerol ph buffer mountant fluid window slide enhanc fluoresc gt tabl gt appendix determin contamin level test surfac area window multispot slide gt num gt gt den gt gt tabl gt surfac area object field gt num gt gt den gt gt tabl gt calcul direct measur formula gt num gt gt den gt gt tabl gt amp radic gt num gt gt den gt amp radic gt num gt gt num gt gt den gt gt den gt gt num gt gt den gt gk count number typic fluoresc cell field calcul number typic fluoresc cell window gt num gt gt den gt calcul number typic fluoresc cell ml pellet gt num gt gt den gt gt tabl gt appendix materi elisa test doubl strength carbon coat buffer ph gt tabl gt dissolv ingredi check ph aliquot consid sterilis autoclav minut sodium sulphit final concentr ad anti oxid extract high fraction aromat molecul phosphat buffer salin pb ph gt tabl gt dissolv ingredi check ph aliquot consid sterilis autoclav minut phosphat buffer salin tween pb gt tabl gt block antibodi buffer freshli prepar gt tabl gt alkalin phosphatas substrat solut ph gt tabl gt mix adjust ph concentr hci make litr distil water add mgcl dissolv phosphatas substrat mg tablet sigma ml solut appendix materi pcr test oligonucleotid primer sequenc gt tabl gt materi seal al appendix materi select plate enrich test smsa select medium engelbrecht modifi elphinston al basal medium gt tabl gt prepar litr volum medium litr flask dissolv ingredi check ph adjust ph autoclav pseudomona solanacearum grow medium ph amp gt sterilis autoclav minut cool add ingredi sigma obtain final concentr gt tabl gt dissolv ingredi ethanol concentr volum medium prepar ingredi polymixin chloramphenicol requir slight warm shake smsa broth elphinston al prepar smsa select medium delet agar dispens ml aliquot ml dispos univers tube refer buddenhagen sequeira kelman descript race pseudomona solanacearum phytopatholog cook elizabeth sequeira genet divers pseudomona solanacearum detect restrict fragment length polymorph dna probe virul hypersensit respon molecular plant microb interact dinesen deboer extract clavibact michiganensi subsp sepedonicu composit sampl potato tuber american potato journal elphinston hennessi wilson stead sensit method detect pseudomona solanacearum smith smith potato tuber extract eppo bulletin engelbrecht modif semi select medium isol quantif pseudomona solanacearum aciar bacteri wilt newslett hayward characterist pseudomona solanacearum journal appli bacteriolog hayward systemat phylogeni pseudomona solanacearum relat bacteria bacteri wilt diseas caus agent pseudomona solanacearum ed hayward hartman cab intern oxford jans detect method pseudomona solanacearum symptomless potato tuber data sensit specif eppo bulletin jans infra intraspecif classif pseudomona solanacearum strain cell fatti acid analysi systemat appli microbiolog kelman relationship pathogen pseudomona solanacearum coloni appear tetrazolium medium phytopatholog lelliot stead method diagnosi bacteri diseas plant preec ed blackwel scientif public oxford pp louw fulbright stephen de bruijn differenti genom structur rep pcr fingerprint rapidli classifi xanthomona campestri pv vesicatoria phytopatholog lozano sequeira differenti race pseudomona solanacearum leaf infiltr techniqu phytopatholog mirza rademak jans akkerman specif ribosom rna target oligonucleotid probe clavibact michiganensi subsp sepedonicu canadian journal microbiolog robinson smith jone elphinston ford product antibodi pseudomona solanacearum caus agent bacteri wilt food agricultur immunolog seal jackson young daniel differenti pseudomona solanacearum syzygii picketti blood diseas bacterium partial rrna sequenc construct oligonucleotid primer sensit detect polymeras chain reaction journal gener microbiolog smith offord holder saddler genet divers burkholderia solanacearum synonym pseudomona solanacearum race kenya appli environment microbiolog stead techniqu detect identifi plant pathogen bacteria techniqu rapid detect plant pathogen ed duncan torranc blackwel scientif public oxford stead group plant pathogen pseudomona spp cellular fatti acid profil intern journal systemat bacteriolog van beuningen derk jans detect identif clavibact michiganensi subsp sepedonicu special attent fluoresc situ hybridis fish rrna target oligonucleotid probe züchtungsforschung