DistributionDistribution andand populationspopulations ofof ArmillariaArmillaria inin thethe regionregion

R. Rippy

JOHN W. HANNA1,2, James A. Moore1, Terry M. Shaw1, Geral I. McDonald2, Mee-Sook Kim2, Ned B. Klopfenstein2

1Department of Forest Resources, University of , Moscow, ID 83844 2USDA Forest Service, RMRS, 1221 S. Main St. Moscow, ID 83843

Hanna et al. 2004 Hanna et al. 2004 nifers Mixed Primary Host Co Hardwoods Hardwoods Hardwoods Hardwoods Mixed Mixed Mixed Mixed athogenicity arely pathogenic R High High High? Moderate? Moderate? Low Low Low Low P ican Biological ican Biological Species Species and Relative Pathogenicity Species and Relative Pathogenicity nnamed U Species A. ostoyae A. mellea A. tabescens A. gemina A. nabsnona A. calvescens A. sinapina A. gallica A. cepistipes a = North Amer = North Amer NABS NABS a II IX III V VII XI X a Armillaria Armillaria NABS I VI IFTNCIFTNC ArmillariaArmillaria speciesspecies summarysummary

DuringDuring thethe 19991999 andand 20002000 fieldfield seasonsseasons overover 8,5008,500 treestrees werewere sampledsampled forfor ArmillariaArmillaria,, 1,3101,310 isolatesisolates ofof ArmillariaArmillaria werewere collectedcollected fromfrom 2323 IFTNCIFTNC sites.sites. BasedBased onon overover 8,0008,000 somaticsomatic--incompatibilityincompatibility tests,tests, thesethese isolatesisolates werewere foundfound toto consistconsist ofof 221221 genetsgenets (individual(individual clones).clones).

OfOf thethe 1010 NorthNorth AmericanAmerican BiologicalBiological SpeciesSpecies 66 werewere associatedassociated withwith thethe IFTNCIFTNC sites.sites.

Hanna et al. 2004 Huckleberry

Soldier Creek

Stanton

Enterprise

Hanna et al. 2004 NABSNABS II –– ArmillariaArmillaria ostoyaeostoyae 6262 genetsgenets collectedcollected fromfrom 1717 IFTNCIFTNC sitessites

™™ SeriousSerious plantplant pathogenpathogen foundfound throughoutthroughout thethe NorthernNorthern HemisphereHemisphere thatthat causescauses rootroot andand buttbutt rotrot onon diversediverse woodywoody plantplant hosts.hosts.

™™ AdverselyAdversely impactsimpacts commercialcommercial timbertimber productionproduction byby causingcausing treetree mortalitymortality andand growthgrowth reduction.reduction.

™™ GeneticallyGenetically diversediverse andand maymay bebe comprisedcomprised ofof severalseveral races.races. IntraspecificIntraspecific hybridshybrids areare common.common.

Hanna et al. 2004 NABSNABS XX –– UnnamedUnnamed ArmillariaArmillaria speciesspecies 6969 genetsgenets collectedcollected fromfrom 1616 IFTNCIFTNC sitessites

™™ UnnamedUnnamed ArmillariaArmillaria speciesspecies foundfound inin thethe northwesternnorthwestern UnitedUnited States.States.

™™ MainlyMainly saprophytic,saprophytic, livingliving onon deaddead organicorganic matter.matter.

Hanna et al. 2004 NABSNABS III,III, V,V, VIIVII complexcomplex –– A.A. calvescenscalvescens,, A.A. sinapinasinapina,, A.A. gallicagallica 5858 genetsgenets collectedcollected fromfrom 1414 IFTNCIFTNC sitessites

™™ GroupGroup ofof similarsimilar speciesspecies ofof ArmillariaArmillaria foundfound throughoutthroughout NorthNorth America.America.

™™ PrimarilyPrimarily saprophyticsaprophytic withwith lowlow pathogenicitypathogenicity (less(less thanthan 5%).5%).

™™ HybridizationHybridization isis commoncommon betweenbetween species.species.

™™ HybridsHybrids areare difficultdifficult toto identify.identify.

Hanna et al. 2004 NABSNABS XX // NABSNABS III,III, V,V, VIIVII complexcomplex hybridshybrids 3131 genetsgenets collectedcollected fromfrom 1212 IFTNCIFTNC sitessites

™™ CommonCommon withinwithin thethe InlandInland Northwest.Northwest.

™™ PreviouslyPreviously groupedgrouped withwith NABSNABS XX untiluntil theirtheir discoverydiscovery usingusing molecularmolecular techniques.techniques.

™™ MainlyMainly saprophyticsaprophytic (less(less thatthat 5%5% pathogenic).pathogenic).

Hanna et al. 2004 NABSNABS IXIX –– ArmillariaArmillaria nabsnonanabsnona 11 genetgenet collectedcollected fromfrom 11 IFTNCIFTNC sitesite

™™ AA rarerare speciesspecies foundfound primarilyprimarily onon hardwoods,hardwoods, suchsuch asas AlnusAlnus species,species, inin riparianriparian areasareas ofof westernwestern NorthNorth America.America.

™™ IFTNCIFTNC’’ss isolationisolation ofof thisthis speciesspecies fromfrom AbiesAbies grandisgrandis isis oneone ofof thethe firstfirst reportsreports ofof thisthis speciesspecies beingbeing isolatedisolated fromfrom aa conifer.conifer.

Thomas J. Volk Thomas J. Volk Hanna et al. 2004 TypesTypes ofof informationinformation collectedcollected fromfrom NutritionNutrition CoCo--opop sitessites

™ Isolate ID ™ Location ™ Host Species ™ Collection (Bark Fan, Rhizomorph, or Wood) ™ Host Collection (Dead or Alive) ™ Habitat Type ™ Fertilization Treatment ™ Species ID ™ Rock Type

Hanna et al. 2004 BarkBark fanfan collectioncollection typetype

BarkBark fansfans fromfrom livelive treestrees indicateindicate highhigh pathogenicitypathogenicity ofof anan ArmillariaArmillaria individualindividual

DeadDead treestrees withwith barkbark fansfans andand resinosusresinosus andand oror woundwound peridermperiderm maymay indicateindicate pathogenicitypathogenicity

DeadDead treestrees withwith barkbark fansfans withoutwithout resinosusresinosus indicateindicate saprophyticsaprophytic behaviorbehavior Hanna et al. 2004 WoodWood collectioncollection typetype

WoodWood samplessamples areare usuallyusually takentaken fromfrom treestrees thatthat havehave recentlyrecently beenbeen killedkilled oror areare infectedinfected withwith aa pathogenicpathogenic ArmillariaArmillaria individual.individual.

OftenOften darkdark lineslines knownknown asas zonezone lineslines cancan bebe seenseen inin

infectedinfected wood.wood. Hanna et al. 2004 RhizomorphRhizomorph collectioncollection typetype

All species ofof Armillaria form rhizomorphs.

Most rhizomorphs are found on the surface of a root, some grow freely through the soil, and others can be found under the bark of a highly infected tree.

Presence of rhizomorphs does not necessarily indicate pathogenicity of an Armillaria individual. Hanna et al. 2004 DNADNA SequencingSequencing

5.8S Nuclear Nuclear Large rDNA 5S Small Nuclear Small rDNA rDNA rDNA

IGS-2 ITS-1 ITS-2 IGS-1 IGS-2

Ribosomal DNA regions currently being investigated for phylogenetic signal include IGS, ITS, and LSU.

Hanna et al. 2004 Variable nucleotide positions of aligned nLSU sequences.

GENET 126bp 129bp 165bp172-173bp 227bp 407bp 470bp 477bp 481bp 500bp 514bp 518bp 524bp 602bp 656bp 750bp 787bp 812bp 837-A***, D82-A***, POR100***, R1299, ST1, ST8*, ST11-A*, U5, U16 ATTCGA T C C C C G A C G A A T T

ST11-B* ATC GA T C T C C CGA C G G A T T

837-B*** A CC GA TC T C C C G AAC G A T T

D82-B*** ATC GA TCT C C C G G C G A A T T

MNF4, NC94, NC540, NC765-A, P4352, R1348-A, R1389, SSF4-A, ST2 ATC GA T C T T CCGAC G AATT

NC765-B, P1404, R1348-B, R1389-B A TCGA T C T T C A G A C G A A T T

NC1126, SSF4-B A T C GA T C T T C C G A C A A A T T

BC18F-A, NC1070-A ATY GA T C TCT CGAC GAATT

BC18F-B, NC1070-B ATY ~~ TCTT C CGAC GAATT

R1329 ATC GA Y C T T C CGAC RAATT

OR1 ATC GA TCTY M C RAC GAATY

OR10 ATC GA TCTY MCR A C G A ATT

PC514 ATC GA T C T Y C CGAY GAATT

NC837, R1330 ATC GA TYYY CCGAY GAATT

R1374 ATC GA T Y Y Y CMGA Y GAATT

NM115, NM120, TS7, U73 ATC GA TCT C C C G A T G A ATT

ST16** G T C GA T C T C C C G A T G AGTT

C21**, M90** A T C GA T C T C C C G A T G A A CT

Hanna et al. 2004 Radial 50% majority rule consensus trees based on 24,000 trees from the Bayesian inference analysis of 850 base-pairs in the nLSU rDNA region. Scale bar = 0.001 substitutions per site.

C21**, M90** NC765-B,404, P1

R1348-B, R1389-B

MNF4, NC94, NC540, NC765-A, P4352, SSF4-A,Pacific ST2, R1389, R1348-A NM115, NM120, TS7, U73 Northwest ST11-B*

NC1126,

SSF4-B

ST16**

NC837,133 0R R1374 837-B*** OR1 0.98 0.64 OR10

37-A***, D82-A***, POR100***, Circumboreal 1299, ST1, ST8*, 0.98 PC514 T11-A*, U5, U16

ST11-B* D82-B*** NM115, NM120,

TS7, U73 MNF4, NC94, NC540, D82-B*** 837-A***, D82-A***, POR100***, NC765-A, P4352, R1348-A, R1299, ST1, ST8*, ST11-A*, U5, U16 R1389, SSF4-A, ST2 BC18F-B, R1329 NC1070-B Rockies BC18F-A, NC1070-A NC1126, SSF4-B 0.001 837-B*** NC765-B,404, P1 C21**, M90** R1348-B, R1389-B 0.001

* NABS II ST16** ** NABS IX *** NABS X

Hanna et al. 2004 Phylogenetic tree showing genetic diversity of Armillaria ostoyae based on the IGS-1 DNA region

NC588 N. Idaho - Soldier Creek NC542A N.E. - Enterprise NC435A N. Idaho - Stanton P416A N. Idaho - Stanton NC403 N. Idaho - Stanton PacificPacific NC666A N. Idaho - Dick’s Creek P2399 N. Idaho - Priest River NorthwestNorthwest AA NC436A N. Idaho - Stanton NC94 N. Idaho - Huckleberry NC435B N. Idaho - Stanton P4352 N. Idaho - Priest River P416B N. Idaho - Stanton PacificPacific NC1126 N. Idaho - Whiskey NC436B N. Idaho - Stanton NorthwestNorthwest BB NA142 S.W. - T.T. Munger NC542B N.E. Oregon - Enterprise NC666B N. Idaho - Dick’s Creek NM120A New.Mexico - Jemez NM115 New.Mexico - Jemez RockiesRockies NM120B New.Mexico - Jemez HUA9242 Japan - Terashima et al. 1998 P255A N. Idaho - Priest River P255B N. Idaho - Priest River Hanna et al. 2004 FactorsFactors associatedassociated withwith geneticgenetic racesraces andand intraspecificintraspecific hybridizationhybridization withinwithin A.A. ostoyaeostoyae

™™ AdaptationAdaptation toto aa varietyvariety ofof environmentsenvironments

™™ MayMay bebe responsibleresponsible forfor differingdiffering levelslevels ofof pathogenicitypathogenicity andand virulencevirulence

™™ “Hybrid“Hybrid vigor”vigor” maymay contributecontribute toto enhancedenhanced growth,growth, survival,survival, and/orand/or pathogenicitypathogenicity

™™ HybridsHybrids maymay adaptadapt betterbetter toto environmentalenvironmental changechange

Hanna et al. 2004 DoesDoes ArmillariaArmillaria affectaffect treetree growth?growth?

Hanna et al. 2004 ImpactImpact ofof ArmillariaArmillaria RootRoot DiseaseDisease onon TreeTree GrowthGrowth

Volume loss has been shown as high as 40% over 4-8 years in eighteen-year-old Douglas-fir (Cruickshank, 2000).

Hanna et al. 2004 AverageAverage mortalitymortality ratesrates inin aa 1010--yearyear periodperiod forfor sitessites coveringcovering northernnorthern Idaho,Idaho, northeasternnortheastern Oregon,Oregon, andand easterneastern WashingtonWashington varyvary fromfrom 8.8%8.8% forfor DouglasDouglas--fir,fir, 4.2%4.2% forfor grandgrand fir,fir, andand 1.5%1.5% forfor redcedarredcedar habitathabitat typestypes basedbased onon 88 sitessites (Moore(Moore etet al.,al., 2000).2000).

GrowthGrowth lossloss andand mortalitymortality duedue toto ArmillariaArmillaria inin BritishBritish ColumbiaColumbia havehave beenbeen citedcited asas betweenbetween 1.41.4 andand 3.83.8 millionmillion mm3 annuallyannually (White(White andand Morrison,Morrison, 1999).1999).

Hanna et al. 2004 DoesDoes ArmillariaArmillaria affectaffect treetree growth?growth?

Influence of Armillaria ostoyae collected adjacent to growth plots on six-year gross- basal-area growth (sq.ft/a)

60 a e r 50 a

l **

sa ** a 40 b n a)

i Without A.ostoyae ft/ .

se 30 q ea (s With A.ostoyae cr

n 20 e i g a 10 er v

A 0 717 7 6 8 4 02 5 7 7 0 # of plots Spirit Lake Whiskey Huckleberry Soldier Creek Skookum Lake Dick's Creek averaged IFTNC Site

* α < .05 ** α < .10 Hanna et al. 2004 Hanna et al. 2004 (!D !( Plot 11 D (!( 300N & 0K 6-year Net- and Gross-Basal-Area Growth (sq.ft/a) in GREEN (!(D (![(! D ´ D 53.1G 53.0N Soldier Creek - Study Site D D(! Intermountain Forest Tree Nutrition Co-op (!D D D(! Plot 10 49.8G 300N & 0K 49.8N Tsuga heterop hylla / Asarum caudatum western hemlock / wild ginger D D Plot corners Plot 9 D (!! 44.4G [(!(!( 200N & 0K D 42.8N [ Bark fans live t rees (![(! D D (! NABS X Plot 7 Control D D (! (! (! !( D NABS X - III-V-VII Hy brid (![(D Plot 8 32.4G D (! 42.5G (! (! 100N & 0K D(! D (! NABS I 32.2N D D (! (! D 42.0N D [(! D D(! (! (!(! D 0170 40280Meters (!D Plot 6 200N & 0K 41.8G Plot 4 D D (! 32.3G 100N & 0K D 41.8N D 1 (!(! D D (! D 23.1N D (! D D D(! Plot 12 60.5G (! D (! 49.4G Plot 5 D 600N & 0K (!D D 58.2N D 45.8N 200N & 0K Plot 3 D D (!(!D D (! 100N & 0K [(! [(! D (!(!(!(! Plot 13 D D 51.3G D D 200N & 0K [(! D D D 35.6G (! D 51.3N D Tsuga heterophylla / Clintonia uniflora (!(! D D[(! D 35.6N D Plot 1 (! D (! western hem lock / queen cup bea dlily D 300N & 200K D[(! [(!D Plot 14 45.5G 55.0G ! D Plot 2 59.7G 300N & 0K [(D 44.4N 55.0N 300N & 200K2SO4 57.5N 2 Influence of Armillaria ostoyae collected adjacent to growth plots on six-year net-basal-area growth (sq.ft/a)

50 45

area * ** 40

asal 35 b n ) 30 a

ft/ Without A.ostoyae . 25 q * (s crease i 20 n With A.ostoyae e i 15 10

Averag 5 07 7 7 68 4710 25 7 0 Spirit Lake Whiskey Huckleberry Soldier Creek Skookum Lake Dick's Creek # of plots IFTNC Site averaged

* α < .05 ** α < .10 Hanna et al. 2004 6-year Net- and Gross-Basal-Area Growth (sq.ft/a) in GREEN Legend

D plot corners Huckleberry Creek - Study Site D plot centers ´ Intermountain Forest Tree Nutrition Co-op (! Genet 1 NABS I A.ostoyae (! Genet 2 NABS X 19.3G (! Genet 3 NABS III,V,VII complex 1.1N (! Abies Grandis / Clintonia uniflora - Physocarpus malvaceus phase Genet 4 NABS III,V,VII complex Plot 14 D 300N & 200K2SO4 Grand fir / queen cup beadlily - ninebark ph ase [_ Barkfan on dead tree Barkfan on live tree (active pathogen) 24.0G (!(! D [ (![ (! (!D 24.0N (![_D (![ 31.6G Plot 13 (!D D Plot 1 D D 30.9N Control (! 27.4G 512.1N & 290.2K (! (! D (![_ D 29.3G (! (! D (![_ 9.9N (! D D 23.9G Plot 5 (! (!D D (!(! (! D D (! 21.8N D 600N & 170K Plot 3 (![_ D D 18.8N D (! Plot 8 (! (![_ 300N & 170K (! D (!(! 300N & 340K (! D D (! D (! (! (! Abies Grandis / Spi raea betulifolia D D D ! D D [_(! ( D D (! D Grand fir / spiraea (! Plot 6 D D (![_ (! 26.6G D 512.1N & 49.8K D ![(! D D ( (! D D (!(! D 27.9G _ (! D (! (![D (! D D 18.0N D 37.2G (! Plot 11 27.9N Plot 2 D(! 300N & 170K (! 31.5N 300N & 170K D ![ D (![( D D D D D D D D D D D D (! (![ (! Plot 7 (! 21.3G Plot 12 (!(![(! (!(! (! 300N & 0K (! [_ Plot 4 87.9N & 290.2K D [ D D(! 9.3N (! D (! 300N & 170K (! Plot 10 Plot 9 (! 87.9N & 290.2K 31.9G (![_ D 29.2G (! 0N & 170K D D 21.9N D 28.1G 27.5N 26.2N 20.2G Abies Grandis / Physocarpus malvaceus - Physocarpus malvaceus phase 15.9N Grand fir / ninebark phase - ninebark phase

02140 80 560Meters

1 inch equals 61.886223 meters Hanna et al. 2004 Thuja plic ata Series 6-year Net- and Gross-Basal-Area Growth (sq.ft/a) in GREEN western redcedar Series D Spirit Lake - Study Site (! Intermountain Forest Tree Nutrition Co-op D D ´ D Legend Plot 1 D D 600N & 0K plot co rners [_ Barkfan on dead tree D plot ce nters [ Barkfan on live tree (active pathogen) 1 27.7G _ Wood Sample (Note: from Holodiscus discolor) 24.2N (! Genet 1 NABS I A.ostoyae [ Genet 2 NABS I A.ostoyae D 17.4G (! D -12.9N (! Genet 3 NABS I A.ostoyae (![ D Plot 2 (! Gene t 4 NABS X (![ 0N & 170K (! D D Tsuga hetero phylla Series western heml ock Series D D D

(! D ![_ Plot 5 [_!(! ! ( D ( ( 300N & 170K D D (! D D 21.3G Plot 3 (! D 200N & 0K D 20.7N D 28.7G 2 (!D 24.7N Plot 4 (! 300N & 0K (! D

D D 18.6G Thuja plic ata Serie s 31.4G D D 16.4N western redcedar Series 29.5N D (! (![_ Plot 7 D D Control

D D Plot 6 D 100N & 0K 0 65 130 260 Meters 19.4G D 14.7N

Hanna et al. 2004 Influence of Armillaria species collected adjacent to growth plots on six-year net- basal-area growth (sq.ft/a)

60 Without Armillaria asal 50 b ) **** a NABS X Only / 40 t f . q ease in s 30 X and I cr n ea ( 20 e i ar NABS I Only ag 10 ver 0002 5 7704217 4046 00223 761 A 0 Spirit Lake Whiskey Huckleberry Soldier Skookum Dick's Creek Creek Lake # of plots averaged IFTNC Site

* α < .05 ** α < .10 Hanna et al. 2004 FactorsFactors thatthat interactinteract withwith ArmillariaArmillaria toto influenceinfluence treetree growthgrowth

InteractingInteracting FactorsFactors ::

HabitatHabitat TypeType SoilSoil CharacteristicsCharacteristics ParentParent MaterialMaterial FertilizationFertilization OtherOther PestsPests

Hanna et al. 2004 FutureFuture StudiesStudies

BetterBetter characterizecharacterize ArmillariaArmillaria speciesspecies withwith molecularmolecular geneticgenetic techniquestechniques toto betterbetter understandunderstand pathogenicitypathogenicity andand virulencevirulence

AssessAssess directdirect effectseffects ofof ArmillariaArmillaria ostoyaeostoyae onon treetree growthgrowth

EvaluateEvaluate effecteffect ofof ArmillariaArmillaria speciesspecies interactionsinteractions onon treetree growthgrowth

InvestigateInvestigate saprophyticsaprophytic ArmillariaArmillaria speciesspecies asas aa possiblepossible biologicalbiological controlcontrol ofof ArmillariaArmillaria ostoyaeostoyae

Hanna et al. 2004 TreeTree growthgrowth isis affectedaffected byby aa multitudemultitude ofof interactinginteracting factorsfactors thatthat includeinclude ArmillariaArmillaria..

ItIt isis difficultdifficult toto assessassess howhow nutritionalnutritional treatmentstreatments affectaffect ArmillariaArmillaria impactsimpacts onon growthgrowth withoutwithout knowledgeknowledge ofof prepre-- existingexisting ArmillariaArmillaria influences.influences.

PretreatmentPretreatment studiesstudies ofof ArmillariaArmillaria distributiondistribution andand impactsimpacts acrossacross aa rangerange ofof environmentsenvironments areare neededneeded asas baselinebaseline information.information. ThisThis baselinebaseline wouldwould helphelp forestforest managersmanagers betterbetter understandunderstand howhow treatmentstreatments affectaffect ArmillariaArmillaria andand overalloverall growthgrowth ofof aa stand.stand.

Hanna et al. 2004 AcknowledgementsAcknowledgements

IntermountainIntermountain ForestForest TreeTree USDAUSDA ForestForest ServiceService NutritionNutrition CoCo--opop JamesJames B.B. DonleyDonley andand thethe RainiRaini C.C. RippyRippy UniversityUniversity ofof IdahoIdaho JaneJane E.E. StewartStewart MariannMariann GarrisonGarrison--JohnstonJohnston BryceBryce A.A. RichardsonRichardson MarkMark KimseyKimsey TroyTroy HensiekHensiek HowardHoward JenningsJennings JoanneJoanne TirockeTirocke BobBob AtwoodAtwood KwanKwan--SooSoo WooWoo PeterPeter MikaMika JonaleaJonalea R.R. TonnTonn LeonardLeonard JohnsonJohnson PaulPaul J.J. ZambinoZambino DebbieDebbie S.S. PagePage--DumroeseDumroese

Hanna et al. 2004