trees (Acer spp.) are very Acer (sycamore ) Acer (Norway maple) pseudoplatanus http://www.hainaultforest.co.platanoidesuk/Leaf%20Norway%20maple5.JPG Tar spot of maple in Canada important throughout Canada (Acer = silver maple) Tom Hsiang and Lynn Tian saccharinum Dept. Environmental Biology University of Guelph Guelph, Ontario, Canada

email: [email protected] www.uoguelph.ca/~thsiang/present/2007tarspot.pdf

Acer (silver maple) Acer (red maple) Acer (sugar maple) Maples in Canada (Acer species) saccharinum rubrum saccharum Q East: saccharum Q West: macrophylla, rubrum Q Introduced: pseudoplatanus, , circinatumsaccharinum Sugar, platanoidesMaple… vs. Norway, spicatum - clear sap in the leaf petiole vs. white…. sap , platanoides , campestre - orange fall color vs. simple yellow) saccharum - shaggy bark on older trees vs. small ... rubrum grooves Silver- leaf lobes more triangular vs. squarish saccharinum lobes 3-6” very deep lobes- seeds more globose vs. flattened seeds silvery underneath http://www.ruhr-uni-bochum.de/boga/html/Acer_saccharinum_ja.jpg http://www.earthworksboston.org/images/Maple%20leaf%20comparison.jpg http://www.laddarboretum.org/images/buttons/leaf%20detail_red%20maple.jpg http://www.forestry.iastate.edu/trees_of_iowa/Trees_of_Iowa_drawings/tsilverm.gif

Tar Spot - Tar Spot - Rhytisma Tar Spot - Rhytisma Presentation Overview

Q DNA sequencing for identity and phylogenetic analyses of tar spot on maples Q Spore release periods and control Q Why does tar spot seem to be increasing?

Q a very showy disease on maple leaves ... Q ... but considered just an aesthetic nuisance

Tar spot of maple - history Rhytisma on maple (Europe) Rhytisma on sycamore maple http://commons.wikimedia.org/wiki/Image:Rhytisma_acerinum.jpg Q In North America, maple tar spot is caused by acerinum ( , Europe) acerinum http://commons.wikimedia.org/wiki/Imagacerinume:Rhytisma_acerinum_brok_2_beentree.jpg Objective 1 Rhytisma but there are differences

from the Europeanacerinum R. acerinum Determine species of Q Hudler et al., Cornell, (1998, Mycotaxon 68:405) Rhytisma present on said that Rhytisma was imported from Europe, and affects acerinum European maple species maples in eastern Q Hudler et al. proposed Rhytisma on Canada native North American maples based americanum on morphology & DNA-RFLP differences . .

? ? ssp ssp

) )

punctatum punctatum

R. R. looks like looks like on striped maple on striped maple

saccharinum saccharinum , Eastern Canada) , Eastern Canada)

Acer saccharum Acer saccharum from Quebec from Quebec ) from Ontario

) from Ontario

Acer Acer on Manitoba maple on Manitoba maple ?? on ?? on punctatum punctatum ?? ??

pensylvanicum pensylvanicum

Acer Acer Acer negundo Acer negundo Rhytisma Rhytisma grandidentata grandidentata silver maple ( silver maple ( (

Rhytisma ( Rhytisma ( Rhytisma Rhytisma ( . . ssp ssp

on silver maple on silver maple

? ? on mountain maple on mountain maple

, Quebec) , Quebec) Eastern Canada) Acer saccharum Eastern Canada) Acer saccharum from Quebec , , from Quebec ) from Ontario

) from Ontario

on Manitoba maple on Manitoba maple ?? on americanum americanum ?? on punctatum punctatum ?? ??

spicatum spicatum

Acer Acer Acer saccharinum Acer negundo Acer negundo Rhytisma Rhytisma grandidentata grandidentata ( ( ( http://www.pbase.com/comafungi/image/51447275

http://www.pbase.com/comafungi/image/51447275

Rhytisma Rhytisma ( Rhytisma Rhytisma ( Rhytisma Rhytisma ( on silver-red on silver-red on silver maple on silver maple

? ? ?? ?? on field maple on field maple on big-leaf maple on big-leaf maple

Ontario) Ontario) , , ) )

americanum americanum americanum americanum punctatum punctatum acerinum? acerinum?

Acer saccharinum Acer saccharinum Acer campestre Acer campestre from B.C.

Rhytisma from B.C. ( maple hybrid from Ontario

Rhytisma ( ( Rhytisma maple hybrid from Ontario Rhytisma Rhytisma Rhytisma ( Rhytisma Rhytisma on red maple on red maple

?? ?? on big-leaf maple on big-leaf maple on field maple on field maple on striped maple on striped maple

, Eastern Canada) , Eastern Canada) , Western Canada) , Western Canada) ) from Ontario ) from Ontario , Ontario) , Ontario)

punctatum punctatum americanum americanum punctatum punctatum acerinum? acerinum?

pensylvanicum pensylvanicum

Acer macrophyllum Acer macrophyllum Acer Acer Acer rubrum Acer rubrum Acer campestre Acer campestre ( ( http://www.flickr.com/photos/johns_pics/278641162/ http://www.flickr.com/photos/johns_pics/278641162/ Rhytisma ( Rhytisma ( ( Rhytisma Rhytisma ( ( Rhytisma Rhytisma ( Rhytisma Rhytisma Acer Acer apothecia apothecia Objective 2 Objective 2 on Norway maple ( on Norway maple ( ) in Canada ) in Canada acerinum acerinum

soaked 24 hrs 2006/5/18 soaked 24 hrs 2006/5/18 Spore Dispersal Period

?? Spore Dispersal Period ?? correlated with climate correlated with climate Tar spot on Norway maple mid September (Guelph) Guelph Norway maple, Guelph Norway maple, Tar spot on Norway maple mid September (Guelph) Rhytisma Rhytisma Rhytisma Rhytisma platanoides platanoides http://www.lincstrust.org.uk/species/fungi/tarspot2_l.jpg

acerinum acerinum

acerinum acerinum

Rhytisma

Rhytisma (on Norway maple) (on Norway maple) maples) maples) Rhytisma Rhytisma England England (on sycamore maple) (on sycamore maple) Guelph Norway maple Guelph Guelph Norway maple Germany Germany red red Guelph Manitoba maple Guelph Manitoba maple species species

maple & striped maple) maple & striped maple) Guelph field maple Guelph field maple 100 100 Guelph sugar maple Guelph sugar maple bigleaf bigleaf (on silver & (on silver &

on Norway maple on Norway maple 84 84 (on (on

100 100 apothecia apothecia ? ? Rhytisma Rhytisma ) ) 100 100

americanum americanum

punctatum punctatum

Rhytisma Rhytisma acerinum acerinum acerinum acerinum Rhytisma Rhytisma

) ) pinastri pinastri ) )

salicinum salicinum distance tree based on NJ-bootstrap of ITS sequences distance tree based on NJ-bootstrap of ITS sequences tarspot tarspot needlecast needlecast

Acer platanoides (pine (pine Lophodermium Lophodermium Lophodermium http://grzyby.strefa.pl/foto04/lophodermium_pinastri.jpg Relationships between Tar spot on Norway maple late July - Niagara Falls early August - Guelph Unrooted (willow Unrooted (willow http://www.floraislands.is/SVEP/rhytisal1s.jpg Relationships between Rhytisma http://www.ambmuggia.it/forum/uploads/post-3-1149259562.jpg 2006/6/2 Italy http://www.ambmuggia.it/forum/uploads/post-3-1149259562.jpg 2006/6/2 Italy Tar spot on Norway maple late July - Niagara Falls early August - Guelph Rhytisma Rhytisma Rhytisma ( Rhytisma Rhytisma ( to confirm identity confirm to to confirm identity confirm to cultured cultured apothecia apothecia & fungal pathogen pathogen & fungal & fungal pathogen pathogen & fungal and herbarium specimens) and herbarium specimens) ? on which maple? ? on which maple? acerinum acerinum

DNA extraction from tar spot (fresh, DNA extraction from tar spot (fresh, Tar spot fungi could not be Samples of various maple species were gathered from across Canada, and two from Europe Qiagen overwintered DNA was amplified with conserved fungal ribosomal DNA ITS primers and sequenced Tar spot fungi could not be Samples of various maple species were gathered from across Canada, and two from Europe Qiagen overwintered DNA was amplified with conserved fungal ribosomal DNA ITS primers and sequenced Tar spot on Norway maple early July (Guelph) DNA extraction and sequencing collected Oct 2007 & 2008 in Guelph http://www.ambmuggia.it/forum/uploads/post-3-1149259507.jpg 2006/6/2 Italy Q Q Q Q DNA tests were used for both plant host DNA tests http://www.ambmuggia.it/forum/uploads/post-3-1149259507.jpg 2006/6/2 Italy collected Oct 2007 & 2008 in Guelph Tar spot on Norway maple early July (Guelph) DNA extraction and sequencing DNA tests were used for both plant host DNA tests Rhytisma Rhytisma Q Q Q Q Rhytisma Rhytisma . . ssp ssp

R. acerinum? R. acerinum? looks like looks like on native maples on native maples

- history of research - history of research on sycamore maple on sycamore maple

on European maples on European maples Acer saccharum Acer saccharum from Quebec from Quebec

on Norway maple?? from on Norway maple?? from ?? on ?? on acerinum acerinum

?? ??

Rhytisma acerinum Rhytisma acerinum Rhystima americanum Rhystima americanum – – – – 1990’s studies at Cornell University (New York) 2006-2008 study at the University of Guelph on tar spot funded by Landscape Ontario & NSERC 1990’s studies at Cornell University (New York) 2006-2008 study at the University of Guelph on tar spot funded by Landscape Ontario & NSERC Tar spot of maple 2004/12/12 Poland 2004/12/12 Poland Q Q Tar spot of maple Rhytisma Rhytisma Rhytisma grandidentata grandidentata Q Q Rhytisma Wisconsin Rhytisma Rhytisma Wisconsin ascus ascus emerging emerging 17-Jul ascospore from tip of ascospore from tip of lynn\20060607queenpark lynn\20060607queenpark

Started by May 25, 100% spore release by July 3 Started by May 25, 100% spore release by July 3 ascus stroma ascus stroma

16-Jun asci asci all entwined all entwined ascospores inside ascus ascospores inside ascus 16-May acerinum acerinum

max Temp (C) max Temp (C) min (mm) rain spore release 1994 (Ottawa) 1954 & 1953 (London) 1947 & 1946 (Ottawa) 1994 (Ottawa) 1954 & 1953 (London) 1947 & 1946 (Ottawa) 15-Apr 5 0 -5 – – – – – – 60 55 50 45 40 35 30 25 20 15 10 The National Herbarium in Ottawa has dried samples of tar spot on Norway maple (& other European maples) Probably in Ontario since the 1940’s The National Herbarium in Ottawa has dried samples of tar spot on Norway maple (& other European maples) Probably in Ontario since the 1940’s Tar spot cross section 2007 climate & spore data When did it arrive in Ontario? http://lh3.google.com/_RZNh8zv5bbU/R0cdSIEG9GI/AAAAAAAAAfo/NmQH2Bf3UTg/s800/100_1236.JPG Q Q http://lh3.google.com/_RZNh8zv5bbU/R0cdSIEG9GI/AAAAAAAAAfo/NmQH2Bf3UTg/s800/100_1236.JPG Tar spot cross section 2007 climate & spore data When did it arrive in Ontario? Rhytisma Rhytisma Q Q Started May 25 100% by June 22 Started May 25 100% by June 22 & the Maritimes) & the Maritimes) Que Que , , asci asci apothecia apothecia ) ) Ont Ont asci asci (percent empty empty (percent (percent empty empty (percent

acerinum acerinum acerinum acerinum

max Temp (C) max Temp min Temp (C) (mm) precip release spore soaked 24 hrs 2006/5/18 soaked 24 hrs 2006/5/18 Norway maple most common shade tree planted Norway maple most after Dutch elm disease in mid-1900’s 1930’s introduced from Europe? 1940’s localized outbreak in Ohio 1983 tar spot outbreak Upstate New York 1990’s tar spot on Norway maple reported from Great Lakes Region ( No reports from the Prairie provinces or B.C. where there are many Norway maples Furthest western sample from Wisconsin 5 0 Norway maple most common shade tree planted Norway maple most after Dutch elm disease in mid-1900’s 1930’s introduced from Europe? 1940’s localized outbreak in Ohio 1983 tar spot outbreak Upstate New York 1990’s tar spot on Norway maple reported from Great Lakes Region ( No reports from the Prairie provinces or B.C. where there are many Norway maples Furthest western sample from Wisconsin 060301 060311 060321 060331 060410 060420 060430 060510 060520 060530 060609 060620 060630 -5 35 30 25 20 15 10 -10 -15 2006 climate & spore data Norway maple tar spot came from where? Guelph Norway maple, http://www.ambmuggia.it/forum/uploads/post-3-1149259629.jpg 2006/6/2 Italy Q Q Q Q Q Q Q Guelph Norway maple, Norway Guelph http://www.ambmuggia.it/forum/uploads/post-3-1149259629.jpg 2006/6/2 Italy 2006 climate & spore data Norway maple tar spot came from where? Rhytisma Rhytisma Rhytisma Rhytisma Q Q Q Q Q Q Q deflated deflated ascospores discharged ascospores discharged m long) cap open m long) cap open μ μ discharing from tip discharing from tip (70 (70 asci asci asci asci apothecia apothecia Canada? Canada? swollen with ascospores swollen with ascospores Objective 3 Objective 3 asci asci

acerinum acerinum soaked 24 hrs 2006/6/5 acerinum acerinum acerinum acerinum Why is the incidence increasing in Eastern Why is the incidence e, soaked 24 hrs 2006/6/5 increasing in Eastern and severity of tar spot and severity of tar spot http://www.bioimages.org.uk/html/P2/P26396.php undifferentiated undifferentiated Niagara Falls Norway mapl http://www.ambmuggia.it/forum/uploads/post-3-1149259612.jpg 2006/6/2 Italy Niagara Falls Norway maple, Rhytisma Rhytisma Rhytisma Rhytisma http://www.ambmuggia.it/forum/uploads/post-3-1149259612.jpg 2006/6/2 Italy Rhytisma Rhytisma ) )

acerinum acerinum

R. R. apothecia ascospores apothecia ascospores observed in late May 06, 07 & 08 observed in late May 06, 07 & 08 asci asci acerinum acerinum acerinum acerinum

kr/img/rhytisma_01.JPG kr/img/rhytisma_01.JPG first empty ascospores first seen in cross sections early May in early May, apothecia would open after soaking first empty ascospores first seen in cross sections early May in early May, apothecia would open after soaking – – – – – – no more spores after 25 June 1985 in New York no more spores after 22 June 2006 in Guelph no more spores after 3 July 2007 in Guelph no more spores after 15 June 2008 in Guelph no more spores after 25 June 1985 in New York no more spores after 22 June 2006 in Guelph no more spores after 3 July 2007 in Guelph no more spores after 15 June 2008 in Guelph Spore release data tar spot of Norway maple ( Spore release data tar spot of Norway maple ( http://www.bio-forum.pl/messages/33/10170.jpg 2005/5/8 Poland http://www.ambmuggia.it/forum/uploads/post-3-1149259752.jpg 2006/6/2 Italy Q Q Q Q http://www.bio-forum.pl/messages/33/10170.jpg 2005/5/8 Poland http://www.ambmuggia.it/forum/uploads/post-3-1149259752.jpg 2006/6/2 Italy Rhytisma Rhytisma Rhytisma Rhytisma Q Q Q Q Tar spot cross section http://www.samwoosc.co. http://www.samwoosc.co. Tar spot cross section Is it getting worse? Why is it getting worse? Sulfate deposition & acid rain Sulfate deposition & tar spot Http://www.ec.gc.ca/cleanair-airpur/caol/canus/report/2006canus/images/figure1_l_e.gif Q Frequent newspaper reports of Frequent newspaper reports of Q Vick & Bevan (1976, Env. Pollut. 11:203) in the Q sulphur dioxide gas emitted by tar spot U.K. found that tar spot (and lichens) are strong power stations & automobiles – St. Catharines Standard (Aug 31 Q reacts with hydroxyl radicals 2005, Nov 7 2003) indicators of pollution ↓ – Barrie Examiner ( Sept 20 2004) Q inhibited by high SO2 Q sulphur trioxide gas which combines – Orillia Packet (Sept 7 2004) sulphur trioxide gas which combines with water – Global National News (Twice in Nov 2007) ↓ 1994? » claimed to be devastating sugar Q sulphuric acid in rain makes acid rain Q Canada has been successful in reducing maple in Montreal, but these turned out to be Norway maple (I got emissions of SO2, major contributor of acid rain samples) Q Less acid rain might allow for more tar spot

images from http://www.windows.ucar.edu/tour/link=/physical_science/chemistry/sulfur_oxides.html http://www.newscientist.com/article/mg15420803.100-science--toxic-chemistry-turns-air-acid.html

Summary of results Implications of the results Thank you for your attention Q current epidemic of tar spot of Norway maple in Q If fungicides are used against tar spot, make an the east is caused by Rhytisma acerinum, also application by mid-May and again by mid-June found on field and sycamore maples (depending on weather & growth) Q R. americanum on silver and red maples Q R. acerinum on sugar maple & Manitoba maples Q The European R. acerinum may be adapting to (need more samples) attack native North American maples, and we Any Questions? Q Punctate spots on Norway, silver, or sugar maple need to monitor the situation resembling bigleaf maple or striped maple multiple black specks are not R. punctatum Q Better air quality, more tar spot? Q Spore release from mid-May to late June