2017 hardwood survey report - wa

2017 Hardwood Survey J. Falacy & C. Looney

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2017 Hardwood Survey Report

Jennifer Falacy & Chris Looney Washington State Department of Agriculture

Year: 2017

State: Washington State

Cooperative Agreement Name: Hardwood Commodity Survey

Cooperative Agreement Number: 17-8550-1997-CA

Project Funding Period: May 1, 2017 – April 30, 2018

Project Report: Farm Bill Survey Report

Project Document Date: July 9, 2017

Cooperators Project Coordinator: Jennifer Falacy & Chris Looney

Name: Jennifer Falacy & Chris Looney

Agency: Washington State Department of Agriculture

Address: 3939 Cleveland Ave. SE

City/ Address/ Zip: Olympia, WA 98501

Telephone: (360) 664-0905

E-mail: [email protected]

2017 Project Report - Plant Protection Division, Pest Program Washington State Department of Agriculture

mailto:[email protected]


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Summary

The WSDA conducted a Hardwood Survey in 2017 to determine the status of invasive pests

and diseases in Washington State affecting deciduous woody plants. No federal target

pathogens were detected during this survey.

Narrative

Washington State Department of Agriculture (WSDA) performed a visual survey of 30

counties for target diseases (see Table 1 & 2). WSDA exceeded its objectives by surveying

above six to eight sites at multiple locations in 30 counties for plant diseases and 14

counties for insects. A total of 670 samples were collected and submitted to the Plant

Pathology and Molecular Diagnostics Lab (PPMDL) for analysis. The Washington State

Department of Natural Resources (DNR) collected 143 samples targeting the symptoms of

Maple Decline (see Table 5). DNR also baited five streams (see table 4) for the target

Phytophthoras in addition to the ten streams they bait for P. ramorum that we repurposed for

this survey. No target Phytophthoras listed in Table 1 were detected from the stream baits.

In addition to taking samples of symptomatic woody plants for the lab to examine, negative

field observations were made of plants that were free from visible symptoms of the diseases

on our target list. The symptomatic sampled material is noted on the map as a yellow dot,

while documentation of the symptom-free plants shows on the map as an orange square.

WSDA deployed 10 translucent cross-vane traps with Megaplatypus mutatus lure and

10 Lindgren funnels with Qercivorol lure starting in May and completed in June

throughout Western Washington as an added trap to their 2016 Exotic Wood Boring

Insect Survey trap lines. Traps were checked on a three-week cycle, and lures were

changed every three to six weeks or as determined by CAPS approved methods and

traps were removed in September. All traps were negative for Megaplatypus mutatus

and Platypus quercivorus. Since no beetles were caught in the beetle traps, no follow-

up tests were performed to determine if they were carrying Raffaelea quercivora.

The insect surveys, covering 14 counties, for the nine insect pests listed in table 6

below were completed with no detections found. Pheromone surveys for four

Lepidoptera pests were conducted at 100 or more sites across the state. The field

trappers for insect surveys were given erroneous instructions by Dr. Chris Looney, the

entomology lab manager in charge of those surveys. He mistakenly re-used the trap

target numbers from FY2016, which were 100 sites for each trapped insect, rather than


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the goal of 120 sites/target specified in the FY2017 workplan. There was no shortage of

traps or lures. This was simply an error that was not realized until compiling the final

report. While this is disappointing, the coverage for those target species was still

relatively thorough across the state. Furthermore, lures for Adoxophyes orana were also

deployed in grape-growing counties in Washington State during FY 2017, and

Thaumatotibia leucotreta lures were used in additional experiments conducted in

western Washington. Nonetheless, traps for Thaumetopoea processionea and Tortix

viridana were deployed at 16 fewer sites than proposed.

Visual surveys for five pests were completed in early autumn and late spring, at 66 total

sites in primarily western WA. No suspect targets were detected. Insects were reared

from acorns collected at 66 sites as a supplement to the chestnut weevil visual survey.

No targets were detected in any of the surveys. Trap location information by county is

provided in Table 7 and noted on the map.

It was our hope that the symptoms of Maple Decline would be associated with only a

handful of organisms. Despite our sampling and testing we were unable to determine a

correlation between Maple Decline and a single or even a handful of organisms. We did

isolate organisms in culture and sequenced the ITS barcoding gene region, but most of the

similar sequences available to compare against in the (National Center for Biotechnology

Information) NCBI and the Barcode of Life Database (BOLD) were not from reference or

voucher specimens. This means that the confidence in determining these organisms to the

species level is not great. The NCBI database can be populated with sequence data that is

incorrectly annotated, has out of date nomenclature, or unpublished sequences which can

lead to incorrect identification of unknown specimens if caution is not used. Also lineage-

specific cutoff values for species determination using the ITS gene region are still being

evaluated and no single value can be applied across all groups (Raja et al. 2017). The

names listed in the table (attached separately Table 3) were not vetted against the two

name system nor verified to see if there were name changes over time. We would urge

caution when interpreting the names of the organisms on the table.

On the table outlining the organisms whose ITS barcoding region was sequenced, there are

a few organisms where both NCBI and BOLD agreed on the identifications (some only at

the genus level) and where BOLD had a match to a voucher specimen. The voucher

matched specimens included: Rhodotorula mucilaginosa (yeast, human pathogen of

immunocompromised patients, potential biocontrol, pathogen antagonist) (Ghani et al.

2014), Epicoccum sp. (plant pathogen), Diaporthe sp. (plant pathogen), Cosmospora sp.

(plant pathogen), and Cadophora melinii (plant pathogen). There were other organisms

where the databases may have agreed but lacked voucher or reference sequences to

increase the confidence in identifying the organisms at the species level. Some of these are

used as biocontrols or are common forest fungi.


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Despite our limited ability to identify these organisms to species using the ITS barcoding

gene region, we were able to determine that the symptoms of Maple Decline seem to be

associated with a very wide variety of organisms and that these organisms may not be

causing the disease but could be secondary opportunistic organisms or are simply in the

forest environment. It would seem that the causes of Maple Decline are complex and are

beyond the funding level of this project.


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6

Table 1 Survey Target Pests

Pathogen Testing Type

Maximum Number of Laboratory

Samples Budgeted

Alder Root and Collar Rot - Phytophthora alni

Morphological 100

Beech Bleeding Canker - Phytophthora kernoviae

ELISA 150

Beech Bleeding Canker - Phytophthora kernoviae

PCR 40

Phytophthora Oak Decline; Root Rot - Phytophthora quercina

Morphological 100

Phytophthora spp. Stream Bait Morphological 20

Phytophthora spp. Stream Bait PCR 10

Apple Proliferation - Candidatus Phytoplasma mali 16SrX-A

PCR 50

Australian Grapevine Yellows - Candidatus Phytoplasma australiense

16SrXII-B PCR 25

Japanese Oak Wilt - Raffaelea quercivora

Morphological 25

Testing to help determine etiology of Bigleaf Maple Decline

Candidatus Phytoplasma asteris PCR 100

Testing to help determine etiology of Bigleaf Maple Decline

Biotic Factor: Fungal Isolation 60


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Table 2 Survey Collection by County

County

Candidatus

Phytoplasma mali 16SrX-A

Phytophthora alni

Phytophthora

kernoviae

Phytophthora quercina

Phytoplasma

australiense 16SrXII-B

Raffaelea

quercivora

Total Samples Collecte

d

Negative Field

Observations

Lewis 5 7 23 6 4 0 59 101

King 0 12 18 11 0 0 50 39

Whatcom 5 1 13 2 13 0 48 22

Mason 11 11 23 7 1 0 48 59

Clallam 3 8 14 4 6 0 33 47

Snohomish 0 3 8 2 0 2 30 71

Jefferson 3 7 5 0 0 0 29 57

Thurston 4 7 7 8 3 0 25 35

Skamania 0 6 1 1 1 0 24 50

Pierce 1 1 7 3 2 3 22 64

Skagit 0 5 4 0 2 0 20 50

Pacific 2 1 11 0 2 0 16 60

Cowlitz 0 13 9 6 0 0 16 59

Clark 3 9 10 10 0 0 14 7

Okanogan 3 3 1 2 1 1 11 27

Douglas 0 0 5 5 0 1 11 37

Grays Harbor 0 0 9 0 0 0 11 48

Wahkiakum 2 1 6 0 0 0 9 43

Chelan 1 1 5 1 0 0 8 49

Kittitas 0 6 4 2 0 0 8 31

Benton 0 0 3 5 0 0 8 31

Yakima 0 0 2 3 0 0 5 32

Franklin 0 1 3 1 0 0 5 15

Ferry 2 1 1 0 0 0 4 22

Kitsap 0 4 4 4 0 0 4 35

Island 0 1 0 0 0 0 2 22

Grant 0 0 1 1 0 0 2 17

Whitman 0 0 2 0 0 0 2 7

Klickitat 0 0 0 2 0 0 2 5

Walla Walla 0 0 0 1 0 0 1 0

Total 45 109 199 87 35 7 527 1142


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Table 4 DNR Phytophthora Stream Bait sites

2017 - 2018 Dept. of Natural Resources Stream Baits

Site Latitude Longitude Number of visits

WDNR17-OP85 47.47522 -122.916 5

WDNR17-RC86 45.80299 -122.434 4

WDNR17-MC87 46.19036 -123.177 5

WDNR17-CR88 46.35173 -122.951 5

WDNR17-JC89 46.31214 -122.556 5

WDNR18-SR07 47.75506 -122.196 4

WDNR18-OP85 47.47522 -122.916 4

WDNR18-GC90 47.52948 -122.708 4

WDNR18-JC91 48.02061 -123.007 4

WDNR18-MC92 48.11047 -123.353 4

WDNR18-SC93 47.96857 -122.885 4

WDNR18-PC94 48.23865 -122.202 3

WDNR18-PR95 48.05369 -122.024 4

WDNR18-TR96 47.63827 -121.918 4

WDNR18-GR97 47.31271 -121.873 4


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Table 5 DNR Phytophthora Maple Decline sites

2017 – 2018 Department of Natural Resource Maple Decline Sites

Plot ID

DNR Plot Number Date

Latitude

Longitude Plot Type

Plot ID

DNR Plot Number Date

Latitude

Longitude Plot Type

1 48.08

769 123.5

6 Roadside Sample 90A 166

9/5/2017

48.90592

121.8804

DNR Resample

2 48.04

945 123.8

152 Roadside Sample 90B 166

9/6/2017

48.90446

121.8754

DNR Resample

3 48.06

51 123.9

118 Roadside Sample 90C 166

9/6/2017

48.90503

121.8879

DNR Resample

4 46.98

576 122.2

176 Roadside Sample

104A 160

9/26/2017

47.42574

121.777

DNR Resample

5 46.75

898 122.0

06 Roadside Sample

104B 160

9/26/2017

47.42724

121.7712

DNR Resample

6 46.62

08 122.2

266 Roadside Sample 61C 162

9/8/2017

48.68824

122.3145

DNR Resample

7 46.59

295 122.4

197 Roadside Sample 2

6/19/2017

47.51688

122.0845

Randomized Plot

8 46.57

153 122.3


6/19/2017

47.51908

122.0814

Randomized Plot

9 48.73

492 121.6


6/20/2017

47.60192

121.9738

Randomized Plot

10 48.53

722 121.2


6/20/2017

47.60716

121.9847

Randomized Plot

11 48.45

71 121.5


6/21/2017

47.36457

121.6517

Randomized Plot

12 48.29

388 121.7


6/26/2017

47.60572

121.9771

Randomized Plot

13 47.73

822 121.3


6/27/2017

47.53992

122.0923

Randomized Plot

14 46.64

638 121.6


6/27/2017

47.53815

122.0924

Randomized Plot

15 46.65

232 121.7


7/3/2017

47.62944

122.2953

Randomized Plot

16 46.64

375 121.7


7/3/2017

47.634

122.2961

Randomized Plot

17 46.62

898 121.6


7/3/2017

47.6379

122.295

Randomized Plot

18 46.52

444 121.9


7/11/2017

47.5368

123.7069

Randomized Plot

19 8/9/20

17 45.67

27 121.9


7/12/2017

47.56741

123.6543

Randomized Plot

20 8/10/2

017 45.62

124 122.0


7/12/2017

47.52768

123.7806

Randomized Plot

21 8/11/2

017 45.55

994 122.2


7/12/2017

47.56683

124.1709

Randomized Plot

22 8/11/2

017 45.56

101 122.2


7/13/2017

47.68095

123.0353

Randomized Plot

23 8/11/2

017 45.56

906 122.2


7/13/2017

47.66627

122.9849

Randomized Plot

24 8/11/2

017 45.58

34 122.1


7/16/2017

47.9633

124.2777

Randomized Plot

25 8/11/2

017 45.59

674 122.1


7/16/2017

47.85182

123.9565

Randomized Plot

26 8/13/2

017 48.10

063 121.1


7/16/2017

47.82878

123.9927

Randomized Plot

27 8/13/2

017 48.16

61 161.4


7/18/2017

47.68393

123.0242

Randomized Plot

28 8/14/2

017 48.26

087 121.2


7/18/2017

47.68137

122.9963

Randomized Plot

29 8/14/2

017 488.3

09 121.4


7/19/2017

48.00805

123.9089

Randomized Plot


10

30 8/14/2

017 46.92

733 122.9


7/19/2017

48.04575

123.9566

Randomized Plot

31 8/24/2

017 46.47

479 123.0


7/23/2017

46.76051

121.9564

Randomized Plot

32 8/24/2

017 46.54

586 123.1


7/23/2017

46.7572

121.9561

Randomized Plot

33 8/26/2

017 46.83

262 123.1


7/23/2017

46.76631

121.9517

Randomized Plot

34 8/26/2

017 47.09

801 123.4


7/26/2017

48.66583

121.2819

Randomized Plot

35 9/7/20

17 46.86

624 122.7


7/27/2017

48.71009

121.6943

Randomized Plot

36 9/7/20

17 48.81

053 122.2


7/27/2017

48.74714

121.5658

Randomized Plot

37 9/7/20

17 47.62

913 122.2


7/28/2017

48.48265

121.2296

Randomized Plot

1A 155 6/13/2

017 47.13

16 122.8

5 DNR Resample 41

7/28/2017

48.50549

121.2471

Randomized Plot

1B 155 6/13/2

017 47.13

29 122.8

467 DNR Resample 43

8/1/2017

47.73165

121.3386

Randomized Plot

1C 155 6/13/2

017 47.12

962 122.8

527 DNR Resample 44

8/1/2017

47.73139

121.3388

Randomized Plot

10A 186 6/30/2

017 47.84

05 121.6

201 DNR Resample 45

8/1/2017

47.67289

121.3088

Randomized Plot

10B 186 6/30/2

017 47.84

224 121.6

144 DNR Resample 46

8/2/2017

46.68532

121.5053

Randomized Plot

10C 186 6/30/2

017 47.84

481 121.6

213 DNR Resample 47

8/2/2017

46.67517

121.5487

Randomized Plot

11A 187 7/1/20

17 47.82

091 12.52

776 DNR Resample 48

8/3/2017

46.67305

121.5743

Randomized Plot

11B 187 7/1/20

17 47.82

105 121.5

212 DNR Resample 50

8/7/2017

46.65896

121.5988

Randomized Plot

15A 816 7/10/2

017 47.54

141 123.6

759 DNR Resample 51

8/8/2017

45.58236

122.1878

Randomized Plot

15B 816 7/11/2

017 47.54

496 123.6

756 DNR Resample 52

8/8/2017

45.58616

122.1839

Randomized Plot

15C 816 7/11/2

017 47.53

729 123.6

774 DNR Resample 54

8/10/2017

45.61042

122.0574

Randomized Plot

22A 190 7/14/2

017 47.49

614 123.2

7 DNR Resample 56

8/13/2017

48.08515

121.3879

Randomized Plot

22B 190 7/14/2

017 47.49

593 123.2

646 DNR Resample 57

8/13/2017

48.10084

121.3791

Randomized Plot

26A 813 7/17/2

017 47.37

671 122.9

728 DNR Resample 58

8/13/2017

48.13361

121.4172

Randomized Plot

26B 813 7/17/2

017 47.37

24 122.9

75 DNR Resample 59

8/14/2017

48.25025

121.1968

Randomized Plot

35A 182 7/24/2

017 46.55

354 122.5

381 DNR Resample 60

8/14/2017

48.25531

121.2783

Randomized Plot

35B 182 7/24/2

017 46.55

006 122.5

439 DNR Resample 74

8/28/2017

47.41633

121.8223

Randomized Plot

35C 182 7/24/2

017 46.55

723 122.5

348 DNR Resample 75

8/28/2017

47.41502

121.8136

Randomized Plot

36A 179 7/24/2

017 46.60

375 122.4

647 DNR Resample 76

8/28/2017

47.40823

121.8035

Randomized Plot

36B 179 7/25/2

017 46.60

495 122.4

708 DNR Resample 77

8/28/2017

47.40766

121.8047

Randomized Plot

36C 179 7/25/2

017 46.60

367 122.4

579 DNR Resample 78

8/29/2017

47.41495

121.8163

Randomized Plot

42A 148 7/29/2

017 48.53

738 121.2

919 DNR Resample 79

8/19/2017

47.4157

121.8137

Randomized Plot

42B 148 7/29/2

017 48.53

871 121.2

851 DNR Resample 80

8/29/2017

47.40529

121.8078

Randomized Plot

42C 148 7/29/2

017 48.53

668 121.2

976 DNR Resample 91

9/7/2017

48.8938

121.9046

Randomized Plot


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49A 176 8/3/20

17 46.53

956 121.8

127 DNR Resample 92

9/7/2017

48.87804

121.9134

Randomized Plot

49B 176 8/3/20

17 46.54

193 121.8

073 DNR Resample 100

9/23/2017

47.86481

121.4771

Randomized Plot

49C 176 8/3/20

17 46.53

728 121.8

16 DNR Resample 102

9/25/2017

47.54823

121.5423

Randomized Plot

53A 143 8/9/20

17 45.63

887 122.0

36 DNR Resample 103

9/25/2017

47.5402

121.5593

Randomized Plot

53B 143 8/9/20

17 45.64

328 122.0

38 DNR Resample 70L

8/24/2017

46.97624

122.7551

Landowner Plot

53C 143 8/9/20

17 45.63

785 122.0

297 DNR Resample 71L

8/23/2017

47.07578

123.2441

Landowner Plot

55A 144 8/10/2

017 45.62

033 122.0


8/25/2017

46.56089

122.6437

Landowner Plot

55B 144 8/10/2

017 45.62

131 122.0


48.371296

48.3713

122.2055

Landowner Plot

61A 162 8/15/2

017 48.69

295 122.3


9/8/2017

48.67667

122.2073

Landowner Plot

61B 162 8/27/2

017 48.69

773 122.3

144 DNR Resample

101L

9/24/2017

48.19994

122.5164

Landowner Plot

29L 7/18/2

017 47.81

447 122.8

808 Landowner Plot


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Table 6 Survey Insect Trap and Visual Survey Summary

Scientific

Name

Common

Name

Survey

Method

Number

proposed

traps/trees

Number

actual

traps/trees

Comments

Adoxophyes

orana

Summer

fruit tortrix

Pheromone

lure, paper

delta trap

120 101 No targets detected

Aeolesthes

sarta

City

longhorned

beetle

Visual

survey

20 66 sites No targets detected

Agrilus

biguttatus

Oak

splendour

beetle

Visual

survey


Curculio

elephas

Chestnut

weevil

Visual

survey /

acorn

rearing

20 66 sites

No targets detected

Scolytus

intricatus

Oak

ambrosia

beetle

Visual

survey


Thaumatotibia

leucotreta

False

codling

moth

Pheromone

lure, wing

trap


Thaumetopoea

processionea

Oak

procession

moth

Pheromone

lure, wing

trap


Tortrix

viridana

Green oak

tortrix

Pheromone

lure, wing

trap


Tremex

fuscicornis

Tremex

woodwasp

Visual

survey



13

Table 7 Insect Trap Location by County 2017

Results

No federal target pests or diseases were detected in this survey. There was no clear front runner in which a single pathogen might be implicated as the causal agent of Maple Decline. Instead there were 43 organisms detected, many of which were not pathogens. The NCBI and BOLD databases frequently did not agree on what the organisms were based on the ITS barcoding region and/or were not matched to a voucher or reference sequence. Expected Benefits

1. The pest free status of Washington State and the Northwest region will be

maintained by early detection and possible eradication of an economically important group of pathogen and insect pests.

2. Washington State and the Northwest region’s fruit, nut, berry, nursery, and forestry

industries will be protected from an important group of destructive pest insects.

3. The survey will help maintain the unregulated export of fruit, nuts, berries, nursery stock, and forestry products from Washington State to international markets.

County Adoxophyes

orana Thaumatotibia

leucotreta Thaumetopoea processionea

Tortrix viridana

Total Trap Sites

Visual Survey Sites

Total Survey Sites

King 11 10 9 9 39 13 91

Spokane 12 10 11 12 45 0 90

Clark 11 11 12 11 45 10 100

Snohomish 10 10 12 12 44 3 91

Thurston 11 10 11 11 43 9 95

Whatcom 9 11 11 9 40 4 84

Yakima 10 10 10 10 40 6 86

Lewis 8 9 9 11 37 5 79

Pierce 6 6 6 6 24 3 51

Grays Harbor 5 5 5 5 20 2 42

Skamania 5 5 5 5 20 1 41

Cowlitz 3 3 3 3 12 4 28

Kittitas 0 0 0 0 0 1 1

San Juan 0 0 0 0 0 5 5

Total 101 100 104 104 409 66 884


14

Acknowledgements We’d like to thank Dan Omdal and Amy Ramsey at Washington State Department of Natural Resources, Jenni Cena at WSDA and the WSDA Molecular Diagnostics & Plant Pathology Lab, for their help in this project. Literature Cited

Ghani, N. A,; Sulaiman, J.; Chan, X., Yin, W., Chan, K. (2014). Rhodotorula

Mucilaginosa, a Quorum Quenching Yeast Exhibiting Lactonase Activity Isolated from a Tropical Shoreline. Sensors, 14, 6463-6473.

Raja, H.A; Miller, A.N; Pearce, C.J.; Oberlies (2017). Fungal Identification Using

Molecular Tools: A Primer for the Natural Products Research Community. American Chemical Society and American Society of Pharmacognosy, 80, 756-770.

Distribution / Content Note This report on the Washington State Hardwood Commodity Survey is provided as a public resource. This entire report, as well as individual graphic images, may be freely copied, distributed, and used in electronic and printed format as long as they are not modified for content or used for commercial purposes. Inquiries regarding the availability of this publication in an alternate format should be directed to the WSDA Receptionist at (360) 902-1976 or Telecommunications Device for the Deaf (360) 902-1996. Jennifer Falacy WSDA, Plant Protection Division 3939 Cleveland Ave. SE Olympia, WA 98501 [email protected]

mailto:[email protected]

Table 3

Sample ID

PCR

Tube

Label

PDA Isolates Notes

BOLD DatabaseSimilarity for

BOLDScore for BOLD

Matched to Voucher

Specimen? Accession # NCBI Database

Indentical for

NCBI

Query

Coverage

NCBI

Matched

to

Voucher

Specimen?

Grunwald Lab USDA-ARS

http://phytophthora-

id.org/seq-id.html

NotesIs the organism reported

in WA?Methodolgy of Detection

DNR11-B 93 Cadophora melinii 100 505.00 No voucher status

AB190403

Cadophora melinii CBS 268 33 ITS region 100% 97% Yes No

Detection of liganase positive fungal cultures through selective cromophoric media, isolation of liganase positive

cultures on PDA to PCR ITS Sequencing

BLM1-A-1-1-

D-600bp 168 Human Pathogen Cladosporium cladosporiodes 100 469.00 No voucher status DQ426525 Cladosporium neolangeronii culture CBS: 797.97 100% 100% Yes Yes, multiple hosts



DNR8-A 148 Colletotrichum acutatum 100 502.00 No voucher status

JQ040073

Colletotrichum godetiae CBS 133.44 100% 99% Yes Numerous Species



BLM -15C-1

7/11/17-JB-

D 44 Cosmospora sp. 4 PC-2011 98.95% 468.00 No voucher status JN995629 Cosmospora viridescens IMI 73377a ITS 99% 100% Yes No



DNR13-C 116 Diaporthe sp. 2 PRJ-2013 99.82 586.00 No voucher status KC145877

Diaporthe alnea CBS 146.46 ITS region: TYPE

material 98% 99% Yes No



DNR13-B 149 Diaporthe sp. 2 PRJ-2013 99.82 541.00 No voucher status

KC145877 Diaporthe alnea CBS 146.46 ITS region: TYPE

material 99% 100% Yes Yes, host: Ilex aquifolium



BLM-AU-i3 293

Pathogenic towards

certain species of

grape, Pyrus and

Malus. Visual match

to Diaporthe viticola. Diaporthe viticola 100 475.00 No voucher status AY485750 Diaporthe rudis strain AR3422 100% 100% Yes

Pathogenic towards certain

species of grape, Pyrus and

Malus No



BLM-AU-i1-

550Bp 300

Fungus is linked to

dieback of urban

London Plane tree in

Geneva.

(https://www.researc

hgate.net/publicatio

n/313356454_Urban

_London_plane_tree

_dieback_linked_to_f

ungi_in_the_Botryos

phaeriaceae) Diplodia mutila 100 465.00 No voucher status

KC789072

Diplodia pyri CBS 121862 100% 100% Yes No



BLM-AU-i2 292

Visually matches

Diplodia seriata

strain. Diplodia mutila 100 465.00 No voucher status KC789072

Diplodia seriata strain CBS 112555 18S

ribosomal RNA gene 97% 100% Yes

Fungus is linked to dieback of

urban London Plane tree in

Geneva.

(https://www.researchgate.net

/publication/313356454_Urban

_London_plane_tree_dieback_l

inked_to_fungi_in_the_Botryos

phaeriaceae) No



DNR2-A 69

visual match is closer

to Drechslera brizae Pyrenophora leucospermi 100% 541.00 No voucher status JN712478 Drechslera brizae CBS 190.29 ITS region 98% 100% Yes No



DNR2-C-1 71

Visual match is

closest to Epicoccum

nigrum. Epicoccum sp. 02TRU72c 100% 461.00 No voucher status

AY305361

Epicoccum rosae strain MFLU 15-3639 99% 95% Yes

Fávaro, Léia & Melo,

Fernando & Aguilar-

Vildoso, Carlos & Araújo,

Welington. (2011).

Polyphasic Analysis of

Intraspecific Diversity in

Epicoccum nigrum

Warrants Reclassification

into Separate Species.

PloS one. 6. e14828.

10.1371/journal.pone.00

14828.

"In fact, based on the

polyphasic approach we

suggest the occurrence of

cryptic species within E. nigrum

and also that many of the

sequences deposited as E.

nigrum in GenBank and culture

collection of microbial strains

should be reclassified, including

the reference strain CBS 161.73

sequenced in this work. In

addition, this study provides

valuable tools for

differentiation of Epicoccum

species." No



DNR2-C-2 72

Visual match is

closest to Epicoccum

nigrum. Epicoccum sp. 02TRU72c 100% 459.00 No voucher status

AY305361

Epicoccum rosae strain MFLU 15-3639 99% 95% Yes

Fávaro, Léia & Melo,

Fernando & Aguilar-

Vildoso, Carlos & Araújo,

Welington. (2011).

Polyphasic Analysis of

Intraspecific Diversity in

Epicoccum nigrum

Warrants Reclassification

into Separate Species.

PloS one. 6. e14828.

10.1371/journal.pone.00

14828.

"In fact, based on the

polyphasic approach we

suggest the occurrence of

cryptic species within E. nigrum

and also that many of the

sequences deposited as E.

nigrum in GenBank and culture

collection of microbial strains

should be reclassified, including

the reference strain CBS 161.73

sequenced in this work. In

addition, this study provides

valuable tools for

differentiation of Epicoccum

species." No



http://phytophthora-id.org/seq-id.html



http://www.ncbi.nlm.nih.gov/nuccore/AB190403

http://www.ncbi.nlm.nih.gov/nuccore/DQ426525

http://www.ncbi.nlm.nih.gov/nuccore/JQ040073

http://www.ncbi.nlm.nih.gov/nuccore/JN995629

http://www.ncbi.nlm.nih.gov/nuccore/KC145877


http://www.ncbi.nlm.nih.gov/nuccore/AY485750






Table 3

Sample ID

PCR

Tube

Label

PDA Isolates Notes


BOLDScore for BOLD

Matched to Voucher


Indentical for

NCBI

Query

Coverage

NCBI

Matched

to

Voucher

Specimen?



id.org/seq-id.html



PM6-D 132 Fusarium oxysporum 100 462.00 No voucher status JX997430 Fusarium inflexum NRRL 20433 ITS region 99% 100% Yes Yes, multiple species/hosts



DNR15-B 129

Visual match is

closest to

Graphostroma sp. Biscogniqiauxia capnodes 86.35 511.00 No voucher status EF026131 Graphostroma sp. SGLMf27 18S ribosomal RNA 98% 99% Yes No



BLM-1-A-1-

1-D(B) 35 Ilyonectria radicicola 100% 505.00 No voucher status HQ840390

Ilyonectria liliigena strain CBS 189.49 18S


Yes, host rhododendron,

Carcissus, Pyrus communis,



PM1-A 128

Visual match is

closest to Ilyonectria

radicicola. Ilyonectria radicicola 100 505.00 No voucher status

HQ840390

Ilyonectria liliigena strain CBS 189.49 18S


Yes, host rhododendron,

Carcissus, Pyrus communis,



DNR13-A 97

Cadophora melinii as

a contaminated ITS

and not isolated

culture Cadophora melinii 100 502.00 No voucher status

DQ404351

Molissia dextrinospora ITS region from TYPE

material 97% 98% Yes No



DNR59A 119

Visual match is

closest to Phoma

herbarum. Paecilomyces sp. MD201 100 499.00 No voucher status DQ512798 Paraboeremia selaginellae CBS 122.93 ITS region 99% 96% Yes Yes, host: Pinus ponderosa



PM2-A 127

Visual match is

closest to

Paraphaesophaeria

neglecta. Paraconiothyrium fuckelii 99.47 557.00 No voucher status

EF540754

Paraphaesophaeria negleceta CBS 124078 ITS

region from TYPE material 99% 99% Yes No



PM2-B-2 126

Visual match is

closest to

Paraconiothyrium

fuckelii. Paraconiothyrium fuckelii 99.47 557.00 No voucher status

EF540754

Paraphaesophaeria neglecta CBS 124078 99% 99% Yes Yes, host: multiple



BLM-1-A-1-

1-D ( C) 36

Rhizomucor variabilis var.

regularior 100% 599.00 No voucher status HM623316

Rhizomuor variablilis var. regularior strain CBS

384.95 18S ribosomal RNA gene 100% 100% Yes No



DNR4-D 78 Rhodotorula mucilaginosa 99.45 536.00 No voucher status

EU177580

Rhodotorula mucilaginosa culture CBS:316 99% 100% Yes No



DNR4-B 76

Visual match is

closest to Cadophora

melinii. Cadophora melinii 100 502.00 No voucher status

DQ404351

Ascomycota sp. Strain A100 small subunit

ribosomal RNA gene 100% 100% No No



DNR11-C 94 Cadophora melinii 99.8 498.00 No voucher status DQ404351

Cadophora spadicis isolate RIC3 internal

transcribed spacer 1 100% 100% No No



DNR4-A 75

Visual match is

closest to

Cosmospora. Cosmospora sp. 99.36 465.00 No voucher status

JN604455

Cetriaceae sp. JCM 28357 genes for 18S rRNA 99% 100% No No



BLM-AU-i4 294

Biopesticide, used as

a forest control

agent. Also naturally

occuring in

forests.https://www.

researchgate.net/pro

file/Ivan_Zmitrovich2

/publication/2733797

11_Wood-

inhabiting_fungi/link

s/54ff441b0cf2741b6

9f5f49d/Wood-

inhabiting-fungi.pdf Chondrostereum purpureum 99.66 578.00 No voucher status

DQ200929

Chondrostereum purpureum isolate 23548 99% 100% No

Biopesticide, used as a forest

control agent. Also naturally

occuring in

forests.https://www.researchg

ate.net/profile/Ivan_Zmitrovich

2/publication/273379711_Woo

d-

inhabiting_fungi/links/54ff441b

0cf2741b69f5f49d/Wood-

inhabiting-fungi.pdf

Yes, host: Abies amabilis,

ACER MACROPHYLLUM



BLM-AU-i5 295

Biopesticide, used as

a forest control

agent. Also naturally

occuring in

forests.https://www.

researchgate.net/pro

file/Ivan_Zmitrovich2

/publication/2733797

11_Wood-

inhabiting_fungi/link

s/54ff441b0cf2741b6

9f5f49d/Wood-

inhabiting-fungi.pdf Chondrostereum purpureum 99.51 603.00 No voucher status No accession # Chondrostereum purpureum isolate 23548 99% 100% No

Biopesticide, used as a forest

control agent. Also naturally

occuring in

forests.https://www.researchg

ate.net/profile/Ivan_Zmitrovich

2/publication/273379711_Woo

d-

inhabiting_fungi/links/54ff441b

0cf2741b69f5f49d/Wood-

inhabiting-fungi.pdf

Yes, host: Abies amabilis,

ACER MACROPHYLLUM



DNR6-A 80 Common forest fungi Coprinellus micaceus 99.39 647.00 No voucher status

JN943115

Coprinellus micaceus voucher DBG: 24506 100% 100% No No






http://www.ncbi.nlm.nih.gov/nuccore/JX997430

http://www.ncbi.nlm.nih.gov/nuccore/EF026131

http://www.ncbi.nlm.nih.gov/nuccore/HQ840390

http://www.ncbi.nlm.nih.gov/nuccore/HQ840390





http://www.ncbi.nlm.nih.gov/nuccore/HM623316

http://www.ncbi.nlm.nih.gov/nuccore/EU177580






Table 3

Sample ID

PCR

Tube

Label

PDA Isolates Notes


BOLDScore for BOLD

Matched to Voucher


Indentical for

NCBI

Query

Coverage

NCBI

Matched

to

Voucher

Specimen?



id.org/seq-id.html



DNR6-B 81

Visually resembles

both Biscogniquxia

and Cryptostroma Biscogniquxia cinereolilacina 95.97 522.00 No voucher status Private? Cryptostroma corticale strain IT01BO 99% 99% No



DNR7-A 84 Epicoccum nigrum 100 511.00 No voucher status FJ424241 Epicoccum nigrum strain CRM-DLRZ2 100% 100% No Yes, multiple hosts



BLM-19-1-

7/12/17-JB-

D 37

Isolates Same Culture

i.e. single sequencing Fomitopsis pinicola 100% 626.00 No voucher status AY854083

Fomitopsis pinicola isolate AFTOL-ID 770 18S

ribosomal RNA gene 100% 100% No Yes, multiple hosts



DNR14 117

DOES NOT MATCH

visual for Fuscoporia

ferrea. Fuscoporia ferrea 99.83 600.00 No voucher status JQ794582

Fuscoporia ferrea isolate FP-133592-Sp 18S




DNR12-A-

600bp 184

DOES NOT MATCH

visual for Fuscoporia

ferrea. Fuscoporia ferrea 99.83 600.00 No voucher status

JQ794582

Fuscoporia ferrea isolate FP-133592-Sp 18S




PM5-B 130 Ganoderma applanatum 99.83 601.00 No voucher status AY884178 Ganoderma applanatum strain 414 99.00% 100 No Yes, multiple hosts



PM5-A 131 Ganoderma applanatum 99.83 602.00 No voucher status AY884178 Ganoderma applanatum strain 414 99% 100% No Yes, multiple hosts



PM3 133 Ganoderma applanatum 100 602.00 No voucher status

AY884178

Ganoderma applanatum strain 414 100% 100% No Yes, multiple hosts



DNR12-B-

650bp 171

No reference image

of Gloeodontia

subasperispora Gloedontia subasperispora 88.85 436.00 No voucher status Private?

Gloeodontia subasperispora voucher

UC2022874 89% 100% No

High quality sequence, Pairwise

identity=96.1%, HQ=99.8% No



DNR15-A 118 Buscogniauxia capnodes 86.09 497.00 No voucher status EF026131 Graphostroma sp. SGLMf27 18S ribosomal 98% 100% No No



DNR4-C-

600bp 170

Cosmospora is a

genera in the

Nectriacea family Cosmospora sp. 99.36 465.00 No voucher status

JN604455

Nectriacea. JCM 28357 99% 100% No No



BLM-AU-i7 297

Can be pathogenic

toward humans.

Visual match to

phialomonium Phialemonipsis ocularis 97.29 413.00 No voucher status

HE599279

Phialemonium sp. Gall 27 18S ribosomal RNA

gene 100% 100% No

Can be pathogenic toward

humans. No



DNR2-B-

600bp 169 Tetracladium sp. CBS 118523 95.73 465.00 No voucher status AJ890435

Uncultured Tetracladium clone LTSP EUKA

P4O10 99% 100% No No



BLM-AU-i6 296 Trametes versicolor 100 362.00 No voucher status

HE599279

Trametes versicolor CFMR FP-135156 99% 100% No

BLASTed forward strand.

Reverse strand very low quality Yes, host: multiple






http://www.ncbi.nlm.nih.gov/nuccore/FJ424241









http://www.ncbi.nlm.nih.gov/nuccore/HE599279

http://www.ncbi.nlm.nih.gov/nuccore/AJ890435

http://www.ncbi.nlm.nih.gov/nuccore/HE599279

2017 hardwood survey report - wa

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