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Sudden Oak Death Archives

Isolating Oomycetes from leaf baits in the lab


Isolating Oomycetes from leaf baits in the lab

5 10 2011 (click on photos to see larger photo).

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Contact: Gary Chastagner, 253-445-4528 | WSU Puyallup Research & Extension Center, 2606 West Pioneer, Puyallup, WA, 98371-4998 USA
Last updated January 2, 2013

Photos “bait in a bottle” method at Clarks Creek


Stream sampling using ”bait in a bottle” method at Clarks Creek

Sample collection 5 3 2011 (click on photos to see larger photo).

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Contact: Gary Chastagner, 253-445-4528 | WSU Puyallup Research & Extension Center, 2606 West Pioneer, Puyallup, WA, 98371-4998 USA
Last updated January 2, 2013

Results of BOB sampling at Clarks Creek


Results of BOB sampling at Clarks Creek

The most commonly isolated group of Oomycetes was Pythium belonging to Clade B2. There was little difference among hosts, except for reed canarygrass from which only one Pythium sp. was isolated. Saprolegnia was isolated from three hosts. Two Phytophthoras belonging to Clade 6 were isolated, and these, as well as Pythiums in Clade B2, were commonly found at the other freshwater sites sampled in this study.

Phylogenetic trees from sequence data obtained in this study:

Phytophthora Pythium Saprolegnia

Host
species isolated
group or clade
Riparian/forest
# isolates
False lily of the valley Maianthemum dilatatum Fusarium spp. fungus
F
2
False lily of the valley Maianthemum dilatatum Phytophthora taxon ‘salixsoil’ 6
F
1
False lily of the valley Maianthemum dilatatum Pythium diclinum/lutarium B2
F
5
Grand fir Abies grandis Phytophthora taxon ‘pgchlamydo’ 6
F
1
Grand fir Abies grandis Pythium aquatile B2
F
1
Grand fir Abies grandis Pythium diclinum/lutarium B2
F
7
Licorice fern Polypodium glycorrhiza Mucor hiernalis zygomycete
F
1
Licorice fern Polypodium glycorrhiza Pythium aquatile B2
F
1
Licorice fern Polypodium glycorrhiza Pythium diclinum/lutarium B2
F
2
Licorice fern Polypodium glycorrhiza Pythium oopapillum B2
F
1
Rhododendron Rhododendron ‘Nova Zembla’ Pythium aquatile B2
F
1
Rhododendron Rhododendron ‘Nova Zembla’ Pythium diclinum/lutarium B2
F
5
Rhododendron Rhododendron ‘Nova Zembla’ Saprolegnia parasitica Saprolegnia
F
1
Red alder Alnus rubra Pythium diclinum/lutarium B2
R
5
Red alder Alnus rubra Pythium oopapillum B2
R
2
Red alder Alnus rubra Pythium undulatum H
R
1
Reed canary grass Phalaris arundinacea Pythium intermedium F
R
8
Salmonberry Rubus spectabilis Pythium diclinum/lutarium B2
R
8
Salmonberry Rubus spectabilis Pythium oopapillum B2
R
1
Salmonberry Rubus spectabilis Saprolegnia parasitica Saprolegnia
R
1
Skunk cabbage Lysichiton americanum Mortierella sp. zygomycete
R
1
Skunk cabbage Lysichiton americanum Pythium diclinum/lutarium B2
R
3
Skunk cabbage Lysichiton americanum Saprolegnia diclina Saprolegnia
R
1

Pierce College BOB 2011_allhosts

Contact: Gary Chastagner, 253-445-4528 | WSU Puyallup Research & Extension Center, 2606 West Pioneer, Puyallup, WA, 98371-4998 USA
Last updated January 2, 2013

Stream sampling using “bait in a bottle” method at Clarks Creek


Stream sampling using ”bait in a bottle” method at Clarks Creek

Oomycetes are fungus-like organisms found in marine, freshwater, and terrestrial environments. Some, such as Phytophthora, Pythium, and Saprolegnia, are parasites of plants and animals. DNA sequence data has revealed that these organisms are not fungi, but are more closely related to brown algae and diatoms. We will be “fishing” for these organisms by using leaf baits from various plant species.

Objectives

  • Is there a relationship between host bait material and Oomycete species isolated?
  • Riparian vs forest plants – are riparian plants resistant to infection?
  • Has P. ramorum made it to Clarks Creek?

Each group will choose a plant species to use for bait. Pick fully expanded leaves that are relatively free of spots or other signs of infection. Punch out 20 leaf discs and put in bottle. Include one intact leaf. Put leaf baits in bottle of creek water and cap tightly. Rest bottle on its side for incubation.

After incubation leaf baits will be cultured on selective media for Oomycetes, transferred to pure culture, and identified using DNA sequencing.

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Students from Pierce College Biology 213 class collecting water samples at Clarks Creek.

Contact: Gary Chastagner, 253-445-4528 | WSU Puyallup Research & Extension Center, 2606 West Pioneer, Puyallup, WA, 98371-4998 USA
Last updated January 2, 2013

Clarks Creek Headwaters 1103


Clarks Creek headwaters

Site #1103

Clarks Creek, near fish hatchery

This site also used for Pierce College “Bait in a Bottle” experiment.


Elevation 48 ft

Stream order 1


Clarks Creek Initiative

Results from 2011 monitoring

2011 sites species data_25436_image018

2011 water temp data_14139_image021

Results from 2010 monitoring

1007

Contact: Gary Chastagner, 253-445-4528 | WSU Puyallup Research & Extension Center, 2606 West Pioneer, Puyallup, WA, 98371-4998 USA
Last updated January 2, 2013

2011-1102 Clarks Creek downstream


Clarks Creek downstream

DSC05321 sm Clarks crkthumb DSC05328 sm Clarks Crkthumb
Land use map 1102_350

Site #1102 Clarks Creek, Puyallup, WA


Elevation 24 ft Stream order 2


Clear/Clarks Creek Basin Plan 2006, Pierce County Public Works & Utilities USGS data for Clark’s Creek

Results from 2011 monitoring
 2011 sites species data_25436_image017
 2011 water temp data_14139_image017
 Results from 2010 monitoring
1001

Contact: Gary Chastagner, 253-445-4528 | WSU Puyallup Research & Extension Center, 2606 West Pioneer, Puyallup, WA, 98371-4998 USA
Last updated January 2, 2013

Clarks Creek – headwaters


Clarks Creek – headwaters

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Site # 1007Baiting at Clarks Creek upstream, near fish hatchery by Biology 213 class from Pierce College. 4/15/10.

see results of the Shoe Experiment here

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Contact: Gary Chastagner, 253-445-4528 | WSU Puyallup Research & Extension Center, 2606 West Pioneer, Puyallup, WA, 98371-4998 USA Last updated January 2, 2013

Clark’s Creek – downstream


Clark’s Creek – downstream

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DSC05863 350 labPHS students learning molecular identification techniques in the lab
DSC05328 sm Clarks Crkthumb Site #1001First bait deployment March 3, 2010 by Puyallup High School students in Dave Wetzel’s class

1001

Baiting period Bait leaf spp. Phytophthora spp. and others identified
1
Rhododendron P. gonapodyides (6), Trichoderma spp. (1)
1
Camellia P. gonapodyides (1), unknown (1)
2
Rhododendron P. gonapodyides (4), unknown (4)
2
Gold dust plant (Aucuba japonica ‘Variegata’) Pythium litorale or Py. sterilum (2)
3
Rhododendron P. gonapodyides (6), unknown (5)
3
Strawberry tree (Arbutus unedo) Pythium sp. or Py. undulatum (1), unknown (1)
4
Rhododendron P. gonapodyides (5), P. gonapodyides “A” (4), P. cryptogea (1), unknown (1)
4
Dwarf cherry laurel P. gonapodyides (2), unknown (2)
5
Rhododendron P. gonapodyides (7), unknown (5)
5
unknown (2)
6
Rhododendron P. gonapodyides (4), unknown (8)
6
Red tip Photinia P. gonapodyides (2)

Viewing Chum Salmon in Clark’s Creek

WDFW 2000 Clear/Clark’s Creek Basin Plan Pierce County 2005

Pink salmon in Clarks Creek near Puyallup NWIFC Blog Sept. 2007

 

Contact: Gary Chastagner, 253-445-4528 | WSU Puyallup Research & Extension Center, 2606 West Pioneer, Puyallup, WA, 98371-4998 USA
Last updated January 2, 2013

2011 Stream Monitoring Results


2011 Stream Monitoring Results

This project provided baseline information about oomycetes in western Washington streams. Some very common species that were ubiquitous in all streams in large numbers were identified, as well as some more rare species only found in certain streams. Several putative new species were also detected. The table below shows the locations of western Washington streams monitored in this study. Simpson’s diversity index (D) was calculated for each site based on the number of isolates and species of oomycetes detected. Values close to 0 indicate higher species diversity.

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Stream ID Name County Land use Total number of isolates Total number of species Simpson’s diversity index (D)
1101 Packard Creek* Clark Agriculture/residential

25

11

0.1333

1102 Clarks Creek Pierce Agriculture

9

6

0.1111

1103 Clarks Creek Pierce Fish hatchery, residential

15

8

0.2095

1104 Woodard Bay Thurston Conservation area, brackish

29

5

0.6823

1105 Goldsborough Creek Mason Forest/residential

31

7

0.428

1106 North Creek Snohomish Suburban/residential

22

9

0.0996

1107 Clear Creek Kitsap Urban/commercial

20

8

0.1526

1108 Peters Creek King Urban/commercial

22

6

0.2035

1109 Vance Creek Mason Undisturbed forest

25

4

0.3833

1110 Thea Foss Waterway Pierce Formerly industrial, heavily polluted, brackish

39

3

0.5196

1112 Wollochet Creek* Pierce Suburban/residential, ditch adjacent to former nursery site

23

5

0.332

1113 Rosedale Creek* Pierce Suburban/residential

16

8

0.1417

* Known to contain Phytophthora ramorum from WSDA or DNR baiting

Contact: Gary Chastagner, 253-445-4528 | WSU Puyallup Research & Extension Center, 2606 West Pioneer, Puyallup, WA, 98371-4998 USA Last updated January 2, 2013

Stream monitoring sites 2011


Stream monitoring sites 2011

Snohomish County

1106 – North Creek

King County

Kitsap County

Mason County

2011 monitoring sites

Contact: Gary Chastagner, 253-445-4528 | WSU Puyallup Research & Extension Center, 2606 West Pioneer, Puyallup, WA, 98371-4998 USA Last updated January 2, 2013