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Mist chamber extraction for improved diagnosis of Meloidogyne spp. from golf course bermudagrass

William T. Crow

William T. Crow

University of FloridaFlorida, US State
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Crow, William T.
, 
Alemayehu Habteweld

Alemayehu Habteweld

University of FloridaFlorida, US State
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Habteweld, Alemayehu
 et 
Thomas Bean

Thomas Bean

University of FloridaFlorida, US State
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Bean, Thomas
  
06 nov. 2020
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Publié en ligne: 06 nov. 2020
Pages: 1 - 12
DOI: https://doi.org/10.21307/jofnem-2020-096
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© 2020 William T. Crow et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

Figure 1:

Symptoms of Meloidogyne graminis on an ultradwarf bermudagrass golf green. Only low numbers of M. graminis vermiform stages were recovered from soil, while staining revealed the roots were infested with large numbers of sedentary stages.
Symptoms of Meloidogyne graminis on an ultradwarf bermudagrass golf green. Only low numbers of M. graminis vermiform stages were recovered from soil, while staining revealed the roots were infested with large numbers of sedentary stages.

Figure 2:

Golf course bermudagrass roots infested by Meloidogyne graminis and M. marylandi typically rot off just below small galls near the soil-thatch interface, creating an extremely abbreviated root system.
Golf course bermudagrass roots infested by Meloidogyne graminis and M. marylandi typically rot off just below small galls near the soil-thatch interface, creating an extremely abbreviated root system.

Figure 3:

The University of Florida Nematode Assay Lab mist chamber.
The University of Florida Nematode Assay Lab mist chamber.

Figure 4:

(A) Funnel hole cut in center shelf. (B) Water flow prevention ring. (C) Drain hole. (D) Shelf support.
(A) Funnel hole cut in center shelf. (B) Water flow prevention ring. (C) Drain hole. (D) Shelf support.

Figure 5:

(A) Misting rail. (B) Superfine mist emitter.
(A) Misting rail. (B) Superfine mist emitter.

Figure 6:

Funnels with sample support rings.
Funnels with sample support rings.

Figure 7:

Samples are placed onto coffee filters on the sample support rings. Funnel necks insert slightly into the 250-ml-flaskes.
Samples are placed onto coffee filters on the sample support rings. Funnel necks insert slightly into the 250-ml-flaskes.

Figure 8:

A washed turf plug including live turf, thatch and roots, ready for mist extraction.
A washed turf plug including live turf, thatch and roots, ready for mist extraction.

Figure 9:

Four washed turf plugs are placed in each funnel in the mist chamber and incubated for 72 hr.
Four washed turf plugs are placed in each funnel in the mist chamber and incubated for 72 hr.

Figure 10:

Average number of Meloidogyne spp. vermiform stages (J2 and males) recovered from diagnostic samples submitted to the University of Florida Nematode Assay Lab using centrifugal flotation extraction from 100 cm3 of soil (Soil) and mist chamber extraction from four 3.8-cm-diam. turf plugs (Mist) from golf course bermudagrasses in Florida (N = 431), golf course bermudagrasses in Texas (N = 36), or golf course bentgrass/bluegrass in California (N = 34).
Average number of Meloidogyne spp. vermiform stages (J2 and males) recovered from diagnostic samples submitted to the University of Florida Nematode Assay Lab using centrifugal flotation extraction from 100 cm3 of soil (Soil) and mist chamber extraction from four 3.8-cm-diam. turf plugs (Mist) from golf course bermudagrasses in Florida (N = 431), golf course bermudagrasses in Texas (N = 36), or golf course bentgrass/bluegrass in California (N = 34).

Figure 11:

Percentage of samples submitted to the University of Florida Nematode Assay Lab from golf course bermudagrass in Florida (N = 431), golf course bermudagrass in Texas (N = 36), and golf course bentgrass/bluegrass in California (N = 34) that were classified as having no risk, low risk, moderate risk, or high risk of damage from Meloidogyne spp. and the percentage of samples that were ‘below threshold’ (no or low risk) and ‘above threshold’ (moderate or high risk) using centrifugal flotation extraction from 100 cm3 of soil (Soil) or mist chamber extraction from four 3.8-cm-diam. turf plugs (Mist). Risk thresholds used per 100 cm3 of soil or per four 3.8-cm-diam. turf plugs were 0 = no risk, 1 ≤ 79 = low risk, 80 ≤ 299 = moderate risk, ≥ 300 = high risk.
Percentage of samples submitted to the University of Florida Nematode Assay Lab from golf course bermudagrass in Florida (N = 431), golf course bermudagrass in Texas (N = 36), and golf course bentgrass/bluegrass in California (N = 34) that were classified as having no risk, low risk, moderate risk, or high risk of damage from Meloidogyne spp. and the percentage of samples that were ‘below threshold’ (no or low risk) and ‘above threshold’ (moderate or high risk) using centrifugal flotation extraction from 100 cm3 of soil (Soil) or mist chamber extraction from four 3.8-cm-diam. turf plugs (Mist). Risk thresholds used per 100 cm3 of soil or per four 3.8-cm-diam. turf plugs were 0 = no risk, 1 ≤ 79 = low risk, 80 ≤ 299 = moderate risk, ≥ 300 = high risk.

Figure 12:

Percentage of samples submitted to the University of Florida Nematode Assay Lab from golf course bermudagrass in Florida (N = 431), golf course bermudagrass in Texas (N = 36), and golf course bentgrass/bluegrass in California (N = 34) from which the resulting risk level was higher, lower, or no different when vermiform Meloidogyne spp. were extracted by mist chamber extraction from four 3.8-cm-diam. turf plugs than by centrifugal flotation extraction from 100 cm3.
Percentage of samples submitted to the University of Florida Nematode Assay Lab from golf course bermudagrass in Florida (N = 431), golf course bermudagrass in Texas (N = 36), and golf course bentgrass/bluegrass in California (N = 34) from which the resulting risk level was higher, lower, or no different when vermiform Meloidogyne spp. were extracted by mist chamber extraction from four 3.8-cm-diam. turf plugs than by centrifugal flotation extraction from 100 cm3.

Comparison of the number of Meloidogyne spp_ J2 and males recovered from either 100 cm3 of soil using centrifugal flotation or from four 3_8-cm-diam_ turf plugs using mist extraction_

J2 Males
Year Cultivar n Soil Mist P < r 2 Soil Mist P < R 2
2017 Jones Dwarf 77a 6b 436 0.0001c 0.014d 1 40 0.0001 0.059
2017 Tifdwarf 49 225 945 0.0001 0.003 100 184 0.0001 0.085
2017 Tifdwarf 45 92 387 0.0001 0.002 21 92 0.0001 0.150
2017 TifEagle 59 69 368 0.0001 0.048 2 25 0.0001 0.059
2018 Tifdwarf 40 130 466 0.0001 0.005 28 129 0.0001 0.000
2019 TifEagle 71 70 723 0.0001 0.332 22 108 0.0001 0.122
2019 TifEagle 100 24 128 0.0001 0.604 2 13 0.0001 0.374
2019 Sunday 120 14  81 0.0001 0.202 <1 <1 0.0001 0.003
2020 Jones Dwarf 35 22 286 0.0001 0.560 5 26 0.0001 0.244
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