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Table 2 from:


Bulluck, R., Shiel, P., Berger, P., Kaplan, D., Parra, G., Li, W., Levy, L., Keller, J., Reddy, M., Sharma, N., Dennis, M., Stack, J., Pierzynski, J., O’Mara, J., Webb, C., Finley, L., Lamour, K., McKemy, J., and Palm, M. 2006. A comparative analysis of detection techniques used in US regulatory programs to determine presence of Phytophthora ramorum in Camellia japonica ‘Nucio’s Gem’ in an infested nursery in Southern California. Online. Plant Health Progress doi:10.1094/PHP-2006-1016-01-RS.



Table 3. Frequency distribution for aboveground disease incidence assays to directly compare symptom observation and the results obtained by the diagnostic methods used in this study.

Group n Percent of appropriate groupingu
Lesion
+
Lesion
-
ELISA
+
ELISA
-
PCR methods Culture
+
Culture
-
Nested
+
Nested
-
Real-Time+ Real-Time-
Lesion+v 240 100.0 0.0 67.5 32.5 87.5 12.5 65.4 34.6 51.7 48.3
Lesion- 60   100.0 25.0 75.0 28.3 71.7 8.3 91.7 3.3 96.7
ELISA +w 177     100.0 0.0 70.5 29.5 74.6 24.4 15.3 84.7
ELISA - 123       100.0 54.5 45.5 24.4 75.6 4.9 95.1
Nested +x 227         100.0 0.0 65.4 34.6 54.2 45.8
Nested - 73           100.0 8.2 91.8 4.1 95.9
Real-Time+y 162             100.0 0.0 76.5 23.5
Real-Time- 138               100.0 1.4 98.6
Culture+z 126                 100.0 0.0
Culture- 174                   100.0

 u Grouped according to rows, each box will add up to 100% of the number n.

 v defined as lesions on camellia leaves characteristic of Phytophthora ramorum.

 w Positive defined as having an OD reading of 2.0 times the known negative tissue samples.

 x Nested PCR is considered positive if a strong band at 293 bp is exhibited in the final gel analysis with consistently performing positive and negative controls.

 y Positive defined as having a Ct of between 17 and 35.99.

 z positive defined as at least one colony of P. ramorum recovered from plated leaf pieces.



Table 4. Pearson’s Correlation matrix of the four different Phytophthora ramorum detection techniques from samples of aboveground tissue of camellia plants (Camellia japonica ‘Nuccio’s Gem’) with and without leaf lesions in a Southern California nursery. (Pearson’s correlation coefficient is provided below each correlation).

  Symp-
toms
(+/-)q
Leaf
Drop
(+/-)r
Moss
(+/-)s
ELISA
(+/-)t
ELISA
(OD)u
Real-
Time
(Ct)v
Real-
Time
(+/-)w
Nested PCR
(+/-)x
Culture
(propor-
tion)y
Culture
(+/-)z
Symp-
tom

(+/-)

1.00 0.06917 0.06073 0.39261 0.30495 -0.3636 0.36003 0.49329 0.29037 0.32418
Leaf Drop

(+/-)

  100 0.0036 0.13298 0.12136 -0.0559 0.07932 0.14132 0.14954 0.10932
Moss     1.00 -0.01198 -0.05992 0.07728 -0.06317 -0.03761 -0.02426 -0.04711
ELISA

(+/-)

      1.00 0.69751 -0.55755 0.58454 0.43898 0.53357 0.53598
ELISA

(OD)

        1.00 -0.67324 0.64639 0.34632 0.68443 0.65051
Real time

(Ct)

          1.00 -0.92348 -0.51389 -0.76036 -0.78473
Real time

(+/-)

            1.00 0.49742 0.7574 0.78133
Nested

(+/-)

              1.00 0.41141 0.43535
Culture(propor-
tion)
                1.00 0.93832
Culture

(+/-)

                  1.00

 q defined as lesions on camellia leaves characteristic of Phytophthora ramorum.

 r Leaf drop is a symptom of premature abscission of leaves due to their infection with P. ramorum.

 s the presence of moss was noted either on the stem of the camellia or on the medium surface.

 t Positive defined as having an OD reading of 2.0 times the known negative tissue samples.

 u The actual OD reading of the ELISA sample.

 v The actual Cycle at which the Threshold value was crossed.

 w Positive defined as having a Ct of between 17 and 35.99.

 x Positive defined as the presence of distinct bands at 293 bp location on an agarose gel of the second running of the nested PCR reaction.

 y the proportion of P. ramorum on PARP-V8 from plated leaf tissue.

 z presence of as at least one colony of P. ramorum recovered from plated leaf pieces.

Significant correlations at the p=0.05 or less level are indicated by the color red.



© 2006 Plant Management Network