Authors
B. J.
Aegerter
,
Department of Plant Pathology, University of California, Davis 95616
;
J. J.
Nuñez
,
University of California Cooperative Extension, Kern County, Bakersfield 93307
; and
R. M.
Davis
,
Department of Plant Pathology, University of California, Davis 95616
ABSTRACT
A technique utilizing the polymerase chain reaction (PCR) was developed to investigate the occurrence and location of Peronospora sparsain dormant, woody rose tissues. PCR primers were designed to amplify the internal transcribed spacer region of the ribosomal DNA of the pathogen. Inhibition of the reaction by plant compounds was minimized by optimizing the reagents used in the extraction of DNA from roses and in the amplification reaction. The PCR assay was capable of detecting as little as 2 pg of DNA from P. sparsa against a background of 4 ng of DNA from rose cane cortex. With this method, DNA of P. sparsa was detected in the cortex of stem and root tissues of symptomatic plants. Pathogen DNA also was detected in the cortex of crown tissues of asymptomatic mother plants used as a source of propagation materials. Epifluorescent and differential interference contrast microscopy were used to confirm the presence of abundant hyphae and oospores within the stem cortex of infected canes. Preplant treatments of dormant rootstock cuttings in fungicides or hot water were evaluated during natural outbreaks of the disease in commercial rose nurseries. In three trials conducted over 2 years, a 10-min preplant dip in the systemic fungicides metalaxyl or mefenoxam at rates of 100 to 10,000 mg a.i./liter reduced the area under the disease progress curve by 63 to 76% relative to nontreated plots. The evidence from PCR assays, microscopy, and fungicide trials all support the occurrence of perennating infections of P. sparsa within rose.
A technique utilizing the polymerase chain reaction (PCR) was developed to investigate the occurrence and location of Peronospora sparsain dormant, woody rose tissues. PCR primers were designed to amplify the internal transcribed spacer region of the ribosomal DNA of the pathogen. Inhibition of the reaction by plant compounds was minimized by optimizing the reagents used in the extraction of DNA from roses and in the amplification reaction. The PCR assay was capable of detecting as little as 2 pg of DNA from P. sparsa against a background of 4 ng of DNA from rose cane cortex. With this method, DNA of P. sparsa was detected in the cortex of stem and root tissues of symptomatic plants. Pathogen DNA also was detected in the cortex of crown tissues of asymptomatic mother plants used as a source of propagation materials. Epifluorescent and differential interference contrast microscopy were used to confirm the presence of abundant hyphae and oospores within the stem cortex of infected canes. Preplant treatments of dormant rootstock cuttings in fungicides or hot water were evaluated during natural outbreaks of the disease in commercial rose nurseries. In three trials conducted over 2 years, a 10-min preplant dip in the systemic fungicides metalaxyl or mefenoxam at rates of 100 to 10,000 mg a.i./liter reduced the area under the disease progress curve by 63 to 76% relative to nontreated plots. The evidence from PCR assays, microscopy, and fungicide trials all support the occurrence of perennating infections of P. sparsa within rose.
