Puccinia jaceae var. solstitialis teliospore priming on yellow starthistle.

Following the introduction of Puccinia jaceae var. solstitialis to California for biological control of yellow starthistle (Centaurea solstitialis, Asteraceae), teliospores, pycnia, and multiple urediniospore generations have been observed in the field. Because urediniospores have a relatively short life span in the field, functioning teliospores are expected to be necessary for the permanent establishment of P. jaceae var. solstitialis in California. To determine if conditions in California were conducive to this, teliospore emergence and priming were evaluated in the field. A factorial experiment in the laboratory with five incubation times and three incubation temperatures was used to determine teliospore priming requirements. Teliospore production coincided with plant senescence in August and September at two sites in 2 years; fewer teliospores were produced in 2006, suggesting inconsistent teliospore production may limit population growth and contribute to local extinctions in some areas. When teliospores were primed in the field, germination was low through the fall and abruptly peaked in January during both years. In the laboratory, teliospore germination increased as incubation time increased from 2 to 6 weeks and temperatures decreased from 12 to 4 degrees C. A degree-hour model derived from laboratory data accurately predicts when teliospores are primed for germination in the field. Based on the results obtained in this study, it is apparent that teliospore germination can occur over a range of priming conditions. However, lower temperatures and longer incubation periods are superior in breaking teliospore dormancy.

Salt Marsh Claviceps purpurea in Native and Invaded Spartina Marshes in Northern California.

The fungal pathogen Claviceps purpurea (subgroup G3) has a worldwide distribution on salt marsh Spartina spp. In Northern California (United States), native Spartina foliosa sustains high rates of infection by G3 C. purpurea in marshes north of the San Francisco Estuary. Invasive populations of S. alterniflora and S. alterniflora × foliosa hybrids are virtually disease free in the same estuary, although S. alterniflora is host to G3 C. purpurea in its native range (Atlantic Coast of the United States). Greenhouse inoculation experiments showed no differences in susceptibility among S. foliosa, S. alterniflora, and Spartina hybrids. Under field conditions, S. foliosa sustained a higher incidence of disease in coastal marshes than in marshes within the bay. This geographic effect may be attributable to environmental differences between the coast and the bay proper, with the former being more conducive to infection by C. purpurea. Seed set of S. foliosa spikelets was 40 to 70% lower on infected than on uninfected inflorescences, but seed germination was not affected. The C. purpurea epidemic on S. foliosa on the coast north of the San Francisco Estuary further reduces the meager competitive ability of this declining native plant species.

First Report of the Pitch Canker Fungus (Fusarium circinatum) in the Sierra Nevada of California.

The pitch canker fungus, Fusarium circinatum (teleomorph Gibberella circinata), was isolated from a branch originating from rootstock of a Douglas-fir (Pseudotsuga menziesii) graft in a breeding orchard at 1,000m elevation in El Dorado County, California. We visited the orchard after the New Zealand Ministry of Agriculture and Forestry reported in November 2003 that a Douglas-fir scion (branch cutting) shipped from there in January-and subsequently grafted and held in a quarantine facility near Christchurch-was infected with the pitch canker fungus. We took samples throughout the orchard of any branches that appeared unhealthy. In addition, asymptomatic branches from the tree alleged to be the source of the New Zealand infestation were collected to assay for propagules of F. circinatum. Wash water from these branches was negative for the pathogen. Likewise, F. circinatum was not recovered from water washings of 20 branches collected randomly throughout the orchard. Fifteen branch samples collected from symptomatic Douglas-fir grafts were cultured on water agar and only one yielded a colony with an appearance consistent with F. circinatum. A single spore subculture of this isolate was confirmed as F. circinatum on the basis of colony morphology and the structure of the microconidiophores (1). The virulence of this isolate was evaluated by inoculating susceptible 2-year-old Monterey pine (Pinus radiata) seedlings with a toothpick to wound the main stem and insert potato dextrose agar colonized by the fungus. Twenty-four days later, pitching and yellow needles were evident at the site of inoculation, and removal of the bark revealed resin-soaked and discolored tissue. Concurrent with the pathogenicity test described above, a culture of the putative F. circinatum isolated in New Zealand was inoculated into Monterey pines with an identical outcome. The fungus was reisolated from lesions from both sets of inoculations and confirmed as F. circinatum based on morphological criteria. Isolates GL285 and GL286 are available from T. R. Gordon upon request. Prior to its discovery in the Sierra Nevada, pitch canker in California was known only from counties on or near the coast. Our report indicates the pathogen can survive and infect trees 110 km east of the previous most-inland site of infestation. It remains to be seen how extensively pitch canker will develop in the Sierra Nevada. Douglas-fir is only moderately susceptible to F. circinatum, which has not been observed to cause significant damage to this species. On the other hand, low-elevation Sierra Nevada pines including P. sabiniana, P. coulteri, and P. ponderosa are substantially more susceptible than are Douglas-fir in greenhouse tests (2). References: (1) T. R. Gordon et al. Mycol. Res. 100:850, 1996. (2) T. R. Gordon et al. Plant Dis. 85:1128, 2001.

Environmental Factors Affecting Rose Downy Mildew and Development of a Forecasting Model for a Nursery Production System.

The effect of various environmental parameters on rose downy mildew caused by Peronospora sparsa was determined under controlled conditions and in the field. In growth chambers, optimal temperatures for infection and colonization of rose leaves were 15 to 20°C and 20 to 25°C, respectively. At optimal temperatures, infection required only 2 h of leaf wetness, although disease severity increased significantly with an increasing duration of leaf wetness up to 10 h. Infection of leaves occurred at temperatures as low as 5°C with 8 h of leaf wetness. The latent period of infection varied from 4 to 7 days. Weather and disease incidence data collected from natural field epidemics were used in the development of a predictive model of rose downy mildew. Logistic regression was used to identify those weather variables that explained the largest portion of the variation in disease incidence. The optimum regression model incorporated three weather variables calculated as cumulative totals over the previous 10 days: (i) hours of leaf wetness when temperatures were less than 20°C (positive correlation); (ii) hours between 15 and 20°C (negative correlation); and (iii) hours when temperatures exceeded 30°C (negative correlation). The simplest model, which was also a good fit, included only the 10-day cumulative number of hours of leaf wetness. The critical number of hours of leaf wetness for disease development was an average of 8.4 h per day over 10 days.

Root and Basal Rot of Leek Caused by Fusarium culmorum in California.

In 1999 and 2000, greenhouse-grown leek (Allium porrum) transplants produced in coastal California (Monterey County) developed a root and basal rot. Affected roots were initially gray and water soaked in appearance and later became pink, soft, and rotted. Basal plates were also affected, becoming water soaked and rotted. Severely affected transplants grew poorly and had chlorotic older leaves; many of these plants collapsed. Disease incidence varied greatly, though some transplant plantings had more than 50% disease. Similar symptoms were found in commercial, field-planted leek crops in the same region. The problem caused significant economic loss to transplant producers because of the loss of plants and the reduction in quality of surviving infected plants. Isolations from transplant and field samples consistently recovered a Fusarium species from both root and basal plate tissues. Single-spore subcultures were grown on carnation leaf agar and incubated under fluorescent light. All isolates produced abundant macroconidia that were stout, thick walled, and had prominent septa. Foot cells were indistinct to slightly notched. Conidiophores were monophialidic. Microconidia were absent and chlamydospores were present. Colonies on potato dextrose agar produced abundant, dense, white, aerial mycelium. The undersurface of these cultures was carmine red. Based on these features, all isolates were identified as Fusarium culmorum. To confirm the identification, a partial sequence (645 bp) of the translation elongation factor (EF-1α) was obtained for one isolate using primers EF-1 and EF-2 (2). The EF-1α sequence from the leek isolate was identical to that of two F. culmorum isolates in Genbank (Accession Nos. AF212462 and AF212463). The next closest match was F. cerealis, which differed from the leek isolate at six nucleotide positions. To test pathogenicity of the leek isolates of F. culmorum, we prepare inocula of four isolates from transplants and three isolates from field plants. A conidial suspension (1 × 105 conidia/ml) of each isolate was applied to the roots of 3-month-old potted leek (cvs. Autumn Giant, Blauwgroene, and Cisco). For the control treatment, leek plants were treated with water. All plants were maintained in a greenhouse at 25°C. After 1 month, inoculated plants showed foliar and root symptoms similar to those observed on the original samples. F. culmorum was reisolated from these symptomatic plants. Control plants did not develop symptoms. Using the same procedures, the seven isolates were inoculated onto other Allium species, but did not cause any symptoms on shallot (A. cepa var. ascalonicum) or eight cultivars of onion (A. cepa). Two of the seven isolates caused slight root symptoms on garlic (A. sativum). All experiments were conducted two times and the results of both tests were similar. To our knowledge, this is the first report of a root and basal rot of leek in California caused by F. culmorum. The occurrence of this disease on transplants grown in a soilless rooting medium and on raised benches in enclosed greenhouses provides circumstantial evidence that the pathogen could possibly be seedborne. This disease was reported recently in Spain (1). References: (1) J. Armengol et al. Plant Dis. 85:679, 2001. (2) K. O'Donnell et al. Proc. Natl. Acad. Sci. 95:2044, 1998.

Detection and Management of Downy Mildew in Rose Rootstock.

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.

Mefenoxam-Resistant Isolates of Pythium irregulare in an Ornamental Greenhouse in California.

Root rot, caused by Pythium species, is a common malady in ornamental greenhouses in Monterey County, CA. In 2001, a root rot of Gerbera daisy (Gerbera jamesonii) and lisianthus (Eustoma grandiflora) caused plant losses in excess of 15 and 75%, respectively, in one greenhouse. Some plantings were total losses. Although mefenoxam was used repeatedly, no disease control was reported. P. irregulare was identified based on morphological structures produced on grass blades in water (1) and on the sequence of the rDNA internal transcribed spacer (ITS) region. Four isolates from each host were tested for sensitivity to mefenoxam in a laboratory bioassay. Using corn meal agar amended with mefenoxam at 0, 0.1, 1, 10, 50, or 100 µg a.i./ml, no inhibition of growth of any isolate occurred at concentrations of 10 µg/ml or less. At 50 and 100 µg/ml, radial growth of colonies was inhibited by approximately 20%. In contrast, 26 isolates of P. ultimum from various agricultural soils in California were completely inhibited by mefenoxam at 100 µg a.i./ml. At 0.1, 1, 10, and 50 µg/ml, growth of these isolates was inhibited by 33, 61, 78, and 96%, respectively. Each treatment was replicated three times, and the experiment was repeated with similar results. Mefenoxam was introduced in 1996 to replace metalaxyl. While metalaxyl contains a mixture of active and inactive enantiomers, mefenoxam contains only the active enantiomer. The intensive use of metalaxyl and mefenoxam in this greenhouse may have resulted in selection for resistant isolates. A resistant isolate (8-007 from Gerbera) has been submitted to the American Type Culture Collection: Manassas, VA. Reference: (1) A. J. van der Plaats-Niterink. Monograph of the Genus Pythium. Stud. Mycol. No. 21. Centraalbureau voor Schimmelcultures, Baarn, the Netherlands, 1981.

Occurrence and Pathogenicity of Fungi Associated with Melon Root Rot and Vine Decline in California.

The occurrence of fungi associated with root rot and vine decline of melon (Cucumis melo) in commercial fields in California was surveyed over 3 years. The fungi most frequently isolated from discolored vascular tissue or root rot were Acremonium cucurbitacearum, Rhizopycnis vagum, Monosporascus cannonballus, Fusarium solani, Macrophomina phaseolina, Pythium spp., and Verticillium dahliae. The frequency of isolation of the various fungi varied with root symptomology. Pythium spp., and M. phaseolina were frequently associated with a wet, brownish root rot, while A. cucurbitacearum, R. vagum, and Rhizoctonia solani were generally associated with a dry, corky root rot. Presence of Monosporascus cannonballus was associated both with a wet, brownish rot as well as with discrete, reddish, corky lesions. The frequency of isolation of a given pathogen varied with geographic location, with M. cannonballus present only in the southern production areas, while A. cucurbitacearum and Rhizopycnis vagum were most common in the northern production areas. In pathogenicity tests in field microplots, M. cannonballus caused vine collapse and severe root rot of cantaloupe, reducing root length density by 93%. California isolates of R. vagum and A. cucurbitacearum, although only weakly pathogenic in field microplots, caused root discoloration and reduced vine growth in greenhouse tests. Reduction in dry weight of greenhouse-grown cantaloupe was 40, 23, and 39% for R. vagum, A. cucurbitacearum, and M. cannonballus, respectively.

First Report of Pythium polymastum on Broccoli and Cauliflower in California.

Damping-off of broccoli (Brassica oleracea var. italica) and cauliflower (B. oleracea var. botrytis) seedlings occurred in several greenhouses in Fresno, CA, in 1997. Symptoms included wilting and root and stem rot. Pythium polymastum was consistently isolated from symptomatic tissues placed on corn meal agar amended with 10 ppm pimaricin, 250 ppm ampicillin, 10 ppm rifampicin, and 25 ppm pentachloronitro-benzene. On grass leaves in water, the fungus produced numerous aplerotic oospores in oogonia 43 to 50 µm in diameter (average 46 µm) with spines about 7 µm long. Spherical sporangia were only rarely observed. In the greenhouse, 4-week-old broccoli and cauliflower seedlings were transplanted into potting mix amended with a colonized vermiculite/rye/V8 juice medium to produce approximately 2,500 CFUs per gram of potting medium. Control plants were transplanted into noninfested potting mix. There were six replicate pots per treatment and three plants per pot. After 12 days, the potting mix was gently washed from the roots and the seedlings were dried and weighed. Symptoms on inoculated plants included wilting, severe root rot, black streaks on the lower stems, and death. The fungus was recovered from symptomatic tissues. There were no symptoms on the control plants. Infection by P. polymastum reduced dry weights of surviving broccoli and cauliflower seedlings by 82 and 58%, respectively. Similar results were obtained in a second experiment. This fungus was previously characterized as a pathogen of both cultivated and wild crucifers in Canada (1). This is the first report of P. polymastum in California. Reference: (1) T. C. Vanterpool. Can. J. Bot. 52:1205, 1974.