Cavity Spot Research Articles

Exploring the Effects of Trichoderma virens Biofungicide on Carrot Cavity Spot and Soil Fungal Community Dynamics

Carrot cavity spot (CCS) has conventionally been managed with fungicides. However, fungicide resistance, their potential risks to human health and the environment, and the increasing demand for organic produce necessitate the exploration of biofungicides as alternatives. In this study, we evaluated varying concentrations of SoilGard (Certis USA, Columbia, MD), a Trichoderma virens-based biofungicide, for efficacy against different CCS-causing Pythium species in vitro. Additionally, its effects on taxonomic and functional diversities of soil fungal communities were studied in vivo in the greenhouse. To our knowledge, this is the first study reporting SoilGard's effectiveness against CCS, with emphasis on its potential as an alternative for fungicide-resistant Pythium isolates. Our in vitro study revealed that SoilGard efficacy was significantly dose-dependent and isolate-specific, thus highlighting the importance of selecting its application rate and the target isolate. Analysis of soil fungal communities using Illumina MiSeq sequencing revealed that SoilGard exerted a significant, albeit temporary, effect on the fungal community structure. It negatively impacted co-occurrence network complexity and alpha diversity in carrot-cultivated soil, whereas bare soil communities remained largely unaffected, thus explaining why preplant applications may yield better results. Our study showed that carrot cultivation without SoilGard enhanced fungal diversity, which was more pronounced late in the season, possibly due to carrot root-associated exudates. Our study sheds light on how complex interactions within soil fungal communities can be impacted by the application of beneficial/pathogenic microbes.

Development of inoculation methods for Pythium violae to evaluate resistance of carrot cultivars and efficacy of crop protection products for cavity spot control

Development of inoculation methods for <i>Pythium violae</i> to evaluate resistance of carrot cultivars and efficacy of crop protection products for cavity spot control

Development of artificial inoculation techniques for cavity spot caused by Pythium violae to identify new management strategies

Development of artificial inoculation techniques for cavity spot caused by <i>Pythium violae</i> to identify new management strategies

First Report of Strains Within the Pythium spinosum Species Complex Causing Carrot Cavity Spot in California.

Carrots (Daucus carota L. subsp. sativus (Hoffm.)) with typical symptoms of cavity spot, i.e., sunken, round to elliptical lesions of 2-5 mm long (Hiltunen and White 2002), were collected from two locations in California, Bakersfield and Riverside, in January and July 2019, respectively. Carrots were rinsed in tap water, 4-mm2 lesion fragments were pressed into selective corn meal agar (CMA-PARP; Schrandt et al. 1994) and incubated at 23ºC in the dark for four days. Identification of pure cultures was performed via amplification and sequence analysis of two genomic regions, the Internal Transcribed Spacer 1-5.8S-ITS2 (ITS) region and the cytochrome C oxidase subunit 1 (COI) gene, using the universal primers UN-UP18S42/UN-LO28S576B (Schroeder et al. 2006) and OomCOXI-Levup/OomCOXI-Levlo (Robideau et al. 2011), respectively. Via BLAST, two isolates from organically grown carrots in Bakersfield (MCIF19) and Riverside (JSCS19), with identical ITS sequences (GenBank Acc. Nos. MZ799354 and MZ799355, respectively), showed 99.61% similarity (1021/1025 bp) to that of Pythium spinosum (AY598701.2). Yet, the COI of MCIF19 (MZ803207) showed 98.72% similarity (692/701 bp) to that of Pythium paroecandrum (GU071818.1), while the COI of JSCS19 (MZ803208) was identical (701/701 bp) to that of Pythium kunmingense (GU071820.1), a rarely isolated species considered within the species complex of P. spinosum (Robideau et al. 2011). According to these results, the isolates were identified as belonging to the P. spinosum species complex, part of Pythium Clade F (Lévesque and De Cock 2004; Robideau et al. 2011). Further research is needed to clarify the exact taxonomic status of both isolates. Koch's postulates were completed using two different assays. Each assay was done twice and with carrots of the cultivar Maverick. Surface-sterilized, freshly harvested, mature carrots, in a plastic box lined with moistened sterile paper towels, were inoculated each with four CMA plugs (5-mm diameter) with actively growing mycelium of each isolate. CMA plugs, non-inoculated or colonized by a known pathogenic P. violae strain, were used as the negative and positive control, respectively. Boxes were closed to maintain humidity and incubated at 23ºC in the dark. Lesions similar to the ones caused by P. violae were observed at day 3 for all plugs of both isolates. No symptoms were observed for the negative control, even after extending the incubation to 7 days. In a more natural assay, four non-treated carrot seeds were planted in tree seedling pots (25 x 6.5 cm) containing sterilized 50/50 peat moss/sand combined with 15-ml V8 broth (Schrandt et al. 1994) with densely grown mycelium. The same inoculation treatments were used as for the carrot disk assay. Plants (one plant/pot, four plants/treatment) were maintained at 23ºC under a 16 h photoperiod with daily watering (20 ml). At 14 weeks, the carrots inoculated with P. violae and the two test isolates showed cavity spot lesions while no symptoms were observed on carrots growing in non-inoculated medium. For both assays, pathogens were re-isolated from rinsed symptomatic tissue and their identity was confirmed using the molecular analysis described above. No oomycetes were recovered from the non-inoculated carrots. Although several Pythium species have been associated with cavity spot before, this is, to our knowledge, the first report of strains within the P. spinosum species complex causing carrot cavity spot in California and elsewhere. Funding: This research was made possible by the California Fresh Carrot Advisory Board (FRA-21).

Carrot biotic stresses: challenges and research priorities

Among the main biotic stresses of carrot crops, the pests such as nematodes and carrot fly and diseases including cavity spot, crown or cottony rot, black rot or leaf blight are targets for scientific researches over the world. Components of integrated control programs for all these organisms are under development. Success of such programs requires systemic approaches including knowledge about genetic diversity of the targets, their epidemiology, host physiology and host resistance mechanisms and compatibility among the different control tools or between programs targeting different goals.

Cropping history shapes fungal, oomycete and nematode communities in arable soils and affects cavity spot in carrot

Fungal, nematode and oomycete communities in arable soil serve diverse ecological functions. However, little is known about the dynamics among these three taxonomic groups. In order to identify the driving factors and networks shaping these communities, and the influence of communities on the development of carrot cavity spot, we collected soil samples from more than sixty fields in which carrot was planned as the next crop, and analyzed the microbial communities using metabarcoding. Crop grown before sampling was best in explaining variation in community composition, but also soil type was shaping fungal and nematode communities, but not oomycete communities. Network analysis revealed that correlations between taxa within taxonomic groups were mostly positive whereas negative correlations dominated between taxa across the different taxonomic groups. Pythium intermedium and P. irregulare from soil samples were found to be associated with cavity spot disease in carrot, and were also found in the lesions of cavity spot. Our study demonstrate that previous crop and soil type significantly affect microbial communities and the development of symptoms of cavity spot in carrot. We also found that differences in co-occurrences within and between taxonomic groups exist.

Development of a disease suppressive soil for managing cavity spot of carrots

Development of a disease suppressive soil for managing cavity spot of carrots

An improved high throughput sequencing method for studying oomycete communities

Culture-independent studies using next generation sequencing have revolutionized microbial ecology, however, oomycete ecology in soils is severely lagging behind. The aim of this study was to improve and validate standard techniques for using high throughput sequencing as a tool for studying oomycete communities. The well-known primer sets ITS4, ITS6 and ITS7 were used in the study in a semi-nested PCR approach to target the internal transcribed spacer (ITS) 1 of ribosomal DNA in a next generation sequencing protocol. These primers have been used in similar studies before, but with limited success. We were able to increase the proportion of retrieved oomycete sequences dramatically mainly by increasing the annealing temperature during PCR. The optimized protocol was validated using three mock communities and the method was further evaluated using total DNA from 26 soil samples collected from different agricultural fields in Denmark, and 11 samples from carrot tissue with symptoms of Pythium infection. Sequence data from the Pythium and Phytophthora mock communities showed that our strategy successfully detected all included species. Taxonomic assignments of OTUs from 26 soil sample showed that 95% of the sequences could be assigned to oomycetes including Pythium, Aphanomyces, Peronospora, Saprolegnia and Phytophthora. A high proportion of oomycete reads was consistently present in all 26 soil samples showing the versatility of the strategy. A large diversity of Pythium species including pathogenic and saprophytic species were dominating in cultivated soil. Finally, we analyzed amplicons from carrots with symptoms of cavity spot. This resulted in 94% of the reads belonging to oomycetes with a dominance of species of Pythium that are known to be involved in causing cavity spot, thus demonstrating the usefulness of the method not only in soil DNA but also in a plant DNA background. In conclusion, we demonstrate a successful approach for pyrosequencing of oomycete communities using ITS1 as the barcode sequence with well-known primers for oomycete DNA amplification.

Incidence and Severity of Cavity Spot of Carrot as Affected by Pigmentation, Temperature, and Rainfall.

Field trials to determine the effect of carrot pigmentation and weather parameters on cavity spot (CS) of carrot were conducted in the Holland/ Bradford Marsh region of Ontario between 2002 and 2009. In all, 23 colored carrot cultivars from the United States Department of Agriculture (USDA) Agricultural Research Service breeding program at the University of Wisconsin (n = 5) and commercial seed companies (n = 18) were seeded in organic soil (pH 6 to 7, 45 to 75% organic matter) in late May to early June and harvested in late October or early November. Carrot roots were assessed for CS severity midseason and postharvest. Evaluations postharvest indicated that the purple pigmented carrot from breeding line 'USDA 106-3' and cultivars 'Purple Rain' and 'Purple Haze' consistently had low CS severity. The orange-pigmented 'USDA 101-23', 'Cellobunch', 'YaYa', and 'Envy' had moderate CS; and the red-pigmented carrot breeding line 'USDA 104-3' and cultivars 'Atomic Red', 'Proline Red', 'Dragon', and an unnamed line from India had high CS. Differences in CS severity in carrot cultivars between evaluations at midseason and postharvest suggest that some carrot cultivars are more susceptible to Pythium spp. inoculum in soil (alloinfection) and others to secondary infection (autoinfection) that can be attributed to the Pythium sp. involved in CS. CS severity was positively correlated with total rainfall 2 and 3 months after seeding, and was negatively correlated with number of days with air temperature ≥30°C 3 and 4 months after seeding. Soil temperature and total rainfall were the best predictors of CS incidence and severity. These results could allow a forecast of disease incidence and severity at harvest.

First Report of Pythium sulcatum Causing Cavity Spot in Processing Carrot Crops in the Columbia Basin of Washington State.

In October 2012, symptoms of cavity spot (1) were observed on roots of two 50 ha, Red Core Chantenay processing carrot (Daucus carota L. subsp. sativus (Hoffm.)) crops in the Columbia Basin of central Washington. Symptoms consisted of sunken, elliptical lesions (3 to 15 mm long) on the root surface. Approximately 6% of the roots in each crop were affected, which was sufficient to present sorting problems for the processor. Symptomatic roots were washed thoroughly in tap water, and then small sections of tissue from the lesion margins were removed aseptically and plated onto water agar (WA) without surface-sterilization. Isolates with morphological characteristics typical of Pythium sulcatum Pratt & Mitchell (2) were obtained consistently from the symptomatic tissue. The genus and species identity of seven isolates was confirmed by sequence analysis of the internal transcribed spacer (ITS) 1-5.8S-ITS2 region of ribosomal DNA (rDNA) using universal eukaryotic primers UN-UP18S42 and UN-LO28S576B with the PCR protocol described by Schroeder et al. (3). The ITS consensus sequences of the seven isolates (Accession Nos. KF509939 to KF509945) were 98 to 99% homologous to ITS sequences of P. sulcatum in GenBank. Pathogenicity of all seven isolates was confirmed by inoculating mature carrot roots of cv. Bolero. Each root was washed with tap water, sprayed to runoff with 70% isopropanol, and dried in a laminar flow hood on sterilized paper toweling. The roots were then placed in plastic bins lined with paper toweling moistened with sterilized, deionized water. Each root was inoculated by placing two 5 mm-diameter agar plugs, taken from the edge of an actively growing WA culture of the appropriate isolate, on the root surface approximately 3 cm apart. Non-colonized agar plugs were used for a non-inoculated control treatment. Four replicate roots were inoculated for each isolate and the control treatment. After inoculation, the roots were misted with sterilized, deionized water, a lid was placed on each bin, and the roots were incubated in the dark at 22°C. Roots were misted daily to maintain high relative humidity. Dark, sunken lesions were first observed 3 days post-inoculation on roots inoculated with the P. sulcatum isolates, and all inoculated roots displayed cavity spot lesions by 7 days. No symptoms were observed on the non-inoculated control roots. Colonies with morphology typical of P. sulcatum were re-isolated from the symptomatic tissue of roots inoculated with the P. sulcatum isolates, and the species identity of the re-isolates was confirmed by ITS rDNA sequence analysis, as described above. Although P. sulcatum is one of several Pythium species that can cause cavity spot of carrot (1), to our knowledge, this is the first report of P. sulcatum causing cavity spot in Washington State, which has the largest acreage of processing carrot crops in the United States (4).

Antifungal activity of aluminium‐containing salts against the development of carrot cavity spot and potato dry rot

AbstractAs an alternative to the use of synthetic chemical fungicides to control plant disease, aluminium‐containing salts were evaluated for their effects on the mycelial growth of various fungal or fungus‐like pathogens and their ability to control carrot cavity spot (Pythium sulcatum) and potato dry rot (Fusarium sambucinum). Results showed that various aluminium‐containing salts provided strong inhibition of all the tested pathogens (Alternaria solani, Botrytis cinerea, F. sambucinum, P. sulcatum and Rhizopus stolonifer) with minimal inhibitory concentration of 1–10 mM. Aluminium chloride and aluminium sulphate were generally the most effective, inhibiting mycelial growth of pathogens by as much as 47% and 100%, respectively, at a salt concentration of 1 mM. Applied at 5 mM, aluminium sulphate also provided 28% and 100% inhibition of dry rot and cavity spot, respectively. Aluminium chloride (5 mM) reduced dry rot by 25% whereas aluminium lactate (5 mM) decreased cavity spot lesions by 86%. These results indicate that various aluminium‐containing salts may provide an alternative to the use of synthetic fungicides to control these pathogens.

First Report of Pythium recalcitrans Causing Cavity Spot of Carrot in Michigan.

During a survey of carrot (Daucus carota L.) cavity spot in Michigan in September 2010, carrot roots with typical cavity spot symptoms were collected from production fields in Fremont Co. The lesions were excised from infected roots, surface-disinfested with 0.62% NaClO for 3 min, rinsed in sterilized, distilled water three times, cut into 0.5 cm long pieces, and then plated on water agar (WA) amended with carbendazim (10 μg/ml), ampicillin (50 μg/ml), rifampicin (50 μg/ml), and pentachloronitrobenzene (10 μg/ml) (cumulatively referred to as CARP). Plates were incubated at 22 ± 1°C in the dark for 3 days. Pure cultures of the isolates were obtained by transferring a single hyphal tip of each colony to potato dextrose agar (PDA) amended with CARP. Among the 33 isolates obtained, M2-05 was identified as a Pythium sp. that differed from the known cavity spot pathogens of carrot. The isolate has spherical hyphal swellings but no other distinguishing morphological characteristics. M2-05 was further classified by analyzing the partial sequences of four genes: the internal transcribed spacer (ITS) region of ribosomal DNA, beta-tubulin (β-tub), cytochrome c oxidase subunit 2 (cox 2), and NADH dehydrogenase subunit 1 (nadh 1) (1,3). A BLAST search of these sequences for M2-05 was conducted using the nucleotide database of GenBank, resulting in 100% similarity to all four sequenced genes of P. recalcitrans (2). The DNA sequences of M2-05 were deposited in GenBank (JQ734349, JQ734229, JQ734289, and JQ734409 for ITS, β-tub, cox 2, and nadh 1, respectively). Koch's postulates were conducted by inoculating mature carrot roots (cv. Nantindo) with mycelial plugs (4 mm in diameter) cut from the margin of actively growing colonies of M2-05 on PDA plates. Two mycelial plugs were placed on each carrot root at 3-cm intervals, with the mycelial side facing the root; and two non-colonized agar plugs were placed similarly for the non-inoculated control treatment. In comparison, carrot roots also were inoculated with an isolate of each of P. sulcatum and P. violae using the same method. There were four replicate carrot roots inoculated for each isolate and each of the control treatments. The inoculated roots were placed on a plastic grid (7 mm in height) in a closed plastic container, with moist paper towels underneath the grids. The container was incubated in the dark at 22 ± 1°C, and the roots were sprayed gently daily with sterilized, distilled water to maintain high humidity. Brown lesions were observed on all inoculated carrot roots 5 days after inoculation. The lesions measured 0.68 ± 0.48, 1.20 ± 0.71, and 0.56 ± 0.31 mm2 averaged over all eight lesions for the isolates of P. recalcitrans, P. sulcatum, and P. violae, respectively. Symptomatic tissues from the inoculated roots were excised and incubated on WA-CARP plates, and the culture from each lesion confirmed as the isolates inoculated using the same molecular methods described above. The carrot tissue under the control agar plugs remained symptomless, and no Pythium was recovered from the control roots. P. recalcitrans was described in 2008 as infecting roots of Beta vulgaris and Vitis vinifera (2). To our knowledge, this is the first published report of P. recalcitrans naturally infecting carrot roots, not only in Michigan, but anywhere in the world.

Fungicide Sensitivity of Pythium spp. Associated with Cavity Spot of Carrot in California and Michigan.

The identity of 172 isolates of Pythium spp. from cavity spot lesions on carrot produced in California and Michigan was determined, and their sensitivity to three fungicides was examined. Pythium violae accounted for 85% of California isolates, with P. irregulare, P. dissotocum (the first report as a carrot pathogen in the United States), P. ultimum, and P. sulcatum making the balance. P. sulcatum, P. sylvaticum, and P. intermedium were the most commonly recovered (85%) species in Michigan; others from Michigan included P. intermedium, P. irregulare, and an unclassified strain, M2-05. On fungicide-amended media, 93% of isolates were sensitive to mefenoxam (inhibition of mycelial growth was >60% at 10 μg active ingredient [a.i.]/ml); however, two of five isolates of P. irregulare from California were highly resistant (≤60% inhibition at 100 μg a.i./ml); about half of the isolates of P. intermedium and P. sylvaticum and a single isolate of P. violae were highly or intermediately resistant to mefenoxam (>60% inhibition at 100 μg a.i./ml, or ≤60% inhibition at 10 μg a.i./ml). P. dissotocum, P. irregulare, P. sulcatum, M2-05, and three of seven isolates of P. intermedium were insensitive to fluopicolide (effective concentrations for 50% growth inhibition [EC50] were >50 μg a.i./ml), while P. sylvaticum, P. ultimum, P. violae, and some isolates in P. intermedium were sensitive (EC50 < 1 μg a.i./ml). All isolates were sensitive to zoxamide (EC50 < 1 μg a.i./ml). Sensitivity baselines of P. violae to zoxamide and fluopicolide were established.

Fiber-Optic Temperature Sensor Based on Difference of Thermal Expansion Coefficient Between Fused Silica and Metallic Materials

In this paper, we report a novel fiber-optic Fabry-Perot interferometric (FFPI) temperature sensor based on the difference of thermal expansion coefficient between fused silica and metallic materials. The sensor head is made by a single-mode fiber (SMF). A gold film and a nickel film are sputtered and electroplated on the surface of the SMF. Then, a microcavity is micromachined by focused ion beam (FIB) milling. Because the thermal expansion coefficient of nickel is about 20 times of fused silica, the different thermal expansions force the sensor head to bend when the temperature is high or low. Its temperature sensitivity is over 14 pm/°C in a wide range from -79°C to +70°C. And the coefficient of determination R <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">2</sup> is excellent (over 0.995). Moreover, the metallic cylinder can reinforce the cavity spot of the fiber sensor, so that this kind of sensor can work in harsh environments. For the first time to the best of our knowledge, we report this type of FFPI temperature sensor based on difference of thermal expansion coefficient between fused silica and metallic materials.

Antifungal activity of sulfur-containing salts against the development of carrot cavity spot and potato dry rot

In the search for alternatives to synthetic fungicides to control postharvest disease, sulfur-containing salts were evaluated for their effects on the mycelial growth of various fungal or fungus-like pathogens and their ability to control carrot cavity spot ( Pythium sulcatum) and potato dry rot ( Fusarium sambucinum). Results showed that metabisulfite-containing salts provided strong inhibition of all the tested fungi. Furthermore, some sulfate-containing salts were also directly inhibitory to P. sulcatum (calcium sulfate and ammonium sulfate) and to F. sambucinum (sodium sulfate). The metabisulfite salts also provided 100% inhibition of cavity spot and dry rot at concentrations of 50 and 200 mM, respectively. Calcium sulfate and sodium sulfate also significantly reduced carrot cavity spot lesions at 50 mM and ammonium sulfate, magnesium sulfate, potassium sulfate and sodium sulfate reduced potato dry rot lesions at 200 mM. These results indicate that various sulfate and metabisulfite salts could be used to control these postharvest microorganisms.

Antimicrobial efficacy of cinnamon, ginger, horseradish and nutmeg extracts against spoilage pathogens

In the search for alternatives to the use of synthetic fungicides, aqueous spice extracts were evaluated for their effects on the mycelial growth of various spoilage pathogens and their ability to control potato dry rot and carrot cavity spot in vivo. Results showed that cinnamon, ginger and nutmeg significantly inhibited the mycelial growth of Aspergillus niger (Ascomycota), Fusarium sambucinum (Ascomycota), Pythium sulcatum (Oomycota) or Rhizopus stolonifer (Zygomycota), whereas horseradish extract did not lead to the inhibition of any microorganism at the tested concentration. Among the most effective extracts, 0.05 g mL‑1 of cinnamon extract completely inhibited A. niger and P. sulcatum, and 0.10 g mL‑1 of cinnamon extract completely inhibited F. sambucinum. A concentration of 0.05 g mL‑1 of ginger extract also caused 100% inhibition of P. sulcatum. In vivo, cinnamon extract significantly reduced lesions of potato dry rot and carrot cavity spot, and ginger extract reduced lesions of carrot cavity spot. These results indicate that aqueous cinnamon and ginger extracts could provide an alternative to the use of synthetic fungicides to control these pathogens.

Lateral roots of carrot have a low impact on alloinfections in cavity spot epidemic caused by Pythium violae

Carrot cavity spot, caused by a complex of Pythium species, is characterized by sunken elliptical lesions on the taproot. Recent epidemiological studies of P. violae have demonstrated the occurrence of both primary and secondary infections, with two types of secondary infection, autoinfection and alloinfection. Investigating the mechanisms underlying alloinfection and the role of carrot lateral roots, we asked whether direct physical root contact plays a role in alloinfection and whether root exudates enhance mycelial growth in soil alone. A rhizobox system was designed to differentiate the effects of each mechanism: a buffer zone created by nylon mesh was used to test the first mechanism, and young carrots with a root system similar to lateral roots were used to test the second. Alloinfections were generated in rhizoboxes via diseased taproots transplanted close to healthy, mature carrots. The nylon mesh had no significant effect on disease intensity (reflecting alloinfection), providing evidence that mycelial growth in soil contributed more to disease spread than did physical contact among roots. Nor did young carrots significantly affect alloinfection; thus root exudates had little effect on mycelial growth.

Pathometric relationships reveal epidemiological processes involved in carrot cavity spot epidemics

Carrot cavity spot (CCS) is one of the most important soilborne diseases affecting the carrot crop. The few epidemiological studies that have investigated the temporal and spatial dynamics of the disease have been based solely on diagrammatic scales or semi-quantitative indices. To reveal epidemiological processes involved in the development of CCS epidemics, we investigated pathometric relationships. To this end, standardised measurements were defined (disease incidence i, lesion density d, conditional lesion density cd, lesion size ls, and total diseased area tda). The evolution of a cohort of CCS lesions according to their size suggested that lesions can expand over time. Two pathometric relationships were tested: a first one, between i and tda, is given by the equation i = 100(1−exp(−a(t)tda)), where t is thermal time, and a second one, between tda, d, and ls, is given by the equation tda = c(t)πd(ls/2)2. These relationships were validated for CCS epidemics in the case of field experiments, a survey in commercial fields, and a controlled-conditions experiment. The temporal linear decrease of the time-dependent parameter a(t) in the first relationship suggested that CCS epidemics followed classical epidemiological phases driven by successive processes: (1) the mobilisation of soil inoculum leading to primary infection, (2) the spread of disease to neighbouring taproots (alloinfection), and (3) the intensification of disease on the taproot (autoinfection). This is consistent with complementary experimental results which demonstrated that auto- and alloinfections occur in CCS epidemics.

PCR-based identification ofPythiumspp. causing cavity spot in carrots and sensitive detection in soil samples

On the basis of ITS sequences PCR primers were designed for the identification of the five Pythium species found to be most important for the development of carrot cavity spot in Norway: P. intermedium, P. sulcatum, P. sylvaticum, P. violae and P. ‘vipa’. The P. ‘vipa’ isolates had a unique ITS sequence, differed morphologically from all other Pythium isolates, and thus probably represent a new species. The PCR primers were species-specific with no cross-reaction to other Pythium species or to fungal isolates from carrot tested. The detection limits varied for the different primer pairs. The two most sensitive assays allowed detection of as little as 5 fg DNA. All five Pythium species could be detected in lesions from diseased carrots. Weak positive signals were obtained from some carrot samples without symptoms. PCR assays allowed detection of pathogens in soil. In samples of soil known to produce cavity spots on cropped carrots, strong signals were obtained. In several soil samples more than one of the five Pythium species could be detected. The utilization of this diagnostic PCR assay in analysis of field soil and carrot tissue might in the future be exploited to reduce the incidence of this serious carrot disease.

Annual Meeting, Saskatoon, Saskatchewan, 2007 / Réunion annuelle, Saskatoon, Saskatchewan, 2007

Annual Meeting, Saskatoon, Saskatchewan, 2007 / Réunion annuelle, Saskatoon, Saskatchewan, 2007