Gary A. Secor

Recent genotypes of Phytophthora infestans in the eastern United States reveal clonal populations and reappearance of mefenoxam sensitivity.

Isolates of Phytophthora infestans (n = 178) were collected in 2002 to 2009 from the eastern United States, Midwestern United States, and eastern Canada. Multilocus genotypes were defined using allozyme genotyping, and DNA fingerprinting with the RG-57 probe. Several previously described and three new mulitilocus genotypes were detected. The US-8 genotype was found commonly on commercial potato crops but not on tomato. US-20 was found on tomato in North Carolina from 2002 through 2007 and in Florida in 2005. US-21 was found on tomato in North Carolina in 2005 and Florida in 2006 and 2007. US-22 was detected on tomato in 2007 in Tennessee and New York and became widespread in 2009. US-22 was found in 12 states on tomato and potato and was spread on tomato transplants. This genotype accounted for about 60% of all the isolates genotyped. The US-23 genotype was found in Maryland, Virginia, Pennsylvania, and Delaware on both tomato and potato in 2009. The US-24 genotype was found only in North Dakota in 2009. A1 and A2 mating types were found in close proximity on potato and tomato crops in Pennsylvania and Virginia; therefore, the possibility of sexual reproduction should be monitored. Whereas most individuals of US-8 and US-20 were resistant to mefenoxam, US-21 appeared to be intermediately sensitive, and isolates of US-22, US-23, and US-24 were largely sensitive to mefenoxam. On the basis of sequence analysis of the ras gene, these latter three genotypes appear to have been derived from a common ancestor. Further field and laboratory studies are underway using simple sequence repeat genotyping to monitor current changes in the population structure of P. infestans causing late blight in North America.

The 2009 late blight pandemic in the eastern United States - causes and results

The tomato late blight pandemic of 2009 made late blight into a household term in much of the eastern United States. Many home gardeners and many organic producers lost most if not all of their tomato crop, and their experiences were reported in the mainstream press. Some CSAs (Community Supported Agriculture) could not provide tomatoes to their members. In response, many questions emerged: How did it happen? What was unusual about this event compared to previous late blight epidemics? What is the current situation in 2012 and what can be done? Its easiest to answer these questions, and to understand the recent epidemics of late blight, if one knows a bit of the history of the disease and the biology of the causal agent, Phytophthora infestans.

Real-time PCR Quantification and Mycotoxin Production of Fusarium graminearum in Wheat Inoculated with Isolates Collected from Potato, Sugar Beet, and Wheat

ABSTRACT Fusarium graminearum causes Fusarium head blight (FHB) in small grains worldwide. Although primarily a pathogen of cereals, it also can infect noncereal crops such as potato and sugar beet in the United States. We used a real-time polymerase chain reaction (PCR) method based on intergenic sequences specific to the trichodiene synthase gene (Tri5) from F. graminearum. TaqMan probe and primers were designed and used to estimate DNA content of the pathogen (FgDNA) in the susceptible wheat cv. Grandin after inoculation with the 21 isolates of F. graminearum collected from potato, sugar beet, and wheat. The presence of nine mycotoxins was analyzed in the inoculated wheat heads by gas chromatography and mass spectrometry. All isolates contained the Tri5 gene and were virulent to cv. Grandin. Isolates of F. graminearum differed significantly in virulence (expressed as disease severity), FgDNA content, and mycotoxin accumulation. Potato isolates showed greater variability in producing different mycotoxins than sugar beet and wheat isolates. Correlation analysis showed a significant (P < 0.001) positive relationship between FgDNA content and FHB severity or deoxynivalenol (DON) production. Moreover, a significant (P < 0.001) positive correlation between FHB severity and DON content was observed. Our findings revealed that F. graminearum causing potato dry rot and sugar beet decay could be potential sources of inoculum for FHB epidemics in wheat. Real-time PCR assay provides sensitive and accurate quantification of F. graminearum in wheat and can be useful for monitoring the colonization of wheat grains by F. graminearum in controlled environments, and evaluating wheat germplasms for resistance to FHB.

Effect of Expression of UDP-Glucose Pyrophosphorylase Ribozyme and Antisense RNAs on the Enzyme Activity and Carbohydrate Composition of Field-Grown Transgenic Potato Plants

Summary The effect of suppression of UDP-glucose pyrophosphorylase (UGPase, UTP ±-D-glucose-1-phosphate uridylyl transferase, EC 2.7.7.9) on carbohydrate metabolism in potato tubers was investigated. Constructions expressing a UGPase-specific ribozyme or a UGPase antisense RNA based on the genomic sequence, each under the control of the 35S promoter of the Cauliflower Mosaic Virus, were tested. Out of 103 plants transformed with the constructs, only 13 showed levels of UGPase activity significantly different from the control. No transgenic plants, carrying either construct, expressed an enzyme activity lower than 50-60 % of the control. Plants of seven clones were grown in the field, and UGPase activity, and the level of sugars were measured in the tubers. After storage at room temperature for one week, tubers of six of these had sucrose levels significantly (P

Phenotypic stability of resistance to late blight in potato clones evaluated at eight sites in the United States

Changes in the fungal pathogenPhytophthora infestans in the United States pose a significant threat to potato production. Sources of resistance to these new genotypes of P.infestans need to be identified for potato breeders to have parental materials for crossing, and the phenotypic stability of late blight resistance in these potato clones needs to be determined. Sixteen potato clones which reportedly have some resistance to late blight were evaluated at eight locations: Florida (FL), Maine (ME), Michigan (MI), Minnesota (MN), North Dakota (ND), New York (NY), Pennsylvania (PA) and Wisconsin (WI) in 1996. Percent infected foliage was recorded at approximately weekly intervals following the onset of the disease at each location. Area under the disease progress curve (AUDPC) was calculated. Clones were ranked for mean AUDPC within location and the nonparametric stability statistics, mean absolute rank differences and variance of the ranks, were analyzed for phenotypic stability. Neither of these statistics was significant, indicating a lack of genotype x environment interaction on the rankings of these clones across locations in 1996. The four clones with lowest AUDPC scores were U.S. clones AWN86514-2, B0692-4, B0718-3 and B0767-2. These clones should be useful parental materials for breeders seeking to incorporate genes for late blight resistance into potatoes.

Variability in Virulence Among Asexual Progenies of Phytophthora infestans

ABSTRACT One hundred two single zoospore isolates of Phytophthora infestans, derived asexually from four parental isolates of US-8 genotype and one isolate of US-1 genotype, were characterized for their virulence phenotypes to determine changes in virulence during asexual reproduction. Potato differentials, each containing a major gene for resistance to P. infestans (R1 to R11), were used to characterize the virulence patterns. Isolates were also characterized for mating type, glucose-6-phosphate isomerase (Gpi) banding pattern, and DNA fingerprints using probe RG57 to determine any genotypic changes in the single zoospore isolates. A subset of these single zoospore isolates was tested for response to mefenoxam to determine any shifts in sensitivity. Results showed that single zoospore isolates derived from parent PI-1 (US-8, 11 isolates) were identical to their parental virulence. Isolates derived from parent PI-191 (US-8, 29 isolates) showed some differences in virulence, mainly toward R8 and R9. Isolates derived from parent PI-126 (US-8, 14 isolates) demonstrated a higher level of virulence diversity. Isolates derived from parents PI-52 (US-1, 28 isolates) and PI-105 (US-8, 20 isolates) showed the highest level of virulence variability among the single zoospore isolates. Mating type, Gpi banding pattern, and DNA fingerprints for the single zoospore isolates were, in most cases, identical to the parental isolates. Single zoospore isolates showed different levels of sensitivity to mefenoxam. Virulence and other genetic changes during asexual reproduction are likely to play a major role in changing the race structure of P. infestans populations. This continuous change in the race structure is a serious problem and now poses a new challenge for utilization of race-specific resistance to manage late blight of potato.

Monitoring Fungicide Sensitivity of Cercospora beticola of Sugar Beet for Disease Management Decisions

Cercospora leaf spot, caused by the fungus Cercospora beticola Sacc., is the most serious and important foliar disease of sugar beet (Beta vulgaris L.) wherever it is grown worldwide. Cercospora leaf spot first caused economic damage in North Dakota and Minnesota in 1980, and the disease is now endemic. This is the largest production area for sugar beet in the United States, producing 5.5 to 6.0 million metric tons on approximately 300,000 ha, which is 56% of the sugar beet production in the United States. This Plant Disease feature article details a cooperative effort among the participants in the sugar beet industry in this growing area and represents a successful collaboration and team effort to confront and change a fungicide resistance crisis to a fungicide success program. As a case study of success for managing fungicide resistance, it will serve as an example to other pathogen-fungicide systems and provide inspiration and ideas for long-term disease management by fungicides.

Identification of the G143A mutation associated with QoI resistance in Cercospora beticola field isolates from Michigan, United States

BACKGROUND Cercospora leaf spot (CLS), caused by the fungus Cercospora beticola, is the most serious foliar disease of sugar beet (Beta vulgaris L.) worldwide. Disease control is mainly achieved by timely fungicide applications. In 2011, CLS control failures were reported in spite of application of quinone outside inhibitor (QoI) fungicide in several counties in Michigan, United States. The purpose of this study was to confirm the resistant phenotype and identify the molecular basis for QoI resistance of Michigan C. beticola isolates. RESULTS Isolates collected in Michigan in 1998 and 1999 that had no previous exposure to the QoI fungicides trifloxystrobin or pyraclostrobin exhibited QoI EC(50) values of ≤ 0.006 µg mL(-1) . In contrast, all isolates obtained in 2011 exhibited EC(50) values of > 0.92 µg mL(-1) to both fungicides and harbored a mutation in cytochrome b (cytb) that led to an amino acid exchange from glycine to alanine at position 143 (G143A) compared with baseline QoI-sensitive isolates. Microsatellite analysis of the isolates suggested that QoI resistance emerged independently in multiple genotypic backgrounds at multiple locations. A real-time PCR assay utilizing dual-labeled fluorogenic probes was developed to detect and differentiate QoI-resistant isolates harboring the G143A mutation from sensitive isolates. CONCLUSION The G143A mutation in cytb is associated with QoI resistance in C. beticola. Accurate monitoring of this mutation will be essential for fungicide resistance management in this pathosystem.

Russet Norkotah: A new russet-skinned potato cultivar with wide adaptation

Russet Norkotah is a long, smooth, shallow-eyed, russet-skinned potato cultivar with wide adaptation. It has a smooth golden russet-skin and produces a high percent of medium sized U.S. No. 1 tubers that are good count-carton size.CompendioRusset Norkotah es un cultivar de papa de amplia adaptación, con tubérculos de forma alargada, ojos superficiales y cubierta lisa de color rojizo. Tiene la cáscara de un tinte rojizo dorado y produce un alto porcentaje de tubérculos US No. 1 de tamaño mediano, apropiados para embalaje reducido.

Sporulation of Helminthosporium solani and infection of potato tubers in seed and commercial storages

Silver scurf has become a major reason for rejection of fresh and processing potatoes in recent years. Control of the disease by chemical or cultural practices or resistant cultivars has been difficult. Observations have shown spread and increase of disease of potatoes in storage, but this has not been extensively studied. The objective of this study was to document Helminthosporium solani conidia production, dispersal, and tuber infection in potato storages. Spore samplers placed in seed, processing, and table stock storages collected conidia ranging from 0 to 12,000 conidia per day in seed and table stock storages (4°C), and from 0 to 24,000 conidia per day in processing storages (10°C). Conidia were detected soon after tubers entered storage and increased progressively during the storage period, with the maximum conidia numbers found during the time of tuber handling. Greenhouse-produced minitubers placed in storages for 1, 2, 3, or 4 weeks were infected by H. solani spores. Infection was significantly higher in those exposed for 4 weeks than in those exposed for 1 week. Results document the buildup of H. solani spores throughout the storage period, and that this inoculum is important in disease epidemiology. Control of this inoculum could lead to disease reduction.