A Comprehensive Characterization of Ecological and Epidemiological Factors Driving Perennation of Podosphaera macularis Chasmothecia on Hop (Humulus lupulus).

Hop powdery mildew, caused by the ascomycete fungus Podosphaera macularis, is a consistent threat to sustainable hop production. The pathogen utilizes two reproductive strategies for overwintering and perennation: (i) asexual vegetative hyphae on dormant buds that emerge the following season as infected shoots; and (ii) sexual ascocarps (chasmothecia), which are discharged during spring rain events. We demonstrate that P. macularis chasmothecia, in the absence of any asexual P. macularis growth forms, are a viable overwintering source capable of causing early season infection two to three orders of magnitude greater than that reported for perennation via asexual growth. Two epidemiological models were defined that describe (i) temperature-driven maturation of P. macularis chasmothecia; and (ii) ascosporic discharge in response to duration of leaf wetness and prevailing temperatures. P. macularis ascospores were confirmed to be infectious at temperatures ranging from 5 to 20°C. The organism's chasmothecia were also found to adhere tightly to the host tissue on which they formed, suggesting that these structures likely overwinter wherever hop tissue senesces within a hop yard. These observations suggest that existing early season disease management practices are especially crucial to controlling hop powdery mildew in the presence of P. macularis chasmothecia. Furthermore, these insights provide a baseline for the validation of weather-driven models describing maturation and release of P. macularis ascospores, models that can eventually be incorporated into hop disease management programs.

Spatiotemporal Distribution of Potato-Associated Alternaria Species in Wisconsin.

Potato early blight caused by Alternaria solani and potato brown spot caused by Alternaria alternata are two common potato foliar diseases. Fungicide applications in Wisconsin target early blight with the expectation of managing brown spot simultaneously. However, distributions of the two pathogens over time and space have not been previously reported in Wisconsin. In this study, six potato fields in Wisconsin were systematically sampled during 2014 to 2017 to investigate the incidences of both pathogens; incidences of the pathogens were compared by location, timing, and year. Incidence of A. alternata had been recorded starting in late June in every field every year, whereas incidence of A. solani started later in some of the fields. At the end of each growing season throughout the four years, the highest incidence (100% most of the years) of A. solani was observed in a field in Plover, located in a concentrated potato production area of central Wisconsin, and a field in Arena, located in an isolated potato production area in southern Wisconsin. The highest incidence (100% every year) of A. alternata was observed in a field in Madison, an isolated potato field that was managed organically. Such distribution patterns could be a result of the different host ranges of the pathogens and abundance of inoculum in different geographic locations. In 2016, disease severity was correlated with the incidence of both pathogens (P < 0.0001 to P = 0.0299), but in 2017, disease severity was correlated with the incidence of A. solani in most of the fields (P < 0.0001 to P = 0.0799). These findings suggest that both Alternaria spp. are important for considering disease management strategies in central and southern Wisconsin.

Investigating potato late blight physiological differences across potato cultivars with spectroscopy and machine learning.

Understanding plant disease resistance is important in the integrated management of Phytophthora infestans, causal agent of potato late blight. Advanced field-based methods of disease detection that can identify infection before the onset of visual symptoms would improve management by greatly reducing disease potential and spread as well as improve both the financial and environmental sustainability of potato farms. In-vivo foliar spectroscopy offers the capacity to rapidly and non-destructively characterize plant physiological status, which can be used to detect the effects of necrotizing pathogens on plant condition prior to the appearance of visual symptoms. Here, we tested differences in spectral response of four potato cultivars, including two cultivars with a shared genotypic background except for a single copy of a resistance gene, to inoculation with Phytophthora infestans clonal lineage US-23 using three statistical approaches: random forest discrimination (RF), partial least squares discrimination analysis (PLS-DA), and normalized difference spectral index (NDSI). We find that cultivar, or plant genotype, has a significant impact on spectral reflectance of plants undergoing P. infestans infection. The spectral response of four potato cultivars to infection by Phytophthora infestans clonal lineage US-23 was highly variable, yet with important shared characteristics that facilitated discrimination. Early disease physiology was found to be variable across cultivars as well using non-destructively derived PLS-regression trait models. This work lays the foundation to better understand host-pathogen interactions across a variety of genotypic backgrounds, and establishes that host genotype has a significant impact on spectral reflectance, and hence on biochemical and physiological traits, of plants undergoing pathogen infection.

Management and Soil Conditions Influence Common Scab Severity on Potato Tubers Via Indirect Effects on Soil Microbial Communities.

Common scab, caused by Streptomyces scabies and related species, is a potato tuber blemish disease that causes reductions in marketable yield worldwide. Evidence of suppression of common scab by indigenous soil microbial populations has been found in several studies. However, we lack a comprehensive understanding of how common scab severity relates functionally to potato varieties, farming systems, soil physical and chemical properties, and soil microbial communities. These factors may affect disease directly or indirectly by affecting one of the other variables. We performed a survey of 30 sampling locations across 12 fields in Wisconsin and used structural equation modeling to disentangle the direct effects of potato market classes, farm management (conventional versus organic), and soil physiochemical properties on common scab severity from their indirect effects mediated through soil bacterial and fungal communities. We found that, although potato market classes affected disease severity directly, the effects of farm management and soil physiochemistry were best explained as indirect, mediated by their impacts on soil bacterial communities. This suggests that evaluating the consequences of specific management practices for soil microbial communities may be useful for understanding disease pressure across fields.

Contact Reflectance Spectroscopy for Rapid, Accurate, and Nondestructive Phytophthora infestans Clonal Lineage Discrimination.

Populations of Phytophthora infestans, the oomycete causal agent of potato late blight in the United States, are predominantly asexual, and isolates are characterized by clonal lineage or asexual descendants of a single genotype. Current tools for clonal lineage identification are time consuming and require laboratory equipment. We previously found that foliar spectroscopy can be used for high-accuracy pre- and postsymptomatic detection of P. infestans infections caused by clonal lineages US-08 and US-23. In this work, we found subtle but distinct differences in spectral responses of potato foliage infected by these clonal lineages in both growth-chamber time-course experiments (12- to 24-h intervals over 5 days) and naturally infected samples from commercial production fields. In both settings, we measured continuous visible to shortwave infrared reflectance (400 to 2,500 nm) on leaves using a portable spectrometer with contact probe. We consistently discriminated between infections caused by the two clonal lineages across all stages of disease progression using partial least squares (PLS) discriminant analysis, with total accuracies ranging from 88 to 98%. Three-class random forest differentiation between control, US-08, and US-23 yielded total discrimination accuracy ranging from 68 to 76%. Differences were greatest during presymptomatic infection stages and progressed toward uniformity as symptoms advanced. Using PLS-regression trait models, we found that total phenolics, sugar, and leaf mass per area were different between lineages. Shortwave infrared wavelengths (>1,100 nm) were important for clonal lineage differentiation. This work provides a foundation for future use of hyperspectral sensing as a nondestructive tool for pathovar differentiation.

Distribution and Stability of Quinone Outside Inhibitor Fungicide Resistance in Populations of Potato Pathogenic Alternaria spp. in Wisconsin.

Quinone outside inhibitor (QoI) fungicides have been an important class in managing potato early blight caused by Alternaria solani and brown spot caused by A. alternata. Because of the single-site mode of action character of QoI fungicides, which are relied on for management of diseases in Wisconsin, and the abundant asexual conidia production of the Alternaria species, pathogen isolates with QoI resistance have been detected after just a few years of QoI fungicide usage in commercial production fields. Resistance to QoIs has been attributed to amino acid substitutions F129L and G143A in cytochrome b of A. solani and A. alternata, respectively, as a result of point mutations. The aim of this study was to assess Alternaria populations in Wisconsin for QoI resistance before and after fungicide applications in order to evaluate resistance stability. A TaqMan single nucleotide polymorphism genotyping assay was designed based on the sequences of the cytochrome b gene from Alternaria isolates collected in Wisconsin to profile QoI resistance in Alternaria populations as well as to explore factors that may influence frequency of QoI resistance in the pathogen populations. This assay successfully identified the mutations conferring QoI resistance in isolates collected from four locations each year from 2015 to 2017. During the course of this study, the frequency of A. solani isolates with the F129L mutation was consistently high and showed primarily the TTA mutation type. The frequency of A. alternata isolates with the G143A mutation started relatively low and increased at the end of the production season in each year (P = 0.0109, P = 0.2083, and P = 0.0159). A potato field managed without use of QoI fungicides showed a significantly lower (P < 0.05) frequency of A. alternata isolates carrying G143A than conventionally managed potato fields. The overall frequency of A. alternata isolates carrying G143A in the four locations was similar over the 3 years (P = 0.2971). The QoI resistance characteristics of the isolates were stable even when QoI selection pressure was removed for at least five subculture transfers, and the mutation types of codons 129 and 143 in the cytochrome b gene in A. solani and A. alternata, respectively, remained the same. This indicated that the application of QoIs in the field is not the sole factor responsible for the variation of the frequency of QoI resistance in the pathogen populations.

Aerial Concentrations of Pathogens Causing Early Blight and Brown Spot Within Susceptible Potato Fields.

Early blight caused by Alternaria solani and brown spot caused by A. alternata are two common foliar diseases of potato, with early blight typically predominating in incidence and severity. Renewed interest in these two diseases has arisen as a result of notable differences in fungicide resistance profiles of the pathogens and inconsistent outcomes of disease management tactics. The pathogens share similar disease cycles, but they differ in the shape and size of their conidia. A. solani has a host range that includes just the Solanaceae, whereas A. alternata has a broad host range spanning numerous plant families. Such differences may result in differences in dispersal of the pathogens and subsequently influence epidemiology and management outcomes. In the commercial potato fields investigated in this study, the aerial conidial concentrations of A. solani and A. alternata differed significantly, with those of A. alternata conidia being higher in number and more variable than those of A. solani. The aerial conidial concentration of A. solani almost always significantly decreased with height (0 to 3 m above the canopy), whereas such a decrease was only observed for 4 of 12 days for A. alternata. The atmospheric concentrations of A. alternata were higher than those of A. solani at both upwind and downwind edges of the field (P < 0.0001), indicating more potential for long-distance dispersal. A higher aerial concentration of conidia at the downwind than the upwind location was observed for A. solani (P < 0.05), whereas overall no such effect was observed for A. alternata. This indicated that the potato fields investigated were likely the source of A. solani conidia, but they may not be the sole source of A. alternata. Results are consistent with inoculum of A. solani coming primarily from within the potato crop, whereas that of A. alternata may be generated from diverse plant sources across the landscape.

Diversity and Virulence of Alternaria spp. Causing Potato Early Blight and Brown Spot in Wisconsin.

Early blight, caused by Alternaria solani, along with brown spot, caused by A. alternata, have the potential to reduce quality and yield in potato production globally. Prior to this study, the incidence, disease impact, and fungicide resistance attributes of A. alternata in Wisconsin were poorly understood. Potato pathogens were isolated from foliar lesions at three commercial locations in Wisconsin in 2012 and 2017 and were initially morphologically identified as A. solani (n = 33) and A. alternata (n = 40). Identifications were further corroborated with the phylogenetic analysis of the internal transcribed spacer (ITS), translation elongation factor 1 (TEF1), gapdh, Alt a 1, and OPA10-2. A multigene phylogeny of ITS, TEF1, gapdh, and Alt a 1 showed five genotypes of A. alternata and one single genotype of A. solani. We demonstrated that the A. alternata isolates were virulent on potato cultivars Russet Burbank (P < 0.013) and Atlantic (P < 0.0073), though they caused less disease than A. solani (P < 0.0001 and P < 0.0001, respectively). A. alternata caused little disease on the breeding line 24-24-12 (P = 0.9929), and A. solani caused fewer disease symptoms on 24-24-12 than on Russet Burbank (P < 0.0001) or Atlantic (P < 0.0001). Breeding line 24-24-12 may be a promising source of potential resistance for the two diseases. There was no significant difference in virulence of different A. alternata genotypes, and no significant difference in virulence or genotype clustering among isolates from the three locations. Isolates of A. alternata that induced chlorosis caused larger lesion areas than isolates that did not in Russet Burbank (P < 0.0001), Atlantic (P < 0.0001), and 24-24-12 (P = 0.0365). There was no significant difference in virulence between quinone outside inhibitor (QoI)-sensitive and QoI-resistant isolates of A. alternata. This study enhanced our understanding of potato early blight and brown spot in Wisconsin, and suggested that A. alternata in addition to A. solani should be carefully monitored and possibly uniquely managed in order to achieve overall disease control.

Genetic Variation within Clonal Lineages of Phytophthora infestans Revealed through Genotyping-By-Sequencing, and Implications for Late Blight Epidemiology.

Genotyping-by-sequencing (GBS) was performed on 257 Phytophthora infestans isolates belonging to four clonal lineages to study within-lineage diversity. The four lineages used in the study were US-8 (n = 28), US-11 (n = 27), US-23 (n = 166), and US-24 (n = 36), with isolates originating from 23 of the United States and Ontario, Canada. The majority of isolates were collected between 2010 and 2014 (94%), with the remaining isolates collected from 1994 to 2009, and 2015. Between 3,774 and 5,070 single-nucleotide polymorphisms (SNPs) were identified within each lineage and were used to investigate relationships among individuals. K-means hierarchical clustering revealed three clusters within lineage US-23, with US-23 isolates clustering more by collection year than by geographic origin. K-means hierarchical clustering did not reveal significant clustering within the smaller US-8, US-11, and US-24 data sets. Neighbor-joining (NJ) trees were also constructed for each lineage. All four NJ trees revealed evidence for pathogen dispersal and overwintering within regions, as well as long-distance pathogen transport across regions. In the US-23 NJ tree, grouping by year was more prominent than grouping by region, which indicates the importance of long-distance pathogen transport as a source of initial late blight inoculum. Our results support previous studies that found significant genetic diversity within clonal lineages of P. infestans and show that GBS offers sufficiently high resolution to detect sub-structuring within clonal populations.

Survival of Isolates of the US-22, US-23, and US-24 Clonal Lineages of Phytophthora infestans by Asexual Means in Tomato Seed at Cold Temperatures.

Survival of Phytophthora infestans, causal agent of potato and tomato late blight, is thought to be negligible when exposed to freezing conditions typical of a Wisconsin winter. However, the persistence of relatively new P. infestans clonal lineages US-22, US-23, and US-24 within a production region during 2010 to 2014 warranted further investigation. We used tomato seed as a culture medium to determine the survival of P. infestans isolates representing the three lineages under temperatures of 18, 4, 0, -3, and -5°C for 11 time points (1 to 112 days postincubation). Survival varied interactively with temperature, duration of time at a temperature, and clonal lineage of the P. infestans isolate. US-22, -23, and -24 isolates survived for 112 days at 18 and 4°C, 84 days at 0°C, and 14 days at -3°C. US-23 survived longer at -3 and -5°C than did US-22 or US-24. The vigor of US-22 and US-24 isolates decreased with increasing exposure to cold temperatures, a trend that was not observed for the US-23 isolate. By calculating the length of time needed to kill the lineage isolates on infested tomato seed at five temperatures, we predicted that P. infestans would survive in 5% of tomato seed for 99, 25, and 16 days at 0, -3, and -5°C, respectively. We further applied a degree-day model to our empirical data to describe P. infestans survival as a function of cooling degree-day accumulations using archived soil temperatures at 5- and 10-cm depths at four Wisconsin locations over 27 years. The model indicated that survival of P. infestans in 5% of infested tomato seed would occur at 35 and 39% of the location-year combinations at 5- and 10-cm soil depths, respectively. Together, these data suggested that P. infestans has the potential to survive over the winter season by asexual means in infested tomato seed in Wisconsin and other Northern latitudes. Our cooling degree-day model for late blight in the tomato production system offers a tool for anticipating and mitigating disease based on integrated pest management concepts previously utilized for insects.

Efficacy of Organic and Conventional Fungicides and Impact of Application Timing on Control of Tomato Late Blight Caused by US-22, US-23, and US-24 Isolates of Phytophthora infestans.

Late blight, caused by Phytophthora infestans, is one of the most economically important diseases of potato and tomato worldwide. Repeated preventative application of fungicides is the primary means of control on susceptible solanaceous host crops. For organic production, fungicide choices are limited, and little efficacy data on noncopper options is available on which to base control recommendations. Twelve fungicides, including organic and conventional selections, were evaluated for both preventative and postinfection control of a single infection cycle of late blight caused by isolates representing three recently identified P. infestans clonal lineages (US-22, US-23, and US-24) using a detached tomato leaf assay. A subset of the most effective fungicides was also tested for preventative control of a single infection cycle of late blight caused by an isolate of US-23 on potted whole tomato plants under laboratory conditions. Fungicide applications made 2 days after inoculation failed to significantly control late blight on detached leaves in all treatments, with the exception of Bravo Ultrex (US-23 only) and Phostrol (US-22 only). Preventative fungicide applications of Bravo Ultrex, Ridomil Gold SL, Revus, Zonix, and low and high rates of EF400 significantly controlled late blight caused by US-22, -23, and -24 isolates. Additionally, preventative application of Phostrol significantly controlled late blight caused by the US-22 isolate; and Phostrol, low rate of Mycostat, and high rate of Champ significantly controlled late blight caused by the US-23 isolate. Late blight caused by the US-24 isolate was significantly reduced compared with US-22 and US-23 isolates for all fungicide treatments applied after inoculation, as well as for all preventative fungicide treatments, with the exception of Bravo, Ridomil, and Revus. In whole-potted-plant assays with the US-23 isolate, late blight was significantly controlled by preventative application of Bravo Ultrex, Ridomil Gold SL, and high rate of EF400; disease was not significantly controlled by Zonix, low rate of EF400, Phostrol, or low and high rates of Champ. Based on these results, it is anticipated that currently available fungicides with suitability to conventional and organic systems can effectively control late blight caused by new clonal lineages of P. infestans when applied preventatively and that late blight caused by the US-24 clonal lineage may require less fungicide use than US-22 or US-23 to mitigate disease.

Effect of Temperature on Growth and Sporulation of US-22, US-23, and US-24 Clonal Lineages of Phytophthora infestans and Implications for Late Blight Epidemiology.

Epidemics of late blight, caused by Phytophthora infestans (Mont.) de Bary, have been studied by plant pathologists and regarded with great concern by potato and tomato growers since the Irish potato famine in the 1840s. P. infestans populations have continued to evolve, with unique clonal lineages arising which differ in pathogen fitness and pathogenicity, potentially impacting epidemiology. In 2012 and 2013, the US-23 clonal lineage predominated late blight epidemics in most U.S. potato and tomato production regions, including Wisconsin. This lineage was unknown prior to 2009. For isolates of three recently identified clonal lineages of P. infestans (US-22, US-23, and US-24), sporulation rates were experimentally determined on potato and tomato foliage and the effect of temperature on lesion growth rate on tomato was investigated. The US-22 and US-23 isolates had greater lesion growth rates on tomato than US-24 isolates. Sporulation rates for all isolates were greater on potato than tomato, and the US-23 isolates had greater sporulation rates on both tomato and potato than the US-22 and US-24 isolates. Experimentally determined correlates of fitness were input to the LATEBLIGHT model and epidemics were simulated using archived Wisconsin weather data from four growing seasons (2009 to 2012) to investigate the effect of isolates of these new lineages on late blight epidemiology. The fast lesion growth rates of US-22 and US-23 isolates resulted in severe epidemics in all years tested, particularly in 2011. The greater sporulation rates of P. infestans on potato resulted in simulated epidemics that progressed faster than epidemics simulated for tomato; the high sporulation rates of US-23 isolates resulted in simulated epidemics more severe than simulated epidemics of isolates of the US-22 and US-24 isolates and EC-1 clonal lineages on potato and tomato. Additionally, US-23 isolates consistently caused severe simulated epidemics when lesion growth rate and sporulation were input into the model singly or together. Sporangial size of the US-23 isolates was significantly smaller than that of US-22 and US-24 isolates, which may result in more efficient release of sporangia from the tomato or potato canopy. Our experimentally determined correlates of fitness and the simulated epidemics resulting from their incorporation into the LATEBLIGHT model suggest that US-23 isolates of P. infestans may have the greatest fitness among currently prevalent lineages and may be the most likely lineage to persist in the P. infestans population. The US-23 clonal lineage has been documented as the most prevalent lineage in recent years, indicating its overall fitness. In our work, US-23 had the highest epidemic potential among current genotypes. Given that epidemic potential is a component of fitness, this may, in part, explain the current predominance of the US-23 lineage.

Novel Resistance in Heirloom Tomatoes and Effectiveness of Resistance in Hybrids to Phytophthora infestans US-22, US-23, and US-24 Clonal Lineages.

Late blight, caused by the oomycete Phytophthora infestans, causes serious losses in tomato production worldwide. Application of fungicides is the primary means of management but cultivar resistance, primarily through Ph resistance genes from Solanum pimpinellifolium, can provide a cost-effective and environmentally sound approach to an overall disease management program. Due to highly adaptable pathogen populations, cultivar resistance against late blight is often short lived and continual assessment of disease response to new pathogen types is necessary. We evaluated the disease response of 11 tomato cultivars to one isolate from each of three clonal lineages (US-22, US-23, and US-24) of P. infestans novel to the United States to determine the efficacy of currently deployed Ph genes in hybrid cultivars and the validity of claims of resistance in heirloom cultivars. Lesion length and pathogen growth were reduced on tomato genotypes 'Plum Regal' (Ph-3) and 'Legend' (Ph-2) compared with the susceptible control 'Brandywine Red' following inoculation with one isolate (US-23) but were not significantly different from the control with an isolate of US-22. 'Mountain Magic' (Ph-2 and Ph-3) and three heirloom cultivars ('Wapsipinicon Peach', 'Matt's Wild Cherry,' and 'Pruden's Purple') had reduced lesion length and pathogen growth to all three isolates. Although the genetics of resistance are not fully understood for many of these, the heirloom cultivars may be useful for future tomato late blight breeding efforts. All of the cultivars investigated in this work are currently available and use of cultivars exhibiting reduced disease development may limit losses to late blight and reduce reliance on fungicides. Resistant cultivars also limit the production of inoculum, reducing overall late blight risk and spread in tomato and potato crops.

Investigating the Host Range of the US-22, US-23, and US-24 Clonal Lineages of Phytophthora infestans on Solanaceous Cultivated Plants and Weeds.

Phytophthora infestans causes late blight, one of the most important diseases of potato and tomato worldwide. Recently in the United States, three newly identified clonal lineages, US-22, US-23, and US-24, have become widespread. While potato and tomato are the most commonly infected solanaceous hosts for P. infestans, new lineages may have a broader or different host range. Under controlled conditions, we determined the host range of isolates representing US-22, US-23, and US-24 genotypes of P. infestans on detached tissues of cultivated solanaceous plants and solanaceous weeds common to the upper midwestern production region. None of the isolates representing the clonal lineages produced late blight symptoms or signs on foliage of selected cultivars of eggplant, pepper, tomatillo, or ground cherry in a detached leaf assay. Symptoms and signs were evident on the potato and tomato cultivars tested, although with the US-24 isolate, infection on tomato was limited. None of the isolates sporulated on the common weed black nightshade, but some sporulation and necrosis was observed with all representatives of the lineages on bittersweet nightshade and petunia. Hairy nightshade supported abundant sporulation and symptoms, and sporangial production was not significantly different than that on tomato for each of the isolates representing the three lineages, indicating the potential for this weed to be a source of inoculum and contribute substantially to late blight epidemics. Interestingly, black nightshade had the highest incidence of sporulation on berries, but the lowest on leaves, suggesting the importance of testing multiple plant organs when determining susceptibility of a species. Our results update knowledge of the host range of the ever-changing P. infestans populations and will help to improve late blight management strategies by targeting these additional hosts.

Characterization of kudzu (Pueraria spp.) resistance to Phakopsora pachyrhizi, the causal agent of soybean rust.

Kudzu (Pueraria spp.) is an accessory host for soybean rust (SBR) (caused by Phakopsora pachyrhizi) that is widespread throughout the southeastern United States. An expanded survey of kudzu sites was conducted in 2008 to determine the proportion of natural resistance in the north-Florida kudzu population. Of the 139 sites evaluated, approximately 18% were found to be free of SBR infection, while 23% had reduced sporulation. Ten accessions of kudzu from north-central Florida were characterized for their response to challenge by a single isolate of P. pachyrhizi under laboratory conditions. Three outcomes were observed: tan lesions with profuse sporulation (susceptible); reddish-brown lesions with delayed, reduced sporulation (resistant); and an immune response in which no lesions developed (immune). Of the 10 accessions, 6 were susceptible, 3 were immune, and 1 was resistant. Cytological examination revealed that resistant interactions were typified by early onset of a multicell hypersensitive response (HR) while typical immune interactions were the result of cell wall depositions that blocked penetration in combination with early onset of the HR. Quantitative real-time polymerase chain reaction was performed to determine the extent of colonization. After 15 days, there was 10-fold less P. pachyrhizi DNA present in resistant compared with susceptible kudzu, while the amount of P. pachyrhizi DNA present in the immune kudzu was below the detection level. Susceptible kudzu had approximately half the amount of P. pachyrhizi DNA present when compared with a susceptible soybean cultivar.