Global Cropland Connectivity: A Risk Factor for Invasion and Saturation by Emerging Pathogens and Pests.

The geographic pattern of cropland is an important risk factor for invasion and saturation by crop-specific pathogens and arthropods. Understanding cropland networks supports smart pest sampling and mitigation strategies. We evaluate global networks of cropland connectivity for key vegetatively propagated crops (banana and plantain, cassava, potato, sweet potato, and yam) important for food security in the tropics. For each crop, potential movement between geographic location pairs was evaluated using a gravity model, with associated uncertainty quantification. The highly linked hub and bridge locations in cropland connectivity risk maps are likely priorities for surveillance and management, and for tracing intraregion movement of pathogens and pests. Important locations are identified beyond those locations that simply have high crop density. Cropland connectivity risk maps provide a new risk component for integration with other factors-such as climatic suitability, genetic resistance, and global trade routes-to inform pest risk assessment and mitigation.

Genotyping of Phytophthora infestans in Eastern Africa Reveals a Dominating Invasive European Lineage.

Strains of Phytophthora infestans, the pathogen causing late blight of potato and tomato, are thought to be moved around the world through infected planting material. Since its first appearance in 1941, late blight has caused important losses to potato production in the eastern-Africa region (EAR). In the current study, the genetic structure of the population in Kenya, Uganda, Tanzania, Burundi, and Rwanda was characterized using 12-plex microsatellite markers with the aim of testing the hypothesis that a strain originating from Europe, 2_A1, has recently dominated the population in EAR. Analyses of 1,093 potato and 165 tomato samples collected between 2013 and 2016 revealed the dominance on potato in all countries of the 2_A1 clonal lineage. On tomato, a host-specialized form of the US-1 lineage appears to persist in Rwanda, Uganda, and Tanzania whereas, in Kenya, most samples from tomato (72.5%) were 2_A1. The US-1 lineage in Tanzania had two private alleles at the Pi02 marker, suggesting a possible independent introduction into the region. US-1 had higher genetic variability than 2_A1, consistent with the earlier establishment of the former. Continuous tracking of P. infestans population changes should help identify new virulent and aggressive strains, which would inform strategic disease management options.

Stacking three late blight resistance genes from wild species directly into African highland potato varieties confers complete field resistance to local blight races.

Considered responsible for one million deaths in Ireland and widespread famine in the European continent during the 1840s, late blight, caused by Phytophthora infestans, remains the most devastating disease of potato (Solanum tuberosum L.) with about 15%-30% annual yield loss in sub-Saharan Africa, affecting mainly smallholder farmers. We show here that the transfer of three resistance (R) genes from wild relatives [RB, Rpi-blb2 from Solanum bulbocastanum and Rpi-vnt1.1 from S. venturii] into potato provided complete resistance in the field over several seasons. We observed that the stacking of the three R genes produced a high frequency of transgenic events with resistance to late blight. In the field, 13 resistant transgenic events with the 3R-gene stack from the potato varieties 'Desiree' and 'Victoria' grew normally without showing pathogen damage and without any fungicide spray, whereas their non-transgenic equivalent varieties were rapidly killed. Characteristics of the local pathogen population suggest that the resistance to late blight may be long-lasting because it has low diversity, and essentially consists of the single lineage, 2_A1, which expresses the cognate avirulence effector genes. Yields of two transgenic events from 'Desiree' and 'Victoria' grown without fungicide to reflect small-scale farm holders were estimated to be 29 and 45 t/ha respectively. This represents a three to four-fold increase over the national average. Thus, these late blight resistant potato varieties, which are the farmers' preferred varieties, could be rapidly adopted and bring significant income to smallholder farmers in sub-Saharan Africa.

Incomplete Infection of Secondarily Infected Potato Plants - an Environment Dependent Underestimated Mechanism in Plant Virology.

The common assumption in potato virus epidemiology is that all daughter tubers produced by plants coming from infected mother tubers (secondary infection) will become infected via systemic translocation of the virus during growth. We hypothesize that depending on the prevalent environmental conditions, only a portion of the daughter tubers of a plant that is secondarily infected by viruses may become infected. To test this hypothesis experimental data from standardized field experiments were produced in three contrasting environments at 112, 3280, and 4000 m a.s.l. in Peru during two growing seasons. In these experiments, the percentage of infected daughter tubers produced by seed tubers that were infected with either potato potexvirus X (PVX), potato Andean mottle comovirus (APMoV), potato potyvirus Y (PVY) (jointly infected with PVX) or potato leafroll luteovirus (PLRV) was determined. Incomplete autoinfection was found in all cases, as the percentage of virus infected daughter tubers harvested from secondarily infected plants was invariably less than 100%, with the lowest percentage of infection being 30%. Changing the growing site to higher altitudes decreased autoinfection for all viruses. Therefore, the assumption of complete autoinfection of secondarily infected plants were rejected, while the hypothesis of environmentally dependent incomplete autoinfection was accepted. The findings help explain the occurrence of traditional seed management practices in the Andes and may help to develop locally adapted seed systems in environments of the world that have no steady access to healthy seed tubers coming from a formally certified seed system. The results obtained almost three decades ago are discussed in light of most recent knowledge on epigenetic regulation of host plant - virus interactions which allow for speculating about the underlying biological principles of the incomplete autoinfection. A research roadmap is proposed for achieving explicit experimental proof for the epigenetic regulation of incomplete autoinfection in the pathosystem under study.

The Irish potato famine pathogen Phytophthora infestans originated in central Mexico rather than the Andes.

Phytophthora infestans is a destructive plant pathogen best known for causing the disease that triggered the Irish potato famine and remains the most costly potato pathogen to manage worldwide. Identification of P. infestan's elusive center of origin is critical to understanding the mechanisms of repeated global emergence of this pathogen. There are two competing theories, placing the origin in either South America or in central Mexico, both of which are centers of diversity of Solanum host plants. To test these competing hypotheses, we conducted detailed phylogeographic and approximate Bayesian computation analyses, which are suitable approaches to unraveling complex demographic histories. Our analyses used microsatellite markers and sequences of four nuclear genes sampled from populations in the Andes, Mexico, and elsewhere. To infer the ancestral state, we included the closest known relatives Phytophthora phaseoli, Phytophthora mirabilis, and Phytophthora ipomoeae, as well as the interspecific hybrid Phytophthora andina. We did not find support for an Andean origin of P. infestans; rather, the sequence data suggest a Mexican origin. Our findings support the hypothesis that populations found in the Andes are descendants of the Mexican populations and reconcile previous findings of ancestral variation in the Andes. Although centers of origin are well documented as centers of evolution and diversity for numerous crop plants, the number of plant pathogens with a known geographic origin are limited. This work has important implications for our understanding of the coevolution of hosts and pathogens, as well as the harnessing of plant disease resistance to manage late blight.

Climate change may have limited effect on global risk of potato late blight.

Weather affects the severity of many plant diseases, and climate change is likely to alter the patterns of crop disease severity. Evaluating possible future patterns can help focus crop breeding and disease management research. We examined the global effect of climate change on potato late blight, the disease that caused the Irish potato famine and still is a common potato disease around the world. We used a metamodel and considered three global climate models for the A2 greenhouse gas emission scenario for three 20-year time-slices: 2000-2019, 2040-2059 and 2080-2099. In addition to global analyses, five regions were evaluated where potato is an important crop: the Andean Highlands, Indo-Gangetic Plain and Himalayan Highlands, Southeast Asian Highlands, Ethiopian Highlands, and Lake Kivu Highlands in Sub-Saharan Africa. We found that the average global risk of potato late blight increases initially, when compared with historic climate data, and then declines as planting dates shift to cooler seasons. Risk in the agro-ecosystems analyzed, varied from a large increase in risk in the Lake Kivu Highlands in Rwanda to decreases in the Southeast Asian Highlands of Indonesia.

Phenotypic stability and genome-wide association study of late blight resistance in potato genotypes adapted to the tropical highlands.

Potato genotypes from a breeding population adapted to tropical highlands were analyzed for the stability of late blight resistance and also for marker-phenotype association. We harmonized the historical evaluation data, consisting of observations spanning 6 years from two field sites utilizing a resistance scale constructed by comparing the area under the disease progress curve (AUDPC) values of 172 genotypes with that of susceptible control 'Yungay'. In total, 70 potato genotypes had a coefficient of variability <0.5 and were considered stable across the environments tested. A principal component analysis demonstrated that the ensemble of experiments formed two distinct groups that reflect the stability of genotype resistance to late blight. Phytophthora infestans isolates present in the experimental fields belonged to the EC-1 clonal lineage and showed variation in virulence beyond the concept of the avirulence determined by the conventionally used R1-R11 differential set. A single-nucleotide polymorphism (SNP) marker on chromosome 9 was associated with late blight resistance and linked to instability. Genotypes with either AACC or AAAC combinations for this SNP were highly resistant only in some environments, while the genotypes with the AAAA combination had more moderate levels of resistance but were stable across environments.

Wide Phenotypic Diversity for Resistance to Phytophthora infestans Found in Potato Landraces from Peru.

The wild and cultivated species of potato have been utilized in potato breeding to good effect but only a very small sample of the available biodiversity has been exploited. In total, 468 accessions of wild and cultivated species of potato were assessed for resistance to the oomycete pathogen Phytophthora infestans using greenhouse assays. Wide phenotypic variation for resistance was found within a species (i.e., among accessions) but not among species which, on average, were similar. Nineteen accessions had resistance levels better than or similar to the variety Chucmarina, which is routinely used by the International Potato Center as a resistant control. Surprisingly, a number of accessions were significantly more susceptible than the susceptible control, Tomasa Condemayta. Frequency histograms of species indicated continuous variation for resistance with little evidence for functional resistance genes.

Use of Phosphonate to Manage Foliar Potato Late Blight in Developing Countries.

Twenty phosphonate products found in the agrochemical market in Ecuador and Peru were evaluated in bioassays for the control of foliar potato late blight, caused by Phytophthora infestans. Eight phosphonate products were evaluated in 16 field experiments done in Peru, Ecuador, Kenya, and Nepal. A meta-analysis across locations involving 71 combinations of potato genotype by site and year demonstrated a significant relationship between phosphonate application rate and efficacy for controlling late blight on potato foliage. The meta-analysis revealed that phosphonate rates of approximately 2.5 g a.i./liter provided efficacy similar to that of the conventional contact fungicides mancozeb and chlorothalonil used at similar rates. At rates higher than 2.5 g a.i./liter, the efficacy of phosphonate was superior to the contact fungicides. Overall, late blight control by phosphonate appeared relatively stable in field experiments across locations. An analysis of field experiments and 64 combinations of potato genotype by site and year showed no correlation between the susceptibility level of potato genotypes and efficacy of phosphonates. The cost of both phosphonate compounds and contact fungicides varied greatly among the countries of the field study; however, in Kenya, control with phosphonate was clearly less expensive than with mancozeb.

The plant pathogen Phytophthora andina emerged via hybridization of an unknown Phytophthora species and the Irish potato famine pathogen, P. infestans.

Emerging plant pathogens have largely been a consequence of the movement of pathogens to new geographic regions. Another documented mechanism for the emergence of plant pathogens is hybridization between individuals of different species or subspecies, which may allow rapid evolution and adaptation to new hosts or environments. Hybrid plant pathogens have traditionally been difficult to detect or confirm, but the increasing ease of cloning and sequencing PCR products now makes the identification of species that consistently have genes or alleles with phylogenetically divergent origins relatively straightforward. We investigated the genetic origin of Phytophthora andina, an increasingly common pathogen of Andean crops Solanum betaceum, S. muricatum, S. quitoense, and several wild Solanum spp. It has been hypothesized that P. andina is a hybrid between the potato late blight pathogen P. infestans and another Phytophthora species. We tested this hypothesis by cloning four nuclear loci to obtain haplotypes and using these loci to infer the phylogenetic relationships of P. andina to P. infestans and other related species. Sequencing of cloned PCR products in every case revealed two distinct haplotypes for each locus in P. andina, such that each isolate had one allele derived from a P. infestans parent and a second divergent allele derived from an unknown species that is closely related but distinct from P. infestans, P. mirabilis, and P. ipomoeae. To the best of our knowledge, the unknown parent has not yet been collected. We also observed sequence polymorphism among P. andina isolates at three of the four loci, many of which segregate between previously described P. andina clonal lineages. These results provide strong support that P. andina emerged via hybridization between P. infestans and another unknown Phytophthora species also belonging to Phytophthora clade 1c.

Assessing the Adequacy of the Simulation Model LATEBLIGHT Under Nicaraguan Conditions.

In this study, the adequacy of the late blight simulation model LATEBLIGHT (version LB2004) was evaluated under Nicaraguan conditions. During 2007 to 2008, five field experiments were conducted in three potato-production regions in northern Nicaragua. Two susceptible ('Cal White' and 'Granola') and one resistant ('Jacqueline Lee') potato cultivars were evaluated without use of fungicides and with three application intervals (4, 7, and 14 days) of the fungicide chlorothalonil. The simulation model was considered adequate because it accurately predicted high disease severity in susceptible cultivars without fungicide protection, and demonstrated a decrease in the disease progress curves with additional fungicide applications, similar to that observed in the plots. The model also generally predicted inadequate fungicide control, even with a 4-day spray interval, which also occurred in the field. Lack of adequate fungicide protection would indicate the need for cultivars with higher levels of durable resistance, and that farmers should consider more effective fungicides applications (higher dosages or different chemistries) if susceptible cultivars are used.

Estimating the level of susceptibility to Phytophthora infestans in potato genotypes.

Resistance and susceptibility are closely related terms but differ in their underlying assumptions and measurement. Standardized methods for determining the level of resistance and susceptibility in potato to Phytophthora infestans, the causal agent of late blight, have traditionally been semiquantitative and are not based on a true interval scale, thus making their use in most mathematical and statistical operations inappropriate. Recently, researchers have attempted to develop interval scales using regression analysis of the direct or transformed area under the disease progress curve (AUDPC). In this article, a similar approach is described based on the relative AUDPC (RAUDPC) of one or two reference cultivars and tested using a data set of field trials involving cultivars with varying levels of susceptibility evaluated in different environments in several countries. The coefficient of variation (CV) among trials of the AUDPC was reduced when the RAUDPC was used and even more so when the RAUDPC was made relative to the RAUDPC of cv. Bintje (RaRAUDPC), which was present in all trials. The RaRAUDPC was used in regression models to estimate scale values for eight potato cultivars in 13 to 15 locations (depending on cultivar). The CVs of scale values measuring variation among sites were similar to those of the RaRAUDPC. Using two cultivars gave a slight improvement in CV, which was statistically significant. The scale developed here has ascending numbers for increasing susceptibility, is simple, and can be constructed as a ratio measure, which permitted the calculation of mean, variance, and CV.

Rainfall Thresholds as Support for Timing Fungicide Applications in the Control of Potato Late Blight in Ecuador and Peru.

Accumulated rainfall thresholds were studied in seven field experiments conducted in Ecuador and Peru for their value in timing applications of fungicide to control potato late blight, caused by Phytophthora infestans. Fungicide regimes based on accumulated rainfall thresholds ranging from 10 to 70 mm were compared with calendar spray schemes of every 5 or 7 days. Very low thresholds of 10 to 20 mm gave similar levels of control and resulted in similar number of sprays as did calendar spraying. However, neither low thresholds nor calendar sprays were effective in protecting susceptible potato genotypes in over half of the experiments. Thresholds of 25 to 50 mm of rainfall led to reduction in the number of sprays needed to protect resistant cultivars but also resulted in high levels of disease on susceptible cultivars. We conclude that timing fungicide sprays based on accumulated rainfall thresholds could be a successful component of integrated management strategies that include cultivars with moderate or high levels of resistance. The simplicity of measuring accumulated rainfall means that the technology can potentially be used by resource-poor farmers in developing countries.

Preemergence Infection of Potato Sprouts by Phytophthora infestans in the Highland Tropics of Ecuador.

Experiments were conducted to determine whether preemergence infection of potato sprouts by Phytophthora infestans occurs in the highland tropics of Ecuador. In three separate experiments in the field, P. infestans was identified on the preemerged sprouts of 49, 5, and 43% of tubers, respectively, which had been removed from soil prior to emergence. Tubers had been planted within 10 m of approximately 300-m2 plots with mature potato plants severely infected with late blight. Infection potential of potato sprouts also was evaluated in the greenhouse by applying 10-ml sporangial suspensions (50 and 250 sporangia/ml) daily for 10 days to the soil surface of pots planted with sprouted seed potato tubers. The daily inoculation rate of 50 sporangia/ml (15.9 × 103 sporangia/m2) resulted in sprout infection in 100% of inoculated pots and roughly corresponded to the sporangial deposition accumulated over 24 h in the field. Deposition had been measured at 1 m from a severely infected potato plot. Our study demonstrated the potential for preemergence infection of potato sprouts by P. infestans in the highlands of Ecuador, where year-round aerial inoculum is present. Preemergence infection is consistent with high levels of disease sometimes seen in recently emerged potato fields. These experiments indicate a need to reconsider disease management approaches.

Simulation of Potato Late Blight in the Andes. I: Modification and Parameterization of the LATEBLIGHT Model.

ABSTRACT LATEBLIGHT, a mathematical model that simulates the effects of weather, host growth and resistance, and fungicide use on asexual development and growth of Phytophthora infestans on potato foliage, was modified so that it can be used in the Andes and, eventually, worldwide. The modifications included (i) the incorporation of improved equations for the effect of temperature on lesion growth rate (LGR) and sporulation rate (SR); (ii) the incorporation of temperature-dependent latent period (LP); and (iii) the use of experimentally measured parameters of LGR, SR, and LP for specific potato cultivars and pathogen lineages. The model was parameterized for three Peruvian potato cultivars (Tomasa, Yungay, and Amarilis) infected with isolates of a new clonal lineage of P. infestans that is currently predominant in Ecuador and Peru (EC-1).

Simulation of Potato Late Blight in the Andes. II: Validation of the LATEBLIGHT Model.

ABSTRACT LATEBLIGHT, a mathematical model that simulates the effect of weather, host growth and resistance, and fungicide use on asexual development and growth of Phytophthora infestans on potato foliage, was validated for the Andes of Peru. Validation was needed due to recent modifications made to the model, and because the model had not been formally tested outside of New York State. Prior to validation, procedures to estimate the starting time of the epidemic, the amount of initial inoculum, and leaf wetness duration were developed. Observed data for validation were from field trials with three potato cultivars in the Peruvian locations of Comas and Huancayo in the department of Junín, and Oxapampa in the department of Pasco in 1999 and 2000 for a total of 12 epidemics. These data had not been used previously for estimating model parameters. Observed and simulated epidemics were compared graphically using disease progress curves and numerically using the area under the disease progress curve in a confidence interval test, an equivalence test, and an envelope of acceptance test. The level of agreement between observed and simulated epidemics was high, and the model was found to be valid according to subjective and objective performance criteria. The approach of measuring fitness components of potato cultivars infected with isolates of a certain clonal lineage of P. infestans under controlled conditions and then using the experimental results as parameters of LATEBLIGHT proved to be effective. Fungicide treatments were not considered in this study.

Qualification of a Plant Disease Simulation Model: Performance of the LATEBLIGHT Model Across a Broad Range of Environments.

ABSTRACT The concept of model qualification, i.e., discovering the domain over which a validated model may be properly used, was illustrated with LATEBLIGHT, a mathematical model that simulates the effect of weather, host growth and resistance, and fungicide use on asexual development and growth of Phytophthora infestans on potato foliage. Late blight epidemics from Ecuador, Mexico, Israel, and the United States involving 13 potato cultivars (32 epidemics in total) were compared with model predictions using graphical and statistical tests. Fungicides were not applied in any of the epidemics. For the simulations, a host resistance level was assigned to each cultivar based on general categories reported by local investigators. For eight cultivars, the model predictions fit the observed data. For four cultivars, the model predictions overestimated disease, likely due to inaccurate estimates of host resistance. Model predictions were inconsistent for one cultivar and for one location. It was concluded that the domain of applicability of LATEBLIGHT can be extended from the range of conditions in Peru for which it has been previously validated to those observed in this study. A sensitivity analysis showed that, within the range of values observed empirically, LATEBLIGHT is more sensitive to changes in variables related to initial inoculum and to weather than to changes in variables relating to host resistance.

Stability in Population of Phytophthora infestans Attacking Tomato in Ecuador Demonstrated by Cellulose Acetate Assessment of Glucose-6-Phosphate Isomerase.

Cellulose acetate electrophoresis was used to examine glucose-6-phosphate (Gpi) isomerase banding patterns of the population of Phytophthora infestans attacking tomato in Ecuador. All but two of 160 sporulating lesions from tomato leaflets collected from 25 tomato fields between January 1998 and March 1999 produced the 86/100 Gpi isozyme electromorph. This isozyme type is characteristic of the US-1 clonal lineage, indicating that no change in the population of P. infestans attacking tomato in Ecuador has occurred since a more exhaustive study was done using isolates collected between 1993 and 1995. The two lesions that produced a different Gpi electromorph in the current study came from a field that was located approximately 80 m from a potato field that had been severely affected by late blight. These two isolates produced a single large band for Gpi with a relative migration distance of 100. This electromorph is characteristic of the clonal lineage EC-1, which was shown previously to be the predominant clonal lineage attacking potato in Ecuador. Therefore, we assume that the two tomato lesions with the EC-1 phenotype were caused by inoculum originating from the potato field. During the current study, 34 infected potato leaflets were collected from five potato fields found in close proximity to blighted tomato fields. All of the potato leaflets produced banding patterns characteristic of EC-1. Our data are consistent with earlier studies indicating that, in Ecuador, tomato and potato are attacked by separate populations of P. infestans, which belong to two different clonal lineages.

A Global Marker Database for Phytophthora infestans.

A marker database was compiled for isolates of the potato and tomato late blight pathogen, Phytophthora infestans, originating from 41 locations which include 31 countries plus 10 regions within Mexico. Presently, the database contains information on 1,776 isolates for one or more of the following markers: restriction fragment length polymorphism (RFLP) "fingerprint" consisting of 23 bands; mating type; dilocus allozyme genotype; mitochondrial DNA haplotype; sensitivity to the fungicide metalaxyl; and virulence. In the database, 305 entries have unique RFLP fingerprints and 258 entries have unique multilocus genotypes based on RFLP fingerprint, dilocus allozyme genotype, and mating type. A nomenclature is described for naming multilocus genotypes based on the International Organization for Standardization (ISO) two-letter country code and a unique number. Forty-two previously published multilocus genotypes are represented in the database with references to publications. As a result of compilation of the database, seven new genotypes were identified and named. Cluster analysis of genotypes from clonally propagated populations worldwide generally confirmed a previously published classification of "old" and "new" genotypes. Genotypes from geographically distant countries were frequently clustered, and several old and new genotypes were found in two or more distant countries. The cluster analysis also demonstrated that A2 genotypes from Argentina differed from all others. The database is available via the Internet, and thus can serve as a resource for Phytophthora workers worldwide.