Genetic structure and diversity of wild sorghum populations (<Emphasis Type="Italic">Sorghum</Emphasis> spp.) from different eco-geographical regions of Kenya

Wild sorghums are extremely diverse phenotypically, genetically and geographically. However, there is an apparent lack of knowledge on the genetic structure and diversity of wild sorghum populations within and between various eco-geographical regions. This is a major obstacle to both their effective conservation and potential use in breeding programs. The objective of this study was to assess the genetic diversity and structure of wild sorghum populations across a range of eco-geographical conditions in Kenya. Sixty-two wild sorghum populations collected from the 4 main sorghum growing regions in Kenya were genotyped using 18 simple sequence repeat markers. The study showed that wild sorghum is highly variable with the Coast region displaying the highest diversity. Analysis of molecular variance showed a significant variance component within and among wild sorghum populations within regions. The genetic structure of wild sorghum populations indicated that gene flow is not restricted to populations within the same geographic region. A weak regional differentiation was found among populations, reflecting human intervention in shaping wild sorghum genetic structure through seed-mediated gene flow. The sympatric occurrence of wild and cultivated sorghums coupled with extensive seed-mediated gene flow, suggests a potential crop-to-wild gene flow and vice versa across the regions. Wild sorghum displayed a mixed mating system. The wide range of estimated outcrossing rates indicate that some environmental conditions may exist where self-fertilisation is favoured while others cross-pollination is more advantageous.     

Fine-scale estimation of outcrossing in western redcedar with microsatellite assay of bulked DNA

Western redcedar (Thuja plicata, Cupressaceae) is a self-fertile conifer with a mixed mating system and significant variation for outcrossing among populations. In this paper, we conducted a fine-scale study of mating system variation to identify correlates of outcrossing in natural populations. We examined variation for outcrossing within and among individual trees, and describe a new method to estimate outcrossing using bulked DNA samples. Bulking (assaying DNA tissues from several individuals simultaneously) increases the experimental power without increasing the experimental effort. We sampled 80 trees from four natural populations in southwestern British Columbia. From each tree, we sampled from up to six crown positions (three heights and inner vs outer branches). From each position, two samples of three seedlings each were bulked before DNA extractions. Using four microsatellite loci, we obtained outcrossing rates for each tree and for each of the six crown positions. We found individual tree selfing rates to increase with tree height in all four populations, but selfing rates did not differ among crown positions. The higher selfing rate of larger trees is probably due to their greater proportional contribution to local pollen clouds. Individual tree outcrossing rates ranged from 22 to 100% and the population outcrossing rates from 66 to 78%. Missed alleles due to bulking and the estimation method used both cause a downward bias in outcrossing rates, so that these estimates are probably lower than the actual outcrossing rates. Nevertheless, the trends we observed are not affected by systematic biases of estimation.     

Fine-scale genetic structure enhances biparental inbreeding by promoting mating events between more related individuals in wild soybean (Glycine soja; Fabaceae) populations

Outcrossing between genetically distant individuals in a plant population enhances allelic heterozygosity-an important source for genetic diversity and adaptive evolution. Fine-scale spatial genetic structure (FSGS) can interfere with outcrossing by promoting mating between more related individuals. To test the influence of FSGS on outcrossing, FSGS and outcrossing rates were analyzed with four wild soybean (Glycine soja) populations from different habitats, using simple sequence repeat (SSR) fingerprints. Spatial autocorrelation analysis indicated variable FSGS (15.44-25.87 m) in all four populations. Multilocus mixed-mating analysis of 1605 progeny indicated substantial variation in single-locus outcrossing (T(s) = 6.3-12.6%) although the total outcrossing rates as estimated by multilocus outcrossing (T(m) = 12.8-17%) did not vary significantly among populations. The comparison between FSGS and outcrossing rates demonstrated that strong FSGS with large genetic patch size can enhance biparental inbreeding by promoting mating between more related individuals in a population. The results suggest that patch size management can aid in situ conservation by avoiding formation of strong FSGS and encouraging true outcrossing among individuals.     

Among- and within-population variation in outcrossing rate of a mixed-mating freshwater snail

Mixed-mating animals self-fertilize a proportion of their offspring. Outcrossing rate may covary with the ecological and historical factors affecting the population. Theory predicts that outcrossing is favored when inbreeding depression is high and when individual heterozygosity is important. Self-fertilization is predicted to be favored when costs of male function, or mate finding are high, for example, when empty patches are colonized by few individuals. In this study, we assessed primary (after hatching) and secondary (after juvenile mortality) outcrossing rates of two mixed-mating snail populations. Our purpose was to assess the variation in mating-system parameters and estimate significance of inbreeding depression for secondary outcrossing rate (the realized outcrossing rate of parents that produce the next generation). Secondary outcrossing rate was higher than the primary outcrossing rate in one of the two populations, suggesting considerable inbreeding depression. In the other study population, secondary outcrossing rates were found to increase when initially low, or decrease when initially high, depending on the family. Moderate outcrossing rates were found to be more stable. Parental inbreeding coefficients were close to zero in both populations. Outcrossing rate was much more variable among families in the population with the lower average outcrossing rate, suggesting that individuals differed considerably in their mating system. Our results add to recent studies suggesting that populations of mixed-mating animals may differ in their mating system parameters and expression of inbreeding depression.     

Pollen pool heterogeneity in jack pine (Pinus banksiana Lamb.): a problem for estimating outcrossing rates?

Summary Pollen pool heterogeneity, which violates an assumption of the mixed-mating model, is a major potential problem in measuring plant mating systems. In this study, isozyme markers were used to examine pollen pool heterogeneity in two natural populations of jack pine, Pinus banksiana Lamb., in northwestern Ontario, Canada. Population multilocus estimates of outcrossing rate ranged from 0.83 to 0.95 and differed significantly between populations. Single-tree multilocus outcrossing rates were found to be homogeneous among trees in both populations. Computer simulation studies indicated that a consanguineous pollen pool (pollen gametes related to the mother tree) was capable of biasing population outcrossing estimates downward. Random pollen pool heterogeneity (uncorrelated with maternal genotypes) did not appear to affect population outcrossing estimates in the simulations. Heterogeneity G-tests and Spearman rank tests showed that pollen pool heterogeneity existed in the two natural populations examined; however, it did not have a major effect on population outcrossing estimates, since the consanguineous pollen pool detected was probably a relatively minor component of the outcross pollen pool in both populations. In addition, heterogeneity G-tests were found to be not sensitive in detecting pollen pool heterogeneity caused by consanguineous pollen pool.     

Estimation of in situ mating systems in wild sorghum (Sorghum bicolor (L.) Moench) in Ethiopia using SSR-based progeny array data: implications for the spread of crop genes into the wild

Because transgenic sorghum (Sorghum bicolor L.) is being developed for Africa, we investigated the potential for transgenes to spread to conspecific wild/weedy sorghum populations in Ethiopia, which is considered the centre of origin of cultivated sorghum. In the current study, the extent of outcrossing, and uniparental and biparental inbreeding were investigated in seven wild/weedy sorghum populations collected at elevations ranging from 631 to 1709 m. Based on allele frequency data of 1120 progenies and 140 maternal plants from five polymorphic microsatellite markers, outcrossing rates were estimated using standard procedures. The average multilocus outcrossing rate was 0.51, with a range of 0.31–0.65 among populations, and the family outcrossing rate was in the extreme range of 0 to 100%. The highest outcrossing (t _m?=?0.65) was recorded in a weedy population that was intermixed with an improved crop variety in Abuare (Wello region). It was also observed that the inbreeding coefficient of the progenies (F _p) tend to be more than the inbreeding coefficient of both their maternal parents (F _m) and the level of inbreeding expected at equilibrium (F _eq), which is a characteristic of predominantly outbreeding species. Biparental inbreeding was evident in all populations and averaged 0.24 (range = 0.10–0.33). The high outcrossing rates of wild/weedy sorghum populations in Ethiopia indicate a high potential for crop genes (including transgenes) to spread within the wild pool. Therefore, effective risk management strategies may be needed if the introgression of transgenes or other crop genes from improved cultivars into wild or weedy populations is deemed to be undesirable.     

普通野生稻小种群的交配系统与遗传多样性

小种群的遗传动态是保育遗传学关注的核心问题之一,而种群遗传动态又与交配系统密切相关.普通野生稻(Oryza rufipogon Griff.)是具有重要经济价值的濒危物种,目前其种群规模都较小,研究其小种群交配系统与遗传变异性对普通野生稻的保护具有重要意义.运用7对SSR引物,对采自江西东乡普通野生稻小种群的36份种茎和其中20个家系共计601份子代进行了分析.结果显示:该种群的表观异交率为0.318,多位点法估计(MLTR)的多位点异交率为0.481;50%以上的子代共享亲本,非随机交配明显;东乡普通野生稻种群交配系统属于混合交配类型.比较亲本和子代种群的遗传变异性显示:子代种群比亲本种群遗传变异性更丰富;子代种群的杂合子不足与种群变小自交比例上升有关;而亲本种群杂合子过剩可能与杂合基因型的选择优势有关.这些结果说明创造条件扩大种群规模对普通野生稻的原生境保护显得尤为重要.     

Conservation implications of the mating system of the Pampa Hermosa landrace of peach palm analyzed with microsatellite markers

Peach palm (Bactris gasipaes) is cultivated by many indigenous and traditional communities from Amazonia to Central America for its edible fruits, and is currently important for its heart-of-palm. The objective of this study was to investigate the mating system of peach palm, as this is important for conservation and breeding. Eight microsatellite loci were used to genotype 24 open-pollinated progenies from three populations of the Pampa Hermosa landrace maintained in a progeny trial for genetic improvement. Both the multi-locus outcrossing rates (0.95 to 0.99) and the progeny level multi-locus outcrossing rates (0.9 to 1.0) were high, indicating that peach palm is predominantly allogamous. The outcrossing rates among relatives were significantly different from zero (0.101 to 0.202), providing evidence for considerable biparental inbreeding within populations, probably due to farmers planting seeds of a small number of open-pollinated progenies in the same plot. The correlations of paternity estimates were low (0.051 to 0.112), suggesting a large number of pollen sources (9 to 20) participating in pollination of individual fruit bunches. Effective population size estimates suggest that current germplasm collections are insufficient for long-term ex situ conservation. As with most underutilized crops, on farm conservation is the most important component of an integrated conservation strategy.     

Inferring recent outcrossing rates using multilocus individual heterozygosity: application to evolving wheat populations

Using multilocus individual heterozygosity, a method is developed to estimate the outcrossing rates of a population over a few previous generations. Considering that individuals originate either from outcrossing or from n successive selfing generations from an outbred ancestor, a maximum-likelihood (ML) estimator is described that gives estimates of past outcrossing rates in terms of proportions of individuals with different n values. Heterozygosities at several unlinked codominant loci are used to assign n values to each individual. This method also allows a test of whether populations are in inbreeding equilibrium. The estimator's reliability was checked using simulations for different mating histories. We show that this ML estimator can provide estimates of outcrossing rates for the final generation outcrossing rate (t(0)) and a mean of the preceding rates (t(p)) and can detect major temporal variation in the mating system. The method is most efficient for low to intermediate outcrossing levels. Applied to nine populations of wheat, this method gave estimates of t(0) and t(p). These estimates confirmed the absence of outcrossing t(0) = 0 in the two populations subjected to manual selfing. For free-mating wheat populations, it detected lower final generation outcrossing rates t(0) = 0-0.06 than those expected from global heterozygosity t = 0.02-0.09. This estimator appears to be a new and efficient way to describe the multilocus heterozygosity of a population, complementary to Fis and progeny analysis approaches.     

Mating system of Brazilian Oryza glumaepatula populations studied with microsatellite markers

BACKGROUND AND AIMS: A knowledge of natural populations' breeding systems is important in order to implement in situ and ex situ management and conservation practices. Using microsatellite markers, three Oryza glumaepatula populations from Brazil were studied to determine the breeding system and genetic structure parameters of this species. METHODS: Each population represented by ten families with ten individuals per family was studied using eight microsatellite primers. Families of the Rio Xingu population (XI) were obtained from the greenhouse, whereas families from Rio Solimoes (SO) and Rio Paraguay (PG) were collected from the wild. Amplified products electrophoresed on non-denaturing polyacrylamide gels were visualized with a silver staining procedure. The mating system parameters were analysed based on the mixed mating model (software MLTR) while genetic structure analyses of the three populations and their families were performed using the FSTAT software. KEY RESULTS: The mean numbers of alleles per loci were 2.5, 3.9 and 2.5, respectively for the XI, PG and SO populations. Compared with their families, higher values for the observed heterozygosity and gene diversity were estimated for the parental populations. The subdivision (based on R(ST)) and inbreeding (F(IS)) in the SO and PG populations had similar effects, while inbreeding was the main effect in the families. Multilocus outcrossing rates varied from 0.011 to 0.223 in the three populations, indicating divergence in the outcrossing rates among O. glumaepatula populations. For the species (considering SO and PG populations together) an intermediate value was observed (tm = 0.116). Biparental inbreeding varied from 0.008 to 0.123, contributing to the selfing rate in these populations. More than 50 % of the outcrossing occurred between related individuals. CONCLUSIONS: The results indicated divergence in the mating system among O. glumaepatula populations, with consequences for conservation practices. The mating system of this species was classified as mixed with a predominance of self-fertilization.     

Flexible mating system in a logged population of <Emphasis Type="Italic">Swietenia macrophylla</Emphasis> King (Meliaceae): implications for the management of a threatened neotropical tree species

Microsatellites were used to evaluate the mating system of the remaining trees in a logged population of Swietenia macrophylla, a highly valuable and threatened hardwood species, in the Brazilian Amazon. A total of 25 open pollinated progeny arrays of 16 individuals, with their mother trees, were genotyped using eight highly polymorphic microsatellite loci. Genotypic data analysis from the progeny arrays showed that 373 out of the 400 seedlings (93.25%) were unambiguously the result of outcrossed matings and that the remaining 6.75% had genotypes consistent with self-fertilisation. Apomixis could be ruled out, since none of the 400 seedlings analysed had a multi-locus genotype identical to its mother tree. The high estimate of the multi-locus outcrossing rate (t _m = 0.938 ± 0.009) using the mixed mating model also indicated that the population in this remnant stand of S. macrophylla was predominantly allogamous. The relatively large difference between the multi-locus and single-locus outcrossing estimates (t _m −t _s = 0.117 ± 0.011) provides evidence that, in spite of the high outcrossing rate, a considerable degree of biparental inbreeding has contributed to the genetic structure of this population. Levels of outcrossing were not evenly distributed among maternal trees (t _m ranging from 0.39 to 1.00), suggesting the occurrence of a variable degree of self-incompatibility and/or dichogamy among individual trees of this monoecious species. Due to its generalist pollination system and some level of tolerance for selfing, S. macrophylla seems to be resilient to environmental disturbances such as those caused by logging, since it sets fruits with predominantly outcrossed seeds even at low stand densities. Therefore, the remaining individuals in logged areas or in relict fragments may be very important for long-term population recovery and genetic conservation programmes.     

Mating system of wild Phaseolus lunatus L. and its relationship to population size

Using isozyme variation in a naturally pollinated seed family for 10 wild Phaseolus lunatus L. (Lima bean) populations, ranging in sizes from 10 to 60 reproductive individuals, we estimated levels of outcrossing (t) and parental inbreeding coefficient (F). We also examined the relationship between outcrossing rate and population size. Average estimates of the single-locus outcrossing rate (ts) ranged from 0.024 to 0.246 (mean=0.091+/-0.065). Estimates of the multilocus outcrossing rate (tm) ranged from 0.027 to 0.268, and averaged 0.096+/-0.071. Inbreeding coefficients based on genotypic frequencies of maternal plants were positive and significantly greater than zero (F=0.504), suggesting an excess of homozygotes in all the populations studied. There was indirect evidence of nonrandom mating for outcrosses and this was mainly attributed to self-fertilisation since the averaged difference between tm and ts, which provides a measure of biparental inbreeding, represents only 1% of the autogamy rate. No significant correlation was observed between outcrossing rate and population size. Estimates of t showed significant heterogeneity among populations and factors explaining this tendency are suggested.     

The interactive effects of herbivory and mixed mating for the population dynamics of Impatiens capensis

In this study, we examine the demographic consequences of mixed mating and explore the interactive effects of vegetative herbivory and mating system for population dynamics of Impatiens capensis, a species with an obligate mixed mating system (i.e., individuals produce both obligately selfing cleistogamous and facultatively outcrossing chasmogamous flowers). In two natural populations, we followed seeds derived from cleistogamous and chasmogamous flowers subject to different herbivory levels throughout their life cycle. Using a mating system-explicit projection matrix model, we found that mating system types differed in important vital rates. Cleistogamous individuals had higher rates of germination than did chasmogamous individuals, whereas chasmogamous individuals expressed a fecundity advantage over cleistogamous individuals. In addition, population growth was most sensitive to changes in vital rates of cleistogamous individuals, indicating the demographic importance of selfing for these populations. Herbivory also had demographic consequences; a 33%-49% reduction in herbivory caused the population growth rates to increase by 104%-132%, primarily because of effects on vital rates of selfed individuals. Our results not only uncover a novel consequence of mating system expression, that is, mating system influences population dynamics, but also shed light on the role of herbivores in maintaining mixed mating.     

The mating system of Hydrophyllum appendiculatum,a protandrous species

Summary The mating system of Hydrophyllum appendiculatum (Hydrophyllaceae), a protandrous, self-compatible, monocarpic plant was examined using progeny arrays assayed at three polymorphic allozyme loci. We were particularly interested in the effect of ecological factors on spatial and temporal variation in outcrossing rates. Multilocus estimates of outcrossing rates in three populations ranged from 0.62–0.81 indicating that the majority of seeds are produced via outcrossing. The population estimates did not differ significantly from each other indicating that there is little or no spatial variation in the mating system of this species. The estimates were, however, significantly less than unity, which demonstrates that although mainly outcrossing, a significant fraction of seeds are produced by selfing. Estimates suggested that biparental inbreeding occurred, although it was statistically significant in only one population. Individuals of H. appendiculatum may remain in anthesis 3–4 weeks and produce up to 30 inflorescences. As a result, the possibility exists for the mating system to vary through the flowering season. Although the mean outcrossing rate was highest in the middle of the flowering phenology than at the beginning or end, there was no evidence for statistically significant temporal heterogeneity. We were also interested in determining if the size of the floral display (number of inflorescences produced by a plant) influenced the outcrossing rate. The results obtained by two different statistical analyses were contradictory; there was a significant positive correlation between size of floral display and outcrossing rate, but the outcrossing rates of large plants (\s> 8 inflorescences) did not differ significantly from small plants (> 8 inflorescences).     

Characterization of 24 microsatellite markers in Primula chungensis(Primulaceae),a distylous-homostylous species,using MiSeq sequencing

Primula chungensis is a species with considerable floral and mating-system variation,including distylous(outcrossing),homostylous(selfing) and mixed populations that contain both outcrossing and selfing forms.We isolated 24 microsatellite markers from P.chungensis using Illumina Mi Seq sequencing.Polymorphism and genetic diversity were then measured based on a sample of 24 individuals from a natural population in southern Tibet.All loci were polymorphic with the number of alleles per locus ranging from 2 to 4.The observed and expected heterozygosity ranged from 0 to 1 and 0.219 to 0.708,respectively.The microsatellite markers we have identified will serve as valuable tools for the investigation of the population genetic structure and phylogeography of P.chungensis and will inform models of the evolutionary history of mating systems in the species.     

Characterization of 24 microsatellite markers in Primula chungensis (Primulaceae), a distylous homostylous species,using MiSeq sequencing

Primula chungensis is a species with considerable floral and mating-system variation, including distylous (outcrossing), homostylous (selfing) and mixed populations that contain both outcrossing and selfing forms. We isolated 24 microsatellite markers from P.chungensis using Illumina MiSeq sequencing. Polymorphism and genetic diversity were then measured based on a sample of 24 individuals from a natural population in southern Tibet. All loci were polymorphic with the number of alleles per locus ranging from 2 to4. The observed and expected heterozygosity ranged from 0 to 1 and 0.219 to 0.708, respectively. The microsatellite markers we have identified will serve as valuable tools for the investigation of the population genetic structure and phylogeography of P.chungensis and will inform models of the evolutionary history of mating systems in the species.     

Comparison of mating system parameters and genetic structure in three natural scenarios of Acacia visco (Leguminosae, Mimosoideae)

Acacia visco is a native South American tree species that has been extensively used for ornamental purposes and in carpentry, bodywork and parquet due to the hardness and durability of its wood. Little is known about genetic diversity and mating system of A. visco. The main aims of this study were to (1) estimate outcrossing rates in natural Argentinean populations using AFLP markers, (2) test for any difference in mating patterns among a large a patchy and relict population, and (3) compare the mating system of A. visco with other Acacia species. The three primer pairs used in the AFLP analysis revealed a total of 569 variable loci. Most genetic variation was observed among individuals within families (61.2 %). The estimate of multilocus outcrossing rate (t _m) was high (≥0.971) in all populations. Average pairwise coancestry between progenies within families for each population ranged from 0.082 to 0.105 or from 0.125 to 0.136, depending on the method θ was estimated. In the three populations studied, the progenies of open pollination were constituted mainly for half-sibs (94.3 %). This work shows a similar mating system in all populations of A. visco in spite of their size differences, hypothesizing that the entire species has a similar mating system of outcrossing preferential. Considering the results obtained here where a high percentage of individuals were half-sibs, sampling large numbers of pods from individual trees for ex situ conservation will result in a genetically diverse sample as a consequence of high outcrossing rates.     

Mixed mating in natural populations of the chestnut blight fungus, Cryphonectria parasitica

As in plants, fungi exhibit wide variation in reproductive strategies and mating systems. Although most sexually reproducing fungi are either predominantly outcrossing or predominantly selfing, there are some notable exceptions. The haploid, ascomycete chestnut blight pathogen, Cryphonectria parasitica, has previously been shown to have a mixed mating system in one population in USA. In this report, we show that both selfing and outcrossing occur in 10 additional populations of C. parasitica sampled from Japan, Italy, Switzerland and USA. Progeny arrays from each population were assayed for segregation at vegetative incompatibility (vic) and DNA fingerprinting loci. Outcrossing rates (t(m)) were estimated as the proportion of progeny arrays showing segregation at one or more loci, corrected by the probability of nondetection of outcrossing (alpha). Estimates of t(m) varied from 0.74 to 0.97, with the lowest rates consistently detected in USA populations (0.74-0.78). Five populations (four in USA and one in Italy) had t(m) significantly less than 1, supporting the conclusion that these populations exhibit mixed mating. The underlying causes of variation in outcrossing rates among populations of C. parasitica are not known, but we speculate that--as in plants--outcrossing is a function of ecological, demographic and genetic factors.     

A novel mating system analysis for modes of self-oriented mating applied to diploid and polyploid arctic Easter daisies (Townsendia hookeri)

We have developed a new model for mating system analysis, which attempts to distinguish among alternative modes of self-oriented mating within populations. This model jointly estimates the rates of outcrossing, selfing, automixis and apomixis, through the use of information in the family structure given by dominant genetic marker data. The method is presented, its statistical properties evaluated, and is applied to three arctic Easter daisy populations, one consisting of diploids, the other two of tetraploids. The tetraploids are predominantly male sterile and reported to be apomictic while the diploids are male fertile. In each Easter daisy population, 10 maternal arrays of six progeny were assayed for amplified fragment length polymorphism markers. Estimates, confirmed with likelihood ratio tests of mating hypotheses, showed apomixis to be predominant in all populations (ca. 70%), but selfing or automixis was moderate (ca. 25%) in tetraploids. It was difficult to distinguish selfing from automixis, and simulations confirm that with even very large sample sizes, the estimates have a very strong negative statistical correlation, for example, they are not independent. No selfing or automixis was apparent in the diploid population, instead, moderate levels of outcrossing were detected (23%). Low but significant levels of outcrossing (2-4%) seemed to occur in the male-sterile tetraploid populations; this may be due to genotyping error of this level. Overall, this study shows apomixis can be partial, and provides evidence for higher levels of inbreeding in polyploids compared to diploids and for significant levels of apomixis in a diploid plant population.     

Genetic diversity of <Emphasis Type="Italic">Phytophthora capsici</Emphasis> recovered from Massachusetts between 1997 and 2014

Phytophthora blight caused by Phytophthora capsici limits the production of cucurbits and peppers in the United States and is a growing threat to sustainable vegetable production in New England. Little is known about the genetic diversity of P. capsici in New England, and a total of 210 P. capsici isolates from 18 sites were genotyped using 46 single nucleotide polymorphism markers, revealing 85 unique and 34 repeated multi-locus genotypes. Both mating types were recovered from 7 of the 18 locations. Isolates with identical genotypes (clonal lineages) ranged from 2 to 16. Three clonal lineages were recovered from multiple sites within the same year, although none were recovered across multiple years. Bayesian clustering revealed individuals with a complex genetic cluster composition. This, coupled with a high outcrossing rate (mean t = 0.87) and no clear clustering in principal coordinates analysis, suggests outcrossing among the populations. Phylogenetic and genetic distance analysis indicate differentiation based on farm location and movement among farms may be infrequent. There was no obvious differentiation based on cucurbit, tomato or pepper hosts.