Although only a few specific pigmentation types are allowed within the Hucul horse registry, accurate determination of particular coat colors can be uncertain due to the presence of variation in color shades and segregation of multiple dun dilution variants. Herein, we genotyped the previously identified polymorphisms within two coat color loci TBX3 (T-box 3) and ASIP (Agouti Signaling Protein) in 462 Hucul individuals and compared the genotype predicted phenotypes with observed pigmentation types provided in the Polish Horse Breeders Association database. We identified disagreement between the predicted and recorded coat color in 157 horses (34%). The most common error was misclassification of horses with the nd1/nd1 and nd1/nd2 genotypes, what may be related with the occurrence of some ‘intermediate’ dilution phenotypes in such individuals. We have also proven that the frequency of the dominant dun dilution allele (D) (0.30) is higher than previously predicted by available studbooks. The D allele(s) is easily ‘hidden’ in various phenotypic groups including dark bay and black, therefore we hypothesized that the dun dilution effect itself is not as strongly epistatic in the Hucul horse as described in other horse breeds. This may be the result of an additional genetic modifier suppressing D allele phenotypic effect.

Few studies on herbicide resistance report data to establish unambiguously the correlation between genotype and phenotype. Here we report on the importance of the EPSPS prolyl106 point mutation to serine (P106S) in conferring resistance to glyphosate in a goosegrass population from Davao, Mindanao Island, the Philippines (Davao). Initial rate-response studies showed clear survivors within the Davao population at glyphosate rates that completely controlled the standard sensitive goosegrass population (STD1). Assessment of potential resistance mechanisms identified the presence of P106S mutant individuals in the Davao population. Polymerase chain reaction (PCR) amplification of specific alleles (PASA) analysis established that the mixed-resistant Davao population was comprised of 39.1% homozygous proline wild-type (PP106), 3.3% heterozygous serine mutant (PS106), and 57.6% homozygous serine mutant (SS106) genotypes. Further rate-response studies on plants with a predetermined genotype estimated the Davao SS106 individuals to be approximately 2-fold more resistant to glyphosate compared to Davao PP106 individuals. Extensive analysis at different goosegrass growth stages and glyphosate rates established strong correlation (P < 0.001) between presence of P106S in EPSPS and the resistant phenotype. Importantly, no differences in the pattern of absorbed or translocated 14C–glyphosate were observed between PP106 and SS106 Davao genotypes or Davao and STD1 individuals, suggesting that glyphosate resistance in the Davao population was attributable to an altered target site mechanism. This study demonstrates that whilst P106S in EPSPS confers a moderate resistance level to glyphosate, the mechanism is sufficient to endow glyphosate failure at the recommended field rates.

Previous research revealed that resistance to cloransulam in at least one population of common ragweed was conferred by an altered herbicide target site, specifically, by a tryptophan-to-leucine amino acid substitution at position 574 (W574L) of acetolactate synthase (ALS). In this study, 22 common ragweed populations, several of which were suspected cloransulam resistant, were assayed to determine if the W574L ALS substitution was correlated with resistance to ALS inhibitors. From each population, 16 greenhouse-grown plants were treated with cloransulam, and another 16 were treated with imazamox. Plant dry weights were recorded 20 d after treatment and individual plants were considered resistant if their dry weight exceeded 50% of that of nonherbicide-treated controls. For each herbicide-treated plant, allele-specific primers were used in polymerase chain reactions to determine whether the ALS alleles contained leucine or tryptophan codons at position 574. Of the 352 plants treated with cloransulam, 70 were determined to be resistant, and all but two contained one or more Leu574 alleles. The frequency of imazamox resistance was much higher than that of cloransulam in the populations, with 149 of 352 plants identified as imazamox resistant. However, only about half (80) of the imazamox-resistant plants contained one or more Leu574 alleles. Correlation of imazamox resistance and Leu574 alleles was population dependent. ALS activity assays confirmed that imazamox resistance in plants from at least one population was due to an altered target site, even though plants from that population did not have a W574L substitution. These results lead to the conclusion that a Leu574 allele is the predominant basis for cloransulam resistance in common ragweed; however, other mechanisms of resistance to ALS inhibitors exist in some populations.

A population of common ragweed not controlled by an acetolactate synthase (ALS)-inhibiting herbicide, cloransulam-methyl, was sampled near Dunkirk, IN, the first year of the herbicide's commercialization in 1998. Resistance in the Dunkirk population was confirmed by treating greenhouse-grown seedlings with cloransulam-methyl. ALS activity assays and DNA sequencing were used to identify the resistance mechanism. ALS isolated from plants of the Dunkirk population exhibited an R/S ratio for cloransulam-methyl of >5,000 when compared to ALS activity of populations from Claire City, SD, and V & J Seed Farms. R/S ratios of 4,100 and 110 were observed for two other ALS-inhibiting herbicides, chlorimuron and imazaquin, respectively. DNA sequencing revealed that an inferred leucine for tryptophan substitution at amino acid position 574 in ALS was responsible for the observed herbicide resistance. Additionally, DNA sequencing revealed significant variability among common ragweed ALS alleles. Two fragments of ALS were sequenced from three plants each of the Claire City and Dunkirk populations, totaling 688 nucleotide base pairs, of which 72 were polymorphic.

Acetolactate Synthase- (ALS) inhibiting herbicides are important components for the control of ryegrass species infesting cereal-cropping systems worldwide. Although resistance to ALS herbicides in ryegrasses has evolved more than 25 yr ago, few studies have been dedicated to elucidate the molecular mechanisms involved. To this end, we have investigated the molecular basis of chlorsulfuron, sulfometuron-methyl, and imazapyr resistance in AUS5 and AUS23, two ryegrass populations from Australia. Comparison between whole-plant herbicide assays and DNA sequencing results showed that resistance to the nonmetabolizable herbicide sulfometuron-methyl was associated with four different proline mutations at ALS codon position 197 (P197) in AUS23. In addition to three P197 amino acid changes impacting on the efficacies of the two sulfonylurea herbicides, the tryptophan to leucine target-site mutation at ALS codon position 574 (W574L) was present in AUS5, conferring resistance to both sulfometuron-methyl and imazapyr. The samples were also characterized by non target-site-based resistance impacting on the metabolizable herbicide chlorsulfuron only. Interestingly, compound mutant heterozygotes threonine/serine at ALS position 197, and plants with double mutations at positions 197 and 574 were detected, thus reflecting the ability of this outcrossing species to accumulate mutant alleles. Whole-plant dose-response assays conducted on predetermined wild-type and mutant genotypes originating from the same populations allowed for a more precise estimation of the dominant and very high levels of resistance associated with the proline to serine target-site mutation at ALS codon position 197 (P197S) and W574L mutations. The two highly efficient polymerase chain reaction- (PCR) based derived cleaved amplified polymorphic sequence (dCAPS) markers developed here will allow for quick confirmation of 197 and 574 ALS target-site resistance in ryegrass species field samples and also contribute to identify populations characterized by other likely resistance mechanisms in this important weed species.

Common ragweed and common cocklebur plants were collected at two sites each in Illinois, Minnesota, and Ohio to analyze intraspecific variability of the gene encoding acetolactate synthase (ALS). A 385-nucleotide fragment within the coding sequence of ALS was compared among 24 plants of each of these two species from the six locations. Common ragweed ALS was highly variable, with polymorphisms observed at 48 (12.5%) of the 385 nucleotides among the 24 plants. Despite the numerous nucleotide polymorphisms, only two inferred amino acid polymorphisms were identified. No apparent population structure was suggested by the ALS sequence data, indicating widespread gene flow consistent with the wind-pollinated nature of common ragweed. In contrast to common ragweed, no ALS polymorphisms were identified among the common cocklebur plants used in this study. As a basis for comparing the extremes observed between common ragweed and common cocklebur, ALS intraspecific variability also was investigated in 10 plants each of tall waterhemp and smooth pigweed. Normalized to the number of plants analyzed, the number of nucleotide polymorphisms for both tall waterhemp and smooth pigweed was greater than that in common cocklebur but less than that observed in common ragweed. Information on variability of herbicide target-site genes may be useful in predicting the likelihood for herbicide-resistance development. However, all four of the species investigated in this study have evolved resistance to ALS-inhibiting herbicides, despite the different levels of ALS variability observed.

A number of redroot pigweed and Powell amaranth populations from various locations in Ontario, Canada, have distinct patterns of resistance to the acetolactate synthase–inhibiting herbicides imazethapyr and thifensulfuron. This suggested the presence of diverse ALS gene mutations among these populations. Seven polymerase chain reaction primer pairs were used to amplify the gene to obtain full sequence information and to determine the identity of resistance-conferring mutations. There was a high degree of similarity in the ALS gene of the two species with only five nucleotides and one amino acid differing. A total of four herbicide resistance-conferring mutations were identified in the two species. The Ala122Thr, Ala205Val, and Trp574Leu amino acid substitutions were found in redroot pigweed whereas Ala122Thr, Trp574Leu, and Ser653Thr were detected in Powell amaranth. The pattern of resistance known to be conferred by the mutations concurred with the resistance level observed at the whole plant level. Distinct mutations being found in geographically separated populations suggest that selection for resistance occurred simultaneously in different locations. It reinforces the fact that resistance to ALS inhibitors is easily selected and that growers need to take this into account when formulating weed management strategies.

Genetic polymorphism was determined among 13 isolates of Taenia solium from various regions using PCR-amplified sequences of 2 mitochondrial genes: cytochrome c oxidase subunit 1 and cytochrome b. The 2 phylogenies obtained were similar to each other regardless of the genes examined. The isolates from Asia (China, Thailand, Irian Jaya and India) formed a single cluster, whereas the isolates from Latin America (Mexico, Peru, Ecuador, Bolivia and Brazil) combined with those from Africa (Tanzania, Mozambique and Cameroon) to form an additional cluster. These results and historical data of swine domestication, distribution of pigs and colonization suggest that T. solium was introduced recently into Latin America and Africa from different regions of Europe during the colonial age, which started 500 years ago, and that the tapeworm of another origin independently spread in Asian countries.