Genetic variation among asexual progeny of Phytophthora infestans detected with RAPD and AFLP markers

Genotypic variation among 32 single-zoospore isolates (SZI) of Phytophthora infestans , derived asexually from two hyphal-tip parental isolates (PI-105 and PI-1) of the US-8 genotype, was assessed with 80 random amplified polymorphic DNA (RAPD) primers and 18 amplified fragment length polymorphic DNA (AFLP) primer pairs. In previous investigations, the SZIs from parental isolate PI-105 showed high levels of virulence variability and were differentiated into 14 races, whereas the SZIs from PI-1 showed identical virulence to the parent. The purpose of this investigation was to determine if phenotypic variation observed among SZIs of P. infestans could be detected at the DNA level in these isolates. Polymorphism was detected with 51 RAPD primers and with all 18 AFLP primer pairs in PI-105 SZIs. In SZIs from PI-1, polymorphism was also detected with 25 RAPD primers and 17 AFLP primer pairs. Cluster analysis using the unweighted pair-group method with arithmetic averages (UPGMA) separated the SZIs from parent PI-105 into six virulence groups, 11 RAPD groups and three AFLP groups. Cluster analysis of PI-1 SZIs, which all belong to the same virulence group, differentiated them into four RAPD groups and six AFLP groups. No close correlation among RAPD, AFLP and virulence groups could be established within the two progenies of SZIs. Results of this study suggest that there is a considerable level of inherent genetic variability among SZIs derived asexually from the same parental isolate. The possible mechanisms and implications of this genetic variation are discussed.     

Chloropicrin Soil Fumigation Reduces <Emphasis Type="Italic">Spongospora subterranea</Emphasis> Soil Inoculum Levels but Does Not Control Powdery Scab Disease on Roots and Tubers of Potato

The effect of chloropicrin fumigation on the soil populations of Spongospora subterranea and the development of powdery scab, formation of root galls and tuber yield was investigated in seven field trials conducted in Minnesota and North Dakota. Sixteen potato cultivars, with different levels of susceptibility to disease on roots and tubers, were planted in plots treated with chloropicrin at rates ranging from zero to 201.8 kg a.i. ha^?1. The amount of S. subterranea DNA in soil was determined using qPCR. Bioassays were conducted to further assess the effect of chloropicrin fumigation on root colonization by S. subterranea in two potato cultivars with contrasting disease susceptibility. In the field, chloropicrin applied at rates between 70.1 to 201.8 kg a.i. ha^?1 significantly decreased S. subterranea initial inoculum in soil but increased the amount of disease observed on roots and tubers of susceptible cultivars. The effect of increasing disease was confirmed in controlled conditions experiments. Although the amount of S. subterranea DNA in roots of bioassay plants increased with increasing chloropicrin rates, it remained similar among potato cultivars. Chloropicrin fumigation significantly increased tuber yield which in cultivars such as Shepody and Umatilla Russet were associated with the amount root galls (r = 0.30; P < 0.03). Results of these studies contradict earlier reports on the use of chloropicrin fumigation for the control of powdery scab. Factors other than inoculum level, such as environmental conditions that affect inoculum efficiency and host susceptibility, may be significant contributors to the development of powdery scab and root gall formation.     

Effect of Oil Overlay on Inhibition Potential of Roscovitine in Sheep Cumulus‐Oocyte Complexes

Inhibitors of cyclin‐dependent kinases, as roscovitine, have been used to prevent the spontaneous resumption of meiosis in vitro and to improve the oocyte developmental competence. In this study, the interference of oil overlay on the reversible arrest capacity of roscovitine in sheep oocytes as well as its effects on cumulus expansion was evaluated. For this, cumulus‐oocyte complexes (COCs) were cultured for 20 h in TCM 199 with 10% foetal bovine serum (Control) containing 75 μm roscovitine (Rosco). Subsequently, they were in vitro matured (IVM) for further 18 h in inhibitor‐free medium with LH and FSH. The culture was performed in Petri dishes under mineral oil (+) or in 96 well plates without oil overlay (?) at 38.5°C and 5% CO₂. At 20 and 38 h, the cumulus expansion and nuclear maturation were evaluated under stereomicroscope and by Hoechst 33342 staining, respectively. No group presented cumulus expansion at 20 h. After additional culture with gonadotrophins, a significant rate of COCs from both Control groups (+/?) exhibited total expansion while in both Rosco groups (+/?) the partial expansion prevailed. Among the oocytes treated with roscovitine, 65.2% were kept at GV in the absence of oil overlay while 40.6% of them reached MII under oil cover (p < 0.05). This meiotic arrest was reversible, and proper meiosis progression also occurred in the Control groups (+/?). So, the culture system without oil overlay improved the meiotic inhibition promoted by roscovitine without affecting the cumulus expansion rate or the subsequent meiosis progression.     

Confirmation of the Carolina slate belt as an exotic terrane

An assemblage of Middle Cambrian Atlantic faunal province trilobites has been found in the rocks of the Carolina slate belt near Batesburg, South Carolina. Geologic and paleomagnetic data suggest that the Carolina slate belt and the adjacent Charlotte belt constitute an exotic terrane that was accreted to North America in early to middle Paleozoic time.     

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

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

An approximately 15,000-year record of El Nino-driven alluviation in southwestern ecuador

Debris flows have deposited inorganic laminae in an alpine lake that is 75 kilometers east of the Pacific Ocean, in Ecuador. These storm-induced events were dated by radiocarbon, and the age of laminae that are less than 200 years old matches the historic record of El Nino events. From about 15,000 to about 7000 calendar years before the present, the periodicity of clastic deposition is greater than or equal to 15 years; thereafter, there is a progressive increase in frequency to periodicities of 2 to 8.5 years. This is the modern El Nino periodicity, which was established about 5000 calendar years before the present. This may reflect the onset of a steeper zonal sea surface temperature gradient, which was driven by enhanced trade winds.     

A comparative molecular field analysis study of obtusifoliol 14α-methyl demethylase inhibitors

This report describes the development of a Comparative Molecular Field Analysis (CoMFA) model from a set of obtusifoliol 14α-methyl demethylase (DM) inhibitors to aid in the design of herbicides targeting sterol biosynthesis. CoMFA is a three-dimensional (3-D) quantitative structure–activity relationship (QSAR) method that is useful in the probing of receptor binding sites when experimental structure data are unavailable. Conformational analysis and SAR of some rigid and active analogs were used to build the initial model using the active analog hypothesis. The model was subsequently used to design compounds that retain the active site shape requirements, but incorporate physical properties that favor soil-applied herbicidal action. In addition, a second-generation CoMFA model incorporating the newly designed inhibitors was developed and represents the current understanding of the DM binding site. This model was derived from a pharmacophore developed from two methods, the active analog approach as well as from the Catalyst program. The fact that two independent methods produced a similar pharmacophore strengthens the validity of the model. © 1999 Society of Chemical Industry     

Genetic relationships among populations of Gibberella zeae from barley, wheat, potato, and sugar beet in the upper Midwest of the United States

Gibberella zeae, a causal agent of Fusarium head blight (FHB) in wheat and barley, is one of the most economically harmful pathogens of cereals in the United States. In recent years, the known host range of G. zeae has also expanded to noncereal crops. However, there is a lack of information on the population genetic structure of G. zeae associated with noncereal crops and across wheat cultivars. To test the hypothesis that G. zeae populations sampled from barley, wheat, potato, and sugar beet in the Upper Midwest of the United States are not mixtures of species or G. zeae clades, we analyzed sequence data of G. zeae, and confirmed that all populations studied were present in the same clade of G. zeae. Ten variable number tandem repeat (VNTR) markers were used to determine the genetic structure of G. zeae from the four crop populations. To examine the effect of wheat cultivars on the pathogen populations, 227 strains were sampled from 10 subpopulations according to wheat cultivar types. The VNTR markers also were used to analyze the genetic structure of these subpopulations. In all populations, gene (H = 0.453 to 0.612) and genotype diversity (GD = or >0.984) were high. There was little or no indication of linkage disequilibrium (LD) in all G. zeae populations and subpopulations. In addition, high gene flow (Nm) values were observed between cereal and noncereal populations (Nm = 10.69) and between FHB resistant and susceptible wheat cultivar subpopulations (Nm = 16.072), suggesting low population differentiation of G. zeae in this region. Analysis of molecular variance also revealed high genetic variation (>80%) among individuals within populations and subpopulations. However, low genetic variation (<5%) was observed between cereal and noncereal populations and between resistant and susceptible wheat subpopulations. Overall, these results suggest that the populations or subpopulations are likely a single large population of G. zeae affecting crops in the upper Midwest of the United States.     

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

BACKGROUND: Cercospora leaf spot (CLS), caused by the fungus Cercospora beticola, is the most serious foliar disease of sugar beet (Beta vulgaris L.) worldwide. Disease control is mainly achieved by timely fungicide applications. In 2011, CLS control failures were reported in spite of application of quinone outside inhibitor (QoI) fungicide in several counties in Michigan, United States. The purpose of this study was to confirm the resistant phenotype and identify the molecular basis for QoI resistance of Michigan C. beticola isolates. RESULTS: Isolates collected in Michigan in 1998 and 1999 that had no previous exposure to the QoI fungicides trifloxystrobin or pyraclostrobin exhibited QoI EC₅₀ values of ?0.006 µg mL^?1. In contrast, all isolates obtained in 2011 exhibited EC₅₀ values of > 0.92 µg mL^?1 to both fungicides and harbored a mutation in cytochrome b (cytb) that led to an amino acid exchange from glycine to alanine at position 143 (G143A) compared with baseline QoI‐sensitive isolates. Microsatellite analysis of the isolates suggested that QoI resistance emerged independently in multiple genotypic backgrounds at multiple locations. A real‐time PCR assay utilizing dual‐labeled fluorogenic probes was developed to detect and differentiate QoI‐resistant isolates harboring the G143A mutation from sensitive isolates. CONCLUSION: The G143A mutation in cytb is associated with QoI resistance in C. beticola. Accurate monitoring of this mutation will be essential for fungicide resistance management in this pathosystem. Copyright © 2012 Society of Chemical Industry