Multiyear Regional Evaluation of Foliar Fungicide Applications for Cotton Target Spot Management in the Southeastern United States.

Fungicide programs for managing target spot of cotton caused by Corynespora cassiicola were evaluated over 15 site-years in the southeastern United States between 2014 and 2016. Two cultivars, hypothesized to vary in target spot susceptibility, PhytoGen 499WRF (PHY499) and Deltapine 1137B2RF (DPL1137), and four fungicides (azoxystrobin, flutriafol, pyraclostrobin, pyraclostrobin + fluxapyroxad) plus nontreated control, were compared. Fungicide programs consisted of 1) a single application at first flower or disease onset and 2) the first application followed by a second 14 days later. Treatments were applied in a factorial, randomized complete block design. Target spot onset and severity varied among site-years. Except when severity was low, target spot-associated defoliation was greater on PHY499 than on DP1137. Fungicides delayed disease development and defoliation, but application number had little impact. Based on a meta-analysis of 15 site-years, pyraclostrobin-based applications resulted in a 4 to 6% yield preservation, and yield preservation was greater at site-years with early disease onset and >40% target spot associated defoliation. Results suggest a single well-timed application of a pyraclostrobin-based fungicide reduces defoliation and protects cotton yield at locations with high target spot severity. Additional research is needed to identify risk factors for target spot-associated yield losses in cotton production systems.

First Report of the NA2 Lineage of Phytophthora ramorum from an Ornamental Rhododendron in the Interior of California.

In July 2012, we collected a rhododendron var. Trilby with twig dieback symptoms in the lower canopy, consistent with the disease "ramorum blight" caused by Phytophthora ramorum. The symptomatic plant had been planted a year earlier to replace a dead rhododendron in a landscape setting in Placer County, California (Lat: 39.036216°; Long: -120.999274°), Sierra Nevada foothills at ~600 m elevation. Isolations yielded a culture with a fast growth rate and overall morphology resembling the P. ramorum NA2 lineage described by Ivors et al. (4). DNA was extracted from the culture as described previously (4) and six SSR loci: MS18, MS39, MS43, MS45, MS64, MS145, were amplified (2,4). Allelic patterns were compared with those of three testers from each of the three lineages NA1, NA2, and EU1 known to be present in ornamental plants in North America, and they unambiguously confirmed the isolate belongs to the NA2 lineage of the pathogen. Although the symptomatic plant was confined to a landscape setting, it had been planted in that location for a year, providing a possible source of inoculum for the surrounding area. This is the first report of P. ramorum from the Sierra Nevada eco-region in the interior of California. It is also the first report of a NA2 isolate from a plant outside of commercial nurseries in California. The mating type of the isolate was not determined, but NA2 isolates are normally A2, the same mating type of NA1 isolates. The only other report of a NA2 isolate found outside of a nursery is from Washington State (1). Although there is no evidence the pathogen may have infected other plants, the infected rhododendron was found at a location situated over 100 km east of the closest known infestation (www.sodmap.org). Additionally, this is the first report of the pathogen outside the coast mountain range of California. Because the three lineages are genetically and phenotypically distinct (3), the escape of NA2 or EU1 isolates, both still absent from plants in natural settings, could have significant implications for California ecosystems. This finding highlights that introductions of P. ramorum via ornamental plants are still possible, in spite of current regulations. References: (1) G. Chastagner et al. Phytopathology 101:S32, 2011. (2) P. P. Croucher et al. Biol. Invasions 15:2281, 2013. (3) N. J. Grünwald et al. Trends Microbiol. 20:131, 2012. (4) K. Ivors et al. Mol. Ecol. 15:1493, 2006.

Influence of the host contact sequence on the outcome of competition among aspergillus flavus isolates during host tissue invasion.

Biological control of aflatoxin contamination by Aspergillus flavus is achieved through competitive exclusion of aflatoxin producers by atoxigenic strains. Factors dictating the extent to which competitive displacement occurs during host infection are unknown. The role of initial host contact in competition between pairs of A. flavus isolates coinfecting maize kernels was examined. Isolate success during tissue invasion and reproduction was assessed by quantification of isolate-specific single nucleotide polymorphisms using pyrosequencing. Isolates were inoculated either simultaneously or 1 h apart. Increased success during competition was conferred to the first isolate to contact the host independent of that isolate's innate competitive ability. The first-isolate advantage decreased with the conidial concentration, suggesting capture of limited resources on kernel surfaces contributes to competitive exclusion. Attempts to modify access to putative attachment sites by either coating kernels with dead conidia or washing kernels with solvents did not influence the success of the first isolate, suggesting competition for limited attachment sites on kernel surfaces does not mediate first-isolate advantage. The current study is the first to demonstrate an immediate competitive advantage conferred to A. flavus isolates upon host contact and prior to either germ tube emergence or host colonization. This suggests the timing of host contact is as important to competition during disease cycles as innate competitive ability. Early dispersal to susceptible crop components may allow maintenance within A. flavus populations of genetic types with low competitive ability during host tissue invasion.

Variation in competitive ability among isolates of Aspergillus flavus from different vegetative compatibility groups during maize infection.

ABSTRACT Aspergillus flavus, the primary causal agent of aflatoxin contamination, includes many genetically diverse vegetative compatibility groups (VCGs). Competitive ability during infection of living maize kernels was quantified for isolates from 38 VCGs. Kernels were inoculated with both a common VCG, CG136, and another VCG; after 7 days (31 degrees C), conidia were washed from kernels, and aflatoxins and DNA were extracted from kernels and conidia separately. CG136-specific single-nucleotide polymorphisms were quantified by pyrosequencing; VCGs co-inoculated with CG136 produced 46 to 85 and 51 to 84% of A. flavus DNA from kernels and conidia, respectively. Co-inoculation with atoxigenic isolates reduced aflatoxin up to 90% and, in some cases, more than predicted by competitive exclusion alone. Conidia contained up to 42 ppm aflatoxin B(1), indicating airborne conidia as potentially important sources of environmental exposure. Aflatoxin-producing potential and sporulation were negatively correlated. For some VCGs, sporulation during co-infection was greater than that predicted by kernel infection, suggesting that some VCGs increase dispersal while sacrificing competitive ability during host tissue colonization. The results indicate both life strategy and adaptive differences among A. flavus isolates and provide a basis for selection of biocontrol strains with improved competitive ability, sporulation, and aflatoxin reduction on target hosts.

Sewage and community shower drains are environmental reservoirs of Fusarium solani species complex group 1, a human and plant pathogen.

In two recent studies, clinical isolates in the Fusarium solani species complex (FSSC) were sequenced; one of the most common lineages was FSSC Group 1 (FSSC 1), a phylogenetic species that is synonymous with F. solani f. sp. cucurbitae race 2, a pathogen of cucurbit fruits. FSSC 1 was also identified in sink and shower drains in two hospitals. The environmental sources of FSSC 1 are important for understanding the epidemiology of both human and plant diseases caused by this organism. FSSC 1 was detected in sewage influent at all six tested urban wastewater treatment plants (WWTPs) in California with a concentration ranging from 75 to 413 colony-forming units (cfu) l(-1), a mean of 246 +/- 52 cfu l(-1) and a median of 254 cfu l(-1). During the treatment process, the concentration of FSSC 1 in the solid and liquid fractions diminished. FSSC 1 was detected in five and six of 14 community shower drains by culturing and polymerase chain reaction, respectively, whereas FSSC DNA was detected in all drains. FSSC accounted for 17 +/- 6% (n = 14) of the total fungal DNA in the drains. FSSC 1 was rarely isolated from post-harvest cucurbit fruits and was not found in cucurbit fields in California.

Identification of Fusarium solani f. sp. cucurbitae Race 1 and Race 2 with PCR and Production of Disease-Free Pumpkin Seeds.

Fusarium solani f. sp. cucurbitae causes a fruit rot of cucurbits and is classified into two races that are actually distinct species: F. solani f. sp. cucurbitae race 1 (Fsc1) and race 2 (Fsc2). Because Fsc1 and Fsc2 are not easily distinguished morphologically, we developed a polymerase chain reaction (PCR) assay for rapid identification. Taxon-specific primers were designed from translation elongation factor 1-α sequences. Because clean seed is critical for disease control, we conducted experiments to determine if we could predict whether seed would be infected based on a visual rating of the fruit at harvest. In two trials in commercial pumpkin fields, eight fruit in each of four categories were selected: asymptomatic fruit, mildly infected fruit, severely infected fruit but without lesions extending into the seed cavity, and severely infected fruit with at least one lesion extending into the seed cavity. Isolates from both lesions and seed were identified as Fsc1 based on the PCR assay. No infected seed were recovered from fruit in which the surface was lesion-free or in which a lesion extended less than midway through the fruit flesh. Consequently, a rapid, visual inspection and exclusion of symptomatic fruit should be sufficient to obtain uninfected seed, even in infested fields.

Validation of a sensitive direct assay for melatonin for investigation of circadian rhythms in different species.

The role of melatonin in animals which do not show marked seasonal changes in reproduction is disputed, in part because of the wide variation in reported concentrations. One reason for this may be the difficulties associated with the measurements of low molar concentrations of melatonin and the presence of a wide variety of potentially cross-reacting substances. The availability of a high affinity antiserum has allowed an assay, with low cross-reactivity and good sensitivity, to be established for the direct measurement of melatonin in a wide range of biological fluids, in particular serum, plasma and follicular fluid from man and rat. The high affinity of the antiserum enabled a tritium label of high specific activity to be used, removing the problems associated with the iodination of a small molecular weight compound. Melatonin concentrations in the assay were evaluated by four different methods: UV absorbance, gas chromatography, comparison of the immunoreactive concentrations of the label with the expected concentration by dilution and by comparison with a previously established assay which uses the same antiserum. Melatonin was measured in serum from twelve healthy women over two 24-h periods; eight women with normal menstrual cycles and four taking the contraceptive pill. Concentrations were found to range from 19.8 to 215 pmol/l during the day in both groups. In women with normal menstrual cycles peak concentrations of 513.2 +/- 54.1 (S.E.M.) pmol/l were recorded at 04.00 h, whereas higher concentrations were found in women taking the pill, reaching a peak of 849.12 +/- 21.8 (S.E.M.) pmol/l at 04.00 h.(ABSTRACT TRUNCATED AT 250 WORDS)

Circadian pattern of plasma melatonin concentrations in the marmoset monkey (Callithrix jacchus).

This study describes the concentrations of melatonin in plasma samples taken from marmoset monkeys (Callithrix jacchus) every 4 h over three 24-h periods. A circadian pattern of secretion was apparent, with higher levels recorded at night (20.00-08.00 h) than during the day (08.00-20.00 h) and a peak concentration at 20.00 h. There was a significant difference in the mean day and night concentrations (32.5 ± 4.5 pg/ml versus 49.0 ± 6.9 pg/ml, respectively) with individual concentrations ranging between<10-60 pg/ml in the day and 15-200 pg/ml at night. Circadian plasma melatonin concentrations were similar over the three 24-h periods, in male (n = 3) and female (n = 3) monkeys, and in dominant (cyclic, n = 5) and subordinate (acyclic, n = 4) females. The results show a less pronounced circadian profile in the marmoset than is seen in the human but a similar profile to that in the seasonally breeding rhesus monkey.

Analysis of molecular hydrogen formation on low-temperature surfaces in temperature programmed desorption experiments.

The study of the formation of molecular hydrogen on low-temperature surfaces is of interest both because it enables the exploration of elementary steps in the heterogeneous catalysis of a simple molecule and because of its applications in astrochemistry. Here, we report results of experiments of molecular hydrogen formation on amorphous silicate surfaces using temperature-programmed desorption (TPD). In these experiments, beams of H and D atoms are irradiated on the surface of an amorphous silicate sample. The desorption rate of HD molecules is monitored using a mass spectrometer during a subsequent TPD run. The results are analyzed using rate equations, and the energy barriers of the processes leading to molecular hydrogen formation are obtained from the TPD data. We show that a model based on a single isotope provides the correct results for the activation energies for diffusion and desorption of H atoms. These results are used in order to evaluate the formation rate of H2 on dust grains under the actual conditions present in interstellar clouds. It is found that, under typical conditions in diffuse interstellar clouds, amorphous silicate grains are efficient catalysts of H2 formation when the grain temperatures are between 9 and 14 K. This temperature window is within the typical range of grain temperatures in diffuse clouds. It is thus concluded that amorphous silicates are good candidates to be efficient catalysts of H2 formation in diffuse clouds.

Picoeukaryotic plankton diversity at the Helgoland time series site as assessed by three molecular methods.

We analyzed picoeukaryote assemblages in the German Bight at the Helgoland time series site by sequencing cloned eukaryotic 18S rRNA genes in six genetic libraries plus one library from the Orkney Islands from a cruise of opportunity. The libraries were constructed from environmental samples collected at different periods of the year. The same samples were also analyzed using a fingerprinting technique, single-strand conformational polymorphism (SSCP), and DNA microarrays with class-level oligonucleotide probes. One hundred unique clones were analyzed from each library, thus insuring over 85% coverage of the library. The V4 region of the 18S rRNA gene was sequenced from each of these clones, thus providing the most discrimination among the clones. The nonphotosynthetic picoeukaryotic component dominated over the photosynthetic one and was represented by the ciliates at 45% and group II alveolates at 42%. Prasinophytes dominated the photosynthetic group at 40%, but other picoplankton groups, such as bolidomonads and chrysophytes, were also present. Totally novel groups were found in the cryptomonads and in the dinoflagellates. A new algal group sister to the cryptophyte nuclear gene and the glaucocystophytes was also found. These three groups have been found in other picoeukaryotic planktonic clone libraries. SSCP analyses at closer time intervals suggest that clone libraries should be made at weekly intervals if succession in the picoeukaryotic plankton community is to be monitored accurately. A comparison of annual samples suggests thatthere appears to be an annual cycle with regard to species composition. Microarray analysis supported the clone library data and offered a faster means of community analysis, which can be performed with similar accuracy and with higher throughput for a more in-depth analysis.

Effects of the triazolopyrimidine trapidil on force of contraction, beating frequency and phosphodiesterase I--IV activity in guinea-pig hearts.

The effects of the triazolopyrimidine trapidil (5-methyl-7-diethylamino-s-triazolo [1,5-alpha]pyrimidine, CAS 15421-84-8) on force of contraction, beating frequency and phosphodiesterase (PDE) activity were investigated in isolated preparations from guinea-pig hearts. The effects of 3-isobutyl-1-methylxanthine (IBMX), theophylline and milrinone were studied for comparison. Trapidil exerted a concentration-dependent (1000-3000 mumol(s)/l) positive inotropic effect (EC50 562.4 mumol(s)/l) in guinea-pig papillary muscles. The positive inotropic effect was accompanied by a shortening of the duration of contraction as described for IBMX, or isoprenaline. The efficacy of trapidil was lower than that of IBMX or milrinone. Both agents maximally enhanced force of contraction to a 3fold (milrinone) or even 6fold greater amount (IBMX). The potency of trapidil was almost in the same order of magnitude as that of milrinone. The positive inotropic effect of trapidil is at least partially due to a cyclic adenosine monophosphate (cAMP)-dependent mechanism because carbachol antagonized the increase in force of contraction. Trapidil concentration-dependently but nonselectively inhibited the activities of cAMP PDE isoenzymes I-IV as did theophylline or IBMX. Based on IC50 values (275 mumol(s)/l on the average) trapidil had a potency similar to that of theophylline while IBMX was about one order of magnitude more potent. Regarding the inhibition of PDE III, IBMX was 49fold and milrinone 114fold more potent than trapidil. Trapidil revealed only a marginal positive chronotropic effect. The frequency of spontaneously beating right auricles was increased by 13% at most. Trapidil did not produce any tachyarrhythmias or contractures. It is concluded that the positive inotropic effect of trapidil is mainly due to PDE inhibition.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of isomazole on force of contraction and phosphodiesterase isoenzymes I-IV in nonfailing and failing human hearts.

The phosphodiesterase (PDE) inhibitor isomazole increased the force of contraction to 278.3 +/- 89.1% (n = 7) of the predrug value in ventricular trabeculae carneae isolated from nonfailing human hearts. This effect can be attributed mainly to a PDE III or a combined PDE III/IV inhibition since at the concentration of the maximal positive inotropic effect of isomazole, PDE III and PDE IV were completely inhibited. In explanted failing human hearts (end-stage myocardial failure, NYHA IV), isomazole increased the force of contraction only marginally to 110.1 +/- 10.7% of the predrug value. The lack of a distinct positive inotropic efficacy of isomazole in failing human hearts could not be explained by an impairment of PDE inhibition since the properties of the PDE I-IV isoenzymes separated by DEAE-Sepharose chromatography and the inhibitory effects of isomazole did not differ in both preparations. The positive inotropic effect of the beta-adrenoceptor agonist isoprenaline was also reduced in failing hearts. However, in the presence of isomazole, the diminished positive inotropic effect of isoprenaline was restored to values obtained with isoprenaline alone in nonfailing hearts. Thus, the decreased effect of inotropic drugs like isoprenaline or isomazole in preparations from failing human heart might be explained mainly by a diminished cAMP formation due to a defect in receptor-adenylate cyclase coupling.

Phosphodiesterase inhibition by enoximone in preparations from nonfailing and failing human hearts.

The effects of enoximone (MDL 17043, Perfan, CAS 77671-31-9) on the activities of the phosphodiesterase (PDE) isoenzymes I-IV and on force of contraction were investigated in ventricular preparations isolated from failing (end-stage myocardial failure, NYHA IV) and non-failing human hearts. In both tissues four PDE isoenzymes (PDE I-IV) with similar properties were separated by DEAE-sepharose chromatography. The effects of enoximone on PDE I-IV activities did not differ between non-failing and failing human hearts. As compared to PDE I (IC50 2100 mumol/l) and II (IC50 2900 mumol/l) enoximone is a selective PDE III (cGMP-inhibited PDE, IC50 5.9 mumol/l) and PDE IV (cGMP-insensitive PDE, IC50 21.1 mumol/l) inhibitor. Milrinone, 3-isobutyl-1-methylxanthine (IBMX) and UD-CG 212 Cl, a derivative of pimobendan, were studied in the failing heart for comparison. Milrinone inhibited PDE I-IV activities similar to enoximone, revealing IC50 values for inhibition of PDE III and IV (1.2 and 3.3 mumol/l) which were about two orders of magnitude lower than that of PDE I and II (173 and 306 mumol/l). UD-CG 212 Cl was the most potent (IC50 0.05 mumol/l) and most selective PDE III inhibitor tested (IC50 for PDE I, II and IV were 175, 181 and 40.8 mumol/l, resp.), whereas IBMX inhibited PDE I-IV nonselectively (IC50 15.3, 26.2, 5.6, 5.8 mumol/l, respectively). In trabeculae carneae from nonfailing and failing human hearts enoximone increased force of contraction only marginally by 18.0 +/- 9.1% (n = 8) and 24.5 +/- 8.7% (n = 9) of the predrug value.(ABSTRACT TRUNCATED AT 250 WORDS)