Distribution and molecular characterization of Corynespora cassiicola isolates resistant to boscalid

A total of 618 isolates of corynespora leaf spot fungus (Corynespora cassiicola) collected from 24 commercial cucumber greenhouses in 12 cities in Ibaraki Prefecture, Japan, were tested for their sensitivity to boscalid. Boscalid‐resistant isolates were detected in 17 out of 19 greenhouses with a history of use of this fungicide and detection frequencies of the resistant isolates exceeded 47% in nine greenhouses. Frequencies of very highly resistant (VHR) isolates with 50% effective concentration (EC₅₀) values of boscalid exceeding 30 μg mL^?1 were higher than those of moderately resistant (MR) isolates with EC₅₀ ranging from 2·0 to 5·9 μg mL^?1 in 11 greenhouses. Additionally, highly resistant (HR) isolates with EC₅₀ from 8·9 to 10·7 μg mL^?1 were first detected. Furthermore, molecular characterization of genes encoding succinate dehydrogenase (SDH) subunits (SdhA, SdhB, SdhC and SdhD) was carried out to elucidate the amino acid substitution responsible for the resistance to boscalid. All 23 VHR isolates had the same mutation from CAC to TAC in the SdhB gene leading to the substitution of histidine with tyrosine at amino acid position 278 (B‐H278Y). At the same position, the substitution to arginine conferred by a mutation to CGC (B‐H278R) was detected in all four HR isolates. Some MR isolates showed a substitution from serine to proline at position 73 in SdhC (C‐S73P), from serine to proline or from glycine to valine at position 89 (D‐S89P) and 109 (D‐G109V), respectively, in SdhD. There was no common mutation in SDH genes of all MR isolates.     

Occurrence of boscalid resistance in cucumber powdery mildew in Japan and molecular characterization of the iron–sulfur protein of succinate dehydrogenase of the causal fungus

A total of 74 mass isolates of cucumber powdery mildew fungus (Podosphaera xanthii) were collected from commercial greenhouses with a history of boscalid use in Ibaraki Prefecture, Japan, and tested in a leaf disk assay for their sensitivity to boscalid. The mildew development of 40 of 74 isolates and five sensitive reference isolates on the disks was completely suppressed at 5 μg boscalid/ml. The minimum inhibitory concentrations (MIC) for the remaining 34 isolates were 50 μg/ml or higher, and 21 of these isolates also grew well at 500 μg/ml. Six single-spore isolates were resistant to boscalid with MIC values higher than 500 μg/ml; four of these were moderately resistant (MR), and two were very highly resistant (VHR) isolates. The growth of MR isolates was almost completely suppressed at 500 μg/ml, whereas two VHR isolates grew vigorously at 500 μg/ml. In foliar inoculation tests of potted cucumber plants, the efficacy of boscalid (500 μg/ml) against both MR and VHR isolates was very low. Partial DNA fragment of the iron–sulphur protein subunit (SdhB) gene of succinate dehydrogenase was PCR-amplified from five sensitive and five resistant isolates and directly sequenced, revealing that VHR isolates possess a substitution from a highly conserved histidine (CAT) to tyrosine (TAT) in a third cysteine-rich center of a putative SdhB, whereas MR isolates so far have not been found to have such substitution in SdhB.     

Analysis of Host Species Specificity of Magnaporthe grisea Toward Foxtail Millet Using a Genetic Cross Between Isolates from Wheat and Foxtail Millet

ABSTRACT Host species specificity of Magnaporthe grisea toward foxtail millet was analyzed using F(1) cultures derived from a cross between a Triticum isolate (pathogenic on wheat) and a Setaria isolate (pathogenic on foxtail millet). On foxtail millet cvs. Beni-awa and Oke-awa, avirulent and virulent cultures segregated in a 1:1 ratio, suggesting that a single locus is involved in the specificity. This locus was designated as Pfm1. On cv. Ki-awa, two loci were involved and one of them was Pfm1. The other locus was designated as Pfm2. Interestingly, Pfm1 was not involved in the pathogenic specificity on cv. Kariwano-zairai. These results suggest that there is no "master gene" that determines the pathogenic specificity on all foxtail millet cultivars and that the species specificity of M. grisea toward foxtail millet is governed by cultivar-dependent genetic mechanisms that are similar to gene-for-gene interactions controlling race-cultivar specificity.     

Apoptosis of bovine neutrophils during mastitis experimentally induced with Escherichia coli or endotoxin

OBJECTIVE: To determine whether apoptosis of neutrophils was accelerated during mastits experimentally induced by use of Escherichia coli or E coli endotoxin and whether differences were apparent in the response to E coli or endotoxin. ANIMALS: 11 healthy lactating Holstein cows. PROCEDURE: Blood samples were collected from cows at various intervals after intramammary inoculation with E coli or endotoxin. Percentage of apoptotic neutrophils detected after in vitro incubation for 3 hours was determined. Fluorescein isothiocyanate-labeled annexin-V in combination with propidium iodide was used to distinguish apoptosis and necrosis of neutrophils. Total and differential circulating leukocyte counts and rectal temperature were determined at the time of collection of blood samples. Milk yield and milk somatic cell counts were determined at the time of milking. RESULTS: Inoculation of endotoxin did not accelerate in vitro induction of neutrophil apoptosis. However, inoculation of E coli increased the percentage of apoptotic neutrophils. At 18 hours after inoculation, 20% of the neutrophils were apoptotic, compared with 5% before inoculation. Milk somatic cell count and rectal temperature increased, milk production and total leukocyte count decreased, and percentage of immature neutrophils increased after inoculation with E coli or endotoxin. However, kinetics of the responses were more rapid, more severe, and of shorter duration during endotoxin-induced mastitis. CONCLUSIONS AND CLINICAL RELEVANCE: In vitro induction of apoptosis of neutrophils was accelerated only during E coli-induced mastitis and not during endotoxin-induced mastitis. Endotoxin inoculation as a model for studying coliform mastitis in dairy cows should be viewed with caution.     

Genetic parameters for somatic cell count (SCC) and milk production traits of Guzerá cows using data normalized by different procedures

Tropical Animal Health and Production - This study aimed to estimate the genetic parameters for somatic cell count (SCC) and the genetic association between SCC and milk production traits using two...     

Genetic parameters for milk,growth, and reproductive traits in Guzerá cattle under tropical conditions

Tropical Animal Health and Production - This study aimed to estimate the genetic parameters of milk (305-day milk yield (MY305)), growth (weaning weight (WW), yearling weight (YW), and weight at...     

Bioelectric phenomena related to protein-fixed charge in a crab nerve fiber

The bioelectrical characteristics of a crab nerve fiber subjected to solutions containing the impermeant anions ferrocyanide, glutamate, or ethanolsulfate substituted for chloride include prolonged slow depolarizations that elicit prolonged trains of impulses sulperimposed on the plateau portion. Propionate and nitrate depress repetitive firing while thiocyancite has only a slighit effect. Nerves treated with ferrocyanide lose potassium and gain sodium, which fact may account for their depolarization. On the other hand, studies with interference microscopy reveal shift by ferrocyanide in the interference fringe pattern of the sheath material; this suggests a reorientation of fixed charges in the protein layers by direct action of the anion on these layers. This may also account for the electrical manifestations observed.