Characterization and PCR-based Typing of Xanthomonas campestris pv. vesicatoria from Peppers and Tomatoes in Serbia

During the last two decades bacterial strains associated with necrotic leaf spots of pepper and tomato fruit spots were collected in Serbia. Twenty-eight strains isolated from pepper and six from tomato were characterized. A study of their physiological and pathological characteristics, and fatty acid composition analysis revealed that all of the strains belong to Xanthomonas campestris pv. vesicatoria. Being non-amylolytic and non-pectolytic, pathogenic on pepper but not on tomato, containing lower amounts of fatty acid 15 : 0 ante–iso, the pepper strains were designated as members of the A group of X. campestris pv. vesicatoria. However, the tomato strains hydrolyzed starch and pectate, caused compatible reactions on tomato but not on pepper, had higher percent of 15 : 0 ante–iso fatty acid, and were classified into B phenotypic group and identified as X. vesicatoria. PCR primers were developed which amplified conserved DNA regions related to the hrp genes of different strains of X. campestris pv. vesicatoria associated with pepper and tomato. Restriction analysis of the PCR product resulted in different patterns and enabled grouping of the strains into four groups. When xanthomonads isolated from pepper and tomato in Serbia were analyzed, they clustered into two groups corresponding to the grouping based on their physiological and pathological characteristics. According to the reaction of pepper and tomato differential varieties, the strains from pepper belong to races P7 and P8 and tomato strains belong to the race T2. All strains were sensitive to copper and streptomycin. Advantages and disadvantages of various bacterial spot management practices are discussed.     

Dopamine Antagonist Affects Luteal Function But Not Cyclicity During the Autumn Transition

The autumn transition is characterized by a number of anomalies affecting the luteal phase of the estrous cycle, such as declining progesterone secretion from the corpus luteum and increased rate of failure of luteolysis. Prolactin also decreases. We theorized that these events may be linked, possibly through dopamine. We administered domperidone, a specific dopamine D2 receptor antagonist from September 12 to anestrus or January 15, either daily (n = 8), or once per day for the first 7 days of each month (n = 7). Controls (n = 7) received no treatment. Mean progesterone and prolactin concentrations were compared with summer cycles. Time to entry into anestrus and incidence of spontaneously prolonged luteal activity was compared between groups during the autumn transition. Prolactin declined from June through October equally in all groups. There was no difference between groups in time to anestrus. Progesterone decline was prevented (daily treatment) or delayed for a prolonged period (weekly group) in autumn. The incidence of spontaneously prolonged corpus luteum activity was reduced to summer levels in both domperidone groups compared with the control. We concluded that autumn prolactin decline was not driven through dopaminergic D2 receptor inhibition of pituitary lactotrophs. Sustained progesterone synthesis through the autumn may be the result of action of the D2 receptor antagonist with dopamine receptors on the corpus luteum. Autumnal luteolytic function appears to have a dopamine-influenced component. There does not appear to be a causative relationship between autumnal progesterone and prolactin secretion.     

Owner compliance with short term antimicrobial medication in dogs

Summary: The degree to which dog owners complied with instructions to administer a 5 to 10 day course of antimicrobial medication to their pets was assessed using microprocessor based monitoring devices. Twenty two clients gave an average of 84% of prescribed doses of amoxycillin-clavulanate. No difference was found between twice and thrice daily dosing regimens in the overall percentage of prescribed doses given. However, timing of doses was far from ideal in many cases and only 34% of doses were given within the designated optimum time period. Adherence to desired dosing intervals tended to be better with twice daily than with thrice daily dosing, although the difference was statistically insignificant.     

Low root‐to‐root transmission of a tobamovirus,yellow tailflower mild mottle virus,and resilience of its virions

Tobamoviruses are serious pathogens because they have extremely stable virions, they are transmitted by contact, and they often induce severe disease in crops. Knowledge of the routes of transmission and resilience of tobamovirus virions is essential in understanding the epidemiology of this group of viruses. Here, an isolate of the tobamovirus yellow tailflower mild mottle virus (YTMMV) was used to examine root‐to‐root transmission in soil and in a hydroponic growth environment. Root‐to‐root transmission occurred rarely, and when it occurred plants did not exhibit systemic movement of the virus from the roots to the shoots over a 30‐day period. The resilience of YTMMV virions was tested in dried leaf tissue over time periods from one hour to one year under temperatures ranging from ?80 to 160 °C. Infectivity was maintained for at least a year when incubated at ?80 or 22 °C, or at fluctuating ambient temperatures of 0.8 to 44.4 °C, but incubation under dry conditions at 160 °C for >4 days eliminated infectivity. Exposure of virions to 0.1 m sodium hydroxide or 20% w/v skimmed milk solution for 30 min, treatments recommended for tobamovirus inactivation, did not abolish infectivity of YTMMV.     

High genotypic and virulence diversity in Ilyonectria liriodendri isolates associated with black foot disease in New Zealand vineyards

A total of 57 Ilyonectria liriodendri isolates were identified by a combination of species‐specific PCR and DNA sequencing from a collection of 174 Ilyonectria‐like isolates recovered from 101 diseased grapevine samples. These samples were representative of the national vineyard, comprising material contributed by 49 grape growers across seven grape growing areas. This species was predominant, representing 33% of the recovered isolates, and has been reported as a major pathogen of grapevines in other countries. The genetic diversity of the 57 New Zealand isolates was compared to that of isolates from Australia and South Africa using universally primed polymerase chain reaction (UP‐PCR). A total of 66 informative loci distinguished 52 genotypes, of which five contained up to four clonal isolates. Four main clades were identified in a neighbour‐joining (NJ) tree. The international isolates (Australia and South Africa) were placed in a clade that did not include New Zealand isolates. There was a high level of intra‐ and inter‐vineyard genetic variation indicating the free movement of isolates between regions. A subset of nine isolates from different branches of the NJ tree produced two vegetative compatibility groups and hyphal fusion was observed between non‐self pairings. Pathogenicity tests using isolates from different genetic groups inoculated onto either detached roots or 1‐year‐old potted vines showed variability in virulence; however, no correlations were detected.     

Predicting effective doses for the joint action of two fungicide applications

A function was derived to predict fungicide efficacy when more than one application of a single active ingredient is made to a crop, given parameters describing the dose–response curves of the component single-spray applications. In the function, a second application is considered to act on that proportion of the total pathogen population which was uncontrollable at the time of the first application (represented by the lower asymptote of the dose–response curve for the first treatment), plus any additional part of the population which survived the first application as a result of a finite dose being applied. Data to estimate the single-spray dose–response curve parameters and validate predictions of two-spray programme efficacy were obtained from separate subsets of treatments in four field experiments. A systemic fungicide spray was applied to wheat at a range of doses, at one or both of two times (t₁ and t₂), in all dose combinations. Observed values of the area under the disease progress curve (AUDPC) for septoria leaf blotch (Mycosphaerella graminicola) were used to construct response surfaces of dose at t₁ by dose at t₂ for each culm leaf layer. Parameters were estimated from single-spray and zero-dose treatment data only. The model predicted a high proportion (R² = 71–95%) of the variation in efficacy of the two-spray programmes. AUDPC isobols showed that the dose required at t₂ was inversely related to the dose at t₁, but the slope of the relationship varied with the relative timings of t₁ and t₂ in relation to culm leaf emergence. Isobols were curved, so the effective dose – the total dose required to achieve a given level of disease suppression – was lower when administered as two applications.     

SSR-based genetic linkage analysis of resistance to crown rust (Puccinia coronata f. sp. lolii) in perennial ryegrass (Lolium perenne)

Crown rust (caused by Puccinia coronata f. sp. lolii) is a serious foliar disease of the pasture and turfgrass perennial ryegrass (Lolium perenne). Previous genetic studies have detected both qualitative and quantitative resistance mechanisms, and interpretation of the genetic system is complicated by variation within the sexually reproducing pathogen. Resistant and susceptible parental genotypes of ryegrass were identified using a composite urediniospore population collected from three geographically distinct locations. A two-way pseudo-testcross mapping population was obtained as the F₁ progeny of the pair-cross between ryegrass parental genotypes Vedette₆ and Victorian₉. Both parents showed intermediate resistance against a pathogen population collected in a single geographical zone (Hamilton, Victoria), but in the F₁ population, significant variation for a range of resistance-associated characters was detected. Statistical analysis of phenotypic data suggested a major gene effect, hence bulked segregant analysis with map-assigned simple sequence repeat (SSR) markers was used to scan the genome. A marker showing strong association with resistance was assigned to linkage group (LG) 2 of perennial ryegrass. Analysis of 11 LG2 SSR markers defined an interval between loci xlpssrh03f03 and xlpssrk02e02 as containing the gene or genes (LpPc1) conferring crown rust resistance. Resistance gene determinants were inherited from both parents, with up to 80% of the total phenotypic variation explained by markers segregating from Vedette₆ and up to 26% of the variation explained by markers segregating from Victorian₉. The two contributions together resulted in an additive increase in effect, with fully resistant individuals requiring determinants from both parents. A conserved syntenic relationship was observed with linkage group B of Avena strigosa, which is the location of a cluster of resistance genes to the oat form of crown rust. The implications of this study for marker-assisted selection of disease resistance in perennial ryegrass are discussed.