Identification of Molecular Markers Linked to a Pyrenophora teres Avirulence Gene

ABSTRACT Genetic control of avirulence in the net blotch pathogen, Pyrenophora teres, was investigated. To establish an appropriate study system, a collection of 10 net form (P. teres f. teres) and spot form (P. teres f. maculata) isolates were evaluated on a set of eight barley lines to identify two isolates with differential virulence on an individual host line. Two net form isolates, WRS 1906, exhibiting avirulence on the cv. Heartland, and WRS 1607, exhibiting high virulence, were mated and 67 progeny were isolated and phenotyped for reaction on Heartland. The population segregated in a 1:1 ratio, 34 avirulent to 33 virulent (chi(2) = 0.0, P = 1.0), indicating single gene control of WRS 1906 avirulence on Heartland. Bulked segregant analysis was used to identify six amplified fragment length polymorphism markers closely linked to the avirulence gene (Avr(Heartland)). This work provides evidence that the P. teres-barley pathosystem conforms to the gene-for-gene model and represents an initial step toward map-based cloning of this gene.     

Bacterial wilt symptoms are impacted by host age and involve net downward movement of <Emphasis Type="Italic">Erwinia tracheiphila</Emphasis> in muskmelon

Cucurbit bacterial wilt, caused by Erwinia tracheiphila, is a devastating disease of cucurbit crops in the Midwest and Northeast U.S. Current management of bacterial wilt relies primarily on insecticide applications to control striped and spotted cucumber beetles (Acalymma vittatum and Diabrotica undecimpunctata howardi, respectively), which vector E. tracheiphila. Development of alternative management strategies is constrained by a lack of understanding of bacterial wilt etiology. The impact of host age on rate on symptom development and extent of bacterial movement in the xylem of muskmelon (Cucumis melo cv. Athena) was evaluated following wound inoculation of 2- to 8-week-old plants in growth chamber experiments. Wilting occurred more rapidly in plants after inoculating E. tracheiphila into 2- or 4-week-old plants than 6- or 8-week-old plants. Recovery of viable cells from stem segments revealed that vascular spread of E. tracheiphila was more extensive below than above the inoculation point. These findings provide experimental evidence that host age impacts the rate of symptom development in cucurbit bacterial wilt and that movement of the xylem-inhabiting pathogen E. tracheiphila within muskmelon plants occurs primarily in the downward direction.     

Efficacies of low- to high-volume (960-10 700 litre ha− 1) citrus sprayers for applying petroleum spray oil to control chinese wax scale

Petroleum spray oil (2, 4 and 6% in water) was applied to Valencia orange, Citrus sinensis (L.) Osbeck, for the control of Chinese wax scale, Ceroplastes sinensis del Guercio, using a low-volume ( <2000 litre ha^?1)air-blast (LV AB) sprayer, a low- to high-volume (L-HV) (up to 7000 litre ha^?1) sprayer with four fan-assisted rotary atomiser (FARA) spray heads mounted on a vertical tower, and a high-volume (>7000 litre ha^?1) oscillating boom (HV OB) sprayer. The most effective sprayer was the L-HV FARA sprayer. The most cost-effective treatment was a 20 ml litre^?1 (60 litre oil ha^?1) spray applied at 3000 litre ha^?1 by the L-HV FARA sprayer. It gave mortality equivalent to a standard 20 ml litre^?1, 10 700 litre ha^?1 spray (214 litre oil ha^?1) applied by the HV OB sprayer but with 72% less spray and significantly less oil deposited per cm² of leaf area. Equivalent or significantly (P = 0·05) higher mortality than that given by the 10 700 litre ha^?1 HV OB spray was given by the 40 ml litre^?1, 3000 (120 litre oil ha^?1) and 60 ml litre^?1, 2180 and 3000 litre ha^?1 (130·8 and 180 litre oil ha^?1) L-HV FARA sprays, but the 60 ml litre^?1 sprays deposited more oil per cm² than the 20 ml litre^?1 HV OB spray and were considered to be potentially phytotoxic. The least effective sprayer was the LV AB sprayer, which applied a 60 ml litre^?1 spray (57·6 litre oil ha^?1) at 960 litre ha^?1. Linear relationships were established for Chinese wax scale mortality, transformed using an angular transformation (arcsin proportion), versus log₁₀ spray volume for the 20, 40 and 60 ml litre^?1 sprays applied by L-HV FARA at 1260,2180 and 3000 litre ha^?1, mortality versus log₁₀ μg oil cm^?2 and log₁₀ μg oil versus log₁₀ volume of oil sprayed.     

Effect of Dimethyl Sulfoxide on Cell Cycle Synchronization of Goldfish Caudal Fin Derived Fibroblasts Cells

Several studies have previously been conducted regarding cell cycle synchronization in mammalian somatic cells. However, limited work has been performed on the control of cell cycle stages in the somatic cells of fish. The aim of this study was to determine the cell cycle arresting effects of several dimethyl sulfoxide (DMSO) concentrations for different times on different cell cycle stages of goldfish caudal fin‐derived fibroblasts. Results demonstrated that the cycling cells or control group (68.29%) yields significantly higher (p < 0.05) arrest in G0/G1 phase compared with the group treated for 24 h with different concentrations (0.5%, 1.0% or 1.5%) of DMSO (64.88%, 65.70%, 64.22% respectively). The cell cycle synchronization in the treatment of cells with 1.0% DMSO at 48 h (81.14%) was significantly higher than that in the groups treated for 24 h (76.82%) and the control group (77.90%). Observations showed that treatment of DMSO resulted in an increase in the proportion of cells at G0/G1 phase for 48 h of culture. However, high levels of apoptotic cells can be detected after 48 h of culture treated with 1% concentration of DMSO.     

Infectivity and persistence of an outbreak strain of Salmonella enterica serotype Typhimurium DT160 for house sparrows (Passer domesticus) in New Zealand

AIM: To examine the infective dose, incubation period and disease progression of an isolate of Salmonella enterica serotype Typhimurium definitive type 160 (DT160) originating from a naturally-infected house sparrow (Passer domesticus) during an outbreak of the disease in New Zealand.

METHODS: Thirty-six house sparrows captured from the wild and free of Salmonella spp were divided into six groups of six birds, housed individually, and inoculated orally with phosphate buffered saline (PBS) or 10¹, 10², 10³, 10⁵, 2 × 10⁸ colony forming units (cfu) of the outbreak strain of S. Typhimurium DT160. The birds were observed for 10 days for clinical signs and/or mortality, and faecal samples were collected to determine excretion of S. Typhimurium. The birds were eutha- nised 11 days post-inoculation (p.i.) and a wide range of tissue samples were collected for histopathological examination, and culture and typing of Salmonella spp. Macro-restriction profiling by pulsed-field gel electrophoresis (PFGE) using XbaI was performed for the epidemiological typing of S. Typhimurium DT160 isolates.

RESULTS: Mortality in house sparrows inoculated with S. Typhimurium DT160 was dose-dependent, and 2/6 birds inoculated with ¹⁰⁵ cfu and all six birds inoculated with 2 × 10⁸ cfu died during the study. Infected sparrows displayed few clinical signs, apart from diarrhoea and/or polyuria, fluffed plumage, and sitting on the floor of the cage. Faecal excretion of DT160 occurred briefly in two birds inoculated with 10² cfu and four birds inoculated with 10³ cfu, on most days in five birds inoculated with 10⁵ cfu, and continuously in six birds inoculated with 2 × 10⁸ cfu. DT160 was isolated from the livers of three birds which received 10³ cfu, five birds dosed with 10⁵ cfu, and all six birds given 2 × 10⁸ cfu. Following necropsy, histopathological lesions similar to those seen in the natural disease were observed in the liver or spleen of three birds which received 10³ cfu, and all birds dosed with ≥10⁵ cfu.

CONCLUSION: The results indicate that an isolate of S. Typhimurium DT60 originating from house sparrows in New Zealand is pathogenic to these birds and that the response is dose- dependent. The persistence and excretion of the pathogen may last for at least 10 days. This confirms that sparrows infected with DT160 could be a source of infection to humans and other in-contact animals.     

Effects of iodine methionine on boar sperm quality during liquid storage at 17°C

Microbial environment is one of the important factors that affect the quality of preserved semen. Iodine methionine (IM), participating in the production and activation of metabolic enzymes, is a new type of amino acid chelate. To date, there has been no report to evaluate the effects of IM on boar semen preservation at 17°C. This study was designed to investigate the effects of IM on boar sperm quality and reproductive performance during liquid storage at 17°C and its antibacterial effect. Semen samples collected from six Yorkshire boars were diluted with basic liquid containing different concentrations of IM (0, 20, 40, 80, 160 and 320 μM). Subsequently, sperm motility, plasma membrane integrity and acrosome integrity were determined. After 6 days of preservation, the difference in microbial composition between control group and 80 μM IM group was compared using 16S rDNA sequencing, and the effects of IM on reproductive performance were also compared and analysed between the two groups. The results demonstrated that 20, 40 and 80 μM IM improved boar sperm motility, plasma membrane integrity and acrosome integrity. 80 μM IM was the optimum concentration. Conversely, 160 and 320 μM IM resulted in deleterious consequences to boar sperm quality compared to the control group and other treatment groups (p < .05). After 6 days of preservation, sperm motility, plasma membrane integrity and acrosome integrity were 56.0%, 51.8% and 59.4%, respectively. There was no significant difference in non‐return rate between the two groups (p > .05). But the litter size of 80 μM IM group was significantly higher than that of control group (p < .05). 80 μM IM inhibited proliferation of the phylum Proteobacteria and the genus Staphylococcus as well as Pseudomonas (p < .05). Further studies are required to understand the antibacterial mechanism of IM in liquid‐preserved boar semen.     

The role of IGFBP-5 in mammary gland development and involution

Insulin-like growth factor-I (IGF-I) plays an important role as a survival factor during mammary gland development and remodelling during involution of the mature/lactating mammary gland, and elevated concentrations have been associated with increased risk of breast cancer. The actions of IGF-I are modulated by a family of binding proteins (IGFBPs) and we have shown that IGFBP-5 is associated with cell death in the mammary gland and more recently provided the first evidence that it is causally related to apoptosis of the mammary gland. A transgenic mouse expressing IGFBP-5 on a mammary-specific promoter led to impaired mammary development involving inhibition of IGF-signalling and involving members of the Bcl-2 family. Subsequent studies in vitro and in vivo using exogenous IGFBP-5 treatment have added support to this concept. Although the effects of IGFBP-5 did appear to involve inhibition of IGF action, a role for IGF-independent effects cannot be ruled out. Such IGF-independent effects involve potential interactions with components of the extracellular matrix involved in tissue remodelling including plasminogen activator inhibitor-1 (PAI-1). In addition, intracellular events involving nuclear localisation of IGFBP-5 have been shown to have the ability to inhibit cell proliferation. Thus, IGFBP-5 seems important for regulating both apoptosis and cell proliferation in the mammary gland during development and post-lactation involution.     

Phosphonomycin: structure and synthesis

Synthesis and resolution of the antibiotic phosphonomycin are described. The structure is (-)(IR, 2S)-1,2-epoxypropylphosphonic acid.