Simultaneous detection of six RNA plant viruses affecting tomato crops using a single digoxigenin-labelled polyprobe

A polyprobe for the simultaneous detection by non-isotopic molecular hybridisation has been developed to detect any of the following six viruses causing important economic losses in tomato crops: Tomato spotted wilt virus, Tomato mosaic virus, Pepino mosaic virus, Cucumber mosaic virus, Potato Y virus and Parietaria mottle virus. The polyprobe detected all six viruses with similar sensitivity to that obtained using individual riboprobes. In addition, we evaluated the possible use of the tissue-printing as a sample preparation technique applied to routine diagnosis of tomato plants with the polyprobe. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Inheritance and heritability of resistance to citrus leprosis

ABSTRACT The genetic inheritance of resistance to leprosis, the most important viral disease of citrus in Brazil, was characterized through the phenotypic assessment of 143 hybrids resulting from crosses between tangor 'Murcott' (Citrus sinensis x C. reticulata) and sweet orange 'Pêra' (C. sinensis), considered to be resistant and susceptible to the disease, respectively. All plants were grafted onto Rangpur lime (C. limonia) and inoculated with Citrus leprosis virus, cytoplasmic type through the infestation with viruliferous mites, Brevipalpus phoenicis. The experiments were arranged in a completely randomized block design with 10 replicates. Incidence and severity of the disease in leaves and stems as well as plant growth parameters (plant height and stem diameter) were recorded for 3 years after the infestation with the viruliferous mites. The average values of all variables were analyzed using principal component analysis, discriminant factorial analysis, estimation of the clonal repeatability coefficients, and frequency of the distributions of the average values for each measured variable. The principal component analysis resulted in the identification of at least two groups with resistance and susceptibility to leprosis, respectively. About 99% of all hybrids were correctly classified according to the discriminant factorial analysis. The broad-sense heritability coefficients for characteristics associated with incidence and severity of leprosis ranged from 0.88 to 0.96. The data suggest that the inheritance of resistance to leprosis may be controlled by only a few genes.     

Genetic Structure of the Population of Pepino mosaic virus Infecting Tomato Crops in Spain

ABSTRACT The population structure of Pepino mosaic virus (PepMV), which has caused severe epidemics in tomato in Spain since 2000, was analyzed. Isolates were characterized by the nucleotide sequence of the triple gene block and coat protein gene and, for a subset of isolates, a part of the RNA-dependent RNA polymerase gene. The full-length sequence of the genomic RNA of a Solanum muricatum isolate from Peru also was determined. In spite of high symptom diversity, the Spanish population of PepMV mostly comprised highly similar isolates belonging to the strain reported in Europe (European tomato strain), which has been the most prevalent genotype in Spain. The Spanish PepMV population was not structured spatially or temporally. Also, isolates highly similar to those from nontomato hosts from Peru (Peruvian strain) or to isolate US2 from the United States (US2 strain) were detected at lower frequency relative to the European strain. These two strains were detected in peninsular Spain only in 2004, but the Peruvian strain has been detected in the Canary Islands since 2000. These results suggest that PepMV was introduced into Spain more than once. Isolates from the Peruvian and US2 strains always were found in mixed infections with the European tomato strain, and interstrain recombinants were detected. The presence of different strains of the virus, and of recombinant isolates, should be considered for the development of control strategies based on genetic resistance.     

Morbidity and mortality associated with a new mycoplasma species from captive American alligators (Alligator mississippiensis).

Nine of 74 American alligators (Alligator mississippiensis) from a captive Florida herd of 3-4-m-long, 200-350-kg, adult males greater than 30 yr of age died within a 10-day period during 1995. Nonspecific clinical signs included anorexia, lethargy, muscle weakness, paraparesis, bilateral white ocular discharge, and various degrees of periocular, facial, cervical, and limb edema. Pneumonia, pericarditis, and arthritis were found on postmortem evaluation of the spontaneously dead and euthanatized alligators. Rapidly growing mycoplasmas were identified by culture, and mycoplasma nucleotide sequences were identified by polymerase chain reaction testing of fresh lung and synovial fluid from an affected alligator. Culture of banked frozen lung from necropsy specimens and fresh lung and fresh synovial fluid from newly affected alligators confirmed the presence of a new mycoplasma species in seven of eight individuals. Oxytetracycline was administered, but related deaths continued for 6 mo until only 14 of the initial alligators remained. An enzyme-linked immunosorbent assay to detect antibody was developed, and the organism was transmitted experimentally to naive juvenile alligators, although the source of the organism, Mycoplasma sp. (ATCC 700619), has not been identified. The alligator isolate is a novel species in the mycoplasma family because its nucleotide sequence does not match those of over 75 characterized mycoplasma species. Such factors as population density, animal age, and mycoplasmal virulence likely contributed to the course of disease.     

Flow analysis of profile extrusion by a modified cross-sectional numerical method

Flow analysis of profile extrusion is essential for design and production of a profile extrusion die. Velocity, pressure, and temperature distribution in an extrusion die are predicted and compared with the experimental results. A two dimensional numerical method is proposed for three dimensional analysis of the flow field within the profile extrusion die by applying a modified cross-sectional numerical method. Since the cross-sectional shape of the die is varied gradually, it is assumed that the pressure is constant within a cross-sectional plane that is perpendicular to the flow direction. With this assumption, the velocity component in the cross-sectional direction is neglected. The exact cross-sectional shape at any position is calculated based on the geometry of standard cross-sections. The momentum and energy equations are solved with proper boundary conditions at a cross-section and then the same calculation is carried out for the next cross-section using the current calculated values. An L-shaped profile extrusion die is produced and employed for experimental investigation using a commercially available polypropylene. Numerical prediction for the varying cross-sectional shape provides better results than the previous studies and is in good agreement with the experimental results.     

Comparison of anther and isolated microspore cultures in barley. Effects of culture density and regeneration medium

Genetic variation in ratoon growth and cane yieldafter mechanical harvesting under wet conditions wasexamined among 26 sugarcane clones of diverse geneticbackground, including materials closely related toSaccharum spontaneum and some commercialcultivars. The clones were evaluated under a controltreatment (harvesting under dry conditions andallowing the cane to ratoon without furtherdisturbance), and a traffic treatment (sprayirrigation and then driving heavy field equipment overthe cane stubble immediately after harvesting). Averaged over all clones, the traffic treatment had alarge adverse effect on early growth and final ratoonyield. There was significant genotype × treatmentinteraction for early growth and canopy development,but not for final cane yield. There was also a highgenetic correlation between genetic performance underthe different treatments, indicating that selectionfor ratooning after dry harvesting conditions would bealso effective for improving performance after wetharvesting conditions. A sugarcane breeding programspecifically targeting better ratoon performance underwet conditions would be difficult to justify.     

The Economics of Animal Disease Control; B.D. Perry,editor

Tropical Animal Health and Production -     

Effect of storage temperature and water activity on the content and profile of isoflavones, antioxidant activity, and in vitro protein digestibility of soy protein isolates and defatted soy flours

The aim of this work was to study the effect of 1 year of storage at different temperatures and 1 month of storage at different water activities on the content and profile of isoflavones, antioxidant activity, and in vitro protein digestibility of defatted soy flours (DSF) and soy protein isolates (SPI). The storage for up to 1 year, at temperatures from -18 to 42 degrees C, had no effect on the total content of isoflavones, but the profile changed drastically at 42 degrees C, with a significant decrease of the percentage of malonylglucosides with a proportional increase of beta-glucosides. A similar effect was observed for SPI stored at a(w) = 0.87 for 1 month. For DSF, however, there was observed a great increase in aglycons (from 10 to 79%), probably due to the action of endogenous beta-glucosidases. The antioxidant activity decreased after storage at 42 degrees C, but the in vitro protein digestibility did not change.     

Induced spawning of hatchery-raised Brazilian catfish, cachara Pseudoplatystoma fasciatum (Linnaeus, 1766)

In the months of January 2001 and 2002, female cachara Pseudoplatystoma fasciatum were selected during their first and second gonadal maturation (2 years and 7 months old and 3 years and 7 months old, respectively) with an of oocyte diameter of 937.5 μm (82.5% with central nuclei and 17.5% with peripheral nuclei). Nine females in first maturation received two doses of carp pituitary extract (CPE), 0.5 mg/kg and 5.0 mg/kg; seven received two doses of human chorionic gonadotropin (hCG), 5 and 10 IU/g; five received doses of 0.5 CPE mg/kg and 5 hCG IU/g (CPE+hCG); and four received 0.9% saline (saline). Nine females from CPE and seven from hCG presented oocytes with the same diameter at the moment of oocyte release (100% with germinal vesicle breakdown and fertilization rate of 53.44±18.3 and 54.81±11.8%; larvae number of 165,330±94.1 and 158,570±20.6, respectively). The five females from CPE+hCG did not respond to the hormonal treatment. The four females from the saline group did not ovulate. In January 2002, 6 of 15 selected females that were going through the second reproductive cycle received CPE (five received hCG and four received saline), showing oocyte diameters similar to the ones in the first maturation. At stripping, CPE females had an oocyte diameter of 1062.5 μm (the hCG females had oocyte diameters ranging from 937.5 to 1125.0 μm; fertilization rates of 56.08±30.9 and 81.90±17.3%; 364,547±244 and 633,129±190, larvae, respectively). The fertilization rates and larvae number were higher in the second gonad maturation, both for CPE and hCG.