Emergence and Partial Characterization of Rice Stripe Necrosis Virus and its Fungus Vector in South America

A new disease of rice, known as entorchamiento (crinkling), was first noticed in the Department of Meta, Colombia, in 1991. Symptoms include seedling death, foliar striping and severe plant malformation. Tissue extracts and purified preparations from diseased rice plants, contained virus-like particles ca. 20nm in diameter, with a bimodal length of 260 and 360nm. Particle aggregates were also observed in the cytoplasm of infected rice leaf cells. Electrophoretic analyses of purified preparations and dsRNA extracts, revealed a single protein species of M 22,500, and four dsRNA bands ca. 6300, 4600, 2700 and 1800bp in size. Cystosori, characteristic of plasmodiophorid fungal vectors of plant viruses, were consistently observed in the roots of diseased rice plants. PCR and sequence analyses of amplified fungal DNA products from infected rice roots, revealed that the putative fungus vector was Polymyxa graminis. A Western blot of tissue extracts obtained from crinkled rice plants from Colombia, using antiserum against a West African isolate of rice stripe necrosis furovirus (RSNV), resulted in the detection of a protein band of approximately M 22,000. The RSNV antiserum recognized the Colombian virus isolate in serologically specific electron microscopy tests. These results confirm the presence of RSNV in the Americas.     

Detection of circulating hypodermin C: an antigen capture ELISA for diagnosis of cattle grub (Diptera: Oestridae) infestations

An antigen capture assay for the detection of circulating hypodermin C was developed for diagnosis of hypodermosis. A murine monoclonal antibody to recombinant hypodermin C was raised using rapid immunization and a one-step hybridization-cloning technique. A highly reactive, specific monoclonal antibody was tested using sera spiked with known quantities of purified, native hypodermin C or with recombinant hypodermin C. Sensitivity of 96.4% and specificity of 95.6% for the antigen capture assay was assessed using a panel of sera from animals unexposed to cattle grubs and from cattle with palpation proven cattle grub infestations. Data from this panel of sera was used to establish the cut-off OD for further testing. The kinetics of circulating hypodermin C was assessed using the assay in three groups of cattle artificially infested with 50, 100 or 200 first instar Hypoderma lineatum. Antigen was first detected approximately 6 weeks after infestation. The amount of antigen detected increased in each group of animals reaching peaks at different times in each group. Levels of antigen fell quickly following arrival of grubs at the back and completion of the molt to second instar.     

Comparison of three monoclonal and three polyclonal antibodies in the immunohistochemical diagnosis of canine autoimmune skin diseases

The aim of this study was to evaluate the utility of three monoclonal antibodies (mAbs), and two anticanine IgG and one anticanine IgM polyclonal antibodies (pAbs) for the immunohistochemical diagnosis of canine autoimmune skin diseases. Skin biopsies from 11 cases of pemphigus (7 foliaceus, 3 vulgaris and 1 erythematosus), 12 cases of discoid lupus erythematosus (DLE) and 12 cases of chronic hyperplasic dermatitis were used. The CA4E7 mAb (IgG1 + IgG2) showed similar sensitivity, but higher specificity and lower background than the two anti-IgG pAbs for the immunohistochemical diagnosis of pemphigus and DLE. The CA4F1 mAb (IgG2) and CA3H1 mAb (IgG2) showed moderate and low interepithelial reactivity, respectively, in autoimmune skin diseases, but strong staining of the cytoplasm of plasma cells of the inflammatory infiltrates. These results suggest that the CA4E7 mAb may be valuable in the immunohistochemical diagnosis of such disorders.     

Copper Resistance in Pseudomonas syringae Strains Isolated from Mango Is Encoded Mainly by Plasmids

ABSTRACT Bacterial apical necrosis of mango, elicited by Pseudomonas syringae pv. syringae, limits fruit production in southern Spain and Portugal. Examination of a collection of P. syringae pv. syringae isolates for copper resistance showed that 59% were resistant to cupric sulfate. The survey of a mango orchard revealed an increase in frequencies of copper-resistant bacteria after repeated treatments with Bordeaux mixture. These data suggest that selection of copper-resistant strains could be a major reason for control failures following management with copper bactericides. Most copper-resistant isolates harbored plasmids, although the majority of them contained a 62-kb plasmid that also was present in copper-sensitive strains. The 62-kb plasmids were differentiated by restriction enzyme analysis and hybridization to copABCD DNA. The most frequently found copper-resistant plasmid type (62.1) was transferable by conjugation. Southern blot hybridizations showed that genetic determinants partially homologous to copABCD were present in all the copper-resistant strains examined, and usually were associated with plasmids; these determinants were not detected in copper-sensitive strains. The selective pressure exerted by copper bactericide sprays on the diversity of copper resistance determinants in bacterial populations of mango is discussed.     

Brassinosteroid analogue effects on the yield of yellow passion fruit plants (Passiflora edulis f. flavicarpa)

The effects of successive brassinosteroid analogue (BR) applications (0.1 mg l⁻¹ of brassinosteroid analogue BB-16) were evaluated on commercial yellow passion fruit (Passiflora edulis f. flavicarpa) orchards in the first year of production. The treatments applied were: control, BR-1 (1 BR application shortly after the first flowers appeared), BR-2, BR-3, BR-4 and BR-5 (BR application in two, three, four and five consecutive weeks after the appearance of the first flowers, respectively). The fruits were collected for seven consecutive weeks (105 fruits treatment⁻¹) and fruit mass, length and diameter, soluble solid contents; pulp mass and peel thickness were evaluated in the laboratory. Multivariate analysis was performed in order to determine whether there were differences among the treatments taking into account all the measurements made. BR-3 was the most promising treatment because it produced the highest number of fruits plant⁻¹ (81.5) compared to the control (53.5) and the soluble solid content was 1 °Brix greater than the control. The BR-3 treatment resulted in a 65% increase in the estimated yield of the passion fruit plants, corresponding to 20 t ha⁻¹ compared to the control yield of 12 t ha⁻¹. The results showed that BB-16 sprayed during a period of reproductive development can increase the number of fruits per plant.     

Unilateral orchitis and epididymitis caused by Salmonella enterica subspecies diarizonae infection in a ram.

A case of unilateral suppurative epididymo-orchitis associated with Salmonella enterica subsp. diarizonae serovar 61:k:1,5,(7) infection is described in a 2-year-old ram. Gross lesions were characterized by severe enlargement of the scrotal contents, fibrous adhesions between testicular layers, coexistence of epididymal abscesses and foci of fibrinous exudate, and testicular atrophy. Microscopically, testicular and epididymal microabscesses and diffuse inflammatory infiltrates with abundant macrophages containing short Gram-negative rods were observed. Superimposed on the chronic lesions were fibrin deposits with clusters of neutrophils, as well as walled-off granulation tissue. Bacterial colonies were also identified in thrombosed spermatic cord vessels, scrotal lymph nodes, lung, and liver. S. enterica subsp. diarizonae serovar 61:k:1,5,(7) was isolated from the affected testis. To the authors' knowledge, this is the first report of infection of the testis and epididymis by Salmonella in rams. This organism must be taken into account in the differential diagnosis of ovine genital infections.     

Evaluation of ‘Hamlin’ sweet orange + ‘Montenegrina’ mandarin somatic hybrid for tolerance to Xanthomonas axonopodis pv. citri and Xylella fastidiosa

Asiatic citrus canker (ACC), caused by Xanthomonas axonopodis Starr & Garces pv. citri (Hasse) Vauterin et al., and citrus variegated chlorosis (CVC), caused by Xylella fastidiosa Wells et al., are considered the main diseases affecting sweet orange scion varieties in Brazil. Among commercial varieties, mandarins and tangerines are recognized as tolerant to these pathogens. We report herein the production of ‘Hamlin’ sweet orange (Citrus sinensis L. Osbeck) + ‘Montenegrina’ mandarin (Citrus deliciosa Ten.) allotetraploid somatic hybrid plants by protoplast fusion with improved disease tolerance that could be used as a donor of resistance genes in interploid hybridisation. Somatic hybridisation was confirmed by leaf morphology, flow cytometry and RAPD analyses. The somatic hybrid was propagated by grafting and cultivated in a screenhouse for tolerance assays. For X. axonopodis pv. citri assays, buds were collected from both ‘Hamlin’ sweet orange and the somatic hybrid and grafted onto ‘Cleopatra’ mandarin (Citrus reshni hort. ex Tanaka). As a negative control, buds from ‘Mexerica Tardia’ mandarin (C. deliciosa) were collected and grafted onto ‘Cleopatra’ mandarin. Two-month old plants with at least one young vegetative flush were individually spray-inoculated with a 10⁶ CFU mL⁻¹X. axonopodis pv. citri suspension and incubated in a growth chamber, at 27 °C, under 16-h photoperiod. The somatic hybrid showed a statistically significant reduction in susceptibility to ACC 30 days after inoculation. Compared to ‘Hamlin’ sweet orange, disease severity was reduced by 70%, with similar tolerance to that of the mandarin negative control. For X. fastidiosa assays, buds were collected from the somatic hybrid and its parental plants and grafted onto ‘Rangpur’ lime (Citrus limonia Osbeck). The developed plants were needle-inoculated with a X. fastidiosa suspension (8.7 × 10¹⁰ CFU mL⁻¹) into the new growth flush stem. Bacterial population was quantified both at 4 (at the inoculation point) and 8 months (50 cm above the inoculation point) after inoculation. The first evaluation detected X. fastidiosa in 63% of ‘Hamlin’ sweet orange and ‘Hamlin’ + ‘Montenegrina’ mandarin samples. In the second evaluation, X. fastidiosa was detected in 47.4% of ‘Hamlin’ sweet orange and 10.5% of ‘Hamlin’ + ‘Montenegrina’ somatic hybrid samples, suggesting that bacterial movement was restricted in the somatic hybrid. X. fastidiosa was not detected in both evaluations in samples collected from leaves of ‘Montenegrina’ mandarin. These results indicate that the ‘Hamlin’ sweet orange + ‘Montenegrina’ mandarin somatic hybrid has potential for improved disease tolerance that should enhance its value regarding future use in citrus breeding programs.     

Juvenile selection of Gmelina arborea clones in the Philippines

Data collected periodically from one of the oldest Gmelina arborea Roxb. (gmelina) clonal trials of Provident Tree Farms Incorporated in the Philippines were analyzed to identify what clones were genetically superior. Age–age correlations were also examined to determine the efficacy of early clonal selection. Results showed that among the 11 clones of Gmelina arborea that were tested, two clones, 25 and 26, were the best performers. At 72 months, these two clones had an average per tree volume of 0.675?m³ and 0.725?m³ and diameter at breast height of 28.9?cm and 29.3?cm, respectively. Analysis of juvenile data also showed that these two clones were consistently the best at a young age. Of the two juvenile traits measured (basal diameter and total height), total height at 9 months of age was the best predictor of performance at 72 months with genetic correlations ranging from 0.50 to 0.90. These results suggests that early selection at 9 months in gmelina clonal plantings is feasible and is a practical means of reducing the breeding and testing cycles.

Palabras clave: Correlaciones de edad–edad, Productividad, Propagación

Resumen. Se analizaron los datos recolectados periódicamente de uno de los ensayos clonales más antiguos de Gmelina arborea (gmelina) localizado en las Filipinas, en la finca de árboles Provident Tree Farm Inc Para identificar los clones genéticamente superiores. Así mismo se examinaron las correlaciones de edad–edad, para determinar la eficacia de la selección clonal temprana. Los resultados mostraron que entre los 11 clones de Gmelina arborea que fueron ensayados, dos clones, los números 25 y 26 fueron los que dieron mejores resultados y a los 72 meses de edad, estos dos clones tuvieron un promedio de volumen por árbol de 0.675?m³ y 0.725?m³ y diámetro a la altura de pecho de 28.9?cm y 29.3?cm, respectivamente. El análisis de datos de edad temprana también mostró a estos dos clones como los mejores consistentemente a edad temprana. De las dos características juveniles medidas (diámetro basal y altura total), la altura total a los 9 meses fué el mejor pronosticador del rendimiento del árbol a los 72 meses de edad, con correlaciones genéticas con un rango de 0.50 a 0.90. Estos resultados sugieren que es posible hacer selecciones tempranas, a los 9 meses de edad, en las plantaciones clonales de gmelina y que este es un medio práctico de reducir los ciclos de mejoramiento y de ensayos.

     

Starch Debranching Enzyme Activity and Its Effects on Some Starch Physicochemical Characteristics in Developing Substituted and Complete Triticales (X Triticosecale Wittmack)

The present work was carried out to make a comparison between the starch debranching enzyme activity and the synthesis of total carbohydrates, starch, amylose, and amylopectin in developing kernels of two sets of triticales differing in genome composition (complete and substituted triticales). The results showed that the carbohydrate and starch accumulation observed in both genotypes of triticales increased rapidly at the early stage, reaching the maximum at the medium stage and decreasing slightly or remaining relatively constant at the last stage of kernel development. At the end of the development stage, the mature grains of complete and substituted triticales accumulated 62.5 and 54.6% starch, respectively. Both sets of triticales showed the same amylose accumulation profile as well as degree of polymerization; however, at the maturity stage there was a small but significant difference between the two sets. The final polymerization values of amylose chains in mature seeds of complete and substituted triticales were DP 1,549 and 1,313. A and B₁ amylopectin chains of the complete triticales were approximately 19 and 33% shorter than those of the substituted triticales. The complete triticales showed higher isoamylase and pullulanase activities than the substituted triticales, which coincided with the highest starch and amylose contents observed in kernels under development. The results of this study demonstrate that the synthesis of starch differs between the triticale genotypes used in the study, which influences their chemical structure and physicochemical properties.