Association of double-stranded RNA with whitefly-mediated silvering in squash

A disorder of unknown aetiology appearing as silvering on squash (Cucurbita pepo) leaves has become prevalent in south Florida. Typical symptoms, observed after 2 weeks of exposure to adults of Bemisia tabaci , the sweet potato whitefly (SPWF) include vein clearing followed by silvering of whole leaves. Fruits from affected plants are blanched or streaked longitudinally. The silverleaf syndrome was not mechanically transmissible when crude extracts of symptomatic leaves were used as inoculum. Two sizes of double-stranded RNA (dsRNA) of c. 4.6 and 4.2 kilobase pairs (kbp) were consistently observed in leaf extracts of caged, whitefly-infested, symptomatic plants. In contrast, caged plants not exposed to whiteflies showed no symptoms and contained no detectable dsRN A. In addition, squash plants exposed to SPWF colonies from California neither developed silverleaf symptoms nor contained any dsRNA. Double-stranded RNA could also be readily detected in whitefly-infested yellow summer squash and zucchini from the field. However, asymptomatic eggplant and watermelon infested with SPWF from nearby fields did not contain detectable dsRNA. Dot-spot hybridization assays using an RNA-specific probe detected homologous sequences in both adults and nymphs of SPWF that induced silverleaf symptoms but not in adults and nymphs of SPWF that could not induce silverleaf symptoms. These results, along with the non-endogenous nature of the detected dsRNA suggest that the dsRNA is or is associated with the causal agent of whitefly-mediated leaf silvering in squash.     

First report of the Q biotype ofBemisia tabaci in Japan by mitochondrial cytochrome oxidase I sequence analysis

The recent upsurgence ofBemisia tabaci (Genn.) as an important insect pest and vector ofTomato yellow leaf curl virus (TYLCV) is directly linked to serious damage to tomato crops grown throughout Japan. The molecular genetic identification and phylogenetic relationships of 12B. tabaci populations collected from representative locations in Japan were determined based on the mitochondrial cytochrome oxidase I (mtCOI) sequence. Phylogenetic analysis of the whitefly mtCOI sequence indicated that both the invasive B and Q biotypes now occur in Japan. The Q biotype was found at four locations: Mihara in Hiroshima, Nishigoshi in Kumamoto, Miyanojo and Okuchi in Kagoshima prefectures; the remaining eight collections were identified as the B biotype. This is the first report of the introduction of Q biotype in Japan. http://www.phytoparasitica.org posting July 21, 2006.     

Genetic and physiological evidence for the production of N-acyl homoserine lactones by Pseudomonas syringae pv. syringae and other fluorescent plant pathogenic Pseudomonas species

N-acyl homoserine lactones (AHLs) function as cell density (quorum) sensing signals and regulate diverse metabolic processes in several gram negative bacteria. We report that strains of Pseudomonas syringae pvs. syringae (Pss), tabaci and tomato as well as P. corrugata and P. savastanoi produce difussible AHLs that activate the lux operons of Vibrio fischeri or the tra::lacZ fusion of Agrobacterium tumefaciens. In Pss strain B3A, AHL production occurs in cell density dependent manner. Nucleotide sequence and genetic complementation data revealed the presence of ahlI_Pss, a luxI homolog within the Ahl⁺ DNA of Pss strain B3A. The DNA expresses in AHL-deficient strains of P. fluorescens and E. carotovora subsp. carotovora (Ecc), and restores extracellular enzyme production and pathogenicity in the Ecc strain. The derivatives of Pss strains B3A and 301D carrying chromosomal ahlI::lacZ do not produce AHL, but like their wild type parents, produce extracellular protease and the phytotoxin syringomycin as well as elicit the hypersensitive reaction in tobacco leaves. While these strains also produce a basal level of -galactosidase activity, the expression of ahlI::lacZ is substantially stimulated in the presence of multiple copies of the DNA or by the addition of cell-free spent cultures containing AHL. The activation of -galactosidase production occurs with spent cultures of some, but not all Pseudomonas strains which produce AHL as indicated by the Lux and tra::lacZ assays. Pss strains deficient in the global regulatory genes, gacA or lemA, produce very low levels of AHL. Since inactivation of ahlI_Pss eliminates AHL production and since Ahl⁺ Pseudomonas strains carry the homolog of ahlI_Pss, we conclude that ahlI_Pss specifies a key step in AHL biosynthesis and it has been conserved in many plant pathogenic pseudomonads.     

Characterization of the first two viruses described from wild populations of hammer orchids (Drakaea spp.) in Australia

Sequences representing the genomes of two distinct virus isolates infecting wild plants of two members of the genus Drakaea (hammer orchids) in Western Australia are described. The virus isolated from Drakaea livida has a bipartite genome of 4490 nt (RNA1) and 2905 nt (RNA2) that shares closest sequence and structural similarity to members of the genus Pecluvirus, family Virgaviridae, described from legumes in the Indian subcontinent and West Africa. However, it differs from pecluviruses by lacking a P39 protein on RNA2 and having a cysteine‐rich protein gene located 3′ of the triple gene block protein genes. It is the first peclu‐like virus to be described from Australia. The name Drakaea virus A is proposed (DVA; proposed member of the family Virgaviridae, genus unassigned). The second virus isolate was identified from Drakaea elastica, a species classed as endangered under conservation legislation. The genome sequence of this virus shares closest identity with isolates of Donkey orchid symptomless virus (DOSV; proposed member of the order Tymovirales, family and genus unassigned), a species described previously from wild Caladenia and Diuris orchids in the same region. These viruses are the first to be isolated from wild Drakaea populations and are proposed to have an ancient association with their orchid hosts.     

The effect of a straw meal on the crude protein and amino acid metabolism and digestibility of the crude nutrients in broiler hen breeds. 2. Digestibility of crude nutrients of rations and 15N from a straw meal and wheat

In experiments with colostomized broiler hens apparent digestibility of the crude nutrients of the ration after straw meal supplements of 20, 30 and 40 g per animal was determined. In addition, the 15N digestibility of straw meal and wheat was ascertained on the basis of straw meal supplements. The digestibility of the crude nutrients of the rations decreased significantly (P less than 0.05) after the straw meal supplement. The adaptation of the test animals to the straw meal intake resulted, at a daily consumption of 20 g straw meal, in an increase of the apparent crude fat digestibility (P less than 0.05) in dependence on the time of straw meal feeding, in which the original values without straw meal supplement were not reached. The digestibility of the 15N excess (15N') of the wheat was, at 86 +/- 1%, largely independent of the straw meal intake. The apparent digestibility of the straw-15N excess in broiler hens of 42 +/- 8 to 55 +/- 2% is surprisingly high.     

In vitro markers for virulence in Yersinia ruckeri

In this study, different traits that have been associated with bacterial virulence were studied in Yersinia ruckeri. Two isolates that had been shown to cause disease and mortality in experimentally infected rainbow trout were compared with five avirulent isolates. Both virulent isolates showed high adhesion to gill and intestinal mucus of rainbow trout, whereas the majority of non‐virulent strains demonstrated significantly lower adhesion. A decrease in adherence capability following bacterial treatment with sodium metaperiodate and proteolytic enzymes suggested the involvement of carbohydrates and proteins. All strains were able to adhere to and invade chinook salmon embryo cell line (CHSE‐214), fathead minnow epithelial cell line (FHM) and rainbow trout liver cell line (R1). One non‐virulent strain was highly adhesive and invasive in the three cell lines, whereas the virulent strains showed moderate adhesive and invasive capacity. The internalization of several isolates was inhibited by colchicine and cytochalasin‐D, suggesting that microtubules and microfilaments play a role. For all strains, intracellular survival assays showed a decrease of viable bacteria in the cells 6 h after inoculation, suggesting that Y. ruckeri is not able to multiply or survive inside cultured cells. Analysis of the susceptibility to the bactericidal effect of rainbow trout serum demonstrated that virulent Y. ruckeri strains were serum resistant, whereas non‐virulent strains were generally serum sensitive.     

Decomposition Dynamics and Nutrient Release Pattern from Leaf Litters of Five Commonly Occurring Homegarden Tree Species in Mizoram,India

Leaf litter decomposition and nutrient release patterns from five common multipurpose tree species—viz., Artocarpus heterophyllus, Mangifera indica, Areca catechu, Citrus sp., and Tamarindus indica, found in homegardens of Mizoram—were evaluated using a litter bag technique. The result of the study indicates a varying pattern of decomposition and nutrient release (N&P) among the species. Citrus sp. and T. indica were found to be the most labile species with comparatively much higher decay constant and faster nutrient release. Initial nitrogen concentration, lignin content, and lignin/N ratio of foliage litter showed significantly higher (p < .01) correlation with the decay coefficient and were found to be the important determinants in the decay process. The initial slow release and immobilization of N in A. heterophyllus and M. indica leaf litter reflect their potential as a source of nitrogen storage and effective mulching material. While litter from T. indica and Citrus sp. can provide the short-term nutrient need, foliage for the other three species may supply the long-term nutrient requirement for the understory crops in such agroforestry systems.     

A unified approach for estimating population size in capture-recapture studies with arbitrary removals

A unified approach is suggested to estimate the population size for a closed population in discrete time. Individuals can be removed after capture at any time during the experiment. The usual recapture and removal experimentsare shown to be particular cases of the general formulation. The capture probability is assumed to have a logistic function that depends on individual covariates and can be time dependent. The unified approach involves a two-step procedure. A conditional likelihood function is used to estimate the covariates coefficients and a Horvitz-Thompson type estimator to estimate the population size. The asymptotic and small-sample properties of the resulting estimators are in vestigated. A real example is given.     

Electron solvation in two dimensions

Ultrafast two-photon photoemission has been used to study electron solvation at two-dimensional metal/polar-adsorbate interfaces. The molecular motion that causes the excess electron solvation is manifested as a dynamic shift in the electronic energy. Although the initially excited electron is delocalized in the plane of the interface, interactions with the adsorbate can lead to its localization. A method for determining the spatial extent of the localized electron in the plane of the interface has been developed. This spatial extent was measured to be on the order of a single adsorbate molecule.