Characterization of Distinct Tombusviruses that Cause Diseases of Lettuce and Tomato in the Western United States

A soilborne disease of lettuce, associated with necrosis and dieback, has been found with increasing frequency in California and Arizona over the last 10 years. An isometric virus, serologically related to Tomato bushy stunt virus (TBSV), was consistently isolated from lettuce plants with these disease symptoms. Back-inoculation to healthy lettuce plants and subsequent reisolation of the virus from symptomatic lettuce leaves suggested that this virus was the causal agent of this disease. A tombusvirus was also associated with a necrosis disease of greenhouse-grown tomatoes in Colorado and New Mexico. Complementary DNA representing the 3' end of viral genomic RNAs recovered from diseased lettuce and tomato plants had identical nucleotide sequences. However, these sequences were divergent (12.2 to 17.1%) from sequences of the previously described strains of TBSV, Petunia asteroid mosaic virus (PAMV), Artichoke mottled crinkle virus, and Carnation Italian ringspot virus. Additional tombusvirus isolates were recovered from diseased lettuce and tomato plants and these were most closely related to the TBSV-cherry strain (synonymous with PAMV) and to Cucumber necrosis virus based on comparison of 3'-end sequences (0.1 to 0.6% and 4.8 to 5.1% divergence, respectively). Western blot analysis revealed that the new tombusvirus isolated from diseased lettuce and tomato plants in the western United States is serologically distinct from previously described tombusvirus species and strains. Based on genomic and serological properties, we propose to classify this virus as a new tombusvirus species and name it Lettuce necrotic stunt virus.     

Isolation and Characterization of Derivative Lines of the Onion Yellows Phytoplasma that Do Not Cause Stunting or Phloem Hyperplasia

ABSTRACT Two lines of onion yellows phytoplasma producing milder symptoms were isolated from the original line (OY-W). One has an additional characteristic, non-insect-transmissibility (OY-NIM), compared with the other (OY-M). OY-M was established after maintaining OY-W for 11 years on a plant host (Chrysanthemum coronarium) with an insect vector (Macrosteles striifrons), and OY-NIM was isolated after subsequent maintenance of OY-M in plants by periodic grafting. Polymerase chain analysis suggested that OY-NIM cannot traverse the gut or survive in the hemolymph of the leafhopper. OY-W results in witches'-broom formation and stunted growth in the host plant. In contrast, OY-M and OY-NIM do not cause stunting in the host plant, although they result in witches'-broom. Histopathological analysis of these lines revealed that the hyperplastic phloem tissue and severe phloem necrosis seen in OY-W did not exist in OY-M and OY-NIM. This was attributed to a reduction in the population of phytoplasma in tissues in both OY-M- and OY-NIM-infected plants. The results suggest that the cause of stunting and phloem hyperplasia may be genetically different from the cause of witches'-broom. Pulsed field gel electrophoresis analysis showed that OY-M had a smaller genome size ( approximately 870 kbp) than OY-W ( approximately 1,000 kbp). Thus, some of the OY-W genes responsible for pathogenicity may not be present in OY-M.     

Bacterial populations associated with rice seed in the tropical environment

ABSTRACT During the 1995 wet season, harvested rice seed was collected from farmers' fields at different locations in Iloilo, Philippines. Bacterial isolations from crushed seed yielded 428 isolates. The isolates were characterized by BOX-polymerase chain reaction fingerprinting of total genomic DNA and represented 151 fingerprint types (FPT). Most FPTs were found on a single occasion, although matching fingerprints for isolates from different samples also were found. Identifications were made by cellular fatty acid methyl ester analysis and additional use of Biolog GN/GP MicroPlates and API 20E/50CHE systems. The predominant bacteria were Enterobacteriaceae (25%), Bacillus spp. (22%), and Pseu-domonas spp. (14%). Other bacteria regularly present were identified as Xanthomonas spp., Cellulomonas flavigena, and Clavibacter michiganense. Of the total number of isolated bacteria, 4% exhibited in vitro antifungal activity against Rhizoctonia solani or Pyricularia grisea. Two percent of isolates were pathogens identified as Burkholderia glumae and Burkholderia gladioli. Five percent of isolates induced sheath necrosis on only 50 to 90% of inoculated plants and were related to Bacillus pumilus, Paenibacillus spp., Pseudomonas spp., and Pantoea spp.     

Risk Analysis for Latent Infection of Prune by Monilinia fructicola in California

ABSTRACT The quantitative relationships between incidence of latent infection (ILI) of prune by Monilinia fructicola and wetness duration (WD) for different bloom and fruit developmental stages and different inoculum concentrations were obtained. Three levels of ILI were considered as criteria for low, moderate, and high risks of latent infection, respectively. Seasonal patterns of WD leading to different risk levels of latent infection were obtained for low (IP(L)) and high (IP(H)) inoculum potential conditions in orchards. Longer WD was needed at a resistant than at a susceptible fruit developmental stage to induce similar levels of latent infection. The curves of WD leading to different levels of ILI over the growing season (risky WD curves) were used in risk analysis for latent infection. Multi-year historical WD data from 10 prune-growing locations in California were compared with risky WD curves. The percentage of days (P) with WD leading to a certain risk level of latent infection was calculated for each month from historical weather data. Under the IP(L) condition, the P distributions for low risk of latent infection were higher in March and April than in May and were the lowest in June for most locations. Under the IP(H) condition, the number of days that WD leading to low risk of latent infection in May increased compared with those under the IP(L) condition. The risk analysis approach was evaluated by using separate experimental data as incidence of fruit brown rot obtained from different prune orchards over years. Consistency between predicted overall risk levels of latent infection and observed incidence of fruit brown rot was obtained. The results demonstrated the usefulness of the risk analysis in decision support system for disease management.     

Classification, nomenclature, and plant pathogenic bacteria - a clarification

ABSTRACT In a recent Letter to the Editor of Phytopathology, proposals were made for endorsement and for rejection of selected names of plant pathogenic Pseudomonas spp. and Xanthomonas spp. We believe that support for, and rejection of, several names was based on misconceptions concerning the Approved Lists of Bacterial Names and entails misinterpretations of several Rules of the International Code of Nomenclature of Bacteria. This letter aims to clarify those misconceptions and misinterpretations.     

Cytogenetic identification of a hybrid owl monkey, Aotus nancymaae x Aotus lemurinus griseimembra.

A neonate male owl monkey (Aotus sp.) was identified cytogenetically as a hybrid after it failed to nurse and died. Phenotypically, the male parent possessed characteristics of the "gray-neck group," and G-banded karyotypes identified him as Aotus lemurinus griseimembra (2n = 53), heterozygous for the centric fusion of chromosomes 13 and 14. The female parent belonged to the "red-neck group" and was identified cytogenetically as Aotus nancymaae (2n = 54). The neonate hybrid had 2n = 54 chromosomes with 13 homologous pairs of autosomes, 26 nonhomologous autosomes, and XY sex chromosomes. Thirteen of the nonhomologous chromosomes represented the paternal complement, and 13 were from the maternal complement. Chromosomal rearrangements occurring between the karyotypes of A. l. griseimembra and A. nancymaae were believed to include two paracentric inversions, a reciprocal translocation, and two complex rearrangements involving pericentric inversion, telocentromeric fusion, and centromeric adjustment. Cytogenetic analyses are necessary to identify most Aotus taxa and thus should be utilized to pair chromosomally compatible animals and avoid interspecies hybridization.     

The influence of lameness on equine stride length consistency

The aim of this study was to assess the influence of orthopaedic pain on the variation of stride length as a kinematic system-parameter in 21 horses with forelimb lameness. Data were collected while the horses were trotting on a treadmill during a minimum of 12 motion cycles, both before and after intra-articular or perineural anaesthesia. Stride length was assessed for each motion cycle, and the mean and standard deviation were calculated for each condition. Forelimb lameness was documented as percentage of asymmetry of vertical head movement. With significant decrease of forelimb lameness after regional anaesthesia, the SD of stride length increased significantly (+0.35%, P< 0.05). Our results show that in the presence of orthopaedic pain horses keep stride variability low, possibly because the lame horse employs an optimum compensatory mechanism to reduce the pain in the affected limb, and every deviation from this pattern increases pain.     

Characterization of mutations in the two-component histidine kinase gene that confer fludioxonil resistance and osmotic sensitivity in the os-1 mutants of Neurospora crassa

Osmotic-sensitive (os-1) mutant alleles in Neurospora crassa exhibit resistance to dicarboximides, aromatic hydrocarbons and phenylpyrroles. We have previously reported that the os-1 mutants can be classified into two groups based on their resistance to fungicides and osmotic stress: type I, which are highly resistant to iprodione and fludioxonil but moderately sensitive to osmotic stress, and type II, which are highly sensitive to osmotic stress but moderately resistant to fungicides. To explain the mechanism of resistance to these fungicides, we cloned and sequenced the mutant os-1 genes that encode putative osmo-sensing histidine kinase. Within the os-1 gene product (Os1p), the type I strains, NM233t and Y256M209, carried a stop codon at amino acid position 308 and a frameshift at amino acid position 294, respectively. These mutation sites were located on the upstream of histidine kinase and the response regulator domains of Os1p, strongly suggesting that type I strains are null mutants. The null mutants, NM233t and Y256M209, were highly resistant to iprodione and fludioxonil; thus Os1p is essential for these fungicides to express their antifungal activity. The amino acid changes in Os1p, 625Pro from Leu, 578Val from Ala, and 580Arg from Gly were found in the type II strains, M16, M155-1 and P5990, respectively. Os1p is novel in having six tandem repeats of 90 amino acids in the N terminal. Each amino acid change of the type II strains was located on the fifth unit of six tandem repeats. Type II strains with single amino acid changes were more sensitive to osmotic stress than the null mutants (type I), indicating that the amino acid repeats of Os1p were responsible for an important function in osmo-regulation.     

Laboratory and field evaluation of the pathogenicity of entomopathogenic nematodes to the red palm weevil, Rhynchophorus ferrugineus (Oliv.) (Col.: Curculionidae)

Ten Egyptian and imported entomopathogenic nematodes were evaluated for their pathogenicity to R. ferrugineus in both the laboratory and the field. In the laboratory, most nematodes were pathogenic to the pest larvae, pupae and adults. Larvae and adults were more susceptible to nematode infection (mostly 100% mortality) than pupae enclosed in their cocoons. In the field however, the highest insect larval mortality was 66.67% and most of nematodes failed in controlling the pest. Such failure could be due to hot weather, the tunnelling behaviour of the pest larvae and the too much sap in the infested sites in the trunks of palm trees.