Protein production encoded by the avirulence gene avrPphB from Pseudomonas syringae pv. phaseolicola was examined. Incorporation of [35S]-labeled methionine into the AvrPphB protein indicated processing of the full-length peptide in Escherichia coli to give a major 28-kDa product. The 28-kDa native peptide was isolated from E. coli following over-expression of avrPphB and found not to elicit the hypersensitive response (HR) after infiltration into bean leaves. Antiserum raised to the 28-kDa peptide allowed expression of avrPphB and processing of AvrPphB protein to be examined in P. syringae pv. phaseolicola; immunoreactive peptides of both 35 and 28-kDa were detected in races 3 and 4 (which contain avrPphB) only after induction in minimal medium + 10 mM sucrose. Antiserum raised to a synthetic peptide, derived from the sequence of the 62 amino acids found to be cleaved from the full-length AvrPphB protein, revealed the accumulation of peptides corresponding to the smaller cleavage products, in both E. coli and P. syringae pv. phaseolicola. Biochemical localization experiments showed that all AvrPphB peptides were cytoplasmic in P. syringae pv. phaseolicola. No AvrPphB peptides were produced in a hrpL mutant unless expression of the gene was directed by a strong vector promoter; induction kinetics similar to wild type were observed in a hrpY- strain, although it also failed to cause a confluent HR. Growth of P. syringae pv. phaseolicola under inducing conditions removed the requirement for rifampicin-sensitive mRNA synthesis by bacteria to allow HR development (the induction time) in bean and lettuce leaves. Constitutive expression of hrpL reduced but did not remove the induction time. Expression of the hrp gene cluster of P. syringae pv. phaseolicola from plasmid pPPY430 in E. coli enabled phenotypic expression of avrPphE (also carried by pPPY430) and avrPphB (if over-expressed from pPPY3031). Despite constitutive expression of the hrp and avr genes in E. coli, a protein synthesis dependent induction time was still required for development of the HR in bean genotypes with matching resistance genes. The significance of processing for the function of AvrPphB peptides and the delivery of elicitors of the HR are discussed.
SummaryConventional methods indicated that Pythium violae was most commonly isolated from carrot cavity spot samples from 14 UK sites. For one site the most frequently isolated species was Pythium sulcatum. Results of similar isolation work were compared with the assay of cavity spot lesions using polyclonal antibodies, raised to P. violae or P. sulcatum, in competition ELISA. Where lesions were artificially induced the test confirmed which pathogen was causal. With cavities developed on the field‐grown carrots P. violae again predominated and the ELISA confirmed this. In one sample P. sulcatum was also isolated from a small number of lesions and was not detected in ELISA. The competition ELISA did not indicate presence of either Pythium in a range of non‐cavity spot lesions from which attempts at isolation were negative.
Two women with primary amenorrhoea and few other stigmata of Turner's syndrome were found to be chromosome mosaics: 45,X/46,X,idic(Y). In Case 1, the dicentric isochromosome Y was found to have a long-arm breakpoint of formation. This structure was interpreted as containing two Y short arms and centromeres separated by a region derived from the proximal Y long arm. One of the centromeres in the Case 1--idic(Y) was suppressed in 80% of cells in blood, and in these cells it appeared as a regular Y-shaped chromosome. In Case 2 the idic(Y) was derived by a short-arm breakpoint of formation. In all the dicentrics of this case with one primary constriction (functional monocentrics) there was a single Cd band. In the 10% of dicentrics with two primary constrictions, there were two Cd bands. It is argued that the instability of sex isochromosomes is due to this functional dicentricity in some cells. These cases are compared with 42 other Y isochromosomes with various short- and long-arm breakpoints of formation. It is suggested that some of the nonheterochromatic, nonfluorescent Y chromosomes previously reported may be explained as dicentric i(Y) with proximal long-arm breakpoints of formation and one suppressed centromere.
An ELISA system was developed to assist with the diagnosis of neurocysticercosis. Antigens were prepared from the fluid, the protoscolex and the cyst wall of cysticerci dissected from infected pigs. The fluid antigen was tested both fresh and after freeze-drying and one year's storage. Sera from patients with proven neurocysticercosis were most reactive using the fluid and least reactive using the cyst wall antigens. Freeze-drying and prolonged storage of fluid antigen did not reduce its reactivity and repeated testing of sera with this antigen gave optical density values within 15% of the original value. Using this ELISA system positive results were obtained from all patients with active neurocysticercosis. Sera from patients with calcified cysts were usually non-reactive or marginally reactive to the fluid antigen.
SUMMARYCytogenetic examination of the Rhodesian population of Praomys natalensis revealed two chromosomally distinct species, one having 32 chromosomes and the other 36 chromosomes. The G-banding patterns and suggested karyotype of the 36 chromosome species is presented here.
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