Ralstonia solanacearum virulence in tomato seedlings inoculated by leaf clipping

Ralstonia solanacearum is a phytopathogenic bacterium that colonizes the xylem vessels of host plants leading to a lethal wilt disease. Although several studies have investigated the virulence of R. solanacearum on adult host plants, infection studies of this pathogen on the seedling stages of hosts are less common. In a preliminary observation, inoculation of R. solanacearum F1C1 on 6‐ to 7‐day‐old tomato seedlings by a simple leaf‐clip strategy resulted in a lethal pathogenic condition in seedlings that eventually killed these seedlings within a week post‐inoculation. This prompted testing of the effect of this inoculation technique in seedlings from different cultivars of tomato and similar results were obtained. Colonization and spread of the bacteria throughout the infected seedlings was demonstrated using gus‐tagged R. solanacearum F1C1. The same method of inoculating tomato seedlings was used with R. solanacearum GMI1000 and independent mutants of R. solanacearum GMI1000, deficient in the virulence genes hrpB, hrpG, phcA and gspD. Wildtype R. solanacearum GMI1000 was found to be virulent on tomato seedlings, whereas the mutants were found to be non‐virulent. This leaf‐clip technique, for inoculation of tomato seedlings, has the potential to be a valuable approach, saving time, space, labour and costs.     

A metalloprotease disintegrin that controls cell migration in Caenorhabditis elegans

In Caenorhabditis elegans, the gonad acquires two U-shaped arms by the directed migration of its distal tip cells (DTCs) along the body wall basement membranes. Correct migration of DTCs requires the mig-17 gene, which encodes a member of the metalloprotease-disintegrin protein family. The MIG-17 protein is secreted from muscle cells of the body wall and localizes in the basement membranes of gonad. This localization is dependent on the disintegrin-like domain of MIG-17 and its catalytic activity. These results suggest that the MIG-17 metalloprotease directs migration of DTCs by remodeling the basement membrane.     

Genotyping Common Bean for the Potyvirus Resistance Alleles I and bc-1(2) with a Multiplex Real-Time Polymerase Chain Reaction Assay

ABSTRACT A multiplex real-time polymerase chain reaction (PCR) assay was developed to simultaneously genotype plants for the I and bc-1(2) alleles, which condition resistance in beans to Bean common mosaic virus and Bean common mosaic necrosis virus. A segregating F(2) population was derived from the cross between pinto bean breeding line P94207-189A (bc-1 bc-1 I I) and Olathe (bc-12 bc-1(2) i i). Real-time PCR assays were developed that were specific for each allele, and a multiplex PCR reaction could unambiguously assign F(2) plants to one of nine genotypes. Remnant F(1) plants were used as a comparative reference sample. PCR results among this sample fit a normal distribution for both real-time PCR assays, and 99% probability distributions were determined for heterozygotes. F(2) plants were genotyped based on results relative to the probability distributions for heterozygotes. F(2) plants also were genotyped for the I and bc-1(2) alleles by performing F(3) family progeny tests for virus resistance. Agreement between the two methods was 100% (198/198) for the bc-1(2) allele, and 92.4% (183/198) for the I allele. Erroneous genotyping was due to recombination between the amplicon and the I allele. Realtime PCR assays provide a robust method for genotyping seedlings and, in some cases, may eliminate the need for progeny testing.     

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Eel acetylcholinesterase inhibition studies with heteroarylphosphinates

The inhibition of eel acetylcholinesterase by the 4-nitrophenyl esters of 2-furyl(methyl)-, methyl(2-thienyl)-, di-2-furyl-, and di-2-thienylphosphinic acid (I, II, III, and IV, respectively) was investigated at pH 6.90 in 0.067 M phosphate buffer (25.0°C) using stopped-flow instrumentation and automated data processing. Our evaluation of the dissociation constant, K_d, the unimolecular bonding rate constant, k₂, and the bimolecular reaction constant, k_i, are the first reported values for these constants for alkyl/heteroaryl and diheteroaryl esters of phosphinic acids. The largest k_i value (19,330 M^?1 sec^?1) was observed for the reaction of I with the enzyme. The order for the remaining three is II > IV > III. There is no direct relationship between the hydrolysis rates of the esters and their anticholinesterase activities on eel acetylcholinesterase. Likewise, there is no direct relationship between their anticholinesterase activities and the LD₅₀ values in rats.     

Association with the Syndrome "Basses Richesses" of Sugar Beet of a Phytoplasma and a Bacterium-Like Organism Transmitted by a Pentastiridius sp

ABSTRACT The syndrome "basses richesses" of sugar beet (SBR) was first observed in 1991 in Burgundy, France. A cixiid planthopper, Pentastiridius beieri, has been proved to be involved in the transmission to sugar beet of a stolbur phytoplasma, which could be detected in some affected plants. In 2000, periwinkle and sugar beet exposed to field-collected cixiids developed symptoms similar to SBR on sugar beet. Use of 4'-6-diamidino-2-phenylindole (DAPI) staining and transmission electron microscopy confirmed the presence of phytoplasma in some of the plants, which were also positive for this pathogen in a polymerase chain reaction (PCR) analysis. A phloem-restricted gram-negative bacteria was seen in all other plants with symptoms but PCR-negative for phytoplasma. Three primer pairs reported as diagnostic for phloem-limited bacteria were tested but only primers specific for 'Candidatus Phlomobacter fragariae' gave a positive signal, which related to the presence of DAPI-stained bacteria-like objects in diseased plants. Although phytoplasma and bacterium-like organisms were associated with the same macroscopic symptoms on sugar beet, histochemical analysis of phloem cells showed that phytoplasma were associated with cell necrosis and cell wall lignification, while bacteria were associated with these same abnormalities as well as deposit of phenolic compounds in the lumen of phloem cells.     

Infection criteria,inoculum sources and splash dispersal pattern of <Emphasis Type="Italic">Colletotrichum acutatum</Emphasis> causing bitter rot of apple in New Zealand

Infection of Malus x domestica cv. Royal Gala fruit by Colletotrichum acutatum causing bitter rot was studied in the temperate climate of New Zealand. Temperatures above 15 °C were required for lesions to develop on detached apple wound-inoculated or inoculated without wounding with C. acutatum spores, regardless of maturity. A wetness period of 72 h was required for infection of mature detached apple fruit without wounding. On wound-inoculated detached apple fruits, sporulation was related to temperature and followed a similar pattern. In the field, a mean temperature above 15 °C for 72 h after wound-inoculation was required for lesions to develop. Buds were a more important source of inoculum than twigs, and it was shown that C. acutatum could be isolated more frequently from outer bud scales than from inner scales. Asymptomatic infection of vegetative and reproductive buds was detected. C. acutatum was detected on asymptomic surface-sterilised petals and fruit, more commonly during summer than spring. Symptomless sterilised leaves generally yielded C. acutatum throughout the season, but isolations were more frequent in summer. Recovery of inoculum using a splash meter to detect vertical dispersal showed that in summer inoculum was primarily splashed up from the ground. In spring, inoculum was recovered in similar quantities from all heights up to a metre, suggesting that splash dispersal occurs from the canopy as well as from the ground. A disease cycle for C. acutatum infecting apples and causing bitter rot in New Zealand is suggested.