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.     

Review and analysis of the potential for glyphosate to interact with the estrogen,androgen and thyroid pathways

Glyphosate was recently evaluated for its potential to interact with the estrogen, androgen and thyroid (EAT) hormone pathways, including steroidogenesis, under the United States Environmental Protection Agency's (USEPA) Endocrine Disruptor Screening Program (EDSP), then by Germany, the rapporteur Member State who led the European Annex 1 renewal for glyphosate, and then by the European Food Protection Agency (EFSA) also as part of the Annex 1 renewal for glyphosate. Under the EDSP, 11 Tier 1 assays were run following the USEPA's validated 890‐series test guidelines and included five in vitro and six in vivo assays to evaluate the EAT pathways. Steroidogenesis was evaluated as part of the estrogen and androgen pathways. An up‐to‐date critical review has been conducted that considered results from the EDSP Tier 1 battery, guideline regulatory studies and an in‐depth analysis of the literature studies that informed an endocrine assessment. A strength of this evaluation was that it included data across multiple levels of biological organization, and mammalian and nonmammalian test systems. There was strong agreement across the in vitro and in vivo Tier 1 battery, guideline studies and relevant literature studies, demonstrating that glyphosate does not interact with EAT pathways including steroidogenesis. Based on an analysis of the comprehensive toxicology database for glyphosate and the literature, this review has concluded that glyphosate does not have endocrine‐disrupting properties through estrogen, androgen, thyroid and steroidogenic modes of action. © 2020 Society of Chemical Industry     

Map- vs. homology-based cloning for the recessive gene ol-2 conferring resistance to tomato powdery mildew

The recessive gene ol-2 confers papilla-associated and race-non-specific resistance to tomato powdery mildew caused by Oidium neolycopersici. In order to facilitate marker assisted selection (MAS) in practical breeding programmes, we identified two simple sequence repeat (SSR) markers and one cleaved amplified polymorphic sequence (CAPS) marker which are linked to the resistance locus and co-dominantly inherited. Aiming to provide a base for ol-2 positional cloning, we used a large segregating F₂ population to merge these markers with all the ol-2 linked amplified fragment length polymorphism (AFLP^®) markers previously identified in an integrated genetic map. By screening a tomato bacterial artificial chromosome (BAC) library, we detected two BAC clones containing two expressed sequence tags (ESTs) homologous to the gene mlo, responsible for powdery mildew resistance in barley, as well as an ol-2-linked marker. Chromosomal mapping by Fluorescence in situ Hybridization (FISH) revealed major signals of the two BAC DNAs in the pericentromeric heterochromatin of the short arm of chromosome 4, in the same region where the ol-2 gene was previously mapped. The genetic and cytogenetic co-localisation between ol-2 and tomato mlo-homologue(s), in addition to the similarity of ol-2 and mlo resistances for both genetic and phytopathological characteristics, suggests that ol-2 is likely a mlo-homologue. Thus, a homology-based cloning approach could be more suitable than positional cloning for ol-2 isolation.     

Molecular tracing of <Emphasis Type="Italic">Bradyrhizobium </Emphasis>strains helps to correctly interpret <Emphasis Type="Italic">Acacia mangium </Emphasis>response to inoculation in a reforestation experiment in Madagascar

A field inoculation experiment using two Bradyrhizobium sp. strains was set up in Madagascar to test the growth response of Acacia mangium and to follow up the survival of inoculant strains using molecular tools. Three months after inoculation, one of the inoculant strains, AUST13c, exhibited a marked growth-promoting effect with a shoot height about 40% higher than that of the uninoculated control plants or TEL8-inoculated plants. The positive effect on tree growth initially observed with AUST13c was no more significant 6 months after transfer to the field and disappeared completely at 13 and 19 months. Analyses of nodule bacterial rRNA by PCR/RFLP displayed an early contamination of the different inoculation treatments by AUST13c 3 months after inoculation, spreading to almost all nodules of the trial 6 months later. This work clearly demonstrated that the progressive reduction of the positive effect of AUST13c inoculation on tree growth after field transplanting was not due to a progressive disappearance of this introduced strain but, on the contrary, was related to the widening spreading of AUST13c in all the plots. This was attributed to a higher competitiveness and effectiveness of AUST13c over the local strains and TEL8.     

Proximal sensor-enhanced soil mapping in complex soil-landscape areas of Brazil

Portable X-ray fluorescence (pXRF) spectrometry and magnetic susceptibility (MS) via magnetometer have been increasingly used with terrain variables for digital soil mapping. However, this methodology is still emerging in many countries with tropical soils. The objective of this study was to use proximal soil sensor data associated with terrain variables at varying spatial resolutions to predict soil classes using the Random Forest (RF) algorithm. The study was conducted on a 316-ha area featuring highly variable soil classes and complex soil-landscape relationships in Minas Gerais State, Brazil. The overall accuracy and Kappa index were evaluated using soils that were classified at 118 sites, with 90 being used for modeling and 28 for validation. Digital elevation models (DEMs) were created at 5-, 10-, 20-, and 30-m resolutions using contour lines from two sources. The resulting DEMs were processed to generate 12 terrain variables. Total Fe, Ti, and SiO₂ contents were obtained using pXRF, with MS determined via a magnetometer. Soil class prediction was performed using the RF algorithm. The quality of the soil maps improved when using only the five most important covariates and combining proximal sensor data with terrain variables at different spatial resolutions. The finest spatial resolution did not always provide the most accurate maps. The high soil complexity in the area prevented highly accurate predictions. The most important variables influencing the soil mapping were MS, Fe, and Ti. Proximal sensor data associated with terrain information were successfully used to map Brazilian soils at variable spatial resolutions.     

Changes in articular cartilage after intra-articular injections of methylprednisolone acetate in horses

Eight mature horses with no prior signs of joint disease or history of intra-articular therapy were treated with 8 weekly intra-articular injections of methylprednisolone acetate. Treatments were given at a dose of 120 mg/joint into the right radiocarpal and intercarpal joints, with the left joints as untreated controls. Articular cartilage samples were obtained at necropsy 1, 4, and 8 weeks after the last injection. Compared with controls, cartilage from injected joints had a loss of hematoxylin basophilia and decreased intensity of staining in safranin O fast green dye. Chondrocyte necrosis and hypocellularity were observed in all samples of cartilage from treated joints. Proteoglycan content and its rate of synthesis were reduced. There was a progressive loss of proteoglycan content, whereas proteoglycan synthesis increased somewhat 4 and 8 weeks after treatment. Collagen content was unchanged, but its rate of synthesis was markedly inhibited. Collagen synthesis did not recover, but remained decreased at 5 to 15% of the values from untreated cartilage. Water percentage was increased, but fibronectin content was not significantly different. A single injection of methylprednisolone acetate was also given into the right metacarpophalangeal joints of 3 of the 8 horses in this group, with the left joints serving as untreated controls. Sixteen weeks after the treatment, cartilage of the treated joints had a loss of histochemical staining and proteoglycan content was reduced to 50% of control values. The mean rate of proteoglycan synthesis and mean fibronectin content were increased, but the differences were not statistically significant (P greater than 0.05). Other variables were essentially unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)     

Retrospective study of cecocolic intussusception (cecal inversion) in nine horses (1982–1998)

We retrospectively evaluated the medical records and obtained follow-up information for nine horses which had been treated for cecocolic intussusception (CCI) between January 1982 and April 1998. During the 16-year study period, CCI was diagnosed in nine of 748 horses in which exploratory celiotomy was undertaken for abdominal pain, representing an incidence of 1.2%. Most affected horses (78%) were less than four years of age (median age was 12 months, age range was five months to 15 years). Cecocolic intussusception affected male horses (78%) more commonly than female horses. The most common clinical presentation was abdominal pain of a severe, acute nature or milder but recurrent signs of abdominal pain persisting in spite of conservative treatment for several days. Correction of CCI by either simple reduction or reduction followed by partial typhlectomy was successful if compromise of the intestine by devitalization and adhesion formation was not found at surgery. Definitive diagnosis of CCI necessitates exploratory celiotomy, although an ultrasonographic examination of the abdomen may confirm the diagnosis in some cases. When recognized early during the course of disease, surgical correction of CCI is associated with a favorable outcome; of the eight horses which underwent surgery in our series, five horses (63%) survived surgical correction of CCI. Handling of compromised gut during reduction of CCI necessitates extreme caution because the risk of intestinal tearing is quite high.     

Channels and ditches as the last shelter for freshwater mussels: the case of Margaritifera auricularia and other naiads inhabiting the mid Ebro River Basin,Spain

• 1. Freshwater mussels or naiads are generally considered to thrive in river habitats, provided the ecological conditions are good. The presence of populations of these bivalves in artificial channels and ditches with natural bottoms has only scarcely been reported. The aim of this paper was to present the idea that these ‘channel’ and ‘ditch’ habitats could in fact be a sanctuary for naiads.
• 2. Approximately 80 km of several of these waterways fed by the mid Ebro River were sampled in Spain to monitor their naiad populations. Observations indicate that these habitats harbour substantial colonies of freshwater mussels (including two populations of adult specimens of the endangered species Margaritifera auricularia), much more so than the corresponding river.
• 3. The authors wish to alert conservation authorities and freshwater mussel experts to the extreme fragility and importance of this kind of habitat for the long‐term conservation of these imperilled molluscs.

Copyright © 2008 John Wiley & Sons, Ltd.