First Report of Bacterial Blight Caused by Pseudomonas viridiflava on Pea in Spain.

Because production of dry peas (Pisum sativum L.) is increasing in Spain, disease surveys were carried out from 2004 to 2006 in Castilla y Leon, the largest pea-producing region. In May of 2004, a leaf and stem blight caused an estimated 25% loss in yield in pea (cv. Messire) fields in El Cerrato (Palencia). Bacteria were isolated on King's B medium from 10 symptomatic plants from different fields (3). Thirty gram-negative isolates produced fluorescent, yellowish mucoid colonies. All isolates showed oxidative glucose metabolism on Hugh-Leifson medium and were levan and oxidase negative, potato soft rot positive, arginine dihydrolase negative, and tobacco hypersensitive positive. They also hydrolyzed esculine and gelatine. These results were different than those expected by Pseudomonas syringae pv. pisi and P. syringae pv. syringae (3). API 50 CH tests (bioMerieux, Marcy l'Etoile, France) revealed that all the isolates used the following carbon sources: glycerol, erythritol, l-arabinose, ribose, d-xylose, galactose, d-glucose, d-fructose, d-manose, inositol, manitol, sorbitol, d-raffinose, d-fucose, and d-arabitol. This nutritional profile is identical with that of P. viridiflava strain CFBP 6730, originally from pea plants in France. Therefore, these isolates were tentatively identified as P. viridiflava (2). Since a preliminary test demonstrated that 9 of the 30 isolates were pathogenic on pea plants, pathogenic isolates P44, P45, and P46 were selected arbitrarily for further tests. These three isolates plus strains HRI-W 1704 (P. syringae pv. pisi type race 6) and CFBP 1769 (P. syringae pv. syringae) were inoculated onto 10 pea seedlings (cv. Messire) each in two identical trials, following a described protocol (1). Seedlings inoculated with sterile distilled water were used as controls. After 10 days of incubation in a growth chamber at 22°C and 80% relative humidity, severe rotting and collapse similar to symptoms observed in fields appeared on pea seedlings inoculated with isolates P44, P45, and P46, while water-soaked leaf spots and necrotic symptoms were caused by P. syringae pv. pisi and P. pv. syringae. No symptoms were observed on plants inoculated with sterile water. Isolates recovered from symptomatic stems showed the same morphological and biochemical features of the original isolates. Sequences of 1,399 bp long from the three isolates (GenBank Accession Nos. GQ398128, GQ398129, and GQ398130) were 100% identical to P. viridiflava 16S rDNA database reference sequences. To our knowledge, this is the first report of P. viridiflava causing a disease of pea in Spain. The disease has been reported in New Zealand (4) and France (2). References: (1) E. M. Elvira-Recuenco et al. Eur. J. Plant Pathol. 109:555, 2003. (2) C. Grondeau et al. Plant Pathol. 41:495, 1992 (3) N. W. Schaad et al., eds. Laboratory Guide for the Identification of Plant Pathogenic Bacteria. 3rd ed. The American Phytopathological Society, St. Paul, MN, 2001. (4) J. D. Taylor et al. N. Z. J. Agric. Res. 5:432, 1972.

First Report of Bacterial Blight Caused by Pseudomonas syringae pv. syringae on Common Vetch in Spain.

Common vetch (Vicia sativa L.) is an important legume crop used for livestock feed in the Mediterranean Area. This crop has an important role for sustainable agriculture in dryland rotations in Spain, where the Castilla y León Region is the major production area. During the springs of 2007 and 2008, necrotic lesions on stems, leaves, and flowers were observed in five different common vetch plots around Medina de Rioseco (Castilla y León). Four of the plots were sown with cv. Buza. No information was available about the cultivar in the fifth plot. In many cases, lesions had expanded into the stems causing complete wilting. Disease incidence was estimated at approximately 20%. Two symptomatic plants per plot were sampled. One section per plant was individually surface disinfested in 0.5% NaOCl for 1 min, followed by three washes in sterile water. Macerates were plated in King's B medium (KB) agar (24°C for 48 h) (2). Colonies from all isolations on KB agar were pale yellow and blue-green fluorescent under UV light, as typical of fluorescent Pseudomonas spp. (2). Ten isolates (one per section) were characterized. These were identified as Pseudomonas syringae by the LOPAT scheme (2) and Hugh-Leifson reaction and all utilized erythritol, l-lactate, and dl-homoserine, but not l(+)-tartrate, as carbon sources. They were positive for aesculin and gelatine hydrolysis. The 10 isolates caused severe necrotic lesions when they were puncture inoculated (108 CFU/ml suspension, 50 µl per wound, and two replicates) on immature lemon fruits (Citrus × limon, cv. Primofioro), bean pods (Phaseolus vulgaris L., cv. Ancha Lisa), and pea pods (Pisum sativum L., cv. Ucero). PCR amplification of a 752-bp syrB fragment (3) was positive for all isolates. On the basis of these tests, the 10 isolates were identified as P. syringae pv. syringae (2). Subsequently, each isolate was inoculated into two sets of 10 plants of V. sativa cv. Buza by injecting 200 µl of a bacterial suspension (108 CFU/ml) into the stem (2); 10 plants were injected with sterile water as controls. Ten days after inoculation, necrotic symptoms were observed on all plants, and 1 week later, all plants were completely wilted and dead. These symptoms were similar to those observed in the field. Control plants remained symptomless. Isolations were made from two inoculated plants per each original isolate, and all reisolates were identical to the original isolates in the above biochemical tests and PCR of the syrB gene. P. syringae pv. syringae reference strains, CFBP1768 and CFBP1769 (Collection Française de Bactéries Phytopathogènes), gave the same results in all biochemical, pathogenicity, and PCR tests. To our knowledge, this is the first report of bacterial blight caused by this pathogen on vetch in Spain. This pathogen had been previously identified in this crop in France (4) and in V. villosa (a closely related species) in the United States (1). Therefore, to prevent the spread of this pathogen, research on efficient preventive and control measures is needed. References: (1) G. L. Ercolani et al. Phytopathology 64:1330, 1974. (2) N. W. Schaad et al., eds. Laboratory Guide for the Identification of Plant Pathogenic Bacteria. 3rd ed. The American Phytopathological Society, St. Paul, MN, 2001. (3) K. N. Sorensen et al. Appl. Environ. Microbiol. 64:226, 1998. (4) C. Tourte and C. Manceau. Eur. J. Plant Pathol. 101:483, 1995.

Stable low molecular weight RNA analyzed by staircase electrophoresis, a molecular signature for both prokaryotic and eukaryotic microorganisms.

Low-molecular weight RNA (LMW RNA) analysis using staircase electrophoresis was performed for several species of eukaryotic and prokaryotic microorganisms. According to our results, the LMW RNA profiles of archaea and bacteria contain three zones: 5S RNA, class 1 tRNA and class 2 tRNA. In fungi an additional band is included in the LMW RNA profiles, which correspond to the 5.8S RNA. In archaea and bacteria we found that the 5S rRNA zone is characteristic for each genus and the tRNA profile is characteristic for each species. In eukaryotes the combined 5.8S and 5S rRNA zones are characteristic for each genus and, as in prokaryotes, tRNA profiles are characteristic for each species. Therefore, stable low molecular weight RNA, separated by staircase electrophoresis, can be considered a molecular signature for both prokaryotic and eukaryotic microorganisms. Analysis of the data obtained and construction of the corresponding dendrograms afforded relationships between genera and species; these were essentially the same as those obtained with 16S rRNA sequencing (in prokaryotes) and 18S rRNA sequencing (in eukaryotes).

[Brief family therapy: an option for the treatment of somatoform disorders in primary care]. / Terapia familiar breve: una opción para el tratamiento de los trastornos somatoformes en atención primaria.

OBJECTIVE: To study the application of the brief family therapy model to the treatment of somatoform disorders in scheduled primary care consultations. DESIGN: Intervention study. SETTING: Cazoña Health Centre, Santander. PATIENTS: 18 patients who had suffered somatoform disorder for at least a year. INTERVENTIONS: A general practitioner trained in short-term family therapy applied this treatment under the supervision of a specially trained psychologist. MEASUREMENTS AND RESULTS: The variables of the research into the results of brief family therapy were collected. These showed 61.1% therapeutic success, 27.8% failure and 11.1% abandonment. There were no relapses at the six-month control. Total number of consultations varied between 1 and 11, average 4.6; the average interval between consultations was 27 days, with an average length of 48 minutes for each consultation. Time between the first and last consultation varied from 1 to 242 days, average 96 days. Brevity of treatment was related to its therapeutic success. CONCLUSIONS: Training in brief family therapy, with supervision at the start, can provide primary care doctors with a treatment alternative for resolving somatoform disorders without the patient having to accept a psychological reason for his/her complaints.

Psicoterapia familiar breve. Cómo facilitar el cumplimiento terapéutico a través de la modalidad de cooperación de los pacientes / Brief family psychotherapy. How to facilitate therapeutic compliance through the modality of cooperation of the patiens

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