Use of two PGPR strains in the integrated management of blast disease in rice (Oryza sativa) in Southern Spain

Biocontrol capacity of two plant growth-promoting rhizobacteria (PGPR) strains, against blast disease in rice paddy fields in Southern Spain was studied in three cropping seasons. Both strains (Pseudomonas fluorescens Aur 6 and Chryseobacterium balustinum Aur 9) had already shown biocontrol capacity against pathogens, ability to induce systemic resistance against leaf pathogens and against salt stress in different plant species. Bacterial treatments were carried out on seeds and/or on leaves. Strains were inoculated individually and in combination. Protection against natural disease incidence was evaluated, and rice production and quality measured in 2005 and 2006 trials. In 2004, natural disease incidence was low (between 0.1% and 0.35% of damaged leaf surface) due to environmental conditions; under these conditions, both strains significantly protected plants against rice blast. In 2005, disease incidence was higher than in 2004, reaching higher values of affected leaf surface in controls. In these conditions, each strain individually protected rice against rice blast, although the combination of both strains was the most effective treatment. All three treatments (Aur 6, Aur 9 and Aur 6 + Aur 9) reached 50% protection in panicles, with Aur 9 being the most effective. In 2006, the most effective treatment was the combination of both strains on leaves in three physiological stages, suggesting a biocontrol mediated protection. On the other hand, when bacteria were applied to seeds, disease incidence decreased up to 50%, suggesting induction of systemic resistance. Finally, a direct relation between protection mediated by the PGPR and the increase in rice productivity (mT/ha) and quality (weight of 1000 seeds and number of intact grains after milling) was found.     

Genetic Structure of Peruvian Populations of Phytophthora infestans

ABSTRACT Isolates of the late blight pathogen Phytophthora infestans (n = 327) from the central to southern Peruvian Andes were systematically collected in 1997 to 1999 and analyzed to determine the pathogen's population structure at its host's center of diversity. No isolates of the A2 mating type were detected. Cluster analysis of DNA fingerprinting data indicated that the collection consisted of five major groups that were interpreted to be clonal lineages. Two of the lineages (US-1 and EC-1) have been previously described, and three (PE-3, 5, and 6) are described here for the first time. Collections from three areas in the central Peruvian Andes, including two key sites used in an international potato breeding program, consisted of isolates of the EC-1 lineage, which has been reported to dominate the pathogen population in Andean countries to the north of Peru. The collections from Cusco and Puno were more diverse. More than one lineage was detected in 10 of the 20 fields sampled in Cusco. Data on virulence, metalaxyl sensitivity, and band data for allozymes, mitochondrial DNA, and ipiB1 suggested that PE-3 may have been produced through recombination events between US-1 and EC-1. Restriction fragment length polymorphism and amplified fragment length polymorphism marker data were not consistent with this hypothesis.     

The detection of CD2+, CD4+, CD8+, and WC1+ T lymphocytes, B cells and macrophages in fixed and paraffin embedded bovine tissue using a range of antigen recovery and signal amplification techniques

In order to develop procedures to label the main bovine leucocyte populations in paraffin embedded sections, the immunoreactivity of 25 monoclonal antibodies (mAbs) to different leucocyte antigens was assessed with formal dichromate (FD5) and 10% formalin fixation, a battery of antigen retrieval (AR) methods, and the biotin-tyramide amplification system. All the leucocyte populations investigated (CD2+, CD4+, CD8+, WC1+ T lymphocytes, B cells and macrophages) were strongly and specifically detectable under an appropriate combination of mAb, AR method and signal amplification system. CD4 and CD8 required the most stringent conditions and could only be demonstrated in FD5 fixed sections. For detection of CD2, WC1+ T lymphocytes, B cells and macrophages, all the mAbs produced immunoreactivity in FD5 or formalin fixed tissues. The need to check a range of different AR methods is stressed, as the method of choice varied for each individual mAb. The incorporation of the signal amplification system was necessary to observe a strong signal and the complete distribution of CD4, CD8 and B cells. Fixation by FD5 proved to be better than formalin for the preservation of surface antigens but it was inferior for the detection of markers which were found to show cytoplasmic immunoreactivity, such as the macrophage marker MAC387 or the B cell markers BAQ155 or IL-A59.     

Silica Urolithiasis in the Dromedary Camel in a Subtropical Climate

In 1998, two cases of silica urolithiasis occurred in castrated male dromedaries on an intensive camel farm in the Canary Islands. The immediate attributable cause was the ingestion of large amounts of silica in the feed, estimated as 84.44 g/day. An associated cause was the low level of salt in the diet. Daily ingestion of salt from feed and water was estimated to be 21.8 g (8.6 g of sodium). Seventy-six castrated males from the same farm were divided into four groups: group A received 30 g of salt daily; group B received 40 g; group C received 60 g; and group D received no added salt in the diet (control). The animals were maintained on these dietary regimes for 2 years. No animals from groups A, B or C suffered overt urinary retention. One animal from group D had an obstructive urinary retention 10 months after the study commenced. Thus, 52 g of salt daily appears to be sufficient to prevent urinary retention in dromedaries raised in a subtropical climate.     

Camel trypanosomosis in the Canary Islands: assessment of seroprevalence and infection rates using the card agglutination test (CATT/T. evansi) and parasite detection tests

Trypanosomosis due to Trypanosoma evansi (surra) is a major enzootic disease of the dromedary camel. Thus, the purpose of the present study was to assess seroprevalence and infection rates in the Canary Islands using antibody(-card agglutination test-CATT/T. evansi) and parasite detection tests (micro-Haematocrit Centrifugation technique, Giemsa stained blood smears, microscopic examination of lymph node aspirates and mouse inoculation). PCV was also determined. 745 dromedary camels (483 females and 262 males) were examined. Trypanosomes were detected in seven animals. 36 animals yielded CATT positive results while 709 animals were negative. All parasitologically positive animals were also CATT positive. Results showed a good correlation between CATT positive and low PCV and a higher seroprevalence in older animals. Trypanocidal drugs have not been registered in Spain and, consequently, if vigilance is not exercised the prevalence could be increased in the future.     

Phylogenetic analysis based on ITS sequences and conditions affecting the type of conidial germination of Bipolaris oryzae

One taxonomic characteristic of Bipolaris species is the bipolar germination of conidia, but conidia of Bipolaris oryzae , the causal pathogen of brown spot in rice, are regularly observed to show intercalary germination, a characteristic of Drechslera species. The effect of selection, culture media and culture age on type of conidial germination was determined for three brown spot isolates from Cavinti, San Pablo and Palawan in the Philippines, obtained from infected leaves showing typical disease symptoms. Based on the analyses of their ITS1, ITS2 and 5·8S rDNA nucleotide sequences, the local isolates were clearly identified as B. oryzae . Selection for colonies of the three isolates derived from single conidia with either bipolar or intercalary germination had no effect on the number of spores showing bipolar germination in subsequent cultures. Germination on seven different culture media was tested; of these, rabbit food agar and water agar increased the percentage of bipolar germination of conidia, although this varied between isolates. Incubation of the cultures of all three isolates for longer periods prior to harvesting conidia increased the percentage of bipolar-germinating conidia from c . 40 to c . 90% with 5-day-old and 30-day-old cultures, respectively.