Molecular analysis of resistance mechanisms to Orobanche cumana in sunflower

Resistance to the dicotyledenous parasite Orobanche cumana in sunflower is characterized by a low number of parasitic attachments and a confinement of the parasite in host tissues leading to its necrosis. To help understand what determines such resistance mechanisms, molecular, biochemical and histological approaches were employed before (early response) and after (late response) attachment of the broomrape parasite to susceptible (2603) and resistant (LR1) sunflower genotypes. The expression patterns of 11 defence-related genes known to be involved in different metabolic pathways (phenylpropanoids, jasmonate, ethylene) and/or in resistance mechanisms against microorganisms were investigated. RT-PCR and cDNA blot experiments revealed that the resistant genotype exhibited a stronger overall defence response against O. cumana than the susceptible one, involving marker genes of the jasmonate (JA) and salicylic acid (SA) pathways. Among them, the SA-responsive gene, def. (defensin), appeared to be characteristic of LR1 sunflower resistance. However, no JA accumulation and similar SA contents (250–300 ng g⁻¹ FW) were measured by GC/MS in both genotypes, parasitized or not. In addition, three cDNAs, isolated by a suppression-subtractive hybridization, were shown to be strongly induced only in the resistant genotype 8 days post-inoculation, when the first O. cumana attachments occurred. These genes, putatively encoding a methionine synthase, a glutathione S-transferase and a quinone oxidoreductase, might be involved in detoxification of reactive oxygen species, suggesting the occurrence of an oxidative burst during the incompatible interaction. Finally, host cell-wall modifications leading to parasite-confinement were correlated with more intense callose depositions in the resistant genotype, concomitant with over-expression of the callose synthase cDNA HaGSL1 .     

rep-PCR-Mediated Genomic Fingerprinting: A Rapid and Effective Method to Identify Clavibacter michiganensis

ABSTRACT The genomic DNA fingerprinting technique known as repetitive-sequence-based polymerase chain reaction (rep-PCR) was evaluated as a tool to differentiate subspecies of Clavibacter michiganensis, with special emphasis on C. michiganensis subsp. michiganensis, the pathogen responsible for bacterial canker of tomato. DNA primers (REP, ERIC, and BOX), corresponding to conserved repetitive element motifs in the genomes of diverse bacterial species, were used to generate genomic fingerprints of C. michiganensis subsp. michiganensis, C. michiganensis subsp. sepedonicus, C. michiganensis subsp. nebraskensis, C. michiganensis subsp. tessellarius, and C. michiganensis subsp. insidiosum. The rep-PCR-generated patterns of DNA fragments observed after agarose gel electrophoresis support the current division of C. michiganensis into five subspecies. In addition, the rep-PCR fingerprints identified at least four types (A, B, C, and D) within C. michiganensis subsp. michiganensis based on limited DNA polymorphisms; the ability to differentiate individual strains may be of potential use in studies on the epidemiology and host-pathogen interactions of this organism. In addition, we have recovered from diseased tomato plants a relatively large number of naturally occurring avirulent C. michiganensis subsp. michiganensis strains with rep-PCR fingerprints identical to those of virulent C. michiganensis subsp. michiganensis strains.     

Examination of the thoracic cavity and lung lobectomy by means of thoracoscopy in dogs.

The feasibility of thoracoscopy for viewing the chest cavity and performing pulmonary lobectomy was assessed in 8 mongrel dogs. Previously, selective intubation had been performed in another group of dogs (n = 8) in order to monitor respiratory physiology and assess its safety. Each hemithorax was intubated using a double-barrelled endotracheal tube with one barrel placed in the left main bronchus and the other in the bifurcation of the trachea. The thoracoscope was introduced through a cannula inserted through a 2-cm incision at the ventral third of the left 5th intercostal space. The cranial, dorsal, and caudal surfaces of the pleura, lobes of the left lung, and the mediastinum were examined. A 2nd cannula was located in the dorsal 3rd of the 5th intercostal space with a prior incision and used for the introduction of forceps to separate the viscera. To biopsy, a 3rd cannula was inserted at the dorsal third of the 8th intercostal space with a prior incision, through which a 12-mm diameter stapler was introduced. Should a lobectomy be necessary, a 4th cannula is located in the middle third of the 4th intercostal space. Excision of the left caudal pulmonary lobe was performed through the incision made for the 12-mm diameter cannula (8th intercostal space); a twisting movement facilitated removal. Thoracoscopy is a procedure that can be used in dogs and is particularly suitable for examination, collection of biopsy specimens, and even lung lobectomies.     

Alveolar osteitis (dry socket) in a dog: a case report

Following extraction of a maxillary left first molar tooth in an eight year-old retriever, the dog re-presented five days later because of oral pain, which did not respond to analgesic therapy. The extraction site contained a foul-smelling fluid, but did not contain a clot or granulation tissue. Alveolar osteitis (dry socket) was diagnosed. The alveolus was curetted and flushed, and the dog was given cefalexine and prednisolone. The alveolus was filling with healthy granulation tissue one week later and the dog was no longer in pain.     

Purification and characterization of a protease from Actinobacillus pleuropneumoniae serotype 1, an antigen common to all the serotypes.

A high molecular-mass proteolytic enzyme of Actinobacillus pleuropneumoniae serotype 1, was purified from culture supernatants (CSN) by using DEAE-cellulose and sepharose-4B-gelatin chromatography. In 10% SDS-polyacrylamide gels copolymerized with porcine gelatin, the protease showed a single band of activity of > 200 kDa. However, minor molecular-mass proteolytic bands were observed when the protease was electrophoresed in the presence of either 5% beta-mercaptoethanol, 50 mM dithiothreitol, or 0.25 M urea. Furthermore, when the > 200-kDa purified protein was passed through a sucrose gradient, several bands with proteolytic activity were found: 62, 90, 190, and 540 kDa. The proteolytic activity was increased in the presence of calcium or zinc and was not affected after being heated at 90 degrees C for 5 min. Proteolytic activities were also observed in CSN from all A. pleuropneumoniae serotypes and biotypes. The purified protease hydrolyzed porcine IgA and IgG in vitro. In addition, by immunoblot the protease was recognized by serum of naturally infected pigs with serotypes 1 and 5, and by serum of pigs experimentally infected with serotypes 1, 2, 8, or 9. Serum of a pig vaccinated with CSN of a serotype 3 strain also recognized the protease, but not sera of pigs vaccinated with a bacterin (serotype 1). Proteins from CSN of all the serotypes, which were precipitated with 70% (NH4)2SO4, were recognized by a polyclonal antibody raised against the purified protease. Taken together these results indicate that an antigenic protease is produced in vivo by all the serotypes of A. pleuropneumoniae. The results indicate that proteases could have a role in the disease and in the immune response of pigs infected with A. pleuropneumoniae.     

贵州茶叶加工业与国内外比较的现状及发展趋势

根据贵州茶叶加工业与国内外比较的现状、存在问题和制约发展的因素,以贵州得天独厚的气候资源优势、生产潜力、良种和品牌为基础。浅析贵州茶叶加工业的发展趋势：认为在“十一五”期间,贵州茶叶栽培面积将稳定增长。加工规模不断扩大;茶叶消费市场需求将持续增长;科学技术的提升将促进茶叶加工业的可持续发展;茶叶深加工是茶产业经济的新增长点;贵州茶叶加工业的发展方向是实现茶叶加工产业化。     

Measurement of vitamin A,vitamin E,selenium, and L-lactate in dogs with and without osteoarthritis secondary to ruptured cranial cruciate ligament

This study compared vitamin A, vitamin E, selenium (Se), and L-lactate in blood and synovial fluid in 2 groups of 6 dogs; a control group (without OA) and an osteoarthritic group with spontaneous cranial cruciate ligament rupture and OA. Concentrations of vitamin E were significantly higher in serum than in synovial fluid in both OA (P = 0.006) and control (P = 0.0008) groups. Vitamin E concentration in synovial fluid was significantly higher in the OA group than in the control group (P = 0.009). Concentrations of Se were significantly higher in serum than in synovial fluid in both OA (P = 0.003) and control (P = 0.0006) groups. There were no significant differences in levels of Se, vitamin A, and L-lactate between the 2 groups. This is the first study to show an increased concentration of vitamin E in the synovial fluid of dogs with OA compared with dogs that did not have OA.     

Some alternatives to calving date and interval as measures of fertility in beef cattle

Records of 594 calving dates and 493 calving intervals collected from a herd of Retinta beef cows in southwest Spain were studied. Their efficacy as reproductive measures were compared when a long breeding season was utilized. Variables used were date of present calving (DOC), date of subsequent calving (DOSC), time from start of breeding season to calving (TBSC), interval to next calving (CI), two adjustments of calving interval by subtracting the time that bulls were not available to the cow from actual CI (ACI1 and ACI2), and three scores. Period score (SCOR) measured the 20-d increment of the 220-d calving season in which the cow calved. Pair score (PAIR) was computed by subtracting the deviation of calving interval from 365 d from the average SCOR of the two calvings involved. Adjusted pair score (APAIR) was calculated in the same way as PAIR, but using ACI1 instead of CI. Heritability estimates for DOC, SCOR, and DOSC and CI were .16, .14, .13 and 0, respectively. Heritabilities of all other measures ranged from .01 to .06. Repeatability estimates for DOC, SCOR, DOSC and CI were .29, .28, .34 and .14, respectively. Repeatability values were .28, .18, .18, .23 and .32 for TBSC, ACI1, ACI2, PAIR and APAIR, respectively. Genetic and phenotypic correlations among CI and its alternative measures were very high and favorable.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of two dietary protein levels on energy balance and digestive capacity of <Emphasis Type="Italic">Octopus maya</Emphasis>

Octopus maya is a carnivorous species and protein is the main energy source. During the present study, two different dietary protein levels (40 and 60% CP) were offered to octopuses as specifically designed artificial diets, to determine protein needs and the effects on metabolism. Frozen crab (Callinectes spp.) was used as control. Results obtained demonstrated that crab remains as one of the best diets for O. maya. The artificial diet with 60% CP produced a low but positive growth rate, and at times, a physiological response similar to that observed in octopuses fed crabs. The present results show the capacity of O. maya juveniles to adjust their digestive enzymes to different types of food and protein level, and this appears to be well correlated with octopus growth. General proteases and trypsin from the pancreas were well correlated with growth rates. A low activity was observed in octopuses fed 40% PC diet (negative growth rate), while a high activity was present in octopuses fed 60% CP diet and crabs (low and high growth rate, respectively). In contrast, these same enzymes were inducted in the salivary glands of octopuses fed with the diet that promoted weight loss (40% CP diet), while a reduced activity was observed in octopuses fed crabs. Energy budget indicates that the animals ingested more than 1,000 kJ week⁻¹ kg⁻¹; with such energy, octopuses should satisfy their physiological demands such as was observed when animals were fed crab (I = 1,300 kJ week⁻¹ kg⁻¹; P = 834 kJ week⁻¹ kg⁻¹). However, a very low digested energy was observed in octopuses the fed artificial diets, indicating that these could have a factor that limits digestibility.