Genetic Structure of the Population of Pepino mosaic virus Infecting Tomato Crops in Spain

ABSTRACT The population structure of Pepino mosaic virus (PepMV), which has caused severe epidemics in tomato in Spain since 2000, was analyzed. Isolates were characterized by the nucleotide sequence of the triple gene block and coat protein gene and, for a subset of isolates, a part of the RNA-dependent RNA polymerase gene. The full-length sequence of the genomic RNA of a Solanum muricatum isolate from Peru also was determined. In spite of high symptom diversity, the Spanish population of PepMV mostly comprised highly similar isolates belonging to the strain reported in Europe (European tomato strain), which has been the most prevalent genotype in Spain. The Spanish PepMV population was not structured spatially or temporally. Also, isolates highly similar to those from nontomato hosts from Peru (Peruvian strain) or to isolate US2 from the United States (US2 strain) were detected at lower frequency relative to the European strain. These two strains were detected in peninsular Spain only in 2004, but the Peruvian strain has been detected in the Canary Islands since 2000. These results suggest that PepMV was introduced into Spain more than once. Isolates from the Peruvian and US2 strains always were found in mixed infections with the European tomato strain, and interstrain recombinants were detected. The presence of different strains of the virus, and of recombinant isolates, should be considered for the development of control strategies based on genetic resistance.     

Drought-induced oxidative stress in Canarian laurel forest tree species growing under controlled conditions

We studied photoprotection and antioxidative protection in the three major species of the Canarian laurel forest (Laurus azorica (Seub.) Franco, Persea indica (L.) K. Spreng and Myrica faya Aiton). Trees were exposed to drought under controlled conditions by withholding water until leaf relative water content (RWC) reached 50-55%. Drought reduced photosynthetic rate (P(N)) and was associated with decreased quantum yield of photosystem II (PSII) electron transport and increased non-photochemical quenching (NPQ) in L. azorica and M. faya, but did not increase NPQ in P. indica. Drought-treated trees of L. azorica had the highest de-epoxidation state (DPS) of the xanthophyll cycle and the highest zeaxanthin (Z) concentration, suggesting that this species had more effective photoprotective mechanisms than M. faya and P. indica. Moreover, beta-carotene remained unaltered in L. azorica trees during drought, suggesting that the chloroplasts of this species are better protected against oxidative stress than those of M. faya and P. indica. Increased antioxidation by ascorbate peroxidase, superoxide dismutase and glutathione reductase in L. azorica removed activated oxygen species (AOS) generated during drought treatment. Although M. faya was able to increase its energy dissipation rate by forming Z and thus increasing the DPS of the xanthophyll cycle, it did not respond to drought-induced oxidative stress with the result that beta-carotene degradation occurred. Persea indica did not activate an energy dissipation mechanism in response to drought treatment, hence formation of AOS was likely high in the drought-treated trees. In general, L. azorica was most resistant and P. indica most sensitive to photoinhibition and oxidative stress during drought.     

Praemachiloides iberica n. sp. from Spain (Insecta Microcoryphia)

Praemachiloides iberica n. sp. is described and compared with the other four known species of the genus: P. janetschecki, P. insularis, P.tarsispina and P. autumnalis. The main difference of P. iberica n. sp. from the other known congenerics is the presence, in the male, of a field of plaited setae on the ventral surface of the maxillary palp.     

Control of root zone pH is not effective in preventing Pythium aphanidermatum disease in cucumber

Journal of Plant Diseases and Protection - Pythium aphanidermatum is the most wide-spread root pathogen in greenhouse cucumber. Although environmental condi-tions are known to affect the disease...     

Tissue distribution of Red Spotted Grouper Nervous Necrosis Virus (RGNNV) genome in experimentally infected juvenile European seabass (Dicentrarchus labrax)

The distribution of viral genome in the tissues of juvenile European seabass (Dicentrarchus labrax) during the course of a Red Spotted Grouper Nervous Necrosis Virus (RGNNV) infection has not yet been described. The present study addresses this and indicates which target organs may be involved in viral replication. This information should enable more accurate detection of virus in asymptomatic carriers, and in turn help to control the spread of the disease. The aim of this study was to examine the pattern of expression of viral genomic segments RNA1 and RNA2, using two absolute real-time PCRs (RT-qPCR), over the course of a RGNNV infection after administering the virus by intramuscular injection. In situ hybridization was also used to locate the RNA2 viral segment in different organs throughout the infection. The experimental challenge provoked an acute form of viral nervous necrosis (VNN), with a resulting cumulative mortality of 37%. The RT-qPCRs designed allowed the detection of both genomic segments in all the organs tested (nervous and non-nervous tissues) at all sampling times examined. The highest viral RNA copy number was found in eyes, although viral replication appeared to begin in the brain. Viral replication was also recorded in pooled internal organs and in caudal fin. However, the increase in the viral RNA copy number in these organs did not result in an increased viral titre, which may indicate that a productive infection does not take place in non-nervous tissues, possibly due to a failure in a viral post-replication step.     

Isolation and characterization of primary canine lens epithelial cells

OBJECTIVE: The purpose of this study was to characterize the proliferation and differentiation of primary canine lens epithelial cells (LEC) under standard culture conditions. PROCEDURE: Canine LEC were isolated by mechanical dissection of the canine globe and enzymatic digestion of the lens capsule from fresh lenses. Isolated capsules and cell suspensions were seeded in laminin-coated culture flasks. Canine LEC proliferated and formed monolayers, which could be passaged and maintained for approximately 2 weeks. Cells were characterized morphologically and cell lysates examined for expression of protein markers of epithelial origin and differentiation. RESULTS: Canine LEC exhibit morphologic characteristics of epithelial cells when cultured on laminin/lysine coated flasks. Expression of epithelial cell marker, cytokeratin 5, was highest at passage 1 and diminished with increasing passage number. Expression of gamma-crystallin, a protein found only in differentiated lens fiber cells, increased at passage 6. A laminin/lysine-coated surface supported optimal proliferation of canine LEC. Both an initial seeding density of 1 x 10(5) cells/cm(2) and culture in Dulbecco's modified essential media (DMEM) supplemented with 10% FBS supported a doubling time of less than 48 h in canine LEC. CONCLUSION: This study demonstrates that primary canine LEC retain the characteristics of lens epithelial cells prior to passage 6 under the described culture conditions and represent a suitable in vitro model for investigating lens physiology and cataractogenesis.     

A contribution to the maintenance of grapevine diversity: The rescue of Tinta Castañal (Vitis vinifera L.), a variety on the edge of extinction

The arrival of powdery mildew, phylloxera, and then downy mildew from America in the late 19th/early 20th century, was one of the most important causes of the loss of grapevine diversity in Europe. Many varieties traditionally grown in small winemaking areas of Europe were substituted by direct producing hybrids or by a small number of varieties from other vine-growing areas. For geographic, economic, sociological, and cultural reasons, the north and northwest of Spain acts as a kind of refuge area where grapevine diversity is still high. At the Misión Biológica de Galicia (CSIC) research station a collection of living vines with a number of dispersed individuals belonging to old varieties was established in 1992. Many of these varieties had practically disappeared from vineyards and the majority existed only as centuries-old individuals.