Expression Analysis of Rice Defense-Related Genes in Turfgrass in Response to Magnaporthe grisea

ABSTRACT Magnaporthe grisea (anamorph = Pyricularia grisea) causes blast on rice (Oryza sativa) and gray leaf spot on turfgrass. Gray leaf spot is a serious disease on St. Augustinegrass (Stenotaphrum secundatum), perennial ryegrass (Lolium perenne), and tall fescue (Festuca arundinacea). Virulence assays performed in this study revealed that M. grisea collected from rice could also cause disease on St. Augustinegrass and tall fescue. One rice isolate, Che86061, caused similar disease reactions on susceptible cultivars of rice and St. Augustinegrass and an incompatible interaction on resistant cultivars of both species. To explore whether similar defense-related genes are expressed in rice and St. Augustinegrass, a rice cDNA library was screened using pooled cDNAs derived from M. grisea-infected St. Augustinegrass. Thirty rice EST (expressed sequence tag) clones showing differential expression in St. Augustinegrass following M. grisea inoculation were identified and classified into six putative functional groups. Northern blot analyses of seven EST clones that collectively represented each putative functional group confirmed that the expression of five out of seven EST clones was similar in both rice and St. Augustinegrass. This study represents one of the first attempts to use a broad-scale genomic approach and resources of a model monocot system to study defense gene expression in St. Augustinegrass following M. grisea infection.     

Evaluation of the API 50CH and API ZYM systems for rapid characterization of Clavibacter michiganensis subsp. sepedonicus, causal agent of potato ring rot

API 50CH and API ZYM systems were used to characterize fifty-three strains of Clavibacter michiganensis subsp. sepedonicus from different geographic locations and several reference strains of the same and different species, including other potato pathogens. Clavibacter michiganensis subsp. sepedonicus strains displayed a high level of homogeneity, both in carbohydrate utilization and in enzymatic activity. Using API 50CH and API ZYM it was possible to differentiate C. michiganensis subsp. sepedonicus strains from the remaining taxa analysed in this study, which included representative strains of the other subspecies of C. michiganensis as well as other bacterial pathogens affecting potatoes. Therefore, these systems could be used as an effective method to characterize C. michiganensis subsp. sepedonicus. Such a procedure would constitute an alternative system to the conventional nutritional and physiological identification tests currently included in the official methods employed in the European Union to detect and identify this bacterium. The results obtained with the API systems agreed with the current taxonomic classification of C. michiganensis, clearly separating sepedonicus from the other subspecies belonging to this species.     

Early detection of Neophysopella tropicalis in grapevine leaves and on spore traps by qPCR

Neophysopella tropicalis, one of the causal agents of Asian grapevine leaf rust (AGLR), can cause severe epidemics in Brazil that lead to yield losses in commercial vineyards. An early detection of the pathogen by air sampling of urediniospores on spore traps or in symptomless leaves would be valuable to multiple studies, such as epidemics modelling, risk forecasting, monitoring of pathogen introductions in rust-free areas, and predicting the beginning of epidemics. This study developed a quantitative PCR (qPCR) protocol to quantify N. tropicalis urediniospores attached to adhesive tapes and in grapevine leaves before symptom appearance. A specific primer pair was designed based on the internal transcribed spacer (ITS) sequence region of the AGLR pathogen. Standard amplification curves using genomic DNA from urediniospores of N. tropicalis and from urediniospores attached to adhesive tapes were established. Grapevine leaves inoculated with N. tropicalis were collected at 2, 5, and 7 days postinoculation (dpi). One primer pair (580F/720R) amplified a 140 bp product in all AGLR isolates but did not amplify products of other rust genera, such as Phakopsora, Puccinia, Hemileia, Tranzschelia, Cerotelium, and Coleosporium. As little as 0.1 pg DNA and 10 urediniospores of N. tropicalis attached to adhesive tapes could be detected. qPCR enabled the detection of the pathogen as early as 2 dpi, before symptom appearance. This method can be used to monitor N. tropicalis inoculum in grapevine-growing areas and to quantify symptomless infections of the AGLR pathogen.     

Piquerol a and diacetyl piquerol act as energy transfer inhibitors of photosynthesis

ATP synthesis and phosphorylating (coupled) electron flow of freshly lysed pea chloroplasts were inhibited by piquerol A and diacetyl piquerol, the latter being more active. H + -uptake, basal and uncoupled electron transport were not affected by either compound, indicating that they act as inhibitors of photophosphorylation at the H + -ATPase complex.     

Characterization of common bean wild populations for their in situ conservation in Northwestern Argentina

In situ conservation of wild species is a method of conservation that allows keeping populations in their natural environments, and set the strategies for maintaining the natural populations. The Active Bank of Northwestern Argentina (BANOA) is in charge of the in situ conservation of wild populations of common bean (Phaseolus vulgaris L.) in Northwestern Argentina (NOA), and has an ex situ collection of 401 landraces and 221 wild accessions from the NOA. We evaluated the phenotypic diversity of 68 common bean wild populations from the NOA both in protected and unprotected areas, finding a moderate variation among them. Ten phenotypic reproductive characteristics related to pod and seed displayed significant differences in the analysis of variance; these traits together with the seed weight were the basis for the multivariate analysis. The cluster analysis ordered the populations in 12 groups but trends in geographical distribution or phenotypical variation were not recognized. For the conservation in situ of the wild bean populations, their diversity should be considered. Two types of populations can be highlighted: (i) candidates for in situ conservation in order to preserve the novel variation generated by convergence with cultivated sympatric germplasm (populations 433, 437, 471, 509, 513 and 517) and (ii) those whose phenotype represents clearly the wild status and should be preserved in situ as such in their current status (populations 480, 495, 496, 525 and 533).     

Sclerotinia resistance in sunflower: I. Genotypic variations of hybrids in three environments of Argentina

Sclerotinia sclerotiorum infections on sunflower capitula produce white rot, one of the most dangerous diseases sunflower bears in all humid areas. Therefore, disease quantification on developed genetic materials is a very important action in sunflower breeding. Given the horizontal type of resistance these evaluations should be made in different environments. Several hybrids obtained after crossing seven female × seven male sunflower inbred lines in a factorial mating design were evaluated for resistance to white rot in two locations in the southeast of the province of Buenos Aires, Argentina. Disease incidence and the symptom appearance index indicated significant genotypic effects and genotype × environment (G × E) interactions. Genotypic effects were a greater source of variance than the G × E interaction effects. The G × E interactions only represented changes in magnitude. These results were used to identify the best genotypes for both resistance factors. Four hybrids showed low disease incidence values under both artificial and natural infections, of which only two had high values of the symptom appearance index at both locations. These hybrids are considered to have adequate resistance attributes for the ascosporic penetration and the mycelium extension in the capitulum tissue and could therefore be sown in the province of Buenos Aires without increasing risks of S. sclerotiorum attacks.     

Improvement of large-seeded common bean cultivars under sustainable cropping systems in Spain

Approaches are needed to broaden the genetic base and improve earliness and yield potential of large-seeded beans under sustainable cropping systems. The objective of this research was to develop adapted dwarf bean populations having a commercial seed quality and yield suitable for the production in the South of Europe. The original base populations were produced from crosses between genotypes within each Mesoamerica, Nueva Granada and Peru races, and between Peru and Nueva Granada, and Mesoamerica and Nueva Granada races. Visual mass selection for plant performance was practised in the F₂ and F₃ generations. In the F₄ and F₅, single plants were harvested under two cropping systems (sole cropping and intercropping with maize). From F₄, selection was based on precocity, combined with seed yield and seed commercial type. The F_4:7 selected lines from each original population were compared with their parents and five checks at four environments and two cropping systems. Differences among environments, populations, parents and checks were observed for all traits. Under intercropping with maize, there was a 50% reduction in seed yield. Yield of parents and checks belonging to Andean South American races, intraracial (Nueva Granada × Nueva Granada) and interracial (Nueva Granada × Peru) populations, was higher than that of those of Middle American origin. Intraracial crosses within large-seeded Andean South American (Peru race) and Middle American gene pools (Mesoamerica race) did not produce lines yielding more than the highest yielding parent. Only two large-seeded lines selected from crosses between small- and large-seeded gene pools out-yielded the best parent and check cultivar.     

Quantitation of selected odor-active constituents in dry fermented sausages prepared with different curing salts

The odor-active compounds of dry-fermented sausages with added nitrite or nitrate as curing agents were identified by gas chromatography-olfactometry (GC-O) applying the detection frequency (DF) method. The quantification of these compounds in the sausage was determined by multiple headspace solid-phase microextraction (multiple HS-SPME). There were no specific odor-active compounds related to the use of nitrite or nitrate although there were differences in the DF value of several compounds. The nitrite-added sausages presented higher DF values for ethanol, 1-hexanol, propanoic acid, 2-heptenal, and nonanal while the nitrate-added sausages had higher DF values for phenylacetaldehyde and 3-methyl-butanal. Eighteen compounds were quantified by multiple HS-SPME. Most of them were above their air detection thresholds, but only hexanal, heptanal, and 1-octen-3-ol were in a concentration higher than their oil threshold values. These compounds would probably be the main contributors to the aroma of fermented sausages.