Identification and Detection of Rosellinia Necatrix by Conventional and Real-time Scorpion-PCR

Several polymerase chain reaction (PCR) primers were designed from the internal transcribed spacer (ITS) regions of the rDNA genes of Rosellinia necatrix to develop a PCR-based identification method. Screening the primers against two isolates of R. necatrix and six other Rosellinia species resulted in the amplification of a single specific product from R. necatrix for most of the primer pairs. Two primer pairs (R2-R8 and R10-R7) confirmed their specificity when tested against 72 isolates of R. necatrix and 93 other fungi from different hosts and geographic areas. The R10 primer was modified to obtain a Scorpion primer for detecting a specific 112bp amplicon by fluorescence emitted from a fluorophore in a self-probing PCR assay. This assay specifically recognised the target sequence of R. necatrix over a large number of other fungal species. In conventional PCR, with primer pairs R2-R8 and R10-R7, 10-fold dilutions of R. necatrix DNA indicated a detection limit of 10pgul^-1 using a single set of primers and 10fgl^-1 in nested-PCR. For Scorpion-PCR, the detection limit was 1pgl^-1 and 1fgl^-1 in nested Scorpion-PCR, i.e. 10 times more sensitive than conventional PCR. A simple and rapid procedure for DNA extraction directly from soil was modified and developed to yield DNA of purity and quality suitable for PCR assays. Combining this protocol with the nested Scorpion-PCR procedure it has been possible to specifically detect R. necatrix from artificially inoculated soils in approximately 6h.     

Effects of light availability and soil flooding on growth and photosynthetic characteristics of Genipa americana L. seedlings

Genipa americana L. (Rubiaceae) is a late successional neotropical fruit tree used in riparian forest restoration programs. We analyze the effects of light availability and soil flooding on the growth and photosynthetic characteristics of G. americana seedlings under nursery and natural light conditions. Two light levels (full sunlight and shade), and two levels of soil water (flooded and control) were used in the experiment. Flooding induced significant changes in the total seedling biomass (P < 0.01). The differences among water treatments were 70 and 10% at full sunlight and shade, respectively. These changes were explained by alterations in the maximum quantum efficiency of the photosystem 2 (Fv/Fm), light-saturated net photosynthetic rates (A _sat) and intrinsic water use efficiency (WUE). G. americana has high physiological plasticity in relation to the light availability and flooding, although significant interactive effects between high light exposure and soil flooding were observed in several photosynthetic and growth variables. The results highlight the importance of the synergistic effects between physical environmental variables on the establishment and growth of G. americana seedlings. In the practical point of view we can indicate that, in degraded riparian forests subjected to soil flooding, seedlings of this species should be planted under partially shaded environments.     

Accuracy of early fetal sex determination by ultrasonic assessment in goats

The objective of this study was to determine the accuracy of early fetal sex determination by ultrasonic assessment of the relative location of the genital tubercle (GT) in goats at different stages of pregnancy as well as by the identification of fetal external genitalia. Pregnant animals were divided into three experimental groups (EI: n=21, EII: n=28, EIII: n=33). In EI, fetuses (n=27) were transrectally monitored daily from days 40 to 60 of pregnancy with a linear transducer (6.0 and 8.0MHz). In EII, fetuses (n=40) were examined once between days 45 and 70 of pregnancy by transrectal ultrasonography. In EIII fetuses (n=52) between days 100 and 120 of pregnancy, were submitted to a single transabdominal ultrasonography using a convex transducer (5.0 and 7.5MHz). Regardless of fetal sex diagnosis, 15/15 (EI), 13/16 (EII) and 9/14 (EIII) of single pregnancies and 10/12 (EI), 20/24 (EII) and 21/38 (EIII) of twin pregnancies were correctly identified. The accuracy of sex identification among EI (92.6%), EII (82.5%) and EIII (57.7%) was not statistically different (P>0.05). Identification of the GT in male fetuses was possible from day 45 onward. Changes in the GT position were not observed between days 53 and 60 of pregnancy. Accuracy of fetal sexing under field conditions is high in goats when ultrasound imaging is properly timed during pregnancy and when it is performed with proper equipment by experienced operators.     

Evaluation of Nile tilapia in monoculture and polyculture with giant freshwater prawn in biofloc technology system and in recirculation aquaculture system

Biofloc technology system (BFT), recirculation aquaculture system (RAS) and polyculture promote efficient use of water, area and nutrient recycling, which are essential practices for sustainable aquaculture development. The aim of this study was to evaluate the growth, feed efficiency, biofloc composition and water quality of Nile tilapia Oreochromis niloticus (Linnaeus, 1758) in monoculture and polyculture with giant freshwater prawn Macrobrachium rosenbergii (De Man, 1906) in BFT and RAS, over a period of 30 days. Fish (n = 128; 7.29 ± 0.67 g) were distributed randomly in 16 experimental tanks (8 fish/tank). Prawn (n = 96; 0.50 ± 0.09 g) were allocated in 8 experimental tanks (12 prawn/tank) in a polyculture. The experimental design was completely randomized with four treatments with four replicates each, in a factorial design 2 × 2 (BFT and RAS vs. monoculture and polyculture). The experimental diet (28% of digestible protein; 3100 kcal kg−1 of digestible energy) was used both to fish and prawn in BFT and RAS. There was significant effect (p < 0.01) of the system and the culture for weight gain, apparent feed conversion and protein efficiency ratio. The average weight gain and apparent feed conversion of tilapia in monoculture (30.04 g and 1.39) and in polyculture (36.44 g and 1.27) were superior (p < 0.01) in BFT than in monoculture (23.64 g and 1.74) and in polyculture (24.14 g and 1.61) in RAS. Weight gain and survival of giant freshwater prawn was superior (p < 0.01) in BFT (0.43 g and 87%) compared to RAS (0.26 g and 79%). The data showed that BFT provides better growth performance responses in monoculture for Nile tilapia and in polyculture with giant freshwater prawn compared to RAS.     

Infestation potential of Dryocosmus kuriphilus Yasumatsu, 1951 (Hymenoptera: Cynipidae) in different natural populations of Castanea sativa Miller: an experimental ex situ test

The Asian chestnut gall wasp was accidentally introduced in Italy in 2002 and spread across Europe in the following years, becoming a serious threat for chestnut cultivations and rural economies of many countries. Exploring the variation in susceptibility of the host genetic resources is crucial to face the spreading of this pest. We used an experimental approach for testing the differential susceptibility within and between populations of European chestnut. For doing this, we compared both the infestation level and the rate of immune individuals in trees from populations of Spain, Italy, and Greece. We found that the level of infestation is not significantly different in the different provenances but that a higher rate of immune trees occur in Greece. Our results suggest that two different contingents of trees compose Greek populations: one major group of trees with the same susceptibility as the other populations and a second minor group of trees resistant to gall wasp infestation. Our data lay the basis for improving the currently adopted measures to mitigate gall wasp impacts.     

Application of Tamarind Waste Extracts to Improve the Antioxidant Properties of Tamarind Nectars

Plant Foods for Human Nutrition - Tamarind fruits are consumed worldwide and their seeds have an underexploited potential. We assessed the effect of the addition of a freeze-dried aqueous of...     

Impact of preharvest application of biological control agents on postharvest diseases of fresh fruits and vegetables

During the storage of harvested commodities, environmental parameters are quite stable. For this and other reasons, it is generally believed that biological control by means of microbial antagonists may have a greater potential for success when applied postharvest. However, one of the major obstacles to the development of postharvest biocontrol agents is that they are unable to control previously established infections, such as latent and quiescent infections and incipient infections occurring through wounds resulting from harvesting operations. Field application of biocontrol agents may enable early colonisation of fruit surfaces, thus protecting from these infections. Moreover, preharvest applications can be an appropriate strategy for fruits and vegetables subject to damage in postharvest handling. To be successful in preharvest applications, putative biocontrol agents must be able to tolerate low-nutrient availability, UV radiation, high temperature and dry conditions. Some reports of postharvest biological control accomplished by preharvest applications, include apples, avocados, sweet cherries, grapes, and strawberries. This paper provides a brief overview on particular aspects of preharvest application of biocontrol agents to reduce postharvest decay. Research areas relevant for the development of this strategy are also indicated.     

Effect of genotype,environment and genotype-by-environment interaction on metabolite profiling in durum wheat (Triticum durum Desf.) grain

The aim of this study was to assess the relative roles of genotype, environment and genotype-by-environment interactions in determining the metabolite profile of durum wheat grain. Four durum wheat cultivars were grown under conventional and organic farming systems over three consecutive years. The use of a high-throughput gas chromatography–mass spectrometry platform allowed the analysis of sets of different polar and non-polar compounds, including amino acids, sugars, organic acids, fatty acids (saturated and unsaturated), and sterols. Statistical analysis of the data showed a small impact of genotype and large effects of both year and genotype-by-environment interaction on the metabolite composition and quality of the wheat grain. Overall, the data from this study highlight the potential role of metabolic profiling in the analysis of durum wheat quality and production.     

Targeting QseC signaling and virulence for antibiotic development

Many bacterial pathogens rely on a conserved membrane histidine sensor kinase, QseC, to respond to host adrenergic signaling molecules and bacterial signals in order to promote the expression of virulence factors. Using a high-throughput screen, we identified a small molecule, LED209, that inhibits the binding of signals to QseC, preventing its autophosphorylation and consequently inhibiting QseC-mediated activation of virulence gene expression. LED209 is not toxic and does not inhibit pathogen growth; however, this compound markedly inhibits the virulence of several pathogens in vitro and in vivo in animals. Inhibition of signaling offers a strategy for the development of broad-spectrum antimicrobial drugs.     

Molecular and Biological Characterization of Tomato chlorotic mottle virus Suggests that Recombination Underlies the Evolution and Diversity of Brazilian Tomato Begomoviruses

ABSTRACT Tomato chlorotic mottle virus (ToCMoV) is an emerging begomovirus species widely distributed throughout tomato-growing regions of Brazil. ToCMoV appears to have expanded its geographic range recently, invading tomato-growing areas that were free of begomovirus infection before 2004. We have determined the first complete genome sequence of an infectious ToCMoV genome (isolate BA-Se1), which is the first begomovirus species isolated in the northeast of Brazil. When introduced by particle bombardment into tomato, the cloned ToCMoV-[BA-Se1] DNA-A and DNA-B components caused typical chlorotic mottle symptoms. The cloned virus was whitefly-transmissible and, although it was infectious in hosts such as Nicotiana benthamiana, pepper, tobacco, and Nicandra physaloides, it was unable to infect Arabidopsis thaliana, bean, N. glutinosa, and Datura metel. Sequence and biological analyses indicate that ToCMoV-[BA-Se1] is a typical New World begomovirus sp. requiring both DNA-A and DNA-B components to establish systemic infections. Although evidence of multiple recombination events was detected within the ToCMoV-[BA-Se1] DNA-A, they apparently occurred relatively long ago, implying that recombination probably has not contributed to the recent emergence of this species.