Identification and characterization of Agrobacterium spp. isolated from apricot in Serbia

Crown gall, caused by tumorigenic strains of Agrobacterium spp., is considered one of the most important diseases in stone fruit nurseries throughout the world. Since the crown gall disease has not been studied extensively in Serbia for more than 30 years, the objective of this study was to isolate, identify and characterize the bacterium associated with crown gall symptoms on one-year-old apricot trees. Samples were collected from a nursery in central Serbia and subjected to laboratory analysis. Bacteria were isolated from tumour tissue on yeast mannitol agar (YMA) and six gram-negative isolates were selected for further study. PCR assay using primers specific for virD2, virC, ipt and tms2 pathogenicity-associated plasmid genes revealed that isolates harbour Ti plasmids. All studied strains carrying Ti plasmids were classified as nopaline-type based on further molecular analysis. Using a multiplex PCR assay, targeting 23S rRNA gene sequences, and physiological and biochemical tests, five strains were assigned as A. rhizogenes/biovar 2 and the remaining one as A. tumefaciens/biovar 1. Identity of the strains was confirmed by sequence analysis of the 16S rRNA gene. In pathogenicity assay, all six strains caused tumour formation on inoculated carrot root discs, young tomato and sunflower plants.     

Analysis of Wild Raspberries (Rubus idaeus L.): Optimization of the Ultrasonic-Assisted Extraction of Phenolics and a New Insight in Phenolics Bioaccessibility

Plant Foods for Human Nutrition - A simple and efficient ultrasonic-assisted extraction (UAE) technique was developed in order to find optimal conditions for the extraction of total phenolic...     

Differentiation of <Emphasis Type="Italic">Erwinia amylovora</Emphasis> strains from Bulgaria by PCR-RFLP analysis

Fifty strains of Erwinia amylovora isolated in Bulgaria from different host plants and locations as well as in different years were analysed by RFLP analysis of the pEA29 PstI amplified fragment with HpaII. All the strains formed three well-resolved fragments (large—from 365 to 440 bp, medium—about 341 bp and small—about 180 bp).The strains were classified into three RFLP groups based on the polymorphism in the length of the largest fragment. This fragment was of intermediate size for 63% of the strains, and it was the longest (from 410 to 440 bp) for 29% of the strains. The variable region was sequenced for five strains. The DNA sequence analysis confirmed the different size of the largest fragment. Ten or more than ten SSRs were found for the strains in the group with the largest size of the largest fragment. Some correlation between the RFLP profiles and the origin of the strains was revealed. The RFLP profiles displayed stability in certain strains isolated from the same trees and orchards, but in different years. The number of SSRs was different in strains isolated from one and the same host plant, orchard and year, and also in strains isolated from the same host plant and orchard, but in different years. This could indicate that under natural conditions the fire blight symptoms might be caused by a mixture of E. amylovora strains with different SSR numbers, and so coexistence of distinguishable strains or a change in the population could be assumed.     

Histological characteristics of sugar beet leaves potentially linked to drought tolerance

Water is becoming more and more limiting factor of sugar beet production and the productivity of the crop can be significantly improved by increased drought tolerance. It is therefore a great challenge to assess the degree of variability of anatomical and morphological traits of breeding material with respect to water use efficiency and drought, that can be used as potential markers for selection of sugar beet genotypes with better tolerance to water shortage. To achieve this, the first step is to assess the degree of genetic variability with respect to anatomical and histological features linked to water management in plants, under optimal water supply. Comparative histiological analysis of lamina and petiole was done on 12 sugar beet genotypes which previously showed divergent responses to lack of water in the field. The plants were grown in semi-controlled conditions of a glasshouse, and watered daily. Mircromorphological analyses were done to assess leaf epidermal characteristics, by both light and SEM, and lamina and petiole histological features. The measurements were used to calculate the percentage of individual tissues in relation to the thickness of the lamina, main vein area and petiole area. The general structure of sample variability was established by principal component analysis (PCA), based on correlation matrix. In majority of genotypes the ratio of the size of cells of spongy parenchyma and palisade cells in average was 80%. Low genotypic variability of the studied histological parameters of the lamina and petiole may reflect the narrow genetic base of tested breeding material. The most significant genotypic difference, considering leaf epidermal tissue, was in % of adaxial and abaxial epidermis. The highest found number of stomata per mm² on both adaxial and abaxial epidermis was 40% higher than the lowest. During water stress, when stomata are closed, plant survival depends on the amount of water lost through the cuticle. SEM analysis of adaxial epidermis of the lamina show that cuticle varies in texture. Considering the observed genotypic variability in cuticle ornamentation and the fact that plants develop various strategies of adaptation to drought, finding genotypes with increased drought tolerance could be based on the characteristics of the cuticle and epidermis.     

Tuning the properties of PVDF or PVDF-HFP fibrous materials decorated with ZnO nanoparticles by applying electrospinning alone or in conjunction with electrospraying

Novel composite nanofibrous materials of poly(vinylidene fluoride) (PVDF) or poly(vinylidene fluoride-cohexafluoropropylene) (PVDF-HFP) and ZnO nanoparticles were prepared by conjunction of electrospinning and electrospraying techniques. Simultaneous electrospinning of concentrated solution of PVDF or PVDF-HFP and electrospraying of suspension of ZnO in diluted PVDF or PVDF-HFP solution enable the preparation of materials consisting of fibers on which ZnO was deposited on the fibers’ surface (design type “on”). These fibrous materials were compared with materials consisting of PVDF or PVDF-HFP fibers in which ZnO was incorporated in the fibers (design type “in”) and which were obtained by one-pot electrospinning of a suspension of ZnO nanoparticles in concentrated PVDF or PVDF-HFP solution. The fiber morphology and the presence of ZnO “in” or “on” the fibers were observed by scanning electron microscopy (SEM) and by transmission electron microscopy (TEM). The effect of the used technique on the type, size and shape of the obtained structures was discussed. The fibrous mats were studied by differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray diffraction (XRD), contact angle measurements and mechanical tests as well. It was found that the decoration of fibers with ZnO resulted in increase of their thermal stability and hydrophobicity. The microbiological tests showed that the materials of design type “on” possessed strong antibacterial activity against the pathogenic microorganism Staphylococcus aureus. The results suggest that, due to their antibacterial activity, the obtained composite materials are suitable for wound dressing applications.     

Ammonium Enables Aluminum-Induced Stimulation of Nitrogen Assimilation in Roots of Al-Tolerant Maize Genotypes

One aluminum (Al)-sensitive (B-73) and two Al-tolerant (F-2 and L-2039) maize genotypes were subjected to Al stress (100 μM Al) under two nitrogen (N) treatments [13.2 mM nitrate (NO₃^?) and 8.3 mM NO₃^? + 4.9 mM ammonium (NH₄⁺)]. Growth parameters, chlorophyll, root N and NO₃^? contents, root nicotinamide adenine dinucleotide (NADH-) and nicotinamide adenine dinucleotide phosphate (NADPH)-nitrate reductase, glutamine synthetase, and glutamate dehydrogenase activities were determined. Aluminum significantly decreased growth and chlorophyll content in Al-sensitive genotype. Nitrate accumulation in roots was increased by Al in tolerant plants. In the sensitive genotype, Al suppressed all enzymes in NO₃^? plants, while in NO₃^?/NH₄⁺ plants the suppression was less severe, and NADPH-nitrate reductase was even stimulated. In tolerant NO₃^?plants, glutamate dehydrogenase was stimulated in F-2 and glutamine synthetase suppressed in L-2039 genotype. In tolerant NO₃^?/NH₄⁺- plants, all enzymes were stimulated by Al, which may be attributed to their participation in defense mechanisms.     

Ex situ evaluation of cultivation potential in wild populations of large-flowered vetch (Vicia grandiflora)

Large-flowered vetch (Vicia grandiflora Scop.) is widely present in many Eurasian wild floras, where it has significance in improving grassland communities and environment-friendly uses as a cover crop and green manure. The main goal of this study was to assess the possibility of transforming large-flowered vetch into a field crop and assess the basics for its breeding. A small-plot trial was carried out in three consecutive growing seasons 2005–2008 in Novi Sad, including twelve large-flowered wild populations collected at various sites in Serbia and France from 2002 to 2004 and multiplied ex situ during subsequent years. The population MM 02/01 of Serbian origin had the highest three-year average proportion of plants surviving winter (0.96). The smallest average number of days from sowing to first flower was in the population MM 03/10 (199 days). The population MM 03/08 produced the highest forage dry matter (8.0 Mg ha^?1) yield stage between full flowering and forming first pods. The highest seed yield was in the population MM 03/10 (2,084 kg ha^?1). Forage dry matter yield was significantly correlated to main stem length (r = 0.932), number of fertile nodes (r = 0.710), number of internodes (r = 0.640) and number of leaves (r = 0.604). Seed yield was significantly correlated to number of pods (r = 0.930), number of seeds (r = 0.883), number of stems (r = 0.882) and number of fertile nodes (r = 0.768). Our study confirmed that large-flowered vetch may represent a stable and quality source of forage in temperate regions such as the Balkans, as well as that it may have its place in various crop rotations and farming systems. At the same time, the existing phenotypic variability offers a solid basis for further improvement of this species.     

Self-assembly of subnanometer-diameter single-wall MoS2 nanotubes

We report on the synthesis, structure, and self-assembly of single-wall subnanometer-diameter molybdenum disulfide tubes. The nanotubes are up to hundreds of micrometers long and display diverse self-assembly properties on different length scales, ranging from twisted bundles to regularly shaped "furry" forms. The bundles, which contain interstitial iodine, can be readily disassembled into individual molybdenum disulfide nanotubes. The synthesis was performed using a novel type of catalyzed transport reaction including C(60) as a growth promoter.     

Relationship between hybrid performance and genetic variation in self-fertile and self-sterile sugar beet pollinators as estimated by SSR markers

Sugar beet hybrid varieties are produced through the crosses between male sterile lines and the multigerm pollinators. The uniformity of pollinators used for hybrid crosses depends on the presence of self-sterility (S ^s ) and self-fertility (S ^f ) genes. The aim of the study was to analyze correlation between hybrid performance and genetic distance or heterozygosity of the sugar beet pollinators. Twelve diploid pollinators classified as self-sterile (S ^s ) or self-fertile (S ^f ) and two cytoplasmic male sterile (CMS) lines were crossed in line × tester scheme, producing 24 F₁ hybrids. The parents and the hybrids were evaluated for root yield and quality traits, from which F₁ performance, combining abilities, mid-parent and high-parent heterosis were calculated. Parental genetic distance and diversity of the pollinators were estimated by SSR markers and, together with GCA and F₁ performance, correlated with the heterosis effects. The S ^f hybrids had better GCA and higher values of root yield, root weight, and root circumference than the S ^s hybrids. Heterosis was recorded in more combinations with the S ^f than with the S ^s pollinators. Parameters of genetic diversity were higher in the S ^s (Na = 3.125; Ne = 2.341; He = 0.555) than in the S ^f pollinators (Na = 3.000; Ne = 2.188; He = 0.510). Genetic distance between the tested pollinators and the CMS lines was low (0.072–0.224) indicating that the genetic base of the investigated germplasm was narrow. Correlation of the heterosis effects with GD and heterozygosity was detected only for the root yield traits.