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Efikasnost bakar-citrata u zaštiti vinove loze od bolesti
The control of Plasmopara viticola and Botrytis cinerea, two of the most dangerous pathogens on grapevine, requires frequent treatments with chemicals based on weather conditions. Numerous applications of fungicides have resulted in developing fungicide resistance. Active ingredients based on copper have been used very successfully for a long time to protect grapevines against these pathogens. Application of a copper citrate formulation with high degree dissociation at a very low concentration (1%) was evaluated in field trials. The efficacy of two concentrations of copper citrate, 0.5 and 1.0%, were tested against P. viticola on grapevine in three locations, and against B. cinerea in two locations during 2018. Our results demonstrated that the concentration of 1.0% copper citrate was highly effective against P. viticola (87.4%) and B. cinerea (63.7%), compared to standard treatment (90.6 and 53.1%), under a high level of infection.Zaštita vinove loze od prouzrokovača plamenjače vinove loze - Plasmopara viticola i sive truleži - Botrytis cinerea, je vrlo kompleksna i zahteva primenu većeg broja hemijskih tretmana, u skladu sa vremenskim uslovima. Učestala primena fungicida uslovljavala je pojavu rezistentnih izolata patogena na fungicide. Različite forme bakarnih jedinjenja primenjuju se u zaštiti vinove loze dugi niz godina prilično uspešno. Primena bakar-citrata - formulacije sa visokim stepenom disocijacije u niskoj koncentraciji (1,0%) ispitivana je u poljskim uslovima. Efikasnost dve koncentracije bakar-citrata - 0.5 i 1.0% je testirana u suzbijanju P. viticola i B. cinerea na vinovoj lozi na tri (dva) lokaliteta, tokom 2018 godine. Naši rezultati pokazuju da je ispitivana koncentracija od 1,0% bakar-citrata ispoljila zadovoljavajući efekat na P. viticola (87,4%) i B. cinerea (63,7%) u odnosu na primenjene standarde (90,6% i 53,1%) u uslovima visokih zaraza
Specificity and sensitivity of three pcr-based methods for detection of erwinia amylovora in pure culture and plant material
Three PCR methods, referred in this study as,conventional" "nested" and,chromosomal" PCR and suggested for routine detection of Erwinia amylovora in pure culture and plant material, were evaluated according to their specificity and sensitivity. Specificity of PCR methods was analyzed by using 42 strains of E. amylovora, originating from different locations and plant species, with diverse PFGE profiles, representing distant populations of the pathogen. Sensitivity of PCR protocols in pure culture was studied by using nine different concentrations of E. amylovora in sterile ultrapure water as a template in PCR reactions. In order to study inhibitory effect of plant DNA and other inhibitors on sensitivity of the three PCR methods bacterial dilutions were mixed with plant macerate of pear, apple and quince prior to the PCR reaction. In specificity assays, tested PCR protocols were able to detect all E. amylovora strains regardless of the host of the strain, its origin or PFGE group, indicating primer specificity. On the other hand, sensitivity among tested methods varied, depending on bacterial concentration and selected plant material used in the PCR. When working with pure cultures nested PCR showed the greatest sensitivity by detecting 1.9 bacterial cells per PCR reaction, followed by detection limit of 9.5 cells per PCR reaction with conventional PCR and 1.9.105 cells/PCR reaction with chromosomal PCR. In spiked samples plant inhibitors either did not affect or they decreased the sensitivity of the PCR reaction, depending on the protocol and/or type of plant macerate. In our experiments, inhibitors from pear and quince macerates did not affect sensitivity of nested PCR, while apple macerate reduced its sensitivity by a factor of 10. Conventional PCR protocol was able to detect 95 cells/PCR reaction in pear and apple macerate, but only 9.5.103 cells/PCR in quince macerate. Greatest decrease in sensitivity of the PCR method was observed in spiked samples with chromosomal PCR since bacterial DNA was not detected in each of the spiked samples. Our research shows that all three PCR protocols are specific for detection of E. amylovora, but nested PCR proved to be most sensitive when working with pure cultures and plant material
Integration of biological and conventional treatments in control of pepper bacterial spot
Bacterial spot caused by Xanthomonas euvesicatoria is one of the most devastating pepper diseases in Serbia. Questionable seed quality, climatic conditions, and frequent irrigation during summer favour the disease occurrence and spread. The available management practices do not provide adequate disease control. Therefore, development of alternative and more sustainable disease management strategies is needed. Integration of classical and biological treatments could be an effective, environmentally safe option for reducing pepper bacterial spot severity. In order to develop an efficient integrated disease management program, we studied efficacy of biocontrol agents (bacteriophage strain KФ1 and two strains of Bacillus subtilis AAac and QST 713), systemic acquired resistance (SAR) inducer (acibenzolar-S-methyl - ASM), a commercial microbial fertilizer (Slavol), copper based compounds (copper hydroxide and copper oxychloride) in combination with or without mancozeb, and antibiotics (streptomycin sulphate and kasugamycin). They were applied as single treatments in two separate field experiments. Based on the single treatment efficacy, various combinations of the treatments were chosen for further testing in three separate field experiments. Additionally, we evaluated potential negative effect of ASM on pepper growth and yield in the growth chamber experiment. All the tested single treatments significantly reduced disease severity compared to the inoculated control (IC), except microbiological fertilizer and the antagonistic strain AAac. Integration of copper hydroxide, ASM and bacteriophages was the most efficient treatment, reducing the disease intensity by 96–98%. The results indicated that this combination may be an adequate alternative program for control of pepper bacterial spot
Enhancement of antioxidant activity and bioactive compound contents in yellow soybean by plant-extract-based products
Abstract: Polyphenols present in different plant cell organelles increase the resistance of plants to various types of environmental
stresses. We investigated the possibility of increasing the content of bioactive compounds in the seed of yellow
soybean variety Laura. The soybean was treated during vegetation with five products based on plant extracts, on the
assumption of enrichment of plants with various nutrients. Soybean flour extracts were screened spectrophotometrically
for total phenolic content and antioxidant activity. The antioxidant activity was evaluated using three methods. The content
of phenolic acids was determined by HPLC, and the raw protein content was estimated by the Kjeldahl method. Depending
on the treatment, variations in the quantity of individual phenolic acids with up to 90% higher concentration as compared
to the control were observed. Controlled usage of certain plant extracts can increase the concentration of the target group of
bioactive compounds in the samples. The synergistic effect of proteins and phenolic compounds on the antioxidant activity
of extracts was detected. The results of this study are not only important from the aspect of plant resistance to various types
of stress, but also when considering soybean as a functional food
The Incidence and Genetic Diversity of Potato virus S in Serbian Seed Potato Crops
It is essential that certified potatoes are free from known viruses which can negatively affect quality and yield. However, very little is known about the distribution and frequency of Potato virus S (PVS) in Serbia. Until 2014, PVS was present sporadically in the domestic seed potato production. The incidence of PVS was studied by a molecular method over 3 years (2014-2016) in four important potato-growing regions (Moravidd. Zlatiborski, Raski and Macvanski) and in different cultivars. The results showed that the incidence of PVS increased steadily over 3 years from 1.52 to 8.84%. The Moravicki region had the highest incidence (13.06%) and Desiree was the most susceptible cultivar with a mean PVS incidence of 8.2% followed by Marabel and Riviera. The highest significant statistical difference was between the cultivars and in the interaction between cultivars and regions. A detailed phylogenic analysis of the tested isolates contained that Serbian PVS belongs to PVSO. Of the 18 Serbian PVS isolates included in this study, eight were grouped into the PVSO cluster and formed a subgroup (O-I) with isolates from the USA, Syria, Korea and Chile. Ten Serbian isolates of PVS together with the isolates from Iran were clustered in a branch of subgroup O-VII. This study constitutes the fast report of PVS isolates in Serbia which are capable of infecting Chenopodium quinoa and inducing the symptoms of local chlorotic lesions
Characterization of Gnomoniopsis idaeicola, the Causal Agent of Canker and Wilting of Blackberry in Serbia
Blackberry cane diseases with the symptoms of necrosis, canker, and wilting are caused by several fungi worldwide. Surveys conducted from 2013 to 2016 in Serbia revealed the occurrence of Gnomoniopsis idaeicola, the causal agent of cane canker and wilting, which was found to be distributed in almost half of the surveyed orchards, in three blackberry cultivars, and with disease incidence of up to 80%. Wide distribution and high disease incidence suggest that G. idaeicola has been present in Serbia for some time. Out of 427 samples, a total of 65 G. idaeicola isolates were obtained (isolation rate of 34.19%). Reference isolates, originating from different localities, were conventionally and molecularly identified and characterized. G. idaeicola was detected in single and mixed infections with fungi from genera Paraconiothyrium, Colletotrichum, Diaporthe, Botryosphaeria, Botrytis, Septoria, Neofusicoccum, and Discostroma, and no diagnostically specific symptoms could be related directly to the G. idaeicola infection. In orchards solely infected with G. idaeicola, blackberry plant mortality was up to 40%, and yield loses were estimated at 50%. G. idaeicola isolates included in this study demonstrated intraspecies diversity in morphological, biological, pathogenic, and molecular features, which indicates that population in Serbia may be of different origin. This is the first record of a massive outbreak of G. idaeicola infection, illustrating its capability of harmful influence on blackberry production. This study represents the initial step in studying G. idaeicola as a new blackberry pathogen in Serbia, aiming at developing efficient control measures
Biological control of Pseudomonas syringae pv. aptata on sugar beet with Bacillus pumilus SS-10.7 and Bacillus amyloliquefaciens (SS-12.6 and SS-38.4) strains
Aim Assessment of biological control of Pseudomonas syringae pv. aptata using crude lipopeptide extracts (CLEs) of two Bacillus amyloliquefaciens strains (SS-12.6 and SS-38.4) and one Bacillus pumilus strain (SS-10.7). Methods and Results The minimum inhibitory concentration (MIC) of CLEs and their combinations against the pathogen and potential interaction between the extracts were determined in vitro. The most effective antibacterial activity was achieved with the CLE from B. amyloliquefaciens SS-12.6, with an MIC value of 0 center dot 63 mg ml(-1). Interactions between CLE combinations were mostly indifferent. The biocontrol potential of CLEs, mixtures of CLEs, and cell culture of B. amyloliquefaciens SS-12.6 was tested on sugar beet plants inoculated with P. syringae pv. aptata P53. The best result in inhibiting the appearance of tissue necrosis (up to 92%) was achieved with B. amyloliquefaciens SS-12.6 cell culture. Conclusion This work demonstrated significant biocontrol potential of the CLE and cell culture of B. amyloliquefaciens SS-12.6 which successfully suppress leaf spot disease severity on sugar beet plants. Significance and Impact of the Study The findings of biocontrol of sugar beet emerging pathogen will contribute to growers in terms of alternative disease control management. This study represents first assessment of biological control of P. syringae pv. aptata
First pentasetacid mite from Australasian Araucariaceae: morphological description and molecular phylogenetic position of Pentasetacus novozelandicus n. sp. (Eriophyoidea, Pentasetacidae) and remarks on anal lobes in eriophyoid mites
A new vagrant eriophyoid mite species of the archaic genus Pentasetacus (Schliesske 1985), P. novozelandicus n. sp., is described with the aid of conventional microscopy, confocal laser scanning microscopy and scanning electron microscopy. It was found on Araucaria heterophylla, which is an araucarian that is endemic to Norfolk Island and introduced to New Zealand. Partial sequences of mitochondrial barcode COI gene and D1–D2 domains of nuclear rDNA of two pentasetacid mites, P. araucariae (MK903025 and MK898944) and P. novozelandicus n. sp. (MK903024 and MK898943) are provided. Molecular phylogenetic analyses of full-length D1–D2 eriophyoid sequences, including GenBank sequences and newly generated sequences of pentasetacids, confirmed the monophyly of Pentasetacidae but failed to resolve the basal phylogeny of Eriophyoidea. This may be because the D1–D2 domains of 28S are hypervariable in Eriophyoidea. Moreover, in pentasetacids D1–D2 sequences are about 20% shorter than in other eriophyoids, and thus harder to align. Two types of anal lobes are described in Eriophyoidea: (1) Eriophyidae s.l. and Phytoptidae s.l. have bilaterally symmetric lobes; (2) pentasetacids have non-divided lobes. The presence of an anal secretory apparatus, comprising internal structures that have previously been described in Eriophyidae s.l. and Phytoptidae s.l., is confirmed in pentasetacid genera. The phylogeny of pentasetacids is also discussed in the context of the paleobiography of Araucariaceae
Identifikacija i filogenetska analiza Fusarium proliferatum prouzrokovača truleži luka Allium ampeloprasum L
Symptoms of cloves rot of A. ampeloprasum were noticed during 2018 in storage conditions. 16 isolates were obtained (AMP1-AMP16) and according to morphological and cultural characteristics they belong to F. proliferatum (sex. stage Gibberella intermedia), species of Gibberella fujikuroi complex. To confirm morphological identification, total genomic DNA was extracted from mycelium of the 16 isolates by amplification of TEF-1a gene, using polymerase chain reaction (PCR) that was performed with the primer pair EF1 and EF2. Results presented in this article clearly indicated that the new host of Fusarium proliferatum as the causal agent of cloves rot is "elephant garlic" Allium ampeloprasum. Pathogenicity test was confirmed on Allium ampeloprasum cloves. Pathogenicity assays revealed that all isolates caused symptoms on tested Allium spp., like naturally infected cloves.Simptomi truleži uskladištenih čenova belog luka na vrsti poznatoj kao "elephant garlic" (A. ampeloprasum) zapaženi su tokom 2018. godine. Prikupljen je veliki broj zaraženih uzoraka i nakon izolacije odabrano je 16 izolata (AMP1-AMP16) za dalja istraživanja. Na osnovu morfoloških i odgajivačkih karakteristika je ustanovljeno da je prouzrokovač truleži čenova gljiva F. proliferatum (telemorf Gibberella intermedia), koja pripada kompleksu Gibberella fujikuroi. Proučavanjem patogenosti svi izolati su prouzrokovali simptome truleži na različitim vrstama roda Allium spp. koji su identični prirodnoj infekciji. U cilju potvrde morfoloških odlika izvršena je molekularna identifikacija metodom lančane reakcije polimeraze (PCR) korišćenjem para prajmera EF1 i EF2 koji amplifikuju TEF-1a gen, sekvencioniranje DNK i filogenetska analiza dobijenih sekvenci. Dobijeni rezultati potvrdili su da je Fusarium proliferatum prouzrokovač truleži na vrsti A. ampeloprasum kao novom domaćinu
Pectobacterium zantedeschiae sp. nov. a new species of a soft rot pathogen isolated from Calla lily (Zantedeschia spp.)
Four Gram-negative, rod-shaped pectinolytic bacterial strains designated as 2M, 9M, DPMP599 and DPMP600 were subjected to polyphasic analyses that revealed their distinctiveness from the other Pectobacterium species. Strains 2M and 9M were isolated from Calla lily bulbs cultivated in Central Poland. DPMP599 and DPMP600 strains were isolated from Calla lily leaves from plants grown in Serbia. Phylogenetic analyses based on nine housekeeping genes (gapA, gyrA, icdA, pgi, proA, recA, recN, rpoA, and rpoS), as well as phylogeny based on the 381 most conserved universal proteins confirmed that Pectobacterium zantedeschiae strains were distantly related to the other Pectobacterium, and indicated Pectobacterium atrosepticum, Pectobacterium betavasculorum, Pectobacterium parmentieri and Pectobacterium wasabiae as the closest relatives. Moreover, the analysis revealed that Pectobacterium zantedeschiae strains are not akin to Pectobacterium aroidearum strains, which were likewise isolated from Calla lily. The genome sequencing of the strains 2M, 9M and DPMP600 and their comparison with whole genome sequences of other Pectobacterium type strains confirmed their distinctiveness and separate species status within the genus based on parameters of in silico DNA–DNA hybridization and average nucleotide identity (ANI) values. The MALDI-TOF MS proteomic profile supported the proposition of delineation of the P. zantedeschiae and additionally confirmed the individuality of the studied strains. Based on of all of these data, it is proposed that the strains 2M, 9M, DPMP599, and DPMP600 isolated from Calla lily, previously assigned as P. atrosepticum should be reclassified as Pectobacterium zantedeschiae sp. nov. with the strain 9M T (PCM2893 = DSM105717 = IFB9009) as the type strain