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Isolation and cultivation of coccoid Cyanobacteria from aerophytic biofilm in Stopić cave
Full text linkCyanobacteria, characterized by unique physiological and biochemical mechanisms to withstand
harsh conditions in different habitats, can thrive in a variety of environments, including extreme ones
such as illuminated parts of caves. The information on diversity, biology and ecology of aerophytic
Cyanobacteria, especially those from cave habitats, is very limited, which is especially true for the
coccoid forms. Phototrophic biofilm was collected from the entrance zone of Stopić Cave to isolate
coccoid Cyanobacteria. The biofilm was inoculated in situ into test tubes containing solid BG11
growth medium, after which the test tubes were kept in a growth chamber under controlled conditions.
From the initial culture, which contained multiple cyanobacterial species, subsamples were extracted
and transferred to Petri dishes. These were kept under optimal conditions until a pure cultures were
obtained. The representative of the coccoid Cyanobacteria, Chroococcus, was isolated and its
characteristics best matched the original description of Chroococcus turgidus
Scaphoideus titanus up-to-the-minute: biology, ecology, and role as a vector
Full text linkNative to the Nearctic region, Scaphoideus titanus Ball (Hemiptera: Cicadellidae) has become a major threat for
grapevine production after being unintentionally introduced into Europe, where it became the main vector of flavescence
dorée phytoplasma, being mainly associated with the genus Vitis. Scaphoideus titanus is a highly efficient vector of the
most important phytoplasma affecting grapevine. For this reason, compulsory insecticide treatments have been introduced
against this pest in many European countries. Moreover, the continuous expansion of its geographical distribution makes
this leafhopper a serious threat for several non-European Countries. In this article, we review the current knowledge about
its taxonomy, morphology, biology, ecology, and its role as a vector. Finally, we point out the main challenges for research
aimed at reducing S. titanus and flavescence dorée expansion across Europe and avoiding spread of the disease outside the
Old World
The FDemic project – Tracking the role of alien and invasive species in the transmission cycle of Flavescence dorée (FD) phytoplasma in Serbia
Full text linkFlavescence dorée (FD) is a quarantine disease of grapevine caused by FD phytoplasma (FDp). This quarantine
disease severely impacts both vineyard productivity and landscape management in Europe’s key
viticultural regions. While the pathogen itself is native to Europe, its transmission cycle is associated with
several wild host plants, both native and alien, and a number of native and alien insect vectors. In addition
to native plant species (Alnus glutinosa, A. incana, Salix spp., Clematis vitalba and Corylus avellana), alien plants
(Ailanthus altissima and Vitis riparia) also act as constituents of the epidemiological cycle, forming part of
the transmission route as plant reservoirs carrying FDp genotypes in riparian areas. Similarly, native and
alien insect vector species constitute the routes of transmission inducing FDp outbreaks, as part of specific
pathosystems. An alien leafhopper of Nearctic origin, Scaphoideus titanus, is well-known to be the main
vector of FDp. Additionally, another alien leafhopper of Asian origin, Orientus ishidae (the mosaic leafhopper),
has been identified as a potential vector spreading FDp genotypes, thus posing an additional risk to
European vineyards. The presence of established populations of O. ishidae have recently been confirmed in
Serbia, where it was found in riparian areas in association with an invasive plant V. riparia. Consequently, the
primary goal of this project will be to focus on natural riparian habitats and transmission cycles between
and among native and exotic plants and planthopper/leafhopper species to understand how they interact
with the vineyard agroecosystem and lead to FD outbreaks. Moreover, given that the Balkans is tentatively
considered as the area of emergence of the FD3 genotypes inducing FDp epidemics, by studying its natural
sources and reservoirs, we also aim to shed light on the monotypic epidemic of FD3 in the vineyards of
Serbia
Hyalesthes obsoletus and nettle-associated 'Ca. Phytoplasma solani' epidemiological cycle in Serbia and the Balkans: Is it closed and specific?
Full text linkThe planthopper Hyalesthes obsoletus Signoret, 1865 (Hemiptera: Cixiidae) is a major vector and
driver of 'Ca. Phytoplasma solani' epidemiology (Maixner, 1994; Jović & Toševski, 2023) and
associated diseases of cultivated plants, including the grapevine-Bois noir (BN) pathosystem. Because
of its strong association with its host plants as a subterranean nymph and short life span as an adult
(Cargnus et al., 2012), the epidemiological cycle is host plant-specific and is determined by the vector's
host range and the pathogen's reservoir range (Imo et al., 2013; Maixner et al., 2014). Although H.
obsoletus is commonly viewed as a polyphagous insect, there is plenty of proof of host plant adaptation
in its populations, which are referred to as host races, biotypes, or cryptic species (Imo et al., 2013;
Maixner et al., 2014; Kosovac et al., 2016; 2018; 2019). The first described and most prevalent
epidemiological cycles linked with distinct strains of 'Ca. P. solani' are driven by nettle and bindweed
as pathogen reservoir plants and H. obsoletus host plants (Langer & Maixner, 2004). The genetic
distinction between the two pathosystems was initially identified on the tuf gene and then verified by
secY, stamp, and vmp1 gene typing (Langer & Maixner, 2004; Johannesen et al., 2012; Aryan et al.,
2014). In Serbia and the Balkans, nettle-associated 'Ca. P. solani' genotypes are not commonly found
in BN-affected grapevine (Atanasova et al., 2015; Kosovac et al., 2016); hence, research on this
epidemiological pathway is neglected and available data are scarce. During more than a decade of
research on 'Ca. P. solani' epidemiology and H. obsoletus biology, ecology, and genetics in the
Balkans, we have frequently found the presence of "wrong" 'Ca. P. solani' genotypes in vector
specimens obtained from nettles. Here, we assess these findings and call into question the geographic
specificity of nettle-associated epidemiology in southeastern Europe
PROJEKAT FDemic – PRAĆENJE ULOGE ALOHTONIH I INVAZIVNIH BILJNIH VRSTA U EPIDEMIOLOŠKOM CIKLUSU Flavescence dorée (FD) FITOPLAZME U SRBIJI
No full textFlavescence dorée (FD) je karantinska bolest vinove loze koju prouzrokuje FD
fitoplazma koju u vinogradima epidemijski prenosi cikada severnoameričkog porekla,
Scaphoideus titanus. Prisustvo FD bolesti u vinogradu ima snažan uticaj na prinos vinove
loze i održivo upravljanje vinogradima u ključnim vinogradarskim regionima Evrope. Iako
je insekt vektor bolesti alohtonog porekla, FD fitoplazma je autohtona za područje
Evrope, usled čega FD epidemiološki ciklus uključuje prirodne biljke domaćine patogena,
od kojih su neke alohtone i invazivne biljne vrste. Ciklus prenošenja FD fitoplazme takođe
uključuje i niz autohtonih i alohtonih insekata vektora. Pored autohtonih biljnih vrsta
(Alnus glutinosa, A. incana, Clematis vitalba, Salix spp. i Corylus avellana) i dve alohtone
i invazivne vrste (Ailanthus altissima i Vitis riparia) takođe igraju važnu ulogu u
epidemiološkom ciklusu FD fitoplazme. Ove biljne vrste predstavljaju biljke rezervoare
patogena, koje su nosioci genotipa FD fitoplazme u riparijalnim oblastima. Vitis riparia je
istovremeno i biljka domaćin za glavnog vektora FD fitoplazme - S. titanus. Kako
epidemiološki ciklus FD-a u velikoj meri zavisi od diverziteta biljaka prirodnih rezervoara
fitoplazme, primarni cilj projekta FDemic je da se fokusiranjem na prirodna riparijalna
staništa i cikluse transmisije između autohtonih i alohtonih vrsta biljaka i insekata ostvari
razumevanje na koji način ove vrste interaguju sa agroekosistemima vinograda i dovode
do FD epidemije i pojave novih žarišta. Balkan se smatra oblašću iz koje prirodno potiče
FD epidemija FD3 genotipa fitoplazme, zbog čega je izučavanje prirodnih izvora i
rezervoara FD fitoplazme usmereno ka razjašnjavanju porekla monotipske epidemije FD3
u vinogradima Srbije
An integrative approach for taxonomic characterization of Aculus spp. pests on stone fruits (Prunus spp.)
Full text linkEriophyids are among the most important pest mites worldwide. By considering the genus Aculus, the most economically important species on Prunus spp. are Aculus fockeui (Nalepa and Trouessart), on plum and several other stone fruits, and A. cornutus (Banks) on peaches. They cause symptoms such as chlorotic spots and curling of the young leaves, bronzing and rusting on the older leaves. Cryptic speciation is common in tiny and morphologically simple taxa such as the eriophyids and could also be expected in Aculus spp. To evaluate this, we applied an integrative approach to taxonomic characterization of pest species belonging to the genus Aculus on different stone fruit hosts. The results of phenotypic variability and COI mtDNA sequence analysis confirmed the present taxonomic status of the species A. cornutus and that A. fockeui represents a complex of cryptic species on different hosts
Unveiling Genetic Diversity Among Bacterial Isolates Using SCoT Markers
No full textThis study aimed to unveil the genetic diversity among 47 bacterial isolates from various species using start codon targeted (SCoT) markers. Six SCoT primers yielded 219 reproducible bands, with 89.04% exhibiting polymorphism. The amplification process generated 28 to 50 fragments per primer, with an average of 36.50. Genetic diversity was quantified using polymorphic information content (PIC) values ranging from 0.11 to 0.14, with SCoT32 showing the highest PIC (0.14) and SCoT23 the lowest (0.11). The resolving power (RP) index, used to assess primer discriminatory power, varied significantly, with SCoT23 demonstrating the highest RP (6.00) and SCoT29 the lowest (4.51). Comparative analysis with conventional markers like M13 and (GTG)5 revealed that certain SCoT primers exhibited superior PIC values, which indicates enhanced utility for interspecies differentiation. The high discrimination level achieved by SCoT primers underscores their effectiveness in genetic differentiation and biodiversity assessment within bacterial populations. This research highlights SCoT markers as powerful tools for microbial genetic studies, which offers valuable insights into bacterial diversity and provides a robust methodological framework for future investigations aimed at elucidating genetic variation and improving species identification. The application of SCoT markers represents a significant advancement in molecular techniques for bacterial characterization and phylogenetic analysis, demonstrating their potential to enhance our understanding of microbial genetics and evolution
INFLUENCE OF BACILLUS SAFENSIS STRAIN P114 ON PEPPER FRUITS IN TERMS OF BIOACTIVE COMPOUNDS AND ANTIOXIDANT CAPACITY
No full textSweet pepper as a commercial type of vegetable is one of the most important economic crop that
includes a large number of different varieties. The nutritional value of pepper fruits is significant due to
the content of bioactive compounds and antioxidants such as ascorbic acid, phenols, lycopene,
carotenoids, chlorophylls and antioxidant activity. Some bacteria from the genus Bacillus are well
known to be effective in controlling plant diseases and are associated with increased yields and plant
resistance. The aim of this work was to test the influence of Bacillus safensis (strain originated from
pepper seed, coded as P114) on total anthocyanins, flavonoid synthesis and antioxidant
capacity. Inoculum of bacterium was prepared in sterile distilled water and adjusted to 108
CFU mL-1
before treatment of pepper plants. Plants were initially treated in growth stage: leaf development and
then periodically, every 10 days until the fruits were formed. No treated pepper plants served as
control. The content of flavonoid was extracted in 70% acetone and measured spectrophotometrically
at 360 nm. Total anthocyanin content was extracted with 2% (v/v) HCl in methanol and analyzed
spectrophotometrically. The effect of strain P114 was significant in anthocyanin synthesis compared
to control. The content of anthocyanin in the fruits was 887.98±32.55 ug CGE/g dm in the treated
peppers, while in control was 744.77 ± 24.75 ug CGE/g dm. The content of flavonoid in fruits was
11.05±0.09 μg/g in the treated pepper, while in control was 11.53 ± 0.66 μg/g and no statistical
difference was found. The antioxidant capacity in pepper fruit was similar from both, treated and
untreated plants and amounted to 85.34 ± 0.78 mmol Trolox/kg. The obtained results indicate that
treatments of pepper plants with B. safiens strain P114 increased significantly anthocyanins contents
in the pepper fruits
