No abstract available

[en] The combination of 2 techniques, i, e., pollination after the destruction of chromosomes in pollen grains with ionizing radiations and in vitro culture of crossed ovules, was applied to interspecific hybridization between N.repanda and N. tabacum in order to investigate the possibility of overcoming hybrid inviability in the cross-combinations. By using these techniques, it became possible to obtain aneuploid hybrids, which had never been reported previously

[en] Content of polonium 210 was studied in the leaves of six varieties of Bulgarian oriental tobacco according to their class. Tobacco that had to be examined was kept for more than three years for equilibrium between polonium 210 and lead 210 to be reached. Tobacco leaves of higher classes contained smaller quantities polonium 210 compared with those of lower classes (III-V classes). This is due to leaves of the upper and middle parts of the plant with a shorter vegetation period belonging to higher tobacco classes; radionuclide penetration by the root. The established divergence from this relationship was due to some other reasons such as mechanical damage, etc. (A.B.)

[en] Infectious cDNA clones were developed for Grapevine leafroll-associated virus 3 (GLRaV-3, genus Ampelovirus, family Closteroviridae). In vitro RNA transcripts generated from cDNA clones showed replication via the production of 3′-coterminal subgenomic (sg) mRNAs in Nicotiana benthamiana protoplasts. The detection of sgRNAs and the recovery of progeny recombinant virions from N. benthamiana leaves agroinfiltrated with full-length cDNA clones confirmed RNA replication and virion formation. The 5′ non-translated region (5′ NTR) of GLRaV-3 was exchangeable between genetic variants and complement the corresponding cognate RNA functions in trans. Mutational analysis of the 5′ NTR in minireplicon cDNA clones showed that the conserved 40 nucleotides at the 5′-terminus were indispensable for replication, compared to downstream variable portion of the 5′ NTR. Some of the functional mutations in the 5′ NTR were tolerated in full-length cDNA clones and produced sgRNAs and virions in N. benthamiana leaves, whereas other mutations affected replication and virion formation.

[en] Highlights: • Generation of infectious clones of BNYVV and BSBMV by in vitro recombination. • The clone characteristics were comparable to the wild-type virus isolates. • Small RNAs displayed interspecies exchangeability. • Reassortant viruses were able to systemically infect different host plants. Two members of the Benyviridae family and genus Benyvirus, Beet soil-borne mosaic virus (BSBMV) and Beet necrotic yellow vein virus (BNYVV), possess identical genome organization, host range and high sequence similarity; they infect Beta vulgaris with variable symptom expression. In the US, mixed infections are described with limited information about viral interactions. Vectors suitable for agroinoculation of all genome components of both viruses were constructed by isothermal in vitro recombination. All 35S promoter-driven cDNA clones allowed production of recombinant viruses competent for Nicotiana benthamiana and Beta macrocarpa systemic infection and Polymyxa betae transmission and were compared to available BNYVV B-type clone. BNYVV and BSBMV RNA1 + 2 reassortants were viable and spread long-distance in N. benthamiana with symptoms dependent on the BNYVV type. Small genomic RNAs were exchangeable and systemically infected B. macrocarpa. These infectious clones represent a powerful tool for the identification of specific molecular host-pathogen determinants.

[en] Highlights: • Nicotiana tabacum harbours a highly complex phyllosphere bacterial community. • Pesticide treatment induces a response by the abundant Alphaproteobacteria and Gammaproteobacteria. • Negative as well as positive responders to the applied broad-spectrum fungicide were identified. • Bacteria that are commonly associated with beneficial contributions in terms of host health and fitness are also affected. Pesticide application can be accompanied by harmful non-target effects that affect humans, animals, as well as whole ecosystems. However, such effects remain mainly unaddressed in connection with microorganisms, and especially bacteria therein, which are essential for ecosystem functioning and host health. We analysed bacterial communities by sequencing 16S rRNA gene fragment amplicons following spray application of a broad-spectrum fungicide based on the active ingredient N-(3,5-dichlorophenyl) succinimide on Nicotiana tabacum L. leaves. The plant's phyllosphere was predominantly colonized by Proteobacteria, with Alphaproteobacteria accounting for up to 33.8% of the indigenous bacterial community. Bioinformatic analyses indicated that pesticide applications had an effect on the core microbiome as well as the rare microbiome. Moreover, the interference of the pesticide with phyllosphere bacteria was found to be selective. We have identified four positive responders including an ASV assigned to the genus Acinetobacter and 12 negative responders mainly assigned to bacterial genera known for beneficial plant-microbe interactions, including Stenotrophomonas, Sphingomonas, Flavobacterium and Serratia. Complementary inference of bacterial functioning on community level indicated that microbes with distinct stress response systems were likely enriched in the conducted treatments. The overall findings confirmed that pesticide treatments can induce measureable shifts in non-target bacterial communities colonizing the plant phyllosphere. They also indicate that potentially beneficial bacteria, which are known for their intrinsic association with plants, are among the most sensitive responders to the employed fungicide and thus highlight the importance of off-target studies in the context of the plant microbiome.

[en] ARGONAUTEs (notably AGO1 and AGO2) are effectors of plant antiviral RNA silencing. AGO1 was shown to be required for the temperature-dependent symptom recovery of Nicotiana benthamiana plants infected with tomato ringspot virus (isolate ToRSV-Rasp1) at 27 °C. In this study, we show that symptom recovery from isolate ToRSV-GYV shares similar hallmarks of antiviral RNA silencing but occurs at a wider range of temperatures (21–27 °C). At 21 °C, an early spike in AGO2 mRNAs accumulation was observed in plants infected with either ToRSV-Rasp1 or ToRSV-GYV but the AGO2 protein was only consistently detected in ToRSV-GYV infected plants. Symptom recovery from ToRSV-GYV at 21 °C was not prevented in an ago2 mutant or by silencing of AGO1 or AGO2. We conclude that other factors (possibly other AGOs) contribute to symptom recovery under these conditions. The results also highlight distinct expression patterns of AGO2 in response to ToRSV isolates and environmental conditions.

[en] Antimicrobial and antioxidant activities of plant Nicotiana plumbaginifolia Viv. Were carried out using various techniques. The petroleum ether, chloroform, methanol and aqueous extracts of the N. plumbaginifolia were obtained by maceration technique. The maximum antibacterial potential was exhibited by chloroform leaves extract (76.3 ± 0.3 mm), methanolic root extract (69 ± 0.8 mm) and petroleum ether root extract (67 ± 1.7 mm) against P. aureginosa. Methanolic root extract possessed 64 ± 2.3 mm zone of inhibition against E. coli, whereas chloroform root extract displayed 49 ± 0.8 mm against B. subtilis. Chloroform root extract showed 48 ±1.2 against S. aureus. The maximum zone of inhibition of antifungal potential was displayed by methanolic extracts of leaves against A. niger (43 ± 0.8 mm) and F. solani (43 ± 1.6 mm). The MIC assay was determine for further analysis which showed the MIC value of methanolic root extract (0.04 ± 0.1 mg/mL) against E. coli and the MIC value was noticed (0.108 ± 0.04 mg/mL) against A. niger by methanolic root extract. Antioxidant potential was determined using four methods i.e. (1,1-diphenyl-2-picrylhydrazyl radical (DPPH) scavenging activity, total antioxidant activity (TAA), total phenolic contents (TPC) and metal chelating activity. The highest value of percent DPPH was observed 90.56 at 1000 microL concentration in petroleum ether extract. The maximum values of TAA, TPC, FRAP and FTC were 1.352 ± 0.01, 1.683 ± 0.09 and 80.66 ± 0.08, respectively. (author)

[en] Native and amino acid (aa) substitution mutants of HCPro from potato virus Y (PVY) were transiently expressed in Nicotiana benthamiana leaves. Properties of those HCPro variants with regard to silencing suppression activities, mediation of viral transmission by aphids, and subcellular localization dynamics, were determined. One mutant failed to suppress silencing in agropatch assays, but could efficiently mediate the transmission by aphids of purified virions. This mutant also retained the ability to translocate to microtubules (MTs) in stressed cells. By contrast, another single aa substitution mutant displayed native-like silencing suppression activity in agropatch assays, but could not mediate transmission of PVY virions by aphids, and could not relocate to MTs. Our data show that silencing suppression by HCPro is not required in the aphid-mediated transmission of purified virions. In addition, since the same single aa alteration compromised both, viral transmission and coating of MTs, those two properties could be functionally related.

[en] The genome of Potato yellow dwarf virus (PYDV; Nucleorhabdovirus type species) was determined to be 12,875 nucleotides (nt). The antigenome is organized into seven open reading frames (ORFs) ordered 3'-N-X-P-Y-M-G-L-5', which likely encode the nucleocapsid, phospho, movement, matrix, glyco and RNA-dependent RNA polymerase proteins, respectively, except for X, which is of unknown function. The ORFs are flanked by a 3' leader RNA of 149 nt and a 5' trailer RNA of 97 nt, and are separated by conserved intergenic junctions. Phylogenetic analyses indicated that PYDV is closely related to other leafhopper-transmitted rhabdoviruses. Functional protein assays were used to determine the subcellular localization of PYDV proteins. Surprisingly, the M protein was able to induce the intranuclear accumulation of the inner nuclear membrane in the absence of any other viral protein. Finally, bimolecular fluorescence complementation was used to generate the most comprehensive protein interaction map for a plant-adapted rhabdovirus to date.