The first report of Tomato brown rugose fruit virus in tomato in Norway

In May 2021, tomato (Solanum lycopersicum L.) plants with necrosis and ringspot symptoms were observed in a farm greenhouse at Sundbyfoss, Norway. This greenhouse production focused on ecological growing of several tomato varieties (i.e., “Blush Tiger”, “Sailor’s Luck”, “Evil Oliver”, etc.) with compost for local customers. The presence of tomato brown rugose fruit virus (ToBRFV) was suspected due to the symptoms in the diseased plants. Sap inoculation was carried out with extracts from 100 mg symptomatic tomato fresh leaves in 0.03 M PBS (phosphate buffered saline, pH 7.0), and inoculation on three Nicotiana tabacum cv. Xanthi plants at the 4-6 leaf stage grown in an experimental greenhouse. The test plants showed local necrotic lesions after 3-5 days. Two symptomatic leaf samples, one from tomato and the other from a test plant, were analysed by transmission electron microscope (TEM) with the help of negative staining. The samples were treated with 0.1 M PBS (pH 7.3) first and placed on carbon-coated EM grids and stained with 2% uranyl acetate before TEM observation. Rigid rod-shaped viral particles, typical of tobamovirus particls (around 300 nm) were observed. Total RNA was isolated from two tomato leaves showing symptoms using a Norgen Plant/Fungi RNA kit (Norgen Biotek, Canada). One-step reverse-transcription (RT)-PCR with specific primers ToBRFV-F/ToBRFV-R for ToBRFV (Alkowni et al. 2019), which amplified a 560-bp fragment, was performed. The sequence obtained by Sanger sequencing from the amplicon (535 nt) showed 99.8% nt identity with ToBRFV isolate PV-1241 (NCBI accession no. MZ202349) from DSMZ (Leibniz Institute, German) and was deposited in the GenBank database under the accession number OK358628. The infection was also confirmed with duplex real-time RT-PCR test for ToBRFV using CaTa28 and CSP1325 primers and probe (ISF 2020; EPPO, PM7/146 2021). In addition, eight tomato samples from the same farm greenhouse were collected according to cultivars, location in the greenhouse and symptoms before eradication and disinfection: four of the tomato samples were confirmed positive for ToBRFV with one-step RT-PCR as described above. A surveillance program for ToBRFV in commercial greenhouse production has been carried out in 2021 in Norway. Around 4000 tomato plants from 18 commercial tomato growers were tested. No positive ToBRFV samples have been detected. However, unregulated tomato seeds from abroad and self-propagation of tomato by private gardeners and hobby growers is a potential threat to commercial production of tomato in Norway. To our knowledge, this is the first report of ToBRFV associated with tomato in Norway and in the Nordic countries.

First report of a mastrevirus (Geminiviridae) transmitted by the corn leafhopper

Maize striate mosaic virus (MSMV; genus Mastrevirus), was recently reported in maize plants in Brazil, and also detected by metagenomic analyses in the corn leafhopper, Dalbulus maidis (DeLong & Wolcott). Although these findings suggested that D. maidis is a potential vector, no transmission studies have been performed. Here, we tested the transmission of MSMV by D. maidis from field-collected infected plants and plants infected with MSMV via leafhopper-mediated transmission in the laboratory; all plants were confirmed positive for MSMV by PCR. In each one of three transmission replicates, aviruliferous D. maidis nymphs and adults were confined together on a source plant during a 4-day acquisition access period (AAP), and subsequently transferred to healthy maize seedlings (10 individuals per test plant) in a series of 4-day inoculation access periods (IAPs). We also tested transmission by the corn aphid, Rhopalosiphum maidis (Fitch) and by mechanical inoculation of healthy maize seedlings. Only D. maidis transmitted MSMV, with overall transmission rates of 29.4 and 39.5% using field-collected infected plants, and 18.5% using infected plants in laboratory. D. maidis transmitted MSMV until the third (8-12 days after the AAP) or fourth successive IAP (12-16 days) with gradual loss in transmission efficiency and rate of viruliferous insects over time, suggesting a persistent, but non-propagative mode of transmission. Infected test plants showed mottling symptoms with mild chlorotic streaks and height reduction. This is the first report of transmission of a mastrevirus by D. maidis, facilitating the completion of Koch’s postulate for MSMV.

EVALUATION OF PHYTOTOXICITY OF NATURAL MINERALS

An assessment of the phytotoxicity of natural (lime, zeolite and phosphorite) minerals and their nanostructured analogues (nanostructured lime, nanostructured water-zeolite suspension (NWZS), nanostructured water-phosphate suspension (NWPS) and a spine) has been carried out. a monocotyledonous test plant (spring wheat variety Ulyanovskaya-105) has a positive effect on all studied minerals. The length of the seedling among native minerals is stimulated by lime and phosphorite, among nanominerals - nanostructured lime and NWZS. All native minerals have a positive effect on biometric indicators, among nanostructural ones – nano-lime and NWPS. Seedling biomass is stimulated only by zeolite and NWZS. Thus, native zeolite has the best data on the length of the seedling (by 48.9 %), the length of the coleoptile (by 53.6 %) and the biomass of the seedling (by 16.1 %), and in NWZS - along the root length (by 12.8 %), root biomass (by 13.5 %) and seedling biomass (by 13.7 %). Thus, according to most indicators, the best are zeolite (among native ones) and NWZS (among nanostructured minerals).

Growth and phytoremediative capacity of Eleusine indica in a typical farmland soil under previous exposure to organochlorine pesticide

This study investigated the growth and phytoremediative response of Eleusine indica in a typical farmland soil that had been exposed to organochlorine pesticides. Different soil treatments were obtained by mixing pesticide polluted soil (P) with control soil (C) in ratios, 1P:99C, 5P:95C, 50P:50C, 25P:75C, 75P:25C, 100P and 100C. Three-leaf tillers of the test plant were sown in all the treatments for 3 months. The results revealed that there were no significant differences in all plant morphological parameters measured between plants in P impacted soils and C-soil. All the pH values were acidic, although an increase in pH and decrease in conductivity was observed with the introduction of the test plant. There was reduction in total pesticide residual (TPR) contents in the soil as a result of the plant activities. Significant reduction in q – BHC, α – chlordane and ϒ – chlordane was observed in the P1:C99 soil mix; a 90% remediation efficiency (1.663 mg kg-1 ) was registered in the all treatments. This study thus presents E. indica as a potential concentration-dependent phytoremediator of pesticide, with no significant morphological changes.

Plant root PET: visualization of photosynthate translocation to roots in rice plant

Roots are essential to plants for uptake of water and nutrients. For the improvement of crop production, it is necessary to understand the elucidation of the root development and its function under the ground. Especially, photosynthate translocation from plant leaves to roots is an important physiological function that affects the root elongation, adaptation to the soil environment and nutrients uptake. To evaluate the translocation dynamics to roots, positron emission tomography (PET) and 11C tracer have been used. However, the spatial resolution is degraded at roots that develop around the peripheral area of field of view (FOV) due to parallax errors. In this study, to overcome this problem, we developed a small OpenPET prototype applying four-layer depth-of-interaction detectors. We demonstrated the imaging capability of 11C-photosynthate translocation to rice roots that develop throughout the entire PET field. We also tried to obtain structural information of roots by high-throughput X-ray computerized tomography (CT) system using the same test plant. As a result, we succeeded in visualizing the root structure that developed around the peripheral region of FOV and imaging the accumulation of 11C-photosynthate to the roots in those areas without degrading the spatial resolution. From obtained images, we also succeeded in evaluating the translocation dynamics varied by roots. The combined use of the high-throughput CT system and the OpenPET prototype was demonstrated to be appropriate for structural and functional analysis of roots.

Antibacterial and phytotoxic activity of the Schiff’s bases of 5-phenyl-4-amino-3-mercapto-4H-1,2,4-triazole with the donor substituents in the 4th position on heterosystem

Background. Triazoles and Schiff’s bases have a high biological activity. For the practical use of the derivatives, their low toxicity is important. The purpose of this work was to investigate the antibacterial and phytotoxic properties of Schiff’s bases of 5-phenyl-4-amino-3-mercapto-4H-1,2,4-triazole with donor substituents in the 4th position of heterosystem. Materials and methods. In the study of antibacterial activity of the derivatives, corrosion-active 4-day association cultures of ammonifying and sulfate-reducing bacteria were used as a test culture of microorganisms. Sensitivity of bacteria to derivatives was determined by diffusion method in agar using sterile paper disks according to the standard method. In the investigation of phytotoxic activity of the derivatives, Lepidium sativum of the “Ajour” cultivar was used as a test plant. Seed germination and biometric indices (length, weight of the aboveground part and roots) of 5-day sprouts were determined, the phytotoxic effect of the derivatives was calculated. Experimental data were processed using methods of mathematical statistics. Results. The introduction of substituents does not provide for an increase in antibacterial properties of the studied compounds in relation to some corrosion active ammonifying and sulfate-reducing bacteria. Low activity was observed regarding the association culture of ammonifying bacteria to the compound without substituents in the phenyl fragment and the compound with fluor as a substituent in the phenyl fragment at a concentration of 2.0%. Derivatives with the methoxyl substituent in the phenyl fragment and with the hydroxyl substituent in the phenyl fragment did not show any antibacterial activity against the association culture of ammonifying bacteria isolated from ferrosphere in meat-peptone broth. Antibacterial action against the association of sulfate-reducing bacteria Desulfovibrio orizae with organic acid-producing bacteria Anaerotignum propionicum for derivatives were not detected. Phytotoxic properties were observed for the compound with the hydroxyl substituent that influenced the processes of growth in the test plant. Conclusion. The introduction of electron-donor substituents into the basic structure did not provide for an increase in antibacterial properties against corrosive bacteria. Phytotoxic properties were observed for the compound with the hydroxyl substituent in the phenyl fragment, which influenced the L. sativum growth processes by inhibiting growth of the above-ground part and roots. Other compounds either did not show any action, or demonstrated a weak stimulating effect on the growth and development of the test plant.

Growth and nitrogen accumulation assessment of banana (Musa acuminata) after exposure to hydroquinone in a charcoal-ultisol mix

The present study investigates plant growth and nitrogen accumulation assessment of young banana plant (Musa acuminata) after exposure to hydroquinone (a mutagen) in a charcoal-ultisol mix. Charcoal was obtained from the wood of Pentaclethra macrophylla aerobically, and was crushed into a smooth powdered form. This was mixed with an ultisol obtained from the University of Benin Botanic garden. Different levels of the soil-charcoal mix were prepared as follows; 100% charcoal, 75% charcoal-25% soil, 50% charcoal-50% soil, 25% charcoal- 75% soil, and 100% sand. The charcoal-ultisol mix was then amended with 5ppm hydroquinone one week before propagating young banana suckers. Results showed that sprouting began fifteen days after propagation. There were also different changes in plant height in the various soil treatments; the lowest was reported in the 75-25 charcoal ultisol mix (10.50cm), compared to 45.83cm in plants sown in 100% charcoals. Foliar yield was lowest in the 75-25 charcoal-ultisol mix (8.55 grams per plant), compared to 29.15 grams per plant in the 100% charcoal. The below ground morphological characteristics test plant revealed that there were significant differences between the growth parameters. Nitrate nitrogen, Ammonia Nitrogen and total nitrogen were significantly accumulated on the leaves of plant. However, higher nitrogen accumulation in the leaves was found in treatment with high charcoal percentage.

Phytoremediation of Crude Oil Polluted Microbial Augmented Soil Using Cyperus esculentus and Phyllanthus amarus

Aim: To assess the phytoremediation potential of Cyperus esculentus and Phyllanthus amarus in crude oil polluted soil and ascertain the enhancement of augmented microbes (fungi). Study Design: The study employs experimental design, statistical analysis of the data and interpretation. Place and Duration of Study: Rivers State University demonstration farmland in Nkpolu- Oroworukwo, Mile 3 Diobu area of Port Harcourt, was used for this study. The piece of land is situated at Longitude 4°48’18.50” N and Latitude 6ᵒ58’39.12” E measuring 5.4864 m x 5.1816 m with a total area of 28.4283 square meter. Phytoremediation process monitoring lasted for 240 days, analyses were carried out weekly at 30 days’ interval. Methodology: Seven (7) experimental plots (two Control (Unpolluted and polluted soil) and five polluted amended/treated plots) employing Randomized Block Design (each having dimensions: 100 x 50 x 30 cm LxBxH) were formed and mapped out on agricultural soil and left fallow for 6 days before contamination on the seventh day; after which it was allowed for 21 days for proper contamination and exposure to natural environmental factors (to mimic soil crude oil spill site); thereafter bioaugmenting organisms were applied. Baseline studies were carried out on the top soil before and after contamination, major parameters monitored and assessed were Total Petroleum Hydrocarbon (TPH) uptake by plant roots and stem, Polycyclic Aromatic Hydrocarbon (PAHs) and TPH reduction in soil. Other physicochemical analyzed in the soil of different plots were pH, Electrical Conductivity, Moisture Content, Total Nitrogen, Available Phosphorus, Potassium, Total Organic Carbon, Plant Height, Iron, Lead at regular intervals; days 1, 60, 90, 120, 150, 180, 210 & 240. Application of augmenting organisms was to enhance phytoremediation by test plant Cyperus esculentus (Cyp) and Phyllanthus amarus (Phy). The rate of phytoremediation was estimated from percentage (%) uptake of Total petroleum hydrocarbon (TPH) in plant roots and stem from day 1 -240; while percentage (%) reduction of TPH and PAHs in soil was estimated from day 1 to the residual at day 240. Results: The test plants decreased significant amount of crude oil as revealed in TPH uptake in their roots and Stem. Mean amount and percentage Total Petroleum Hydrocarbon (TPH) uptake by Cyperus esculentus roots and stem were; 152.33±50.34mg/kg, 12.57±4.16% and 201.13±8.80mg/kg, 13.27±0.58% respectively; while that of Phyllanthus amarus roots and stem were 141.50±35.62mg/kg, 11.68±2.94% and 174.44±19.98mg/kg, 11.51±1.32% respectively. Similar trend was observed in the control plots were TPH uptake by Cyperus esculentus roots and stem were; 24.2mg/kg, 2.00% and 20.01mg/kg, 1.32% respectively while in control plot of Phyllanthus amarus TPH uptake by roots and stem were 23.19mg/kg, 1.91% and 19.80mg/kg, 1.31% respectively. Comparatively, uptake of TPH was higher in plant stem than roots. From the initial TPH contamination value of 5503.00mg/kg , Total Petroleum Hydrocarbon Reduction and % Hydrocarbon Reduction in soil at 240 days in the different treatment plots in a decreasing order were as follows: PS+AN+MR+SMS+Phy (5470.9mg/kg; 99.43%) >PS+MR+SMS+Phy (5460.60mg/kg; 99.23%) >PS+AN+MR+Phy (5451.30mg/kg; 99.06%) >PS+MR+Cyp (5448.30mg/kg; 99.01%) >PS+AN+MR+Cyp (5440.00mg/kg; 98.86%) >PS+AN+Phy (5422.905mg/kg; 98.54%) >PS+Cyp (no amendment) (5380.90mg/kg; 97.78%). Comparative evaluation revealed higher reduction of PAHs in soil (plot) planted with Phyllanthus amarus. Highest PAHs reduction in soil was seen in PS+AN+MR+SMS+Phy (31.3mg/kg; 65.89%) while least was recorded in PS+ Cyp (no amendment) (23.4mg/kg, 49.26%). Conclusion: it was observed that plots planted with Cyperus esculentus (TPH 5492.75±76.36mg/kg) showed higher reduction of TPH from soil than those planted with Phyllanthus amarus (TPH 5449.72±18.27mg/kg); while PAHs degradation/reduction in plots planted with Phyllantus amarus (PAHs 28.72±2.74mg/kg; 60.46±5.77%) was higher than plots planted with Cyperus esculentus (PAHs 25.77±2.12mg/kg, 54.24±4.47%). More so, plots amended with augmentating microbes showed significant higher percentage reduction in hydrocarbon in the polluted soil than unamended polluted soil. It is therefore recommended that Cyperus esculentus is a suitable plant species for phytoremediation of crude oil contaminated soil with high TPH value while Phyllanthus amarus is the best option for phytoremediation of polluted soil with high PAHs value, in combination with augmenting microbes.

Perbanyakan dan Aplikasi PGPR (Plant Growth Promoting Rhizobactheria) di Laboratorium Pengamatan Hama dan Penyakit Banyumas

PGPR (Plant Growth Promoting Rhizobacteria) is a substance that helps plant growth with the help of rhizosphere microorganisms. PGPR propagation can be done with liquid media. This PGPR propagation needs to be done because this substance has many benefits for agricultural cultivation. The application of PGPR to the test plant, namely the pakcoy plant, proved that there was an effect of giving PGPR to the plant. Observations on the test plants were carried out by observing several observation variables such as plant height, root length, number of leaves, wet weight and dry weight. The results of these observations showed that the effect on the test plants was seen in the variables of root length, wet weight, and dry weight. Meanwhile, the variable plant height and number of leaves did not show a visible difference.

Short-Term Decomposition and Nutrient-Supplying Ability of Sewage Sludge Digestate, Digestate Compost, and Vermicompost on Acidic Sandy and Calcareous Loamy Soils

Organic waste and the compost and vermicompost derived from it may have different agronomic values, but little work is available on this aspect of sewage sludge. A 75-day pot experiment with perennial ryegrass (Lolium perenne) as the test plant aimed to investigate the fertiliser value and organic matter replenishment capacity of digested sewage sludge (DS) and the compost (COM) and vermicompost (VC) made from it, applied in 1% and 3% doses on acidic sand and calcareous loam. The NPK content and availability, changes in organic carbon content and plant biomass, and the efficiency of the amendments as nitrogen fertilisers were investigated. The final average residual carbon content for DS, COM, and VC was 35 ± 34, 85 ± 46, and 55 ± 46%, respectively. The organic carbon mineralisation rate depended on the soil type. The additives induced significant N mineralisation in both soils: the average increment in mineral N content was 1.7 times the total added N on acidic sand and 4.2 times it on calcareous loam for the 1% dose. The agronomic efficiency of COM and VC as fertilisers was lower than that of DS. In the short term, DS proved to be the best fertiliser, while COM was the best for organic matter replenishment.