Double-Stranded RNA Targeting Dicer-Like Genes Compromises the Pathogenicity of Plasmopara viticola on Grapevine

Downy mildew caused by Plasmopara viticola is one of the most devastating diseases of grapevine, attacking all green parts of the plant. The damage is severe when the infection at flowering stage is left uncontrolled. P. viticola management consumes a significant amount of classical pesticides applied in vineyards, requiring efficient and environmentally safe disease management options. Spray-induced gene silencing (SIGS), through the application of exogenous double-stranded RNA (dsRNA), has shown promising results for the management of diseases in crops. Here, we developed and tested the potential of dsRNA targeting P. viticola Dicer-like (DCL) genes for SIGS-based crop protection strategy. The exogenous application of PvDCL1/2 dsRNA, a chimera of PvDCL1 and PvDCL2, highly affected the virulence of P. viticola. The reduced expression level of PvDCL1 and PvDCL2 transcripts in infected leaves, treated with PvDCL1/2 dsRNA, was an indication of an active RNA interference mechanism inside the pathogen to compromise its virulence. Besides the protective property, the PvDCL1/2 dsRNA also exhibited a curative role by reducing the disease progress rate of already established infection. Our data provide a promising future for PvDCL1/2 dsRNA as a new generation of RNA-based resistant plants or RNA-based agrochemical for the management of downy mildew disease in grapevine.

Download Full-text

Plant extracts - importance in sustainable agriculture

Italian Journal of Agronomy ◽

10.4081/ija.2021.1851 ◽

2021 ◽

Vol 16 (2) ◽

Author(s):

Katarzyna Godlewska ◽

Domenico Ronga ◽

Izabela Michalak

Keyword(s):

Natural Products ◽

Sustainable Agriculture ◽

Bioactive Compounds ◽

Plant Extracts ◽

Crop Protection ◽

Cultivated Plants ◽

Positive Effects ◽

Environmentally Safe ◽

New Generation ◽

The Impact

Plants due to the high content of various bioactive compounds are the main raw material for production of valuable, and useful bio-products (e.g., food, cosmetics, medicines, biostimulants, biopesticides, and feed). Different plant parts, for instance: seeds, fruits, flowers, stems, leaves, and roots can be used for their manufacture. Nowadays, there is a clear need to develop new, efficient, and environmentally safe methods of stimulation of plant, growth and crop protection. Plant-based extracts are new, natural, and multi-compounds products that could be used for these purposes. They possess antifungal, antimicrobial, antiparasitic, antiprotozoal, antioxidant, medicinal, aromatic, and anti-inflammatory properties. This group of natural products has the potential to become a new generation of bio-products suitable for use in sustainable agriculture. The purpose of this review is to provide an overview of the literature describing the impact of plant-derived extracts/biostimulants (PDBs) on crops grown in controlled, and real conditions as well as under various abiotic and biotic stresses; the extraction methods used to obtain PDBs, and the specific constituents responsible for their biostimulating activity. The application of these bio-products could be beneficial for sustainable production, due to several advantages, such as low toxicity to humans and the environment, enhanced resistance of cultivated plants to biotic and abiotic stress, increased yields and quality of crops, as well as the reduction in the use of mineral fertilisers and pesticides. However, deeper cooperation between industrial and academic research is required to accelerate the development of new environmentally safe solutions for future agriculture. Highlights - Higher plants constitute a rich source of various bioactive compounds for the production of useful natural products. - The importance of the proper choice of extraction method and solvent to process and preserve the desired substances.- Plant extracts as biostimulants and plant protection products for use in modern and sustainable agriculture. - The positive effects of plant-based extracts on plants cultivated under normal and unfavourable conditions.- Plant extracts as a new generation of eco-friendly products for the increment of the production of high-quality food.

Download Full-text

Photosynthesis of grapevine leaves infected by downy mildew

OENO One ◽

10.20870/oeno-one.1998.32.3.1049 ◽

1998 ◽

Vol 32 (3) ◽

pp. 121

Author(s):

Simone Orlandini ◽

Alberto Giuntoli

Keyword(s):

Downy Mildew ◽

Crop Protection ◽

Net Photosynthesis ◽

Plasmopara Viticola ◽

Plant Responses ◽

Physiological Alterations ◽

Grapevine Leaves ◽

Normal Trend ◽

Source Sink ◽

The Impact

A correct approach to crop protection must consider the effect of disease on plant activity and its impact in terms of economical losses. These evaluations can drive the farmers to a quantification of plant responses to pathogen attack and to the application of economical thresholds for fungicide sprays. For grapevine (Vitis vùlÎfera L.), the knowledge of plant responses to disease is still very limited independently from the level of investigation (physiology, growth, development, etc.). The aim of this research was to evaluate the impact of downy mildew (Plasmopara viticola Berl. et De Toni) on grapevine photosynthesis under controlled and field conditions. Downy mildew was shown to reduce to a negative value net photosynthesis in the oilspot, as well as to affect the physiology of gas exchanges of the tissues surrounding the oilspot. The daily trend under field condition pointed out a normal trend of assimilation, without differences between green portions of healthy and diseased leaves. The consequences on source-sink relationships were discussed, as well as the relation between visible symptoms and physiological alterations.

Download Full-text

DECONTAMINATION OF VETERINARY SURVEILLANCE OBJECTS BY NEW GENERATION MEDICINE

Vestnik of the Russian agricultural science ◽

10.30850/vrsn/2019/2/64-67 ◽

1970 ◽

pp. 64-67

Author(s):

М. S. Saypullaev ◽

А. U. Koychuev ◽

Т. B. Mirzoeva

Keyword(s):

Staphylococcus Aureus ◽

Inhalation Exposure ◽

Gram Negative Bacteria ◽

Hazard Class ◽

E Coli ◽

Highly Efficient ◽

Mucous Membranes ◽

Environmentally Safe ◽

New Generation ◽

And Staphylococcus Aureus

The successful conduct of disinfection measures largely depends on the availability of veterinary practice a highly efficient, environmentally safe disinfectants. In this regard, finding new highly efficient disinfectant remains relevant. Studies found that the "Polied" (OOO "Razvitie XXI Vek, Russia) can be attributed to the highly efficient and environmentally friendly means. Solutions "Polied" have a high disinfectant activity against smooth and rough surfaces in the laboratory against gram-positive, gram-negative bacteria, mycobacteria and spores of microorganisms. Studies have established that solutions should be "Polied" obezzarajivatmi E. coli (EA 1257) concentrations of 0.1% on smooth surfaces and Staphylococcus aureus concentration of 0.05% in 1 hour from the calculation of 0.25-0.3 litres/m2. Disinfection of rough test surfaces against Escherichia coli and Staphylococcus aureus occurred after treatment with 0,3% solution of 3-hour exposure, at a rate of 0.5 l/m2. It was also found that 1.0% solution "Polied" fully obezzarazhivatel test the surface of mycobacteria (PCs-5) and at double the 0.6% concentration for 24 hours. Disinfection of rough test surfaces contaminated with spores of B. cereus (PCs 96) was achieved with a 4.0% solution at twice the irrigation rate of 0.5 l/m2 at an exposure of 24 hours. Toxicity solutions of the drug "Polied" refer to "moderate" threat (hazard class 3) and low-hazard substances (4 hazard class) when applied to the skin, mucous membranes of the eyes, and inhalation exposure on the respiratory system.

Download Full-text

Content of trans-resveratrol in leaves and berries of interspecific grapevine (Vitis sp.) varieties

Czech Journal of Food Sciences ◽

10.17221/238/2008-cjfs ◽

2009 ◽

Vol 26 (Special Issue) ◽

pp. S13-S17 ◽

Cited By ~ 4

Author(s):

P. Bábíková ◽

N. Vrchotová ◽

J. Tříska ◽

M. Kyseláková

Keyword(s):

Powdery Mildew ◽

Downy Mildew ◽

Plasmopara Viticola ◽

Fungal Diseases ◽

Uncinula Necator ◽

Botryotinia Fuckeliana

The aim of this project was to study changes in the content of trans-resveratrol in berries and leaves of grapevine (Vitis sp.) infested by fungal diseases, especially by Botryotinia fuckeliana Whetzel, called as grey mildew, Plasmopara viticola (Berk. & M.A. Curtis) Berl & De Toni, called downy mildew and Uncinula necator (Schw.) Burr, called powdery mildew. In our experiments two white and two blue varieties were used. Contents of trans-resveratrol were determined in healthy and infested leaves and in healthy berries. Infested leaves of white varieties contained more trans-resveratrol than those of blue varieties. The content of trans-resveratrol in berries was lower than that in leaves.

Download Full-text

Investigation of long non-coding RNAs as regulatory players of grapevine response to powdery and downy mildew infection

BMC Plant Biology ◽

10.1186/s12870-021-03059-6 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Garima Bhatia ◽

Santosh K. Upadhyay ◽

Anuradha Upadhyay ◽

Kashmir Singh

Keyword(s):

Downy Mildew ◽

Plasmopara Viticola ◽

Defense Responses ◽

Protein Coding ◽

Functional Roles ◽

Real Time Quantitative Pcr ◽

Transcriptional Reprogramming ◽

Sequencing Technologies ◽

Non Coding Rnas ◽

Fungal Phytopathogens

Abstract Background Long non-coding RNAs (lncRNAs) are regulatory transcripts of length > 200 nt. Owing to the rapidly progressing RNA-sequencing technologies, lncRNAs are emerging as considerable nodes in the plant antifungal defense networks. Therefore, we investigated their role in Vitis vinifera (grapevine) in response to obligate biotrophic fungal phytopathogens, Erysiphe necator (powdery mildew, PM) and Plasmopara viticola (downy mildew, DM), which impose huge agro-economic burden on grape-growers worldwide. Results Using computational approach based on RNA-seq data, 71 PM- and 83 DM-responsive V. vinifera lncRNAs were identified and comprehensively examined for their putative functional roles in plant defense response. V. vinifera protein coding sequences (CDS) were also profiled based on expression levels, and 1037 PM-responsive and 670 DM-responsive CDS were identified. Next, co-expression analysis-based functional annotation revealed their association with gene ontology (GO) terms for ‘response to stress’, ‘response to biotic stimulus’, ‘immune system process’, etc. Further investigation based on analysis of domains, enzyme classification, pathways enrichment, transcription factors (TFs), interactions with microRNAs (miRNAs), and real-time quantitative PCR of lncRNAs and co-expressing CDS pairs suggested their involvement in modulation of basal and specific defense responses such as: Ca2+-dependent signaling, cell wall reinforcement, reactive oxygen species metabolism, pathogenesis related proteins accumulation, phytohormonal signal transduction, and secondary metabolism. Conclusions Overall, the identified lncRNAs provide insights into the underlying intricacy of grapevine transcriptional reprogramming/post-transcriptional regulation to delay or seize the living cell-dependent pathogen growth. Therefore, in addition to defense-responsive genes such as TFs, the identified lncRNAs can be further examined and leveraged to candidates for biotechnological improvement/breeding to enhance fungal stress resistance in this susceptible fruit crop of economic and nutritional importance.

Download Full-text

The Bacterial Enzyme Cas13 Interferes with Neurite Outgrowth from Cultured Cortical Neurons

Toxins ◽

10.3390/toxins13040262 ◽

2021 ◽

Vol 13 (4) ◽

pp. 262

Author(s):

Qin-Wei Wu ◽

Josef P. Kapfhammer

Keyword(s):

Rna Editing ◽

Cortical Neurons ◽

High Efficiency ◽

Primary Cultures ◽

Cultured Neurons ◽

Therapeutic Tool ◽

Bacterial Enzyme ◽

Rna Targeting ◽

Cultured Cortical Neurons ◽

New Generation

The CRISPR-Cas13 system based on a bacterial enzyme has been explored as a powerful new method for RNA manipulation. Due to the high efficiency and specificity of RNA editing/interference achieved by this system, it is currently being developed as a new therapeutic tool for the treatment of neurological and other diseases. However, the safety of this new generation of RNA therapies is still unclear. In this study, we constructed a vector expressing CRISPR-Cas13 under a constitutive neuron-specific promoter. CRISPR-Cas13 from Leptotrichia wadei was expressed in primary cultures of mouse cortical neurons. We found that the presence of CRISPR-Cas13 impedes the development of cultured neurons. These results show a neurotoxic action of Cas13 and call for more studies to test for and possibly mitigate the toxic effects of Cas13 enzymes in order to improve CRISPR-Cas13-based tools for RNA targeting.

Download Full-text

The Grapevine E3 Ubiquitin Ligase VriATL156 Confers Resistance against the Downy Mildew Pathogen Plasmopara viticola

International Journal of Molecular Sciences ◽

10.3390/ijms22020940 ◽

2021 ◽

Vol 22 (2) ◽

pp. 940

Author(s):

Elodie Vandelle ◽

Pietro Ariani ◽

Alice Regaiolo ◽

Davide Danzi ◽

Arianna Lovato ◽

...

Keyword(s):

Downy Mildew ◽

Ubiquitin Ligase ◽

Control Measure ◽

Durable Resistance ◽

Genetic Resistance ◽

E3 Ubiquitin Ligase ◽

Plasmopara Viticola ◽

Vitis Vinifera L ◽

Vitis Species ◽

Vitis Riparia

Downy mildew, caused by Plasmopara viticola, is one of the most severe diseases of grapevine (Vitis vinifera L.). Genetic resistance is an effective and sustainable control strategy, but major resistance genes (encoding receptors for specific pathogen effectors) introgressed from wild Vitis species, although effective, may be non-durable because the pathogen can evolve to avoid specific recognition. Previous transcriptomic studies in the resistant species Vitis riparia highlighted the activation of signal transduction components during infection. The transfer of such components to V. vinifera might confer less specific and therefore more durable resistance. Here, we describe the generation of transgenic V. vinifera lines constitutively expressing the V. riparia E3 ubiquitin ligase gene VriATL156. Phenotypic and molecular analysis revealed that the transgenic plants were less susceptible to P. viticola than vector-only controls, confirming the role of this E3 ubiquitin ligase in the innate immune response. Two independent transgenic lines were selected for detailed analysis of the resistance phenotype by RNA-Seq and microscopy, revealing the profound reprogramming of transcription to achieve resistance that operates from the earliest stages of pathogen infection. The introduction of VriATL156 into elite grapevine cultivars could therefore provide an effective and sustainable control measure against downy mildew.

Download Full-text

A High-Quality Grapevine Downy Mildew Genome Assembly Reveals Rapidly Evolving and Lineage-Specific Putative Host Adaptation Genes

Genome Biology and Evolution ◽

10.1093/gbe/evz048 ◽

2019 ◽

Vol 11 (3) ◽

pp. 954-969 ◽

Cited By ~ 12

Author(s):

Yann Dussert ◽

Isabelle D Mazet ◽

Carole Couture ◽

Jérôme Gouzy ◽

Marie-Christine Piron ◽

...

Keyword(s):

Downy Mildew ◽

Genome Assembly ◽

Population Genomics ◽

Plasmopara Viticola ◽

Plant Diseases ◽

Host Specialization ◽

Grapevine Downy Mildew ◽

Downy Mildews ◽

Genes Encoding ◽

High Level

Abstract Downy mildews are obligate biotrophic oomycete pathogens that cause devastating plant diseases on economically important crops. Plasmopara viticola is the causal agent of grapevine downy mildew, a major disease in vineyards worldwide. We sequenced the genome of Pl. viticola with PacBio long reads and obtained a new 92.94 Mb assembly with high contiguity (359 scaffolds for a N50 of 706.5 kb) due to a better resolution of repeat regions. This assembly presented a high level of gene completeness, recovering 1,592 genes encoding secreted proteins involved in plant–pathogen interactions. Plasmopara viticola had a two-speed genome architecture, with secreted protein-encoding genes preferentially located in gene-sparse, repeat-rich regions and evolving rapidly, as indicated by pairwise dN/dS values. We also used short reads to assemble the genome of Plasmopara muralis, a closely related species infecting grape ivy (Parthenocissus tricuspidata). The lineage-specific proteins identified by comparative genomics analysis included a large proportion of RxLR cytoplasmic effectors and, more generally, genes with high dN/dS values. We identified 270 candidate genes under positive selection, including several genes encoding transporters and components of the RNA machinery potentially involved in host specialization. Finally, the Pl. viticola genome assembly generated here will allow the development of robust population genomics approaches for investigating the mechanisms involved in adaptation to biotic and abiotic selective pressures in this species.

Download Full-text

First Report of Downy Mildew Caused by Plasmopara viticola on Vitis ficifolia var. sinuata in Korea

Plant Disease ◽

10.1094/pdis-04-17-0571-pdn ◽

2017 ◽

Vol 101 (11) ◽

pp. 1958-1958

Author(s):

Y. J. Choi ◽

S. E. Cho ◽

H. D. Shin

Keyword(s):

Downy Mildew ◽

Plasmopara Viticola ◽

First Report

Download Full-text

Toward a Weather-Based Forecasting System for Fire Blight and Downy Mildew

Atmosphere ◽

10.3390/atmos9120484 ◽

2018 ◽

Vol 9 (12) ◽

pp. 484 ◽

Cited By ~ 2

Author(s):

Ana Firanj Sremac ◽

Branislava Lalić ◽

Milena Marčić ◽

Ljiljana Dekić

Keyword(s):

Downy Mildew ◽

Fire Blight ◽

Meteorological Data ◽

Weather Prediction ◽

Weather Forecast ◽

Atmospheric Model ◽

Plasmopara Viticola ◽

Forecasting System ◽

Nwp Model ◽

The Impact

The aim of this research is to present a weather-based forecasting system for apple fire blight (Erwinia amylovora) and downy mildew of grapevine (Plasmopara viticola) under Serbian agroecological conditions and test its efficacy. The weather-based forecasting system contains Numerical Weather Prediction (NWP) model outputs and a disease occurrence model. The weather forecast used is a product of the high-resolution forecast (HRES) atmospheric model by the European Centre for Medium-Range Weather Forecasts (ECMWF). For disease modelling, we selected a biometeorological system for messages on the occurrence of diseases in fruits and vines (BAHUS) because it contains both diseases with well-known and tested algorithms. Several comparisons were made: (1) forecasted variables for the fifth day are compared against measurements from the agrometeorological network at seven locations for three months (March, April, and May) in the period 2012–2018 to determine forecast efficacy; (2) BAHUS runs driven with observed and forecast meteorology were compared to test the impact of forecasted meteorological data; and (3) BAHUS runs were compared with field disease observations to estimate system efficacy in plant disease forecasts. The BAHUS runs with forecasted and observed meteorology were in good agreement. The results obtained encourage further development, with the goal of fully utilizing this weather-based forecasting system.

Download Full-text