scholarly journals Rpv Mediated Defense Responses in Grapevine Offspring Resistant to Plasmopara viticola

Plants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 781
Author(s):  
Tyrone Possamai ◽  
Daniele Migliaro ◽  
Massimo Gardiman ◽  
Riccardo Velasco ◽  
Barbara De Nardi

Downy mildew, caused by the biotrophic oomycete Plasmopara viticola, is one of the most serious grapevine diseases. The development of new varieties, showing partial resistance to downy mildew, through traditional breeding provides a sustainable and effective solution for disease management. Marker-assisted-selection (MAS) provide fast and cost-effective genotyping methods, but phenotyping remains necessary to characterize the host–pathogen interaction and assess the effective resistance level of new varieties as well as to validate MAS selection. In this study, the Rpv mediated defense responses were investigated in 31 genotypes, encompassing susceptible and resistant varieties and 26 seedlings, following inoculation of leaf discs with P. viticola. The offspring differed in Rpv loci inherited (none, one or two): Rpv3-3 and Rpv10 from Solaris and Rpv3-1 and Rpv12 from Kozma 20-3. To improve the assessment of different resistance responses, pathogen reaction (sporulation) and host reaction (necrosis) were scored separately as independent features. They were differently expressed depending on Rpv locus: offspring carrying Rpv3-1 and Rpv12 loci showed the strongest resistance response (scarce sporulation and necrosis), those carrying Rpv3-3 locus showed the highest levels of necrosis while Rpv10 carrying genotypes showed intermediate levels of both sporulation and necrosis.

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Garima Bhatia ◽  
Santosh K. Upadhyay ◽  
Anuradha Upadhyay ◽  
Kashmir Singh

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.


2018 ◽  
Vol 22 (6) ◽  
pp. 703-707 ◽  
Author(s):  
E. T. Ilnitskaya ◽  
M. V. Makarkina ◽  
S. V. Tokmakov ◽  
L. G. Naumova

Downy mildew is one of the most common fungal diseases of the vine, caused by Plasmopara viticola. An effective way to control the spread of the pathogen is to cultivate resistant varieties. Cultivars of Vitis vinifera, being the basis of high-quality viticulture, practically do not possess genetic resistance to P. viticola, so screening for resistance donors is an important stage in breeding. One of the major resistance loci to downy mildew, the Rpv3 gene, was identified in the genotype of a complex interspecific hybrid of grapes Bianca. Later, it was found that this gene had seven haplotypes of resistance inherited from North American grape species, and that it was possible to identify the allelic status of the gene using DNA-markers UDV305, UDV737. However, only two haplotypes can be combined in one diploid form. To determine the Rpv3 gene in the grape gene pool we, using these markers, studied 35 different genotypes of grapevines, most of which are interspecies cultivars. Three varieties with known allelic status of the Rpv3 gene (Dunavski lazur, Noah, Seyve Villard 12-375) were included in the study as reference genotypes. The genotypes were studied through polymerase chain reaction with separation of amplification products by capillary electrophoresis in automatic genetic analyzer ABI Prism 3130. In the studied grape cultivars DNA marker analysis indentified the Rpv3 gene in sixteen genotypes of interspecific origin, including haplotype Rpv3299-279 found in twelve varieties, Rpv3321-312 – in three, and haplotype Rpv3null-271 – in one variety. Seyve Villard 12-375 turned out to be the donor of resistance gene in the most of the genotypes carrying Rpv3 in this study. The obtained data can be useful in selection of mildew resistant grape varieties and screening for hybridization pairs.


2018 ◽  
Vol 53 (5) ◽  
pp. 602-610 ◽  
Author(s):  
Luciano Saifert ◽  
Fernando David Sánchez-Mora ◽  
Wilson Taybar Assumpção ◽  
Jean Alberto Zanghelini ◽  
Renan Giacometti ◽  
...  

Abstract: The objective of this work was to use a marker-assisted selection for pyramiding the resistance loci Rpv1 and Rpv3.1 in grapevine (Vitis vinifera), and to evaluate their conferred resistance against Brazilian downy mildew (Plasmopara viticola) isolates. A progeny of 23 plants, segregating for the two resistance loci, was obtained by the cross between the Gf 2000-305-122 and Gf.Ga-52-42 genotypes. The progeny was genotyped with four microsatellite markers and phenotyped for resistance to P. viticola using a bioassay with leaf discs. Six plants containing the Rpv1 and Rpv3.1 pyramided loci were identified by the molecular analysis. Plants harboring the Rpv1 + Rpv3.1, Rpv3.1, and Rpv1 loci showed 12.8, 30.0, and 33.1 sporangiophores per leaf disc, respectively. Plants with no resistance loci showed a dense sporulation. The phenotypic analysis of the expression of the two pyramided loci was only confirmed for four plants that showed the highest resistance level, i.e., mean value of 1.8 sporangiophores. A high-throughput method for pyramiding the Rpv1 and Rpv3.1 loci was developed, which confirmed the increased resistance to P. viticola. The selected elite genetic material shows a high resistance to downy mildew and elevated enological potential for grapevine breeding in Brazil.


2008 ◽  
Vol 98 (7) ◽  
pp. 776-780 ◽  
Author(s):  
A. M. Díez-Navajas ◽  
S. Wiedemann-Merdinoglu ◽  
C. Greif ◽  
D. Merdinoglu

Following inoculation of host and nonhost plants with Plasmopara viticola, the grapevine downy mildew, a histological survey was undertaken to identify the stage where its development is contained in nonhosts and in resistant host plants. Three herbaceous nonhost species, Beta vulgaris, Lactuca sativa, and Capsicum annuum, and three grapevine species displaying different level of resistance (Vitis vinifera [susceptible], Vitis riparia [partially resistant] and Muscadinia rotundifolia [totally resistant]) where inoculated by P. viticola using a controlled leaf disk inoculation bioassay. During the early steps of infection, defined as encystment of zoospores on stomata, penetration of the germ tube, and production of the vesicle with the primary hypha, there was no evidence of a clear-cut preference to grapevine tissues that could attest to host specificity. The main difference between host grapevine species and nonhosts was observed during the haustorium formation stage. In nonhost tissues, the infection was stopped by cell wall-associated defense responses before any mature haustorium could appear. Defense responses in resistant grapevines were triggered when haustoria were fully visible and corresponded to hypersensitive responses. These observations illustrate that, for P. viticola, haustorium formation is not only a key stage for the establishment of biotrophy but also for the host specificity and the recognition by grapevine resistance factors.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Chantal Wingerter ◽  
Birgit Eisenmann ◽  
Patricia Weber ◽  
Ian Dry ◽  
Jochen Bogs

Abstract Background The high susceptibility of European grapevine cultivars (Vitis vinifera) to downy mildew (Plasmopara viticola) leads to the intensive use of fungicides in viticulture. To reduce this input, breeding programs have introgressed resistance loci from wild Vitis species into V. vinifera, resulting in new fungus-resistant grapevine cultivars (FRC). However, little is known about how these different resistance loci confer resistance and what the potential reduction in fungicide applications are likely to be if these FRCs are deployed. To ensure a durable and sustainable resistance management and breeding, detailed knowledge about the different defense mechanisms mediated by the respective Rpv (Resistance to P. viticola) resistance loci is essential. Results A comparison of the resistance mechanisms mediated by the Rpv3–1, Rpv10 and/or Rpv12-loci revealed an early onset of programmed cell death (PCD) at 8 hours post infection (hpi) in Rpv12-cultivars and 12 hpi in Rpv10-cultivars, whereas cell death was delayed in Rpv3-cultivars and was not observed until 28 hpi. These temporal differences correlated with an increase in the trans-resveratrol level and the formation of hydrogen peroxide shortly before onset of PCD. The differences in timing of onset of Rpv-loci specific defense reactions following downy mildew infection could be responsible for the observed differences in hyphal growth, sporulation and cultivar-specific susceptibility to this pathogen in the vineyard. Hereby, Rpv3- and Rpv12/Rpv3-cultivars showed a potential for a significant reduction of fungicide applications, depending on the annual P. viticola infection pressure and the Rpv-loci. Furthermore, we report on the discovery of a new P. viticola isolate that is able to overcome both Rpv3- and Rpv12-mediated resistance. Conclusion This study reveals that differences in the timing of the defense reaction mediated by the Rpv3-, Rpv10- and Rpv12-loci, result in different degrees of natural resistance to downy mildew in field. Vineyard trials demonstrate that Rpv12/Rpv3- and Rpv3-cultivars are a powerful tool to reduce the dependence of grape production on fungicide applications. Furthermore, this study indicates the importance of sustainable breeding and plant protection strategies based on resistant grapevine cultivars to reduce the risk of new P. viticola isolates that are able to overcome the respective resistance mechanism.


2022 ◽  
Vol 12 ◽  
Author(s):  
Ying Wang ◽  
Xiao Cao ◽  
Yulei Han ◽  
Xing Han ◽  
Zhilei Wang ◽  
...  

Downy mildew is a major threat to viticulture, leading to severe yield loss. The use of traditional copper-based fungicides is effective, but has adverse effects on the environment and human health, making it urgent to develop an environmentally friendly disease management program. Multi-functional kaolin particle film (KPF) is promising as an effective and safer treatment strategy, since this material lacks chemically active ingredients. In this study, ability of Kaolin particle film (KPF) pretreatment to protect grapevine leaves from Plasmopara viticola was tested and the mode of action of KPF was analyzed. KPF application reduced the disease severity and the development of intercellular hyphae. Additionally, there was reduced accumulation of H2O2 and malondialdehyde (MDA) with pretreatment. The observation of ultrastructure on the leaf surface showed KPF deposition and stomatal obstruction, indicating that KPF protected plants against disease by preventing the adhesion of pathogens to the leaf surface and blocking invasion through the stomata. KPF pretreatment also activated host defense responses, as evidenced by increased activities of anti-oxidative enzymes [superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT)] and defense-related enzymes [phenylalanine ammonia-lyase (PAL), chitinases, and β-1,3-glucanases], increased phytohormone signals [abscisic acid (ABA), salicylic acid (SA), and jasmonic acid (JA)] and the up-regulation of defense genes related to plant defense. Overall, these results demonstrate that KPF treatment counters grapevine downy mildew by protecting leaves and enhancing plant defense responses.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Jun Kang ◽  
Peijie Gong ◽  
Mengqing Ge ◽  
Ehsan Sadeghnezhad ◽  
Zhongjie Liu ◽  
...  

Abstract Background Papain-like cysteine proteases (PLCPs), a large group of cysteine proteases, are structurally related to papain. The members belonging to PLCPs family contribute to plant immunity, senescence, and defense responses in plants. The PLCP gene family has been identified in Arabidopsis, rice, soybean, and cotton. However, no systematic analysis of PLCP genes has been undertaken in grapevine. Since Plasmopara viticola as a destructive pathogen could affect immunity of grapes in the field, we considered that the members belonged to PLCPs family could play a crucial role in defensive mechanisms or programmed cell death. We aimed to evaluate the role of PLCPs in 2 different varieties of grapevines and compared the changes of their expressions with the transcriptional data in response to P. viticola. Results In this study, 23 grapevine PLCP (VvPLCP) genes were identified by comprehensive bioinformatics analysis. Subsequently, the chromosomal localizations, gene structure, conserved domains, phylogenetic relationship, gene duplication, and cis-acting elements were analyzed. Numerous cis-acting elements related to plant development, hormone, and stress responses were identified in the promoter of the VvPLCP genes. Phylogenetic analysis grouped the VvPLCP genes into nine subgroups. The transcription of VvPLCP in different inoculation time points and varieties indicated that VvPLCP may have vital functions in grapevine defense against Plasmopara viticola. According to transcriptome data and qPCR analysis, we observed the increasing expression levels of VvRD21–1 at 72 h after inoculation in resistant variety, inferring that it was related to grape downy mildew resistance. Meanwhile, 3 genes including VvXBCP1, VvSAG12–1, and VvALP1 showed higher expression at 24 h after pathogen inoculation in the susceptible variety and might be related to the downy mildew phenotype. We nominated these four genes to function during hypersensitive response (HR) process, inferring that these genes could be associated with downy mildew resistance in grapes. Conclusions Our results provide the reference for functional studies of PLCP gene family, and highlight its functions in grapevine defense against P. viticola. The results help us to better understand the complexity of the PLCP gene family in plant immunity and provide valuable information for future functional characterization of specific genes in grapevine.


2020 ◽  
Vol 107 ◽  
pp. 1-4
Author(s):  
Sendhilvel v ◽  
◽  
Marimuthu T ◽  
Geetha Lakshmi V ◽  
◽  
...  

Grapevine downy mildew caused by Plasmopara viticola is an explosivedisease that causes severe damage to leaf, flowers and fruits of the vine.In the present study, an epidemiological model was developed to supportthe vine-growers to reduce the number of fungicidal sprays and protect theenvironmental hazards besides saving the input cost. To record the weatherdata, an automatic weather station was installed at Mathampatti Village,Coimbatore, India and the weather parameters such as relative humidity,minimum, maximum and night temperatures, rainfall, leaf wetness, dew fall,solar radiation etc., were recorded at once in a 10 minutes interval during2014-17. A logistic equation was designed based on the biological data(disease incidence, weather parameter and crop age) and its mathematicalanalysis was shown in this paper. The critical weather parameters werestudied using logistic equation with the support of “Curve expert programme”.The mode described the development of the infection and infection ratethrough computation-based analysis. The Decision Support Systems (DSS)were arrived using the disease progress curve obtained from the logisticequation. The DSS indicates the first two sprays during the congenial phaseof the disease from 25 to 45 Days After Pruning (DAP). Then the secondphase, ie rapid growth stage was termed as ‘Exponential phase’, in which twosprays on 55 and 65 DAP (flowering and fruiting stage) were carried out. Inthe final plateau phase (66 DAP to till harvest), the management strategiesapplied would not be cost-effective and no spray was recommended. Thefield experiment was carried out using the findings of the logistic equationwith different management modules for validation. The number of spraysof fungicide was reduced from 15 to 5 times in the logistic equation-basedtreated plot.


2009 ◽  
Vol 26 (Special Issue) ◽  
pp. S13-S17 ◽  
Author(s):  
P. Bábíková ◽  
N. Vrchotová ◽  
J. Tříska ◽  
M. Kyseláková

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


2021 ◽  
Vol 22 (2) ◽  
pp. 940
Author(s):  
Elodie Vandelle ◽  
Pietro Ariani ◽  
Alice Regaiolo ◽  
Davide Danzi ◽  
Arianna Lovato ◽  
...  

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.


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