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

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.

2021 ◽  
Vol 34 ◽  
pp. 02001
Author(s):  
Elena Ilnitskaya ◽  
Marina Makarkina ◽  
Valeriy Petrov

Downy mildew (Plasmopara viticola) and powdery mildew (Erysiphe necator) are the most common and economically significant fungal diseases in vineyards. The task of this work is to study the genotypes of new promising hybrid forms of table grapes for the presence of resistance genes to downy mildew (Rpv10 and Rpv3) and powdery mildew (Ren9) using DNA-markers. The study was carried out on table grape hybrids under the working names Agat dubovskiy, Akelo, Arabella, Artek, Dubovskiy rozovyi, Gamlet, Ispolin, Kishmish dubovskiy, Kurazh, Pestryi, Valensiya and registered variety Liviya. The studied genes were analyzed using markers UDV305 and UDV737 (Rpv3), GF09-46 (Rpv10), CenGen6 (Ren9). The following cultivars were used as reference genotypes: Saperavi severnyi (carries Rpv10 gene) and Regent (Rpv3 and Ren9). It was established that Rpv3 gene is carried by hybrids Kishmish dubovskiy, Agat dubovskiy, Kurazh, Valensiya, Akelo, Gamlet, Dubovskiy rozovyi, Pestryi. Ren9 gene was found in Artek, Agat dubovskiy, Kurazh, Ispolin, Valensiya, Arabella, Gamlet, Dubovskiy rozovyi, Pestryi. The Rpv10 gene was not detected in any of the analyzed grapevine samples. genotypes Agat dubovskiy, Kurazh, Gamlet, Dubovskiy rozovyi, Pestryi, Valensiya carry Rpv3 and Ren9 genes simultaneously. These grapevines have an elegant bunch and large berries that are attractive to consumers.


2008 ◽  
Vol 35 (1) ◽  
pp. 77 ◽  
Author(s):  
Giovanni Agati ◽  
Zoran G. Cerovic ◽  
Anna Dalla Marta ◽  
Valentina Di Stefano ◽  
Patrizia Pinelli ◽  
...  

The role of flavonoids in the response of plants to Plasmopara viticola, the phytopathogen agent of downy mildew, was studied in the Vitis vinifera L. cultivar Sangiovese. Grapevines in the vineyard were exposed to two light regimes, 100% and 35% of full sunlight in order to induce differences in total leaf polyphenolic content. Epidermal leaf phenolic compounds were assessed optically, using the Dualex chlorophyll fluorescence-based portable leaf-clip. Dualex data were calibrated by means of HPLC analysis of extracts from the same measured leaves. Good correlations were obtained with total flavonoid contents, which consist mainly of quercetin 3-O-glucuronide. From the Dualex non-destructive measurements, we showed that full-sun exposed leaves contained 75% more flavonoids than shaded leaves. Inoculation of leaves with P. viticola sporangia resulted in a significantly lower infected leaf area in sun-lit leaves compared with shaded ones, as seen from subsequent analysis of the downy mildew severity. These results indicated an inverse relationship between preformed flavonoids and the susceptibility of grapevines to downy mildew. The rapid optical method for the non-destructive assessment of flavonoids presented here could be useful for large scale screening and predicting V. vinifera susceptibility to P. viticola.


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.


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.


Plant Disease ◽  
2021 ◽  
Author(s):  
Bora Kim ◽  
Jae Sung Lee ◽  
Young-Joon Choi

Vitis davidii (Rom.Caill.) Foëx, commonly known as spine grape, is a deciduous climber native to China. Its fruits are consumed fresh or used to make wine in South and Central China. In recent years, spine grape has been cultivated in Korea. In July 2020, downy mildew was detected on spine grape vines in Jeongeup (35°42′17″N, 126°54′02″E), Korea, with a disease incidence of 70%. The symptoms appeared as yellowish, brownish, or reddish, vein-limited, poly-angular adaxial leaf spots, correspond to dense, white downy growth abaxially. A representative specimen was deposited in the Kunsan National University Herbarium (KSNUH679). Sporangiophores were tree-like, hyaline, mostly straight, and monopodially branched in orders of three to six; they measured (219.4–)273.2 to 435.1(–546.6) × (4.8–)6.7 to 9.0(–10.0) μm (n = 50). Ultimate branchlets were bi or trifurcate, straight to slightly curved, with truncate or, rarely, a swollen tip and measured 2.9 to 9.7 μm long and 0.8 to 2.5 μm wide at the base (n = 50). Sporangia were hyaline, ovoidal or lemon-shaped; they measured (16.8–)20.0 to 28.8(–34.2) × (11.4–)13.1 to 17.0(–20.1) μm with a length to width ratio of (1.28–)1.46 to 1.78(–2.07) (n = 50). This morphology was as described for Plasmopara viticola (Berk. & M. A. Curtis) Berl. & De Toni (Hall, 1989). Genomic DNA was extracted directly from infected V. davidii leaves. Three regions were PCR-amplified and sequenced: cox2 mtDNA with primers cox2F and cox2-RC4 (Choi et al., 2015), actin with primers pve04815-F and pve04815-R, and beta-tubulin with primers pvc389-F3 and pvc389-R4 (Rouxel et al., 2013). The resulting sequences were deposited in GenBank (accession nos. MT834527 for cox2, MT834525 for actin, and MT834526 for beta-tubulin). A BLASTn search revealed that the Korean sample was identical to P. viticola clade aestivalis originating from Vitis species: MK215072 for cox2 sequence, KY933800 for actin, and MK358393 for beta-tubulin. In all phylogenetic analyses of the three genes (cox2, actin, and beta-tubulin), KSNUH679 came out as phylogenetically place within P. viticola clade aestivalis, which has recently been reported on V. coignetiae and V. ficifolia var. sinuata in Korea (Kim et al., 2019). A pathogenicity test was performed twice by inoculating the leaves of 10 healthy spine grape plants with a sporangial suspension (~1 × 106 sporangia·mL-1) and incubating them in a growth chamber at 25 °C, 12-h day/night cycle, and 90% relative humidity; five non-inoculated plants served as controls. After two weeks, all inoculated plants developed typical downy mildew symptoms could be observed, whereas the controls remained symptomless. Morphology and molecular features confirmed the identity of the pathogen of spine grape to be P. viticola. To the best of our knowledge, this is the first report of downy mildew caused by P. viticola on V. davidii in Korea. Recently, downy mildew outbreaks caused by P. viticola have been recorded in spine grape plantations in southern China (Yi et al., 2019). Considering the potential of spine grape as a novel crop for Korea, P. viticola appears to represent a significant threat to this industry.


Plant Disease ◽  
1999 ◽  
Vol 83 (3) ◽  
pp. 301-301 ◽  
Author(s):  
S. J. McKirdy ◽  
I. T. Riley ◽  
I. J. Cameron ◽  
P. A. Magarey

Despite the suitability of climate, Western Australia was one of the few grape (Vitis vinifera L.) growing areas free of grapevine downy mildew (Plasmopara viticola (Berk. & M. A. Curtis) Berl. & De Toni in Sacc.). Area freedom had been maintained by restricting the movement of host material and machinery from outside the state and fungicide use in Western Australia vineyards had been considerably less. P. viticola was detected in 1997 in 14 of 15 vines growing at Kalumburu, a remote community in the semi-arid tropics of Western Australia, and was eradicated. In October 1998, grape leaves with oilspots typical of downy mildew were received from a grower in the Swan Valley near Perth, one of the main production areas of Western Australia. Sporangia were hyaline and ellipsoid (14 × 11 μm), were borne on treelike sporangiophores, and were consistent with those described for P. viticola (1). This is the first record of P. viticola in commercial viticulture in Western Australia. A response plan for exotic diseases was activated and after 2 weeks of surveillance the disease was found in 45 of 70 vineyards surveyed of the 280 vineyards in the Swan Valley. Given the extent of spread, eradication of downy mildew was not considered possible. Weather data for August to October 1998 indicated the likelihood of several infection periods from budburst to flowering when the disease was first detected. Crop loss will be considerable in many vineyards. P. viticola was also found in bench-grafted cuttings in pots in leaf consigned from the Swan Valley to several other areas in August 1998. Downy mildew was found in other areas only in association with these consigned vines. An industry code of practice, including hygiene, is being developed to slow the rate of spread of P. viticola in Western Australia. Reference: (1) Anon. C.M.I. Descriptions of Pathogenic Fungi and Bacteria No. 980, 1989.


2021 ◽  
Vol 12 ◽  
Author(s):  
Gaoqing Xiang ◽  
Xiao Yin ◽  
Weili Niu ◽  
Tingting Chen ◽  
Ruiqi Liu ◽  
...  

Grapevine downy mildew is an insurmountable disease that endangers grapevine production and the wine industry worldwide. The causal agent of the disease is the obligate biotrophic oomycete Plasmopara viticola, for which the pathogenic mechanism remains largely unknown. Crinkling and necrosis proteins (CRN) are an ancient class of effectors utilized by pathogens, including oomycetes, that interfere with host plant defense reactions. In this study, 27 CRN-like genes were cloned from the P. viticola isolate YL genome, hereafter referred to as PvCRN genes, and characterized in silico and in planta. PvCRN genes in ‘YL’ share high sequence identities with their ortholog genes in the other three previously sequenced P. viticola isolates. Sequence divergence among the genes in the PvCRN family indicates that different PvCRN genes have different roles. Phylogenetic analysis of the PvCRN and the CRN proteins encoded by genes in the P. halstedii genome suggests that various functions might have been acquired by the CRN superfamily through independent evolution of Plasmopara species. When transiently expressed in plant cells, the PvCRN protein family shows multiple subcellular localizations. None of the cloned PvCRN proteins induced hypersensitive response (HR)-like cell death on the downy mildew-resistant grapevine Vitis riparia. This was in accordance with the result that most PvCRN proteins, except PvCRN11, failed to induce necrosis in Nicotiana benthamiana. Pattern-triggered immunity (PTI) induced by INF1 was hampered by several PvCRN proteins. In addition, 15 PvCRN proteins prevented Bax-induced plant programmed cell death. Among the cell death-suppressing members, PvCRN17, PvCRN20, and PvCRN23 were found to promote the susceptibility of N. benthamiana to Phytophthora capsici, which is a semi-biotrophic oomycete. Moreover, the nucleus-targeting member, PvCRN19, promoted the susceptibility of N. benthamiana to P. capsici. Therefore, these PvCRN proteins were estimated to be virulent effectors involved in the pathogenicity of P. viticola YL. Collectively, this study provides comprehensive insight into the CRN effector repertoire of P. viticola YL, which will help further elucidate the molecular mechanisms of the pathogenesis of grapevine downy mildew.


2017 ◽  
Vol 18 (3) ◽  
pp. 192-195
Author(s):  
David S. Jones ◽  
Patricia S. McManus

Downy mildew, caused Plasmopara viticola, is a major disease affecting grape production worldwide. While the symptoms and signs of downy mildew on European wine grapes (Vitis vinifera) are well documented, minimal information is available regarding the symptoms and signs on cold-hardy cultivars that are crosses of V. vinifera and Vitis species native to North America. Referred to as “cold-climate cultivars,” these hybrids have been released over the past two decades and form the basis of a burgeoning wine industry in the northern United States. Most research on downy mildew of grape has involved V. vinifera, and consequently, many images used for educational purposes illustrate “classic” symptoms and signs on highly susceptible cultivars of V. vinifera. However, we have observed that such images can be inconsistent with symptomology in the cold-climate grape hybrids, and there are differences in the symptoms and signs of downy mildew among cold-climate cultivars. These inconsistencies can result in incorrect diagnosis, which can lead to poor management decisions, including application of products that are not appropriate for the pathogen(s) present. Thus, the objective of the current guide is to describe downy mildew symptoms and signs on cold-climate cultivars, with special attention to features that are not documented in resources widely used for grape disease diagnosis.


2012 ◽  
Vol 102 (11) ◽  
pp. 1094-1101 ◽  
Author(s):  
Ying Yu ◽  
Yali Zhang ◽  
Ling Yin ◽  
Jiang Lu

The resistance and susceptibility of grapevines to downy mildew (DM) disease caused by Plasmopara viticola were compared among different cultivars/accessions belonging to Vitis vinifera, V. rotundifolia, and 10 oriental Vitis species. After inoculation with P. viticola pathogen, no symptom was found in V. rotundifolia grapevines at all, while oriental species V. davidii and V. piasezkii, like V. vinifera, were susceptible to DM disease. The other eight oriental Vitis species showed various resistance levels to DM disease. Intraspecific resistant variations were also observed in V. amurensis. Microscopy studies were conducted on various time courses after pathogen infection on grape leaves. P. viticola hyphae were not observed in V. rotundifolia cultivars, while symptoms with varying degrees of severity were observed among the Euvitis species. In general, the DM resistant oriental species showed a slower development of hypha and less formation of haustoria than DM susceptible V. vinifera grapevines. Cells with distinctive fluorescence were observed in V. rotundifolia and the oriental species V. pseudoreticulata, and callose deposits were observed in V. rotundifolia, V. pseudoreticulata, and V. amurensis grapevines. Based on the results of morphological observations and microscopy studies, we concluded that there were five levels of grapevine resistance to P. viticola pathogen: (i) immune, (ii) extremely resistant, (iii) resistant, (iv) partly resistant, and (v) susceptible.


Author(s):  
Leilani M. Chirino ◽  
Suresh Kumar ◽  
Mariko Okumura ◽  
David E. Sterner ◽  
Michael Mattern ◽  
...  

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