CRISPR/Cas9-mediated VvPR4b editing decreases downy mildew resistance in grapevine (Vitis vinifera L.)

Li, Mengyuan; Jiao, Yuntong; Wang, Yuting; Zhang, Na; Wang, Bianbian; Liu, Ruiqi; Yin, Xiao; Liu, Guotian

doi:10.1038/s41438-020-00371-4

Cited by 87 publications

(53 citation statements)

References 56 publications

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“…Traditionally, the contribution of candidate genes to salt tolerance in grapevine is assessed by comparing their expression between cultivars with different degrees of tolerance; however, this approach is only correlative and does not take into account additional genotypic variation that contributes towards salt tolerance mechanisms. This obstacle might be overcome using gene editing through CRISPR/Cas9, which has been used to produce mutant grapevines with specific genes knocked out (Osakabe et al 2018, Li et al 2020b, Ren et al 2020); hence, the contribution of candidate genes to salt tolerance could potentially be studied more accurately. Previous studies have proposed many signalling, biosynthesis, metabolism and transport processes involved in grapevine salt tolerance, that would overload functional testing.…”

Section: Present and Future Perspectivesmentioning

confidence: 99%

Grapevine salt tolerance

Zhou‐Tsang

Henderson

et al. 2021

Australian Journal of Grape and Wine Research

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Salinity, which is predominantly an issue for agricultural systems in arid and semi-arid regions, has the potential to impair grape production and wine quality, and its impact on the grape and wine industries is predicted to increase with climate change. Research on the physiological and molecular changes that occur in salt-affected vines has unveiled complex osmotic and ionic responses that include oxidative stress, water loss, photoinhibition, growth inhibition and necrosis. Proposed salt tolerance mechanisms include elevated antioxidant production, hydric regulation and salt exclusion from shoots and berries. These later of these mechanisms is found in certain Vitis genotypes that, when grafted as rootstocks, can protect fruit-bearing scions from accumulating significant amounts of saline ions from soils, most notably through the presence of specific transport proteins that are involved in regulating the transfer of ions from root to shoot via the xylem. Significant gaps in knowledge remain, however, regarding salt tolerance mechanisms for Vitis species, with many mechanisms inferred from other species or documented only at the level of gene expression. A better understanding of the mechanisms that confer salt tolerance in Vitis species is needed to improve the production of new germplasm that is locally adapted and better suited to the challenges of a changing climate. Hence, this review covers the current knowledge on the characteristics that are associated with salt damage and tolerance in grapevine cultivars and rootstocks and highlights possible future avenues that will enable development of new options for the industry to combat salinity.

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Section: Present and Future Perspectivesmentioning

confidence: 99%

Grapevine salt tolerance

Zhou‐Tsang

Henderson

et al. 2021

Australian Journal of Grape and Wine Research

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“…The photographs were obtained by a fluorescence microscope (Olympus bx-51). The wavelengths used for excitation were 400-440 nm (blue/purple) 56,57 .…”

Section: Trypan Blue Staining and Aniline Blue Stainingmentioning

confidence: 99%

Importin-αs are required for the nuclear localization and function of the Plasmopara viticola effector PvAVH53

et al. 2021

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Plant pathogenic oomycetes deliver a troop of effector proteins into the nucleus of host cells to manipulate plant cellular immunity and promote colonization. Recently, researchers have focused on identifying how effectors are transferred into the host cell nucleus, as well as the identity of the nuclear targets. In this study, we found that the RxLR effector PvAVH53 from the grapevine (Vitis vinifera) oomycete pathogen Plasmopara viticola physically interacts with grapevine nuclear import factor importin alphas (VvImpα and VvImpα4), localizes to the nucleus and triggers cell death when transiently expressed in tobacco (Nicotiana benthamiana) cells. Deletion of a nuclear localization signal (NLS) sequence from PvAVH53 or addition of a nuclear export signal (NES) sequence disrupted the nuclear localization of PvAVH53 and attenuated its ability to trigger cell death. Suppression of two tobacco importin-α genes, namely, NbImp-α1 and NbImp-α2, by virus-induced gene silencing (VIGS) also disrupted the nuclear localization and ability of PvAVH53 to induce cell death. Likewise, we transiently silenced the expression of VvImpα/α4 in grape through CRISPR/Cas13a, which has been reported to target RNA in vivo. Finally, we found that attenuating the expression of the Importin-αs genes resulted in increased susceptibility to the oomycete pathogen Phytophthora capsici in N. benthamiana and P. viticola in V. vinifera. Our results demonstrate that importin-αs are required for the nuclear localization and function of PvAVH53 and are essential for host innate immunity. The findings provide insight into the functions of importin-αs in grapevine against downy mildew.

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“…PR4, which is a chitinase and chitin-binding protein, plays a significant role in pathogen responses in many plants. Transgenic overexpression of PR4 in grapevine conferred increased resistance to powdery mildew, while loss-of-function PR4 mutations increased susceptibility to downy mildew in grapevine 70 , 71 . In our study, PR4 was induced upon infection in LB but not PN (Table 3 ).…”

Section: Discussionmentioning

confidence: 99%

Proteomic analysis of early-stage incompatible and compatible interactions between grapevine and P. viticola

et al. 2021

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Wild grapevines can show strong resistance to the downy mildew pathogen P. viticola, but the associated mechanisms are poorly described, especially at early stages of infection. Here, we performed comparative proteomic analyses of grapevine leaves from the resistant genotype V. davidii “LiuBa-8” (LB) and susceptible V. vinifera “Pinot Noir” (PN) 12 h after inoculation with P. viticola. By employing the iTRAQ technique, a total of 444 and 349 differentially expressed proteins (DEPs) were identified in LB and PN, respectively. The majority of these DEPs were related to photosynthesis, respiration, cell wall modification, protein metabolism, stress, and redox homeostasis. Compared with PN, LB showed fewer downregulated proteins associated with photosynthesis and more upregulated proteins associated with metabolism. At least a subset of PR proteins (PR10.2 and PR10.3) was upregulated upon inoculation in both genotypes, whereas HSP (HSP70.2 and HSP90.6) and cell wall-related XTH and BXL1 proteins were specifically upregulated in LB and PN, respectively. In the incompatible interaction, ROS signaling was evident by the accumulation of H2O2, and multiple APX and GST proteins were upregulated. These DEPs may play crucial roles in the grapevine response to downy mildew. Our results provide new insights into molecular events associated with downy mildew resistance in grapevine, which may be exploited to develop novel protection strategies against this disease.

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CRISPR/Cas9-mediated VvPR4b editing decreases downy mildew resistance in grapevine (Vitis vinifera L.)

Cited by 87 publications

References 56 publications

Grapevine salt tolerance

Grapevine salt tolerance

Importin-αs are required for the nuclear localization and function of the Plasmopara viticola effector PvAVH53

Proteomic analysis of early-stage incompatible and compatible interactions between grapevine and P. viticola

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