terclim by ICS banner
IVES 9 IVES Conference Series 9 Functional characterization of grapevine MLO genes to define their roles in Powdery mildew susceptibility by CRISPR/Cas9 genome editing

Functional characterization of grapevine MLO genes to define their roles in Powdery mildew susceptibility by CRISPR/Cas9 genome editing

Abstract

Successful powdery mildew (PM) infection in plants relies on Mildew Resistance Locus O (MLO) genes, which encode susceptibility factors essential for fungal penetration. In Arabidopsis, loss-of-function mutations in three clade-V MLOs, AtMLO2, 6, and 12 confer complete resistance to PM infection. Since then, efforts are on to discover MLO genes contributing to PM susceptibility in many species to introduce mlo-based PM-resistance. Earlier studies in tomato and grapevine, using the RNAi approach, attributed PM susceptibility to SlMLO1, 5, and 8 and VvMLO3, 13, and 17, respectively indicating likely functional redundancy among MLOs. Here, we disrupted the closest grapevine orthologues, VvMLO3, 4, 13, and 17 through CRISPR/Cas9-mediated mutagenesis in the microvine model with the goal of identifying the candidate MLO genes to introduce mlo-based PM resistance. Individual mutants mlo3, mlo4, mlo13 and mlo17 showed 8 to 50% less infection to E. necator, whereas double mutants, mlo3/4, mlo3/13 andmlo13/17 and triple mutant mlo3/13/17 showed 60 to 90% less infection. But the quadruple mlo3/4/13/17 mutant plants showed near complete PM resistance. Considerable differences were observed in the resistance level of clones among the triple and quadruple mutants due to the differences in editing efficiency of individual guide RNAs. Some mutants showed pleiotropic effects in the growth and development, ranging from early senescence and stunted growth to non-flowering phenotypes, which also seemed to depend on the percentage of gene-edited cells in the plant. The overarching goal is to excise the genome-integrated T-DNA cassette from the mutants using CRISPR Ribonucleoproteins for transgene-free PM resistance.

Download the abstract

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Article

Authors

Satyanarayana Gouthu1*, Laurent Deluc2,3, Samuel Talbot1

1,2 Department of Horticulture, Oregon State University, Corvallis, OR, USA
3 Oregon Wine Research Institute, Oregon State University, Corvallis, OR, USA

Contact the author*

 

Keywords

Powdery mildew, Grapevine MLO, mildew-resistance, Gene Editing

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

Using GIS to assess the terroir potential of an Oregon viticultural region

Deciding to grow grapes in Oregon is complex issue due to our diverse geography, climate, and relatively short history of grape growing. For any potential grape grower, vineyard site selection is the single most important decision they will face.

Read More

Vine responses to two irrigation systems in the region of Vinhos Verdes

In this work we try to know the influence of two irrigation systems (Drip and Micro – jet ) with the same levels of water applied in an experimental vineyard in the region of Felgueiras.

Read More

Aroma profile of Tempranillo tropical red wines from different seasons in the São Francisco valley, northeast of Brazil

Aromatic characteristics of wines are strongly influenced by agronomical and enological factors, depending of the climate, cultivar and winemaking process. Tropical wines are a new concept of vitiviniculture that is being developped in the Northeast of Brazil since the 80’s, located between 8-9º latitude of the South Hemisphere, where the second most important cultivar used for reds is Tempranillo. In this condition, vines produce grapes and enologists elaborate wines twice a year, because high temperatures, solar radiation and water availability for irrigation.

Read More

Impact of oenological tannins on microvinifications affected by downy mildew

AIM: Vine diseases are still responsible for economic losses. Previous study in our laboratory, have shown effects of oenological tannins against Botrytis cinerea1,2. According to this, the aim was to evaluate the wine protection by oenological tannins against an another disease, the downy mildew. METHODS: During the 2020 vintage, infected grapes by downy mildew (Vitis vinifera cv. Merlot) were collected from the dispositive ResIntBio. The 100 kg were crushed, destemmed and dispatch into 10 aluminium tanks. SO2 was added at 3 g/hL. Oenological tannins (grape, quebracho, ellagitannin or gallotannin) were added at 100 g/hL into eight different tanks (4×2 tanks). The two last tanks were considered as control without addition of oenological tannins. Alcoholic fermentation was achieved with Actiflore 33® at 20 g/hL. Malolactic fermentation was achieved with Lactoenos B7at 1 g/hL. Finished wines were sulfited to obtain 45 mg/L of total SO2.

Read More

The effects of calcite silicon-mediated particle film application on leaf temperature and grape composition of Merlot (Vitis vinifera L.) vines under different irrigation conditions

This study examined whether the application of calcite-silicon mediated particle film (CaPF) at veraison can mitigate a drought-induced increase in leaf temperature on grapevine, thus contributing to improved leaf functionality, yield and grape composition traits. A total of 48 five-year-old Merlot (Vitis vinifera L.)

Read More