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.

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…

Drought responses in Chardonnay and Sauvignon blanc grapevine cultivars: Mechanistic insights and varietal contrasts

This study explored the responses of Chardonnay and Sauvignon blanc grapevine cultivars to water deficit across four years, uncovering their shared patterns and distinctive coping mechanisms. The research was conducted in a commercial vineyard located in Isla de Maipo, Chile. Various characterization approaches were employed including plant water potentials (), gas exchange measurements, shoot vulnerability curves, productivity assessments, and leaf cell water relations. Linear mixed models and sensitivity analyses were performed using various statistical methods to evaluate cultivar responses to water deficit. As the water deficit progressed, both cultivars displayed a parallel reduction in stomatal conductance, leaf turgor, and increased shoot embolism.

Aroma quality of fortified wines from different Moscato cv. Cultivated in sicily

AIM: Vitis vinifera L. cv. Moscato includes different varieties, mainly white grapes with a medium-sized berry, spheroidal or slightly flattened in shape, yellow greenish color which becomes golden yellow or amber when exposed to the sun. Moscato varieties are mainly used for the production of sweet aromatic wines

Effect of the winemaking technology on the phenolic compounds, foam parameters in sparklig wines

Contribution Sparkling wines elaborated following the traditional method undergo a second fermentation in closed bottles of base wines, followed by aging of wines with lees for at least 9 months. Most of the sparkling wines elaborated are white and rosé ones, although the production of red ones is highly increasing. One of the initial problems in red sparkling wine processing is to obtain suitable base wines that should have moderate alcohol content and astringency and adequate color intensity; which is difficult to obtain when grapes must be harvested at low phenolic and industrial maturity stage. The low phenolic maturity degree in the red grapes makes essential to choose an adequate winemaking methodology to obtain the base wines because the extracted polyphenols will vary according the winemaking technique: carbonic maceration or destemmed-crushed grapes.

La zonazione viticola e i compiti dell’amministrazione regionale

Solo attraverso un adeguato intervento di estirpazione e reimpianto dei vigneti è possibile preservare, adeguare e valorizzare il patrimonio viticolo e le produzioni che da esso derivano.
Il reimpianto dei vigneti è pertanto da intendersi come una normale pratica agricola, alla pari della rimonta di stalla in campo zootecnico, ma può assumere toni problematici quando, come si verifica adesso in Toscana per una serie di circostanze legate alla profonda trasfor­mazione della viticoltura avvenuta negli ultimi 30 anni, troppi impianti giungono contem­poraneamente a fine ciclo produttivo e devono essere rinnovati.

Vitamin content of grape musts and yeast nutrition: A review

The management of yeast nutrition is an essential approach for a better control over wine fermentation process. Most of the researches on this subject in the last decades focused on nitrogen nutrition. However, vitamins, while being key compounds for yeast metabolism as co-factors for numerous enzymatic activities, were left mostly unexplored.