ROLE OF PLANT MLO PROTEINS DURING MARCHANTIA – PHYTOPHTHORA INTERACTIONS

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INRAE presentation

The French National Research Institute for Agriculture, Food, and Environment (INRAE) is a major player in research and innovation. It is a community of 12,000 people with 272 research, experimental research, and support units located in 18 regional centres throughout France. Internationally, INRAE is among the top research organisations in the agricultural and food sciences, plant and animal sciences, as well as in ecology and environmental science. It is the world’s leading research organisation specialising in agriculture, food and the environment. INRAE’s goal is to be a key player in the transitions necessary to address major global challenges. Faced with a growing world population, climate change, resource scarcity, and declining biodiversity, the Institute has a major role to play in building solutions and supporting the necessary acceleration of agricultural, food and environmental transitions.

Work environment, missions and activities

Work environment, missions and activities 

You will join the IPO team (Plant-Oomycete Interactions) at INRAE's Sophia Agrobiotech Institute. The team investigates the dynamics between plants and oomycete pathogens from sporulation through late infection. It comprises 6 researchers, 2 research engineers, 2 assistant engineers, and 5 international PhD students. Project Background The plant Mildew Locus O (MLO) loci were discovered more than 80 years ago in barley as the first plant susceptibility S factors and have since been used successfully in agriculture to provide in different crops broad and robust resistance to the filamentous powdery mildew fungi such as Erysiphe spp.1. The role of MLO proteins, recently proposed to function as calcium channels2, during interactions with filamentous oomycetes such as Phytophthora spp. remains largely unexplored in flowering plants and unstudied in bryophytes. This limits our understanding of the conserved host processes that determine disease outcome in these devastating and difficult-to-control oomycete pathogens. This knowledge gap persists despite the similarities between the parasitic hemibiotrophic Phytophthora and the biotroph Erysiphe, including intimate host interactions, formation of specialized host interfaces, effector secretion, and suppression of plant immunity. There are 15 MLOs in the flowering plant model Arabidopsis thaliana and 3 of them, AtMLO2/6/12, have been shown to promote susceptibility to Erysiphe3 with AtMLO2 being the most important one, while there are only 4 MLOs (MpMLO1-4) in the early diverging bryophyte Marchantia polymorpha whose function during plant-pathogen interaction is completely unknown.

Project overview In close collaboration with a 2nd year PhD researcher, and focusing on the unpublished role of MpMLO2 in the Marchantia polymorpha–Phytophthora interaction, the intern will: 

1. Generate transgenic M. polymorpha Mpmlo2 knockout lines expressing Marchantia MpMLO2 or Arabidopsis AtMLO4 proteins fused to the fluorescent reporter Citrine under the constitutive 35S promoter;

2. Use confocal microscopy to characterize the subcellular localization of the MLO-Citrine fusion proteins in Marchantia. Localization patterns will be compared between MpMLO2 and AtMLO4 and with previous observations made in transiently transformed tobacco leaves; 

3. Challenge selected transformed lines with Phytophthora infection to determine whether the MpMLO2-Citrine and AtMLO4-Citrine fusion proteins can complement the infection phenotype of Mpmlo2 knockout mutants. This will allow assessment of whether the previously identified role of MpMLO2 in the Phytophthora interaction is retained by the fluorescently tagged protein and whether this function can be conserved by the evolutionarily divergent Arabidopsis MLO4 protein;

4. Investigate, during the growth and propagation of primary transformants, protein–protein interactions between Marchantia MLO proteins and conserved MLO-associated proteins, such as MARIS, EXO70 or calmodulin, using at least one protein–protein interaction approach. Your work will determine the conservation of MLO-associated interactions across land plant evolution and provide insights into the molecular mechanisms underlying MLO functions.

Training and skills

Required skills and profile 

• MSc 1 or 2 students (or equivalent). 

• Internship duration: 5-6 months. 

• Recommended background: degree in molecular biology, plant physiology, or similar. 

• Desired knowledge: basic knowledge of plant-pathogen interactions; of molecular biology and in vitro culture techniques. 

• Desired skills: Ability to work in a team, to communicate in English.

INRAE's life quality

By joining our teams, you benefit from (depending on the type of contract and its duration):

- up to 30 days of annual leave + 15 days "Reduction of Working Time" (for a full time);
- parenting support: CESU childcare, leisure services;
- skills development systems: training, career advise;
- social support: advice and listening, social assistance and loans;
- holiday and leisure services: holiday vouchers, accommodation at preferential rates;
- sports and cultural activities;
- collective catering.

How to apply

Please send your CV and a motivation letter to aurelien.boisson-dernier@inrae.fr