Doctorate thesis of University of Montpellier

Wednesday, November 15, 2017


Plant adaptation to salinity stress: characterisation of ecotypic variants and knock-out lines for Na+ transport systems in rice

Fouad Al Shiblawi
BPMP, “Ion transport and adaptation to environment in cereals” team


Jury :
Mme Sophie FILLEUR, MCU, Université Paris 7-Diderot, Rapporteur
Mr Renaud BROUQUISSE, DR, INRA, Sophia Antipolis, Rapporteur
Mr Pascal GANTET, Professeur, Université Montpellier, Reviewer
Mme Anne-Aliénor VERY, CR, CNRS, Montpellier, Thesis Director


Abstract :
The salinity of irrigation water and soils is one of the main abiotic constraints in agriculture. Among the 130 mha of rice grown around the world, about 30% are in areas where salinity is too high to allow for good yields. In order to better understand the role of Na+ transport systems in the compartmentalization of Na+ in the plant to maintain a high K+/Na+ ratio in sensitive tissues during salt stress, I focused on two transporters from the HKT family, OsHKT1;1 and OsHKT1;3, in rice, the model cereal species. My main project combined a “promoter::GUS” strategy to specify the expression pattern of both genes, and a reverse genetics approach by producing loss-of-function mutants using the CRISPR-CAS9 technology, which was confirmed to be an efficient tool for generating indel mutations after targeted DNA breaks, stably transmitted in rice. Histochemical tests of GUS activity showed predominant expression of OsHKT1;1 and OsHKT1;3 in vascular tissues (xylem parenchyma and/or phloem), mainly in the aerial parts. Phenotypic analysis of “CRISPR” plants revealed changes in leaf Na+ accumulation profiles in mutants: lack of Na+ accumulation in leaf sheaths for oshkt1;1 plants and altered distribution of Na+ between old and young leaves in oshkt1;3 plants, leading to an increase in the leaf blade Na+ content in both types of mutants. Overall, these results suggest that OsHKT1 and OsHKT1.3 contribute to the desalination of young leaf blades during salt stress, via different mechanisms, controlling transports of Na+ through the xylem and phloem. In addition to this study, I collaborated with a group of geneticists who identified a strong association between ecotypic differences in root Na+ and K+/Na+ ratio, and a region of chromosome 4 of rice including the OsHKT1;1 gene. By comparing electrophysiologically the two major variants of OsHKT1;1, differences in Na+ transport capacities that could explain the root trait were observed. All the data obtained underline the interest of analyzing all HKT genes of rice in the mechanisms of plant tolerance to salinity.