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1.
Plant Biotechnol J ; 21(10): 2140-2154, 2023 10.
Article in English | MEDLINE | ID: mdl-37448155

ABSTRACT

The hyperosmolality-gated calcium permeable channel 4.1 (OSCA4.1) belongs to an evolutionarily conserved small family of mechano-sensitive channels. OSCA members may represent key players in plant resistance to drought and to pathogen infection but are scarcely studied. After screening for resistance to pepino mosaic virus (PepMV) a collection of 1000 mutagenized tomato families, we identified a mutant showing no symptoms and reduced virus accumulation. Resistance was mapped to chromosome 2 between positions 46 309 531 to 47 044 163, where a missense mutation caused the putative truncation of the OSCA4.1 protein. A CRISPR/Cas9 slosca4.1 mutant was resistant to PepMV, but not to tobacco mosaic virus or potato virus X. Inoculation of mutant and wild type tomato protoplasts showed that resistance was expressed in single cells, suggesting a role for SlOSCA4.1 in early viral function(s); congruently, SlOSCA4.1 re-localized to structures reminiscent of viral replication complexes. We propose that SlOSCA4.1 contributes to the correct regulation of the Ca2+ homeostasis necessary for optimal PepMV infection. PepMV is a pandemic virus that causes significant losses in tomato crops worldwide. In spite of its importance, no tomato-resistant varieties have been deployed yet; the mutant identified here has great potential to breed tomato varieties resistant to PepMV.


Subject(s)
Potexvirus , Solanum lycopersicum , Solanum , Solanum lycopersicum/genetics , Potexvirus/genetics , Potexvirus/metabolism , Calcium/metabolism , Plant Breeding , Plant Diseases/genetics
2.
Plant Cell Environ ; 43(7): 1707-1721, 2020 07.
Article in English | MEDLINE | ID: mdl-32275780

ABSTRACT

Root K+ acquisition is a key process for plant growth and development, extensively studied in the model plant Arabidopsis thaliana. Because important differences may exist among species, translational research supported by specific studies is needed in crops such as tomato. Here we present a reverse genetics study to demonstrate the role of the SlHAK5 K+ transporter in tomato K+ nutrition, Cs+ accumulation and its fertility. slhak5 KO lines, generated by CRISPR-Cas edition, were characterized in growth experiments, Rb+ and Cs+ uptake tests and root cells K+ -induced plasma membrane depolarizations. Pollen viability and its K+ accumulation capacity were estimated by using the K+ -sensitive dye Ion Potassium Green 4. SlHAK5 is the major system for high-affinity root K+ uptake required for plant growth at low K+ , even in the presence of salinity. It also constitutes a pathway for Cs+ entry in tomato plants with a strong impact on fruit Cs+ accumulation. SlHAK5 also contributes to pollen K+ uptake and viability and its absence produces almost seedless fruits. Knowledge gained into SlHAK5 can serve as a model for other crops with fleshy fruits and it can help to generate tools to develop low Cs+ or seedless fruits crops.


Subject(s)
Cesium/metabolism , Plant Proteins/physiology , Plant Roots/metabolism , Potassium Channels/physiology , Potassium/metabolism , Solanum lycopersicum/metabolism , CRISPR-Associated Protein 9 , CRISPR-Cas Systems , Flowers/metabolism , Fruit/growth & development , Gene Editing , Solanum lycopersicum/physiology , Plant Proteins/metabolism , Plants, Genetically Modified , Pollen Tube/growth & development , Potassium Channels/metabolism , Reproduction , Seeds/growth & development
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