Abstract
Receptor-like cytoplasmic kinases (RLCKs) are receptor kinases that lack extracellular ligand-binding domains and have emerged as a major class of signaling proteins that regulate plant cellular activities in response to biotic/abiotic stresses and endogenous extracellular signaling molecules. We have identified a rice RLCK (OsRLCK311) that was significantly higher in transgenic pSARK-IPT rice (Oryza sativa) that exhibited enhanced growth under saline conditions. Overexpression of OsRLCK311 full-length protein (RLCK311FL) and the C-terminus of OsRLCK311 (ΔN) in Arabidopsis confirmed its role in salinity tolerance, both in seedlings and mature plants. Protein interaction assays indicated that OsRLCK311 and ΔN interacted in-vivo with the plasma membrane AQP AtPIP2;1. The RLCK311-PIP2;1 binding led to alterations in the stomata response to ABA, which was characterized by more open stomata of transgenic plants. Moreover, OsRLCK311-ΔN effect in mediating enhanced plant growth under saline conditions was also observed in the perennial grass Brachypodium sylvaticum, confirming its role in both dicots and monocots species. Lastly, OsRLCK311 interacted with the rice OsPIP2;1. We suggest that the rice OsRLCK311 play a role in regulating the plant growth response under saline conditions via the regulation of the stomata response to stress. This role seems to be independent of the RLCK311 kinase activity, since the overexpression of the RLCK311 C-terminus (ΔN), which lacks the kinase full domain, has a similar phenotype to RLCK311FL.
Highlights
Crop productivity is severely limited by soil salinity
For co-immunoprecipitation and mass spectrometry analysis, three-week-old seedlings of wild-type and transgenic plants overexpressing OsRLCK311-∆N /OsRLCK311-FL were ground in liquid N2, and incubated on an end-over-end rotator at 4 ◦ C for 4 h with lysis buffer provided in the μMACS Isolation Kit (Miltenyl Biotec, Bergisch Gladbach, Germany), containing protease inhibitor cocktail
For co-immunoprecipitation by transient expression and western blots, OsRLCK31-∆NGFP/OsRLCK311FL-green fluorescent protein (GFP) was co-infiltrated with AtPIP2;1-red fluorescent protein (RFP), OsPIP2;1-RFP or free RFP as described above
Summary
Nir Sade 1,2, *,† , Fei Weng 1,3,† , Hiromi Tajima 1 , Yarden Zeron 2 , Lei Zhang 1 , Maria del Mar Rubio Wilhelmi 1 , George Day 1 , Zvi Peleg 4 and Eduardo Blumwald 1, *.
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