The ArathEULS3 Lectin Ends up in Stress Granules and Can Follow an Unconventional Route for Secretion.

Stress granules are cytoplasmic compartments, which serve as mRNA storage units during stress, therefore regulating translation. The Arabidopsis thaliana lectin ArathEULS3 has been widely described as a stress inducible gene. This study aimed to examine in detail the localization of ArathEULS3 lectin in normal and stressed cells. Colocalization experiments revealed that the nucleo-cytoplasmic lectin ArathEULS3 relocates to stress granules after stress. The ArathEULS3 sequence encodes a protein with a EUL lectin domain and an N-terminal domain with unknown structure and function. Bioinformatics analyses showed that the N-terminal domain sequence contains intrinsically disordered regions and likely does not exhibit a stable protein fold. Plasmolysis experiments indicated that ArathEULS3 also localizes to the apoplast, suggesting that this protein might follow an unconventional route for secretion. As part of our efforts we also investigated the interactome of ArathEULS3 and identified several putative interaction partners important for the protein translation process.

Arabidopsis Lectin EULS3 Is Involved in ABA Signaling in Roots.

The Arabidopsis thaliana lectin ArathEULS3 is upregulated in particular stress conditions and upon abscisic acid (ABA) treatment. ABA is a plant hormone important for plant growth and stress responses. During stress ABA is perceived by PYR/PYL/RCAR receptors, inhibiting protein phosphatases PP2Cs thereby enabling SNRK2s kinases to start downstream phosphorylation cascades and signaling. PYL9, one of the ABA receptors was identified as an interacting partner for ArathEULS3. Promoter::GUS activity studies revealed the expression of ArathEULS3 in the central root cylinder and the cells flanking young lateral root primordia, and showed enhanced expression in root tips after ABA treatment. Transcript levels for ArathEULS3 increased after exposure to ABA and osmotic treatments. ArathEULS3 CRISPR KO mutants served as a tool to expand the knowledge on the role of ArathEULS3 in plant development. KO lines revealed a longer root system compared to WT plants, and showed reduced sensitivity to ABA, salt, and osmotic conditions. Additionally it was noted that the KO mutants had more emerged lateral roots when grown in high osmotic conditions. Together these data suggest that ArathEULS3 may be an important player in ABA responses in roots.

Involvement of OsRIP1, a ribosome-inactivating protein from rice, in plant defense against Nilaparvata lugens.

Rice is the most important staple food in the world, but rice production is challenged by several biotic stress factors like viruses, bacteria, fungi and pest insects. One of the most notorious pest insects is Nilaparvata lugens, commonly known as the brown planthopper, which feeds on rice phloem sap and can cause serious damage to rice fields. In order to protect themselves, plants express a wide array of defense proteins such as ribosome-inactivating proteins (RIPs). This study shows that the expression of 'OsRIP1' is highly induced in rice plants infested with N. lugens, with transcript levels more than 100-fold upregulated in infested plants compared to non-infested plants. Furthermore, recombinant OsRIP1 was toxic for brown planthoppers when administered through liquid artificial diet. OsRIP1 inactivated insect ribosomes in vitro, suggesting that its toxicity relates to the enzymatic activity of OsRIP1. Over-expression of OsRIP1 in transgenic rice plants did not affect the performance of insects reared on these plants, most likely due to insufficient concentrations of OsRIP1 in the phloem. The data obtained in this research indicate that OsRIP1 can play a role in plant defense against herbivorous insects.

OsEUL Lectin Gene Expression in Rice: Stress Regulation, Subcellular Localization and Tissue Specificity.

The Euonymus lectin (EUL) family is a unique group of carbohydrate-binding proteins that is omnipresent in plants. Sequences encoding EUL-related lectins have been retrieved from all completely sequenced plant genomes. The rice (Oryza sativa) genome contains 5 functional EUL genes referred to as OsEULS2, OsEULS3, OsEULD1a, OsEULD1b, and OsEULD2. In this study we focused on the tissue specific expression, stress inducibility and subcellular localization of the rice EULs. Even though the EUL domain sequence is highly conserved among the rice EULs (at least 80% sequence similarity) different biotic and abiotic stress treatments yielded unique responses for the different EULs. Transcript levels for OsEULs were differentially affected by drought and salt stress, ABA treatment, pathogen infection or insect infestation. Analysis of promoter activity revealed differential expression and tissue specificity for the 5 OsEUL genes, with most expression observed in the vascular system of roots and shoots, as well as in the root tips and seeds. At cell level, all OsEULs are located in the nucleus whereas OsEULD1b and OsEULD2 also locate to the cytoplasm. This paper contributes to the functional characterization of the EULs and provides insight in the biological importance of this family of proteins for rice.