Aquaporins are implicated in a wide variety of plant physiological processes, although the mechanisms involved in their regulation are not fully understood. To gain further insight into the regulatory factors involved in this process, we used a yeast two-hybrid system to screen for potential binding partners to the Arabidopsis tonoplast intrinsic protein (TIP) AtTIP1;1. This was the first protein identified to be associated with high water permeability in vacuolar membranes from Arabidopsis thaliana. Using AtTIP1;1 as bait, a novel binding protein was identified in both yeast and plant cells. This prey protein, named AtSM34, was a 309 aa polypeptide with a predicted molecular mass of 34 kD and contained a single MYB/SANT-like domain. AtSM34 promoter:: GUS histochemical staining analysis detected AtSM34 expression in flowers, stems and leaves, particularly in the vascular tissues, in response to osmotic stress. AtSM34 expression was localized in the endoplasmic reticulum membrane, and sequence deletion analysis revealed that the N-terminal coding region (amino acids 1-83) was critical for this localization. Overexpression of AtSM34 resulted in hypersensitivity to exogenous mannitol, sorbitol and abscisic acid, and caused a significant delay in germination. AtSM34 interacted with AtTIP1;2 and AtTIP2;1, which are essential proteins for modulation of tonoplast permeability and highly expressed in germinating seedlings. These data indicate AtSM34 is a novel TIPs binding protein involved in the osmotic stress response of seedlings at an early stage of development.
LI LiJuanREN FeiWEI PengChengCHEN QiJunCHEN JiaWANG XueChen
Boron (B) toxicity to plants is responsible for low crop productivity in many regions of the world. Here we report a novel and effective means to alleviate the B toxicity to plants under high B circumstance. Functional characterization of AtTIP5;1, an aquaporin gene, re-vealed that overexpression of AtTIP5;1 (OxAtTIP5;1) in Arabidopsis significantly increased its tolerance to high B toxicity. Compared to wild-type plants, OxAtTIP5;1 plants exhibited longer hypocotyls, accelerated development, increased silique production under high B treatments. GUS staining and quantitative RT-PCR (qRT-PCR) results demonstrated that the expression of AtTIP5;1 was induced by high B concentration treatment. Subcellular localization analysis revealed that the AtTIP5;1-GFP fusion protein was localized on the tonoplast membrane, which was consistent with the prediction based on bioinformatics. Taken together, our results suggest that AtTIP5;1 is in-volved in B transport pathway possibly via vacuolar compartmentation for B, and that overexpression of AtTIP5;1 in plants may provide an effective way to overcome the problem resulting from high B concentration toxicity.
Yongqi Pang, Lijuan Li, Fei Ren, Pingli Lu, Pengcheng Wei, Jinghui Cai, Lingguo Xin, Juan Zhang, Jia Chen, Xuechen Wang State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100193, China
