Abstract

Aquaporins (AQPs) are major intrinsic proteins (MIPs) that mediate bidirectional flux of water and other substrates across cell membranes, and play critical roles in plant-water relations, dehydration stress responses and crop productivity. However, limited data are available as yet on the contributions of these proteins to the physiology of the major crop barley (Hordeum vulgare). The present work reports the identification and expression analysis of the barley MIP family. A comprehensive search of publicly available leaf mRNA-seq data, draft barley genome data, GenBank transcripts and sixteen new annotations together revealed that the barley MIP family is comprised of at least forty AQPs. Alternative splicing events were likely in two plasma membrane intrinsic protein (PIP) AQPs. Analyses of the AQP signature sequences and specificity determining positions indicated a potential of several putative AQP isoforms to transport non-aqua substrates including physiological important substrates, and respond to abiotic stresses. Analysis of our publicly available leaf mRNA-seq data identified notable differential expression of HvPIP1;2 and HvTIP4;1 under salt stress. Analyses of other gene expression resources also confirmed isoform-specific responses in different tissues and/or in response to salinity, as well as some potentially inter-cultivar differences. The work reports systematic and comprehensive analysis of most, if not all, barley AQP genes, their sequences, expression patterns in different tissues, potential transport and stress response functions, and a strong framework for selection and/or development of stress tolerant barley varieties. In addition, the barley data would be highly valuable for genetic studies of the evolutionarily closely related wheat (Triticum aestivum L.).

Highlights

  • Aquaporins (AQPs) belong to the superfamily of membrane channels called the major intrinsic proteins (MIPs)

  • Four main subfamilies (PIPs, tonoplast intrinsic proteins (TIPs), nodulin-26-like intrinsic proteins (NIPs), SIPs) have been identified in both primitive and higher plants, while hybrid intrinsic proteins (HIPs) and GIPs have only been identified in mosses [3] and X intrinsic proteins (XIPs) identified in mosses and several dicotyledonous plants, e.g., tobacco, potato and tomato [5] and soybean [6], but not monocots

  • A genomic sequence identified in Bowman and Morex genomes was not present in the CDS database, and a BLASTn against NCBI revealed that both contigs corresponded to HvTIP5;1 cDNA (AB540227)

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Summary

Introduction

Aquaporins (AQPs) belong to the superfamily of membrane channels called the major intrinsic proteins (MIPs). The plant MIPs (often generically called ‘aquaporins’) are typically divided into seven subfamilies: the plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs), nodulin-26-like intrinsic proteins (NIPs), small, basic intrinsic proteins (SIPs) [1]. Barley Aquaporins Form a Large and Diverse Family and the novel, less common subfamilies of GlpF-like intrinsic protein (GIP1;1) [2], hybrid intrinsic proteins (HIPs) and the uncategorized X intrinsic proteins (XIPs) [3]. Four main subfamilies (PIPs, TIPs, NIPs, SIPs) have been identified in both primitive and higher plants, while HIPs and GIPs have only been identified in mosses [3] and XIPs identified in mosses and several dicotyledonous plants, e.g., tobacco, potato and tomato [5] and soybean [6], but not monocots. All MIPs exhibit a number of characteristic features (reviewed in [1]), including (i) six transmembrane helices (TM1-TM6) and five inter-helical loops (LA-LE); (ii) two short helices (HB, HE) that contain the highly conserved Asn-Pro-Ala (NPA) motifs that form the pore that allows a single-file passage of water molecules; (iii) the ar/R selectivity filter, comprised of four residues (one from TM2, one from TM5, two from LE), and shown to significantly influence the nature of transport substrates; (iv) five positions (P1-P5) likely important for discriminating between AQPs and glycerol transporters and (v) a conserved motif Ala-Glu-Phe (AEF or AEFXXT) in TM1, of unclear function

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