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The PIP and TIP aquaporins in wheat form a large and diverse family with unique gene structures and functionally important features

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Abstract

Aquaporins, members of major intrinsic proteins (MIPs), transport water across cellular membranes and play vital roles in all organisms. Adversities such as drought, salinity, or chilling affect water uptake and transport, and numerous plant MIPs are reported to be differentially regulated under such stresses. However, MIP genes have been not yet been characterized in wheat, the largest cereal crop. We have identified 24 PIP and 11 TIP aquaporin genes from wheat by gene isolation and database searches. They vary extensively in lengths, numbers, and sequences of exons and introns, and sequences and cellular locations of predicted proteins, but the intron positions (if present) are characteristic. The putative PIP proteins show a high degree of conservation of signature sequences or residues for membrane integration, water transport, and regulation. The TIPs are more diverse, some with potential for water transport and others with various selectivity filters including a new combination. Most genes appear to be expressed as expressed sequence tags, while two are likely pseudogenes. Many of the genes are highly identical to rice but some are unique, and many correspond to genes that show differential expression under salinity and/or drought. The results provide extensive information for functional studies and developing markers for stress tolerance.

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Acknowledgements

KF is supported by a Grains Research and Development Corporation (GRDC, Australia) Ph.D. scholarship. We gratefully acknowledge Dr. Greg Grimes (Australian Winter Cereals Collection, Tamworth, Australia) for providing the seed samples, Dr. Paula Moolhuijzen (State Agricultural Biotechnology Centre, Murdoch University, Western Australia) for advice on EST searches, and Prof. Rudi Appels (State Agricultural Biotechnology Centre, Western Australia) for advice on the project. We thank Damian Cockfield for confirming some of the DNA sequences. We are also thankful to the anonymous referees for their critical comments and suggestions, which helped in the revision of the manuscript greatly.

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  1. Environment and Biotechnology Centre, Faculty of Life and Social Sciences, Swinburne University of Technology, P.O. Box 218, John St, Hawthorn, Victoria, 3122, Australia

    Kerrie L. Forrest & Mrinal Bhave

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  1. Kerrie L. Forrest
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Correspondence to Mrinal Bhave.

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Supplementary Table S1

Major differences between PIP1 and PIP2 isoforms of rice (DOC 21.5 kb)

Supplementary Table S2

Summary of PIP and TIP genes obtained through amplification of gDNA (DOC 30.0 kb)

Supplementary Table S3

Sequence identities of PIPs and TIPs of wheat with those of other plants (DOC 50 kb)

Supplementary Table S4

Highest sequence identity scores from ClustalW alignments {score for Cranbrook sequence}: entries indicate poor identity score for TC or EST, therefore, score from cDNA for the gene isolated from cv. Cranbrook was considered also (DOC 31.0 kb)

Supplemental Figure S1

Primer positions on wheat PIP and TIP cDNA sequences from Genbank. Black shading indicates positions of NPA motifs. Gray shading indicates positions of TMHs (DOC 26.0 kb)

Supplemental Figure S2

(DOC 21.0 kb)

Supplemental Figure S3

Sequence alignment of predicted PIP cDNAs (exon contigs) of genes cloned from gDNA of wheat (DOC 64.5 kb)

Supplemental Figure S4

Sequence alignment of predicted TIP cDNAs (exon contigs) of genes cloned from gDNA of wheat (DOC 74.5 kb)

Supplemental Figure S5

Alignment of putative amino acids sequences of wheat PIPs with aquaporins, a glycerol facilitator, and an aquaglyceroporin (DOC 119 kb)

Supplemental Figure S6

Alignment of putative amino acids sequences of wheat TIPs with other aquaporins, a glycerol facilitator, and an aquaglyceroporin (DOC 81.0 kb)

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Forrest, K.L., Bhave, M. The PIP and TIP aquaporins in wheat form a large and diverse family with unique gene structures and functionally important features. Funct Integr Genomics 8, 115–133 (2008). https://doi.org/10.1007/s10142-007-0065-4

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