title: Abscisic acid prevents the coalescence of protein storage vacuoles by upregulating expression of a tonoplast intrinsic protein gene in barley aleurone.
abstract: Tonoplast intrinsic proteins (TIPs) are integral membrane proteins that are known to function in plants as aquaporins. Here, we propose another role for TIPs during the fusion of protein storage vacuoles (PSVs) in aleurone cells, a process that is promoted by gibberellic acid (GA) and prevented by abscisic acid (ABA). Studies of the expression of barley (Hordeum vulgare) TIP genes (HvTIP) showed that GA specifically decreased the abundance of HvTIP1;2 and HvTIP3;1 transcripts, while ABA strongly increased expression of HvTIP3;1. Increased or decreased expression of HvTIP3;1 interfered with the hormonal effects on vacuolation in aleurone protoplasts. HvTIP3;1 gain-of-function experiments delayed GA-induced vacuolation, whereas HvTIP3;1 loss-of-function experiments promoted vacuolation in ABA-treated aleurone cells. These results indicate that TIP plays a key role in preventing the coalescence of small PSVs in aleurone cells. Hormonal regulation of the HvTIP3;1 promoter is similar to the regulation of the endogenous gene, indicating that induction of the transcription of HvTIP3;1 by ABA is a critical factor in the prevention of PSV coalescence in response to ABA. Promoter analysis using deletions and site-directed mutagenesis of sequences identified three cis-acting elements that are responsible for ABA responsiveness in the HvTIP3;1 promoter. Promoter analysis also showed that ABA responsiveness of the HvTIP3;1 promoter is likely to occur via a unique regulatory system distinct from that involving the ABA-response promoter complexes.
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul 143-701, Korea.
authors.forename: Sung-eun
authors.initials: SE
authors.lastname: Lee
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul 143-701, Korea.
authors.forename: Hui-kyung
authors.initials: HK
authors.lastname: Yim
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul 143-701, Korea.
authors.forename: Mi-na
authors.initials: MN
authors.lastname: Lim
authors.affiliation: Molecular Breeding Division, Natural Academy of Agricultural Science, Rural Development Adminstration, Suwon 441-857, Korea.
authors.forename: In sun
authors.initials: Is
authors.lastname: Yoon
authors.affiliation: Department of Biology, Kyungbook National University, Daegu 702-701, Korea.
authors.forename: Jeong hoe
authors.initials: Jh
authors.lastname: Kim
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul 143-701, Korea yshwang@konkuk.ac.kr.
authors.forename: Yong-sic
authors.initials: YS
authors.lastname: Hwang
mesh_terms: D000040:Abscisic Acid / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D001467:Hordeum / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D010937:Plant Growth Regulators / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011523:Protoplasts / Q000378:metabolism; D015854:Up-Regulation; D014617:Vacuoles / Q000235:genetics / Q000378:metabolism*
chemical_list: D008565:Membrane Proteins; D010937:Plant Growth Regulators; D010940:Plant Proteins; C077491:tonoplast intrinsic protein, plant; D000040:Abscisic Acid
keywords: Abscisic acid; aleurone; cis-acting element; coalescence; protein storage vacuole; tonoplast intrinsic protein.