title: Tissue-specific expression, developmentally and spatially regulated alternative splicing, and protein subcellular localization of OsLpa rice.
abstract: The OsLpa1 gene (LOC_Os57400) was identified to be involved in phytic acid (PA) metabolism because its knockout and missense mutants reduce PA content in rice grain. However, little is known about the molecular characteristics of OsLpa rice and of its homologues in other plants. In the present study, the spatial pattern of OsLpa1 expression was revealed using OsLpa1 promoter::GUS transgenic plants (GUS: β-glucuronidase); GUS histochemical assay showed that OsLpa1 was strongly expressed in stem, leaf, and root tissues, but in floral organ it is expressed mainly and strongly in filaments. In seeds, GUS staining was concentrated in the aleurone layers; a few blue spots were observed in the outer layers of embryo, but no staining was observed in the endosperm. Three OsLpa1 transcripts (OsLpa1.1, OsLpa1.2, OsLpa1.3) are produced due to alternative splicing; quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis revealed that the abundance of OsLpa1.3 was negligible compared with OsLpa1.1 and OsLpa all tissues. OsLpa1.2 is predominant in germinating seeds (about 5 times that of OsLpa1.1), but its abundance decreases quickly with the development of seedlings and plants, whereas the abundance of OsLpa1.1 rises and falls, reaching its highest level in 45-d-old plants, with abundance greater than that of OsLpa both leaves and roots. In seeds, the abundance of OsLpa1 continuously increases with seed growth, being 27.5 and 15 times greater in 28-DAF (day after flowering) seeds than in 7-DAF seeds for OsLpa1.1 and OsLpa1.2, respectively. Transient expression of chimeric genes with green fluorescence protein (GFP) in rice protoplasts demonstrated that all proteins encoded by the three OsLpa1 transcripts are localized to the chloroplast.
authors.affiliation: State Key Laboratory of Rice Biology, Institution of Crop Science, Zhejiang University, Hangzhou 310029, China.
authors.forename: Hai-ping
authors.initials: HP
authors.lastname: Lu
authors.affiliation: State Key Laboratory of Rice Biology, Institution of Crop Science, Zhejiang University, Hangzhou 310029, China.
authors.forename: Wei-qin
authors.initials: WQ
authors.lastname: Pang
authors.affiliation: State Key Laboratory of Rice Biology, Institution of Crop Science, Zhejiang University, Hangzhou 310029, China.
authors.forename: Wen-xu
authors.initials: WX
authors.lastname: Li
authors.affiliation: State Key Laboratory of Rice Biology, Institution of Crop Science, Zhejiang University, Hangzhou 310029, China.
authors.forename: Yuan-yuan
authors.initials: YY
authors.lastname: Tan
authors.affiliation: Wuxi Hupper Bioseed Technology Academy Ltd., Wuxi 214000, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Rice Biology, Institution of Crop Science, Zhejiang University, Hangzhou 310029, China.
authors.forename: Hai-jun
authors.initials: HJ
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Rice Biology, Institution of Crop Science, Zhejiang University, Hangzhou 310029, China.
authors.forename: Qing-yao
authors.initials: QY
authors.lastname: Shu
mesh_terms: D017398:Alternative Splicing / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D018506:Gene Expression Regulation, Plant / Q000502:physiology; D012275:Oryza / Q000502:physiology* / Q000648:ultrastructure; D010833:Phytic Acid / Q000378:metabolism*; D010940:Plant Proteins / Q000502:physiology*; D030821:Plants, Genetically Modified / Q000502:physiology; D062211:Spatio-Temporal Analysis; D013347:Subcellular Fractions / Q000502:physiology* / Q000648:ultrastructure; D014018:Tissue Distribution
chemical_list: D010940:Plant Proteins; D010833:Phytic Acid
keywords: Alternative splicing; Expression pattern; Low phytic acid; OsLpa1; Subcellular localization
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