Tissue-specific expression, developmentally and spatially regulated alternative splicing, and protein subcellular localization of OsLpa1 in rice

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• 作者：Hai-ping Lu ; Wei-qin Pang ; Wen-xu Li…
• 关键词：OsLpa1 ; Low phytic acid ; Expression pattern ; Alternative splicing ; Subcellular localization ; OsLpa1 ; 低植酸
表达模式 ; 可变剪接 ; 亚细胞定位 Q946.2
• 刊名：Journal of Zhejiang University SCIENCE B
• 出版年：2016
• 出版时间：February 2016
• 年：2016
• 卷：17
• 期：2
• 页码：100-109
• 全文大小：1,371 KB
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• 作者单位：Hai-ping Lu (1)
  Wei-qin Pang (1)
  Wen-xu Li (1)
  Yuan-yuan Tan (1)
  Qing Wang (2)
  Hai-jun Zhao (1)
  Qing-yao Shu (1)
  
  1. State Key Laboratory of Rice Biology, Institution of Crop Science, Zhejiang University, Hangzhou, 310029, China
  2. Wuxi Hupper Bioseed Technology Academy Ltd., Wuxi, 214000, China
  
• 刊物主题：Biomedicine general;
• 出版者：Springer Berlin Heidelberg
• ISSN：1862-1783

文摘

The OsLpa1 gene (LOC_Os02g57400) 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 OsLpa1 in 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 reversetranscriptase polymerase chain reaction (RT-PCR) analysis revealed that the abundance of OsLpa1.3 was negligible compared with OsLpa1.1 and OsLpa1.2 in 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 OsLpa1.2 in 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. Keywords OsLpa1 Low phytic acid Expression pattern Alternative splicing Subcellular localization