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Heterologous Expression and Functional Analysis of Rice GLUTAMATE RECEPTOR-LIKE Family Indicates its Role in Glutamate Triggered Calcium Flux in Rice Roots
BACKGROUND: Tremendous progress has been made in understanding the functions of the GLUTAMATE RECEPTOR-LIKE (GLR) family in Arabidopsis. Still, the functions of OsGLRs in rice, especially the ion channel activities, are largely unknown. RESULTS: Using the aequorin-based luminescence imaging system,...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer US
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4783324/ https://www.ncbi.nlm.nih.gov/pubmed/26956369 http://dx.doi.org/10.1186/s12284-016-0081-x |
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author | Ni, Jun Yu, Zhiming Du, Guankui Zhang, Yanyan Taylor, Jemma L. Shen, Chenjia Xu, Jing Liu, Xunyan Wang, Yifeng Wu, Yunrong |
author_facet | Ni, Jun Yu, Zhiming Du, Guankui Zhang, Yanyan Taylor, Jemma L. Shen, Chenjia Xu, Jing Liu, Xunyan Wang, Yifeng Wu, Yunrong |
author_sort | Ni, Jun |
collection | PubMed |
description | BACKGROUND: Tremendous progress has been made in understanding the functions of the GLUTAMATE RECEPTOR-LIKE (GLR) family in Arabidopsis. Still, the functions of OsGLRs in rice, especially the ion channel activities, are largely unknown. RESULTS: Using the aequorin-based luminescence imaging system, we screened the specificity of amino acids involved in the induction of Ca(2+) flux in rice roots. Of all the amino acids tested, glutamate (Glu) was the only one to trigger Ca(2+) flux significantly in rice roots. Detailed analysis showed a dose response of Ca(2+) increase to different concentrations of Glu. In addition, the Ca(2+) spike response to Glu was rapid, within 20 s after the application. A desensitization assay and pharmacological tests showed that the Glu-triggered Ca(2+) flux is mediated by OsGLRs. Whole genome analysis identified 13 OsGLR genes in rice, and these genes have various expression patterns in different tissues. Subcellular localization studies showed that all the OsGLRs examined are likely localized to the plasma membrane. Bacteria growth assays showed that at least OsGLR2.1 and OsGLR3.2 have the potential to mediate ion uptake in bacteria. Further analysis using Fura-2-based Ca(2+) imaging revealed a Glu-triggered Ca(2+) increase in OsGLR2.1-expressing human embryonic kidney (HEK) cells. CONCLUSIONS: Our work provides a molecular basis for investigating mechanisms of Glu-triggered Ca(2+) flux in rice. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12284-016-0081-x) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4783324 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Springer US |
record_format | MEDLINE/PubMed |
spelling | pubmed-47833242016-04-09 Heterologous Expression and Functional Analysis of Rice GLUTAMATE RECEPTOR-LIKE Family Indicates its Role in Glutamate Triggered Calcium Flux in Rice Roots Ni, Jun Yu, Zhiming Du, Guankui Zhang, Yanyan Taylor, Jemma L. Shen, Chenjia Xu, Jing Liu, Xunyan Wang, Yifeng Wu, Yunrong Rice (N Y) Original Article BACKGROUND: Tremendous progress has been made in understanding the functions of the GLUTAMATE RECEPTOR-LIKE (GLR) family in Arabidopsis. Still, the functions of OsGLRs in rice, especially the ion channel activities, are largely unknown. RESULTS: Using the aequorin-based luminescence imaging system, we screened the specificity of amino acids involved in the induction of Ca(2+) flux in rice roots. Of all the amino acids tested, glutamate (Glu) was the only one to trigger Ca(2+) flux significantly in rice roots. Detailed analysis showed a dose response of Ca(2+) increase to different concentrations of Glu. In addition, the Ca(2+) spike response to Glu was rapid, within 20 s after the application. A desensitization assay and pharmacological tests showed that the Glu-triggered Ca(2+) flux is mediated by OsGLRs. Whole genome analysis identified 13 OsGLR genes in rice, and these genes have various expression patterns in different tissues. Subcellular localization studies showed that all the OsGLRs examined are likely localized to the plasma membrane. Bacteria growth assays showed that at least OsGLR2.1 and OsGLR3.2 have the potential to mediate ion uptake in bacteria. Further analysis using Fura-2-based Ca(2+) imaging revealed a Glu-triggered Ca(2+) increase in OsGLR2.1-expressing human embryonic kidney (HEK) cells. CONCLUSIONS: Our work provides a molecular basis for investigating mechanisms of Glu-triggered Ca(2+) flux in rice. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12284-016-0081-x) contains supplementary material, which is available to authorized users. Springer US 2016-03-08 /pmc/articles/PMC4783324/ /pubmed/26956369 http://dx.doi.org/10.1186/s12284-016-0081-x Text en © Ni et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Original Article Ni, Jun Yu, Zhiming Du, Guankui Zhang, Yanyan Taylor, Jemma L. Shen, Chenjia Xu, Jing Liu, Xunyan Wang, Yifeng Wu, Yunrong Heterologous Expression and Functional Analysis of Rice GLUTAMATE RECEPTOR-LIKE Family Indicates its Role in Glutamate Triggered Calcium Flux in Rice Roots |
title | Heterologous Expression and Functional Analysis of Rice GLUTAMATE RECEPTOR-LIKE Family Indicates its Role in Glutamate Triggered Calcium Flux in Rice Roots |
title_full | Heterologous Expression and Functional Analysis of Rice GLUTAMATE RECEPTOR-LIKE Family Indicates its Role in Glutamate Triggered Calcium Flux in Rice Roots |
title_fullStr | Heterologous Expression and Functional Analysis of Rice GLUTAMATE RECEPTOR-LIKE Family Indicates its Role in Glutamate Triggered Calcium Flux in Rice Roots |
title_full_unstemmed | Heterologous Expression and Functional Analysis of Rice GLUTAMATE RECEPTOR-LIKE Family Indicates its Role in Glutamate Triggered Calcium Flux in Rice Roots |
title_short | Heterologous Expression and Functional Analysis of Rice GLUTAMATE RECEPTOR-LIKE Family Indicates its Role in Glutamate Triggered Calcium Flux in Rice Roots |
title_sort | heterologous expression and functional analysis of rice glutamate receptor-like family indicates its role in glutamate triggered calcium flux in rice roots |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4783324/ https://www.ncbi.nlm.nih.gov/pubmed/26956369 http://dx.doi.org/10.1186/s12284-016-0081-x |
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