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Functional characterization and reconstitution of ABA signaling components using transient gene expression in rice protoplasts

The core components of ABA-dependent gene expression signaling have been identified in Arabidopsis and rice. This signaling pathway consists of four major components; group A OsbZIPs, SAPKs, subclass A OsPP2Cs and OsPYL/RCARs in rice. These might be able to make thousands of combinations through int...

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Autores principales: Kim, Namhyo, Moon, Seok-Jun, Min, Myung K., Choi, Eun-Hye, Kim, Jin-Ae, Koh, Eun Y., Yoon, Insun, Byun, Myung-Ok, Yoo, Sang-Dong, Kim, Beom-Gi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4524894/
https://www.ncbi.nlm.nih.gov/pubmed/26300907
http://dx.doi.org/10.3389/fpls.2015.00614
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author Kim, Namhyo
Moon, Seok-Jun
Min, Myung K.
Choi, Eun-Hye
Kim, Jin-Ae
Koh, Eun Y.
Yoon, Insun
Byun, Myung-Ok
Yoo, Sang-Dong
Kim, Beom-Gi
author_facet Kim, Namhyo
Moon, Seok-Jun
Min, Myung K.
Choi, Eun-Hye
Kim, Jin-Ae
Koh, Eun Y.
Yoon, Insun
Byun, Myung-Ok
Yoo, Sang-Dong
Kim, Beom-Gi
author_sort Kim, Namhyo
collection PubMed
description The core components of ABA-dependent gene expression signaling have been identified in Arabidopsis and rice. This signaling pathway consists of four major components; group A OsbZIPs, SAPKs, subclass A OsPP2Cs and OsPYL/RCARs in rice. These might be able to make thousands of combinations through interaction networks resulting in diverse signaling responses. We tried to characterize those gene functions using transient gene expression for rice protoplasts (TGERP) because it is instantaneous and convenient system. Firstly, in order to monitor the ABA signaling output, we developed reporter system named pRab16A-fLUC which consists of Rab16A promoter of rice and luciferase gene. It responses more rapidly and sensitively to ABA than pABRC3-fLUC that consists of ABRC3 of HVA1 promoter in TGERP. We screened the reporter responses for over-expression of each signaling components from group A OsbZIPs to OsPYL/RCARs with or without ABA in TGERP. OsbZIP46 induced reporter most strongly among OsbZIPs tested in the presence of ABA. SAPKs could activate the OsbZIP46 even in the ABA independence. Subclass A OsPP2C6 and -8 almost completely inhibited the OsbZIP46 activity in the different degree through the SAPK9. Lastly, OsPYL/RCAR2 and -5 rescued the OsbZIP46 activity in the presence of SAPK9 and OsPP2C6 dependent on ABA concentration and expression level. By using TGERP, we could characterize successfully the effects of ABA dependent gene expression signaling components in rice. In conclusion, TGERP represents very useful technology to study systemic functional genomics in rice or other monocots.
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spelling pubmed-45248942015-08-21 Functional characterization and reconstitution of ABA signaling components using transient gene expression in rice protoplasts Kim, Namhyo Moon, Seok-Jun Min, Myung K. Choi, Eun-Hye Kim, Jin-Ae Koh, Eun Y. Yoon, Insun Byun, Myung-Ok Yoo, Sang-Dong Kim, Beom-Gi Front Plant Sci Plant Science The core components of ABA-dependent gene expression signaling have been identified in Arabidopsis and rice. This signaling pathway consists of four major components; group A OsbZIPs, SAPKs, subclass A OsPP2Cs and OsPYL/RCARs in rice. These might be able to make thousands of combinations through interaction networks resulting in diverse signaling responses. We tried to characterize those gene functions using transient gene expression for rice protoplasts (TGERP) because it is instantaneous and convenient system. Firstly, in order to monitor the ABA signaling output, we developed reporter system named pRab16A-fLUC which consists of Rab16A promoter of rice and luciferase gene. It responses more rapidly and sensitively to ABA than pABRC3-fLUC that consists of ABRC3 of HVA1 promoter in TGERP. We screened the reporter responses for over-expression of each signaling components from group A OsbZIPs to OsPYL/RCARs with or without ABA in TGERP. OsbZIP46 induced reporter most strongly among OsbZIPs tested in the presence of ABA. SAPKs could activate the OsbZIP46 even in the ABA independence. Subclass A OsPP2C6 and -8 almost completely inhibited the OsbZIP46 activity in the different degree through the SAPK9. Lastly, OsPYL/RCAR2 and -5 rescued the OsbZIP46 activity in the presence of SAPK9 and OsPP2C6 dependent on ABA concentration and expression level. By using TGERP, we could characterize successfully the effects of ABA dependent gene expression signaling components in rice. In conclusion, TGERP represents very useful technology to study systemic functional genomics in rice or other monocots. Frontiers Media S.A. 2015-08-05 /pmc/articles/PMC4524894/ /pubmed/26300907 http://dx.doi.org/10.3389/fpls.2015.00614 Text en Copyright © 2015 Kim, Moon, Min, Choi, Kim, Koh, Yoon, Byun, Yoo and Kim. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Kim, Namhyo
Moon, Seok-Jun
Min, Myung K.
Choi, Eun-Hye
Kim, Jin-Ae
Koh, Eun Y.
Yoon, Insun
Byun, Myung-Ok
Yoo, Sang-Dong
Kim, Beom-Gi
Functional characterization and reconstitution of ABA signaling components using transient gene expression in rice protoplasts
title Functional characterization and reconstitution of ABA signaling components using transient gene expression in rice protoplasts
title_full Functional characterization and reconstitution of ABA signaling components using transient gene expression in rice protoplasts
title_fullStr Functional characterization and reconstitution of ABA signaling components using transient gene expression in rice protoplasts
title_full_unstemmed Functional characterization and reconstitution of ABA signaling components using transient gene expression in rice protoplasts
title_short Functional characterization and reconstitution of ABA signaling components using transient gene expression in rice protoplasts
title_sort functional characterization and reconstitution of aba signaling components using transient gene expression in rice protoplasts
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4524894/
https://www.ncbi.nlm.nih.gov/pubmed/26300907
http://dx.doi.org/10.3389/fpls.2015.00614
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