TaCIPK19-3D Improves Photosynthetic Machinery, Growth, Yield, and Salt Tolerance in Transgenic Rice

Abstract Calcineurin B-like interacting protein kinases (CIPKs) are central regulators of plant development and stress adaptation. However, the specific roles of individual CIPK family members remain largely unexplored in major crops like wheat and rice. In this study, we characterized the function...

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Main Authors: Muhammad Arif, Dingli Hong, Ruhong Xu, Mo Xian Chen, Hafiz Mamoon Rehman, Zhongni Wang, Luhua Li
Format: Article
Language:English
Published: SpringerOpen 2025-07-01
Series:Rice
Subjects:
Online Access:https://doi.org/10.1186/s12284-025-00827-y
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author Muhammad Arif
Dingli Hong
Ruhong Xu
Mo Xian Chen
Hafiz Mamoon Rehman
Zhongni Wang
Luhua Li
author_facet Muhammad Arif
Dingli Hong
Ruhong Xu
Mo Xian Chen
Hafiz Mamoon Rehman
Zhongni Wang
Luhua Li
author_sort Muhammad Arif
collection DOAJ
description Abstract Calcineurin B-like interacting protein kinases (CIPKs) are central regulators of plant development and stress adaptation. However, the specific roles of individual CIPK family members remain largely unexplored in major crops like wheat and rice. In this study, we characterized the function of TaCIPK19-3D through overexpression in transgenic rice and CRISPR-Cas9-mediated oscipk19 knockout lines. Expression profiling and subcellular localization analyses revealed that TaCIPK19-3D is associated with chloroplast development and metabolic activity. Overexpression lines exhibited enhanced chloroplast structure, increased chlorophyll biosynthesis, stomatal conductance, net photosynthetic rate, transpiration, and elevated levels of K⁺/Na⁺, Ca²⁺, and Mg²⁺, resulting in improved growth and yield compared to wild-type and mutant lines. Notably, TaCIPK19-3D overexpression conferred increased salt tolerance by upregulating ABA signaling, antioxidant responses, and proline biosynthesis. Key genes involved in chlorophyll synthesis (OsCAO, OsCHLH) and salt stress responses (OsAPX2, OsP5CS, OsABA2) were significantly upregulated in transgenic plants. Protein interaction studies using yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays demonstrated that TaCIPK19-3D interacts with TaFBA-4D and four CBL proteins (TaCBL1, TaCBL3, TaCBL4, and TaCBL7). Collectively, our findings reveal that TaCIPK19-3D positively regulates photosynthesis, ion homeostasis, and stress-responsive signaling pathways, highlighting its potential for improving crop productivity and stress resilience in wheat and rice.
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institution Kabale University
issn 1939-8425
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publishDate 2025-07-01
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spelling doaj-art-65c55b6938e8488889babb51a061f0422025-08-20T03:46:27ZengSpringerOpenRice1939-84251939-84332025-07-0118111510.1186/s12284-025-00827-yTaCIPK19-3D Improves Photosynthetic Machinery, Growth, Yield, and Salt Tolerance in Transgenic RiceMuhammad Arif0Dingli Hong1Ruhong Xu2Mo Xian Chen3Hafiz Mamoon Rehman4Zhongni Wang5Luhua Li6College of Agriculture, Guizhou UniversityCollege of Agriculture, Guizhou UniversityCollege of Agriculture, Guizhou UniversityState Key Laboratory of Green Pesticide, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Center for R&D of Fine, Ministry of Education, Chemicals of Guizhou UniversityCentre of Agricultural Biochemistry and Biotechnology, University of Agriculture FaisalabadGuizhou Rice Research Institute, Guizhou Provincial Academy of Agricultural ScienceCollege of Agriculture, Guizhou UniversityAbstract Calcineurin B-like interacting protein kinases (CIPKs) are central regulators of plant development and stress adaptation. However, the specific roles of individual CIPK family members remain largely unexplored in major crops like wheat and rice. In this study, we characterized the function of TaCIPK19-3D through overexpression in transgenic rice and CRISPR-Cas9-mediated oscipk19 knockout lines. Expression profiling and subcellular localization analyses revealed that TaCIPK19-3D is associated with chloroplast development and metabolic activity. Overexpression lines exhibited enhanced chloroplast structure, increased chlorophyll biosynthesis, stomatal conductance, net photosynthetic rate, transpiration, and elevated levels of K⁺/Na⁺, Ca²⁺, and Mg²⁺, resulting in improved growth and yield compared to wild-type and mutant lines. Notably, TaCIPK19-3D overexpression conferred increased salt tolerance by upregulating ABA signaling, antioxidant responses, and proline biosynthesis. Key genes involved in chlorophyll synthesis (OsCAO, OsCHLH) and salt stress responses (OsAPX2, OsP5CS, OsABA2) were significantly upregulated in transgenic plants. Protein interaction studies using yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays demonstrated that TaCIPK19-3D interacts with TaFBA-4D and four CBL proteins (TaCBL1, TaCBL3, TaCBL4, and TaCBL7). Collectively, our findings reveal that TaCIPK19-3D positively regulates photosynthesis, ion homeostasis, and stress-responsive signaling pathways, highlighting its potential for improving crop productivity and stress resilience in wheat and rice.https://doi.org/10.1186/s12284-025-00827-yTaCIPK19-3DRiceWheatSalt stressGene functionOsCIPK19
spellingShingle Muhammad Arif
Dingli Hong
Ruhong Xu
Mo Xian Chen
Hafiz Mamoon Rehman
Zhongni Wang
Luhua Li
TaCIPK19-3D Improves Photosynthetic Machinery, Growth, Yield, and Salt Tolerance in Transgenic Rice
Rice
TaCIPK19-3D
Rice
Wheat
Salt stress
Gene function
OsCIPK19
title TaCIPK19-3D Improves Photosynthetic Machinery, Growth, Yield, and Salt Tolerance in Transgenic Rice
title_full TaCIPK19-3D Improves Photosynthetic Machinery, Growth, Yield, and Salt Tolerance in Transgenic Rice
title_fullStr TaCIPK19-3D Improves Photosynthetic Machinery, Growth, Yield, and Salt Tolerance in Transgenic Rice
title_full_unstemmed TaCIPK19-3D Improves Photosynthetic Machinery, Growth, Yield, and Salt Tolerance in Transgenic Rice
title_short TaCIPK19-3D Improves Photosynthetic Machinery, Growth, Yield, and Salt Tolerance in Transgenic Rice
title_sort tacipk19 3d improves photosynthetic machinery growth yield and salt tolerance in transgenic rice
topic TaCIPK19-3D
Rice
Wheat
Salt stress
Gene function
OsCIPK19
url https://doi.org/10.1186/s12284-025-00827-y
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