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Key message <p>We identified the characteristics of CBL and CIPK families of <i>Magnolia biondii</i> and the <i>MbCBL4</i>-overexpressed <i>Arabidopsis</i> plants conferred salt tolerance.</p> Abstract <p>Calcineurin B-like protein (CBLs) and CBL-interacting protein kinase (CIPKs) are important components of the Ca<sup>2</sup>⁺-mediated signal pathway. These proteins play a key role in plant growth, development, and response to environmental stress. <i>Magnolia biondii</i> is a woody plant valued for ornamental and medicinal uses and is frequently exposed to abiotic stresses during its growth cycle. Nevertheless, there are still gaps in the study of CBL and CIPK gene families in <i>M. biondii</i>. In this study, 6 CBL and 20 CIPK genes were identified from the <i>M. biondii</i> genome. Phylogenetic analysis divided these genes into 4 CBL and 7 CIPK subgroups, and cross-species comparisons across 34 plants indicated that monocotyledons generally harbor more CBLs/CIPKs than Magnoliaceae. Quantitative real-time PCR (qRT-PCR) analysis revealed that <i>MbCBLs</i> and <i>MbCIPKs</i> showed different transcription levels under drought, cold, and salt stress. Protein–protein interaction assays (Y2H and LCI) verified physical interaction of MbCBL1/MbCIPK18 and MbCBL4/MbCIPK18. Functionally, <i>MbCBL4</i> overexpression in <i>Arabidopsis</i> conferred enhanced salt tolerance: primary root length and chlorophyll content increased by 2.74-fold and 2.71-fold relative to wild type; fresh weight increased by up to 60%, SOD and CAT activities rose by 47% and 28%, while H₂O₂ and O₂⁻ levels declined by 46% and 38%. These results indicate that <i>MbCBL4</i> enhances salt tolerance by promoting growth, antioxidant capacity, and reactive oxygen species scavenging. These findings provide important insights into the functional roles of MbCBL and MbCIPK genes and the regulation of <i>MbCBL4</i> under salt stress.</p>

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Comprehensive analysis of CBL and CIPK gene families in Magnolia biondii and functional characterization of MbCBL4 under salt stress

  • 期刊论文

  • 发表日期：

    2026年1月26日

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  • Yongchao Ke,
  • Liyong Sun,
  • Yao Chen,
  • Lina Xu,
  • Liming Yang,
  • Zengfang Yin

摘要

Key message

We identified the characteristics of CBL and CIPK families of Magnolia biondii and the MbCBL4-overexpressed Arabidopsis plants conferred salt tolerance.

Abstract

Calcineurin B-like protein (CBLs) and CBL-interacting protein kinase (CIPKs) are important components of the Ca2⁺-mediated signal pathway. These proteins play a key role in plant growth, development, and response to environmental stress. Magnolia biondii is a woody plant valued for ornamental and medicinal uses and is frequently exposed to abiotic stresses during its growth cycle. Nevertheless, there are still gaps in the study of CBL and CIPK gene families in M. biondii. In this study, 6 CBL and 20 CIPK genes were identified from the M. biondii genome. Phylogenetic analysis divided these genes into 4 CBL and 7 CIPK subgroups, and cross-species comparisons across 34 plants indicated that monocotyledons generally harbor more CBLs/CIPKs than Magnoliaceae. Quantitative real-time PCR (qRT-PCR) analysis revealed that MbCBLs and MbCIPKs showed different transcription levels under drought, cold, and salt stress. Protein–protein interaction assays (Y2H and LCI) verified physical interaction of MbCBL1/MbCIPK18 and MbCBL4/MbCIPK18. Functionally, MbCBL4 overexpression in Arabidopsis conferred enhanced salt tolerance: primary root length and chlorophyll content increased by 2.74-fold and 2.71-fold relative to wild type; fresh weight increased by up to 60%, SOD and CAT activities rose by 47% and 28%, while H₂O₂ and O₂⁻ levels declined by 46% and 38%. These results indicate that MbCBL4 enhances salt tolerance by promoting growth, antioxidant capacity, and reactive oxygen species scavenging. These findings provide important insights into the functional roles of MbCBL and MbCIPK genes and the regulation of MbCBL4 under salt stress.