Functional Characterization of Pomegranate CAMTA3 in Cold Stress Responses
Cold stress is a significant factor limiting plant growth and development. Pomegranate is particularly susceptible to low temperatures. Calmodulin-binding transcriptional activators (CAMTAs) are key regulators of cold stress tolerance in plants. In this study, we conducted a comprehensive analysis o...
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MDPI AG
2025-03-01
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| Series: | Plants |
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| author | Shuangshuang Zhao Rui Lu Lijuan Feng Mengyu Zheng Han Zhang Yanlei Yin Ling Zheng |
| author_facet | Shuangshuang Zhao Rui Lu Lijuan Feng Mengyu Zheng Han Zhang Yanlei Yin Ling Zheng |
| author_sort | Shuangshuang Zhao |
| collection | DOAJ |
| description | Cold stress is a significant factor limiting plant growth and development. Pomegranate is particularly susceptible to low temperatures. Calmodulin-binding transcriptional activators (CAMTAs) are key regulators of cold stress tolerance in plants. In this study, we conducted a comprehensive analysis of the CAMTA family proteins across 12 species, including <i>Punica granatum</i> (pomegranate), using bioinformatic methods. Pomegranate <i>CAMTA3</i> (<i>PgCAMTA3</i>) was isolated and characterized, and it demonstrated enhanced cold tolerance when expressed in <i>Arabidopsis thaliana</i>. Quantitative real-time PCR (qRT-PCR) analysis showed that the expression of <i>PgCAMTA3</i> was up-regulated under cold and ABA treatments in pomegranates. Two <i>A. thaliana</i> transgenic lines, OE1 and OE2, which overexpress PgCAMTA3, were generated through genetic transformation. The overexpression of <i>PgCAMTA3</i> enhanced the cold stress tolerance in transgenic <i>A. thaliana</i>. OE1 and OE2 exhibited higher survival rates under cold stress. Furthermore, enzymatic activity assays revealed enhanced peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD) in OE lines. These antioxidant enzymatic activities collectively contribute to better cold stress tolerance by providing more effective reactive oxygen species (ROS) scavenging and cellular protection mechanisms, which was confirmed by lower levels of malondialdehyde (MDA) and ROS production. In addition, the overexpression of <i>PgCAMTA3</i> led to the upregulation of the expression levels of <i>AtCBF2</i>, <i>AtNCED3</i>, and <i>AtWRKY22</i>, which were modulated by <i>CAMTA3</i>. In summary, we report the significant role of <i>PgCAMTA3</i> in plant cold tolerance. Our findings provide valuable insights into the CAMATA family in plants and offer new perspectives on the molecular mechanisms underlying cold tolerance in pomegranates. |
| format | Article |
| id | doaj-art-eaca0cfeec3e4cb5bc5101eb6a58559b |
| institution | DOAJ |
| issn | 2223-7747 |
| language | English |
| publishDate | 2025-03-01 |
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| series | Plants |
| spelling | doaj-art-eaca0cfeec3e4cb5bc5101eb6a58559b2025-08-20T02:53:19ZengMDPI AGPlants2223-77472025-03-0114581310.3390/plants14050813Functional Characterization of Pomegranate CAMTA3 in Cold Stress ResponsesShuangshuang Zhao0Rui Lu1Lijuan Feng2Mengyu Zheng3Han Zhang4Yanlei Yin5Ling Zheng6Shandong Provincial Key Laboratory of Plant Stress, Life Science College, Shandong Normal University, Jinan 250014, ChinaShandong Provincial Key Laboratory of Plant Stress, Life Science College, Shandong Normal University, Jinan 250014, ChinaShandong Institute of Pomology, Taian 271000, ChinaShandong Provincial Key Laboratory of Plant Stress, Life Science College, Shandong Normal University, Jinan 250014, ChinaShandong Provincial Key Laboratory of Plant Stress, Life Science College, Shandong Normal University, Jinan 250014, ChinaShandong Institute of Pomology, Taian 271000, ChinaShandong Provincial Key Laboratory of Plant Stress, Life Science College, Shandong Normal University, Jinan 250014, ChinaCold stress is a significant factor limiting plant growth and development. Pomegranate is particularly susceptible to low temperatures. Calmodulin-binding transcriptional activators (CAMTAs) are key regulators of cold stress tolerance in plants. In this study, we conducted a comprehensive analysis of the CAMTA family proteins across 12 species, including <i>Punica granatum</i> (pomegranate), using bioinformatic methods. Pomegranate <i>CAMTA3</i> (<i>PgCAMTA3</i>) was isolated and characterized, and it demonstrated enhanced cold tolerance when expressed in <i>Arabidopsis thaliana</i>. Quantitative real-time PCR (qRT-PCR) analysis showed that the expression of <i>PgCAMTA3</i> was up-regulated under cold and ABA treatments in pomegranates. Two <i>A. thaliana</i> transgenic lines, OE1 and OE2, which overexpress PgCAMTA3, were generated through genetic transformation. The overexpression of <i>PgCAMTA3</i> enhanced the cold stress tolerance in transgenic <i>A. thaliana</i>. OE1 and OE2 exhibited higher survival rates under cold stress. Furthermore, enzymatic activity assays revealed enhanced peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD) in OE lines. These antioxidant enzymatic activities collectively contribute to better cold stress tolerance by providing more effective reactive oxygen species (ROS) scavenging and cellular protection mechanisms, which was confirmed by lower levels of malondialdehyde (MDA) and ROS production. In addition, the overexpression of <i>PgCAMTA3</i> led to the upregulation of the expression levels of <i>AtCBF2</i>, <i>AtNCED3</i>, and <i>AtWRKY22</i>, which were modulated by <i>CAMTA3</i>. In summary, we report the significant role of <i>PgCAMTA3</i> in plant cold tolerance. Our findings provide valuable insights into the CAMATA family in plants and offer new perspectives on the molecular mechanisms underlying cold tolerance in pomegranates.https://www.mdpi.com/2223-7747/14/5/813pomegranatecold stress<i>CAMTA</i> genes |
| spellingShingle | Shuangshuang Zhao Rui Lu Lijuan Feng Mengyu Zheng Han Zhang Yanlei Yin Ling Zheng Functional Characterization of Pomegranate CAMTA3 in Cold Stress Responses Plants pomegranate cold stress <i>CAMTA</i> genes |
| title | Functional Characterization of Pomegranate CAMTA3 in Cold Stress Responses |
| title_full | Functional Characterization of Pomegranate CAMTA3 in Cold Stress Responses |
| title_fullStr | Functional Characterization of Pomegranate CAMTA3 in Cold Stress Responses |
| title_full_unstemmed | Functional Characterization of Pomegranate CAMTA3 in Cold Stress Responses |
| title_short | Functional Characterization of Pomegranate CAMTA3 in Cold Stress Responses |
| title_sort | functional characterization of pomegranate camta3 in cold stress responses |
| topic | pomegranate cold stress <i>CAMTA</i> genes |
| url | https://www.mdpi.com/2223-7747/14/5/813 |
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