Identification, Characterization, Expression Profiling and Functional Analysis of Tobacco <i>CalS</i> Gene Family
Callose plays an important role in plant development and in response to a wide range of biotic and abiotic stresses. However, the systematic identification of callose synthase (CalS), the major enzyme for callose biosynthesis, has been delayed in crops, especially in <i>Solanaceae</i>. I...
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2025-03-01
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| author | Hong Wang He Meng Xiaohan Qi Yi Pan Bailu Ji Liuying Wen Yanjun Zan Huan Si Yuanying Wang Dan Liu Aiguo Yang Zhengwen Liu Lirui Cheng |
| author_facet | Hong Wang He Meng Xiaohan Qi Yi Pan Bailu Ji Liuying Wen Yanjun Zan Huan Si Yuanying Wang Dan Liu Aiguo Yang Zhengwen Liu Lirui Cheng |
| author_sort | Hong Wang |
| collection | DOAJ |
| description | Callose plays an important role in plant development and in response to a wide range of biotic and abiotic stresses. However, the systematic identification of callose synthase (CalS), the major enzyme for callose biosynthesis, has been delayed in crops, especially in <i>Solanaceae</i>. In the current research, 18 <i>CalS</i> genes (<i>NtCalS1</i>–<i>NtCalS18</i>) were identified in <i>Nicotiana tabacum</i> and classified into four subfamilies. A comprehensive analysis of their physicochemical properties, gene structure, and evolutionary history highlighted their evolutionary conservation. We also identified a number of <i>NtCalSs</i> that responded to the infection with <i>Phytophthora nicotianae</i> and <i>Ralstonia solanacearum</i>, as well as to drought and cold treatments, by analyzing RNA-seq data. <i>NtCalS1</i> and <i>NtCalS12</i>, a highly homologous gene pair, were selected to create mutants using the CRISPR-Cas9 technology for their drastic response to <i>Phytophthora nicotianae</i> infection as well as the strong expression levels in roots. The mutants with the simultaneous knockout of <i>NtCalS1</i> and <i>NtCalS12</i>, compared with the control plants, displayed more resistance to tobacco black shank caused by <i>Phytophthora nicotianae</i>. Furthermore, the real-time quantitative PCR (qRT-PCR) assay showed that the knockout of <i>NtCalS1</i> and <i>NtCalS12</i> activated the signaling pathways mediated by plant hormones salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) before and after the infection of <i>Phytophthora nicotianae</i> and thus may have contributed to tobacco immunity against black shank. These findings contribute valuable information for further understanding the roles of <i>CalS</i> genes in tobacco stress responses and provide alternative genes for resistance improvement. |
| format | Article |
| id | doaj-art-d60eed4a2b7748ca959b0bd0038fb63b |
| institution | OA Journals |
| issn | 2073-4395 |
| language | English |
| publishDate | 2025-03-01 |
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| spelling | doaj-art-d60eed4a2b7748ca959b0bd0038fb63b2025-08-20T02:24:42ZengMDPI AGAgronomy2073-43952025-03-0115488410.3390/agronomy15040884Identification, Characterization, Expression Profiling and Functional Analysis of Tobacco <i>CalS</i> Gene FamilyHong Wang0He Meng1Xiaohan Qi2Yi Pan3Bailu Ji4Liuying Wen5Yanjun Zan6Huan Si7Yuanying Wang8Dan Liu9Aiguo Yang10Zhengwen Liu11Lirui Cheng12Key Laboratory for Tobacco Gene Resources, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, ChinaKey Laboratory for Tobacco Gene Resources, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, ChinaSchool of Agriculture, Yunnan University, Kunming 650091, ChinaKey Laboratory for Tobacco Gene Resources, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, ChinaKey Laboratory for Tobacco Gene Resources, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, ChinaKey Laboratory for Tobacco Gene Resources, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, ChinaKey Laboratory for Tobacco Gene Resources, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, ChinaKey Laboratory for Tobacco Gene Resources, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, ChinaKey Laboratory for Tobacco Gene Resources, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, ChinaKey Laboratory for Tobacco Gene Resources, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, ChinaKey Laboratory for Tobacco Gene Resources, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, ChinaKey Laboratory for Tobacco Gene Resources, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, ChinaKey Laboratory for Tobacco Gene Resources, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao 266101, ChinaCallose plays an important role in plant development and in response to a wide range of biotic and abiotic stresses. However, the systematic identification of callose synthase (CalS), the major enzyme for callose biosynthesis, has been delayed in crops, especially in <i>Solanaceae</i>. In the current research, 18 <i>CalS</i> genes (<i>NtCalS1</i>–<i>NtCalS18</i>) were identified in <i>Nicotiana tabacum</i> and classified into four subfamilies. A comprehensive analysis of their physicochemical properties, gene structure, and evolutionary history highlighted their evolutionary conservation. We also identified a number of <i>NtCalSs</i> that responded to the infection with <i>Phytophthora nicotianae</i> and <i>Ralstonia solanacearum</i>, as well as to drought and cold treatments, by analyzing RNA-seq data. <i>NtCalS1</i> and <i>NtCalS12</i>, a highly homologous gene pair, were selected to create mutants using the CRISPR-Cas9 technology for their drastic response to <i>Phytophthora nicotianae</i> infection as well as the strong expression levels in roots. The mutants with the simultaneous knockout of <i>NtCalS1</i> and <i>NtCalS12</i>, compared with the control plants, displayed more resistance to tobacco black shank caused by <i>Phytophthora nicotianae</i>. Furthermore, the real-time quantitative PCR (qRT-PCR) assay showed that the knockout of <i>NtCalS1</i> and <i>NtCalS12</i> activated the signaling pathways mediated by plant hormones salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) before and after the infection of <i>Phytophthora nicotianae</i> and thus may have contributed to tobacco immunity against black shank. These findings contribute valuable information for further understanding the roles of <i>CalS</i> genes in tobacco stress responses and provide alternative genes for resistance improvement.https://www.mdpi.com/2073-4395/15/4/884callose synthase<i>CalS</i> gene family<i>Nicotiana tabacum</i>expression analysisgene editingstress response |
| spellingShingle | Hong Wang He Meng Xiaohan Qi Yi Pan Bailu Ji Liuying Wen Yanjun Zan Huan Si Yuanying Wang Dan Liu Aiguo Yang Zhengwen Liu Lirui Cheng Identification, Characterization, Expression Profiling and Functional Analysis of Tobacco <i>CalS</i> Gene Family Agronomy callose synthase <i>CalS</i> gene family <i>Nicotiana tabacum</i> expression analysis gene editing stress response |
| title | Identification, Characterization, Expression Profiling and Functional Analysis of Tobacco <i>CalS</i> Gene Family |
| title_full | Identification, Characterization, Expression Profiling and Functional Analysis of Tobacco <i>CalS</i> Gene Family |
| title_fullStr | Identification, Characterization, Expression Profiling and Functional Analysis of Tobacco <i>CalS</i> Gene Family |
| title_full_unstemmed | Identification, Characterization, Expression Profiling and Functional Analysis of Tobacco <i>CalS</i> Gene Family |
| title_short | Identification, Characterization, Expression Profiling and Functional Analysis of Tobacco <i>CalS</i> Gene Family |
| title_sort | identification characterization expression profiling and functional analysis of tobacco i cals i gene family |
| topic | callose synthase <i>CalS</i> gene family <i>Nicotiana tabacum</i> expression analysis gene editing stress response |
| url | https://www.mdpi.com/2073-4395/15/4/884 |
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