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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| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-03-01
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| Series: | Agronomy |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4395/15/4/884 |
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| Summary: | 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. |
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| ISSN: | 2073-4395 |