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: 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
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Agronomy
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Online Access:https://www.mdpi.com/2073-4395/15/4/884
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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.
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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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