Kasavajhala V. S. K. Prasad, Amira A. E. Abdel-Hameed, Anireddy S. N. Reddy
{"title":"CAMTA3 转录因子中的天然单核苷酸多态性可调节其功能及其靶基因的转录","authors":"Kasavajhala V. S. K. Prasad, Amira A. E. Abdel-Hameed, Anireddy S. N. Reddy","doi":"10.1111/mpp.13428","DOIUrl":null,"url":null,"abstract":"CAMTA3, a Ca<sup>2+</sup>/calmodulin-binding transcription factor, is a key regulator of plant immunity in <i>Arabidopsis</i>. Here, we identified a novel naturally occurring single-nucleotide polymorphism that results in a missense nonconservative mutation (<i>CAMTA3</i><sub><i>H386D</i></sub>) in many <i>Arabidopsis</i> ecotypes. This region of CAMTA3 is not part of any previously characterized regulatory domains. To study the consequence of this change on the function of CAMTA3, we introduced the <i>CAMTA3</i><sub><i>H386D</i></sub> into <i>camta3</i>, a loss-of-function mutant that exhibits a constitutive cell death phenotype, chlorotic lesions on leaves, and reduced plant size. Phenotypic and molecular analysis of these lines indicated that the expression of <i>CAMTA3</i><sub><i>H386D</i></sub> in the <i>camta3</i> mutant did not complement the mutant phenotypes. Also, the ecotypes containing the <i>CAMTA3</i><sub><i>H386D</i></sub> exhibited <i>camta3</i> phenotypes. Marker genes associated with salicylic acid biosynthesis and pathogen response were upregulated in the <i>CAMTA3</i><sub><i>H386D</i></sub> lines and the <i>Arabidopsis</i> accessions 7127 (Est-1) and 9941 (Fei-0), as in <i>camta3</i>, indicating that H386D mutation alters CAMTA3 activity in regulating the expression of known target genes. In <i>Nicotiana benthamiana</i> transient expression assays, <i>CAMTA3</i><sub><i>H386D</i></sub> failed to induce the expression of a luciferase reporter gene driven by the rapid stress-responsive elements (RSRE) that contain the known binding sites of CAMTA3, suggesting that CAMTA3<sub>H386D</sub> mutation impairs its ability to activate its target genes. Transgenic lines and tested natural accessions expressing CAMTA3<sub>H386D</sub> showed enhanced levels of H<sub>2</sub>O<sub>2</sub> and increased resistance to the bacterial pathogen <i>Pseudomonas syringae</i> pv. <i>tomato</i> DC3000. Collectively, our results indicate that the H386D mutation in a previously unknown regulatory region of CAMTA3 is essential for its function.","PeriodicalId":18763,"journal":{"name":"Molecular plant pathology","volume":"14 1","pages":""},"PeriodicalIF":4.8000,"publicationDate":"2024-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"A natural single-nucleotide polymorphism in the CAMTA3 transcription factor regulates its function and transcription of its target genes\",\"authors\":\"Kasavajhala V. S. K. Prasad, Amira A. E. Abdel-Hameed, Anireddy S. N. Reddy\",\"doi\":\"10.1111/mpp.13428\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"CAMTA3, a Ca<sup>2+</sup>/calmodulin-binding transcription factor, is a key regulator of plant immunity in <i>Arabidopsis</i>. Here, we identified a novel naturally occurring single-nucleotide polymorphism that results in a missense nonconservative mutation (<i>CAMTA3</i><sub><i>H386D</i></sub>) in many <i>Arabidopsis</i> ecotypes. This region of CAMTA3 is not part of any previously characterized regulatory domains. To study the consequence of this change on the function of CAMTA3, we introduced the <i>CAMTA3</i><sub><i>H386D</i></sub> into <i>camta3</i>, a loss-of-function mutant that exhibits a constitutive cell death phenotype, chlorotic lesions on leaves, and reduced plant size. Phenotypic and molecular analysis of these lines indicated that the expression of <i>CAMTA3</i><sub><i>H386D</i></sub> in the <i>camta3</i> mutant did not complement the mutant phenotypes. Also, the ecotypes containing the <i>CAMTA3</i><sub><i>H386D</i></sub> exhibited <i>camta3</i> phenotypes. Marker genes associated with salicylic acid biosynthesis and pathogen response were upregulated in the <i>CAMTA3</i><sub><i>H386D</i></sub> lines and the <i>Arabidopsis</i> accessions 7127 (Est-1) and 9941 (Fei-0), as in <i>camta3</i>, indicating that H386D mutation alters CAMTA3 activity in regulating the expression of known target genes. In <i>Nicotiana benthamiana</i> transient expression assays, <i>CAMTA3</i><sub><i>H386D</i></sub> failed to induce the expression of a luciferase reporter gene driven by the rapid stress-responsive elements (RSRE) that contain the known binding sites of CAMTA3, suggesting that CAMTA3<sub>H386D</sub> mutation impairs its ability to activate its target genes. Transgenic lines and tested natural accessions expressing CAMTA3<sub>H386D</sub> showed enhanced levels of H<sub>2</sub>O<sub>2</sub> and increased resistance to the bacterial pathogen <i>Pseudomonas syringae</i> pv. <i>tomato</i> DC3000. Collectively, our results indicate that the H386D mutation in a previously unknown regulatory region of CAMTA3 is essential for its function.\",\"PeriodicalId\":18763,\"journal\":{\"name\":\"Molecular plant pathology\",\"volume\":\"14 1\",\"pages\":\"\"},\"PeriodicalIF\":4.8000,\"publicationDate\":\"2024-02-07\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Molecular plant pathology\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://doi.org/10.1111/mpp.13428\",\"RegionNum\":1,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"PLANT SCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular plant pathology","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1111/mpp.13428","RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"PLANT SCIENCES","Score":null,"Total":0}
A natural single-nucleotide polymorphism in the CAMTA3 transcription factor regulates its function and transcription of its target genes
CAMTA3, a Ca2+/calmodulin-binding transcription factor, is a key regulator of plant immunity in Arabidopsis. Here, we identified a novel naturally occurring single-nucleotide polymorphism that results in a missense nonconservative mutation (CAMTA3H386D) in many Arabidopsis ecotypes. This region of CAMTA3 is not part of any previously characterized regulatory domains. To study the consequence of this change on the function of CAMTA3, we introduced the CAMTA3H386D into camta3, a loss-of-function mutant that exhibits a constitutive cell death phenotype, chlorotic lesions on leaves, and reduced plant size. Phenotypic and molecular analysis of these lines indicated that the expression of CAMTA3H386D in the camta3 mutant did not complement the mutant phenotypes. Also, the ecotypes containing the CAMTA3H386D exhibited camta3 phenotypes. Marker genes associated with salicylic acid biosynthesis and pathogen response were upregulated in the CAMTA3H386D lines and the Arabidopsis accessions 7127 (Est-1) and 9941 (Fei-0), as in camta3, indicating that H386D mutation alters CAMTA3 activity in regulating the expression of known target genes. In Nicotiana benthamiana transient expression assays, CAMTA3H386D failed to induce the expression of a luciferase reporter gene driven by the rapid stress-responsive elements (RSRE) that contain the known binding sites of CAMTA3, suggesting that CAMTA3H386D mutation impairs its ability to activate its target genes. Transgenic lines and tested natural accessions expressing CAMTA3H386D showed enhanced levels of H2O2 and increased resistance to the bacterial pathogen Pseudomonas syringae pv. tomato DC3000. Collectively, our results indicate that the H386D mutation in a previously unknown regulatory region of CAMTA3 is essential for its function.
期刊介绍:
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