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2021
Amorim, Maurício Silva; Toyosumi, Iumi Silva; Lopes, Wilson Amorim; Camillo, Luciana Rodrigues; Ferreira, Laiane Nascimento; Junior, Dilson Souza Rocha; Filho, Walter Santos Soares; Gesteira, Abelmon Silva; Costa, Marcio Gilberto Cardoso; Filho, Maurício Antônio Coelho
Em: https://doi.org/10.1080/14620316.2021.1900750, vol. 96, iss. 5, pp. 663-672, 2021, ISSN: 14620316.
Resumo | Links | BibTeX | Tags: Citrus, Fruit quality, Gene expression, irrigation, water deficit, Yield
@article{Amorim2021,
title = {Partial rootzone drying and regulated deficit irrigation can be used as water-saving strategies without compromising fruit yield and quality in tropically grown sweet orange},
author = {Maurício Silva Amorim and Iumi Silva Toyosumi and Wilson Amorim Lopes and Luciana Rodrigues Camillo and Laiane Nascimento Ferreira and Dilson Souza Rocha Junior and Walter Santos Soares Filho and Abelmon Silva Gesteira and Marcio Gilberto Cardoso Costa and Maurício Antônio Coelho Filho},
url = {https://www.tandfonline.com/doi/abs/10.1080/14620316.2021.1900750},
doi = {10.1080/14620316.2021.1900750},
issn = {14620316},
year = {2021},
date = {2021-01-01},
journal = {https://doi.org/10.1080/14620316.2021.1900750},
volume = {96},
issue = {5},
pages = {663-672},
publisher = {Taylor & Francis},
abstract = {We examined the effects of partial rootzone drying (PRD) and regulated deficit irrigation (RDI) on some physiological and agronomical traits in tropically grown sweet orange (Citrus sinensis (L.) O...},
keywords = {Citrus, Fruit quality, Gene expression, irrigation, water deficit, Yield},
pubstate = {published},
tppubtype = {article}
}
We examined the effects of partial rootzone drying (PRD) and regulated deficit irrigation (RDI) on some physiological and agronomical traits in tropically grown sweet orange (Citrus sinensis (L.) O...
2020
Corte, L. Erpen Dalla; Mendes, B. M. J.; Filho, F. A. A. Mourão; Grosser, J. W.; Dutt, M.
Functional characterization of full-length and 5′ deletion fragments of Citrus sinensis-derived constitutive promoters in Nicotiana benthamiana Journal Article
Em: In Vitro Cellular and Developmental Biology - Plant, vol. 56, iss. 3, pp. 280-289, 2020, ISSN: 14752689.
Resumo | Links | BibTeX | Tags: Cis-regulatory elements, Gene expression, genetic transformation, sweet orange
@article{Corte2020,
title = {Functional characterization of full-length and 5′ deletion fragments of Citrus sinensis-derived constitutive promoters in Nicotiana benthamiana},
author = {L. Erpen Dalla Corte and B. M. J. Mendes and F. A. A. Mourão Filho and J. W. Grosser and M. Dutt},
url = {https://link.springer.com/article/10.1007/s11627-019-10044-0},
doi = {10.1007/S11627-019-10044-0/FIGURES/7},
issn = {14752689},
year = {2020},
date = {2020-01-01},
journal = {In Vitro Cellular and Developmental Biology - Plant},
volume = {56},
issue = {3},
pages = {280-289},
publisher = {Springer},
abstract = {Full-length and 5′ deletion fragments of three constitutively expressed gene promoters identified from the Citrus sinensis L. genome (cyclophilin (CsCYP), glyceraldehyde-3-phosphate dehydrogenase C2 (CsGAPC2), and elongation factor 1-alpha (CsEF1)) were evaluated for their ability to express the uidA gene in leaf, stem, and root tissues of transgenic N. benthamiana plants. According to the fluorometric GUS assays, the CsGAPC2 promoter activity was significantly reduced in the leaves when the sequence between position − 1090 and − 497 was removed, while the activity remained unchanged in the roots. The CsCYP promoter activity was not affected by the different deletions, and even the smallest evaluated fragment was sufficient to maintain the expression of the uidA gene in N. benthamiana leaves and roots. Truncated promoter fragments of CsEF1 conferred higher GUS activity in leaves compared with the full-length promoter, while GUS activity was not affected in the roots. The removal of the intron in the 5′ UTR of the CsEF1 promoter reduced gene expression in the roots, although it did not cause any reduction in the gene expression level in the leaves. These results indicate that any of the three deletions of the CsCYP promoter can be utilized for efficient gene expression. In addition, the full-length CsGAPC2 promoter and two of the truncated CsEF1 promoter fragments were sufficient for efficient uidA gene expression.},
keywords = {Cis-regulatory elements, Gene expression, genetic transformation, sweet orange},
pubstate = {published},
tppubtype = {article}
}
Full-length and 5′ deletion fragments of three constitutively expressed gene promoters identified from the Citrus sinensis L. genome (cyclophilin (CsCYP), glyceraldehyde-3-phosphate dehydrogenase C2 (CsGAPC2), and elongation factor 1-alpha (CsEF1)) were evaluated for their ability to express the uidA gene in leaf, stem, and root tissues of transgenic N. benthamiana plants. According to the fluorometric GUS assays, the CsGAPC2 promoter activity was significantly reduced in the leaves when the sequence between position − 1090 and − 497 was removed, while the activity remained unchanged in the roots. The CsCYP promoter activity was not affected by the different deletions, and even the smallest evaluated fragment was sufficient to maintain the expression of the uidA gene in N. benthamiana leaves and roots. Truncated promoter fragments of CsEF1 conferred higher GUS activity in leaves compared with the full-length promoter, while GUS activity was not affected in the roots. The removal of the intron in the 5′ UTR of the CsEF1 promoter reduced gene expression in the roots, although it did not cause any reduction in the gene expression level in the leaves. These results indicate that any of the three deletions of the CsCYP promoter can be utilized for efficient gene expression. In addition, the full-length CsGAPC2 promoter and two of the truncated CsEF1 promoter fragments were sufficient for efficient uidA gene expression.
Curtolo, Maiara; Granato, Laís Moreira; Soratto, Tatiany Aparecida Teixeira; Curtolo, Maisa; Gazaffi, Rodrigo; Takita, Marco Aurélio; Cristofani-Yaly, Mariângela; Machado, Marcos Antonio
Em: Genetics and Molecular Biology, vol. 43, iss. 2, pp. 1-16, 2020, ISSN: 1415-4757.
Resumo | Links | BibTeX | Tags: Gene expression, Molecular markers, polymorphism
@article{Curtolo2020,
title = {Expression Quantitative Trait Loci (eQTL) mapping for callose synthases in intergeneric hybrids of \textit{Citrus} challenged with the bacteria \textit{Candidatus} Liberibacter asiaticus},
author = {Maiara Curtolo and Laís Moreira Granato and Tatiany Aparecida Teixeira Soratto and Maisa Curtolo and Rodrigo Gazaffi and Marco Aurélio Takita and Mariângela Cristofani-Yaly and Marcos Antonio Machado},
url = {http://www.scielo.br/j/gmb/a/N8vX5CTGhkQhD8KGxdWPpTH/?lang=en},
doi = {10.1590/1678-4685-GMB-2019-0133},
issn = {1415-4757},
year = {2020},
date = {2020-01-01},
journal = {Genetics and Molecular Biology},
volume = {43},
issue = {2},
pages = {1-16},
publisher = {Sociedade Brasileira de Genética},
abstract = {Abstract Citrus plants have been extremely affected by Huanglongbing (HLB) worldwide, causing economic losses. HLB disease causes disorders in citrus plants, leading to callose deposition in the phloem vessel sieve plates. Callose is synthesized by callose synthases, which are encoded by 12 genes (calS1– calS12)in Arabidopsis thaliana. We evaluated the expression of eight callose synthase genes from Citrus in hybrids between Citrus sunki and Poncirus trifoliata infected with HLB. The objective of this work was to identify possible tolerance loci combining the expression quantitative trait loci (eQTL) of different callose synthases and genetic Single-Nucleotide Polymorphism (SNP) maps of C. sunki and P. trifoliata. The expression data from all CscalS ranged widely among the hybrids. Furthermore, the data allowed the detection of 18 eQTL in the C. sunki map and 34 eQTL in the P. trifoliata map. In both maps, some eQTL for different CscalS were overlapped; thus, a single region could be associated with the regulation of more than one CscalS. The regions identified in this work can be interesting targets for future studies of Citrus breeding programs to manipulate callose synthesis during HLB infection.},
keywords = {Gene expression, Molecular markers, polymorphism},
pubstate = {published},
tppubtype = {article}
}
Abstract Citrus plants have been extremely affected by Huanglongbing (HLB) worldwide, causing economic losses. HLB disease causes disorders in citrus plants, leading to callose deposition in the phloem vessel sieve plates. Callose is synthesized by callose synthases, which are encoded by 12 genes (calS1– calS12)in Arabidopsis thaliana. We evaluated the expression of eight callose synthase genes from Citrus in hybrids between Citrus sunki and Poncirus trifoliata infected with HLB. The objective of this work was to identify possible tolerance loci combining the expression quantitative trait loci (eQTL) of different callose synthases and genetic Single-Nucleotide Polymorphism (SNP) maps of C. sunki and P. trifoliata. The expression data from all CscalS ranged widely among the hybrids. Furthermore, the data allowed the detection of 18 eQTL in the C. sunki map and 34 eQTL in the P. trifoliata map. In both maps, some eQTL for different CscalS were overlapped; thus, a single region could be associated with the regulation of more than one CscalS. The regions identified in this work can be interesting targets for future studies of Citrus breeding programs to manipulate callose synthesis during HLB infection.
Granato, Laís Moreira; Oliveira, Tiago S.; Boscariol-Camargo, Raquel L.; Galdeano, Diogo M.; Silva, Nicholas V.; Máximo, Heros J.; Dalio, Ronaldo J. D.; Machado, Marcos A.
Em: Tropical Plant Pathology, vol. 45, iss. 6, pp. 646-657, 2020, ISSN: 19832052.
Resumo | Links | BibTeX | Tags: ‘Candidatus Liberibacter asiaticus’, Citrus HLB, Gene expression, Secretion, Virulence factors
@article{Granato2020,
title = {‘Candidatus Liberibacter asiaticus’ putative effectors: in silico analysis and gene expression in citrus leaves displaying distinct huanglongbing symptoms},
author = {Laís Moreira Granato and Tiago S. Oliveira and Raquel L. Boscariol-Camargo and Diogo M. Galdeano and Nicholas V. Silva and Heros J. Máximo and Ronaldo J. D. Dalio and Marcos A. Machado},
url = {https://link.springer.com/article/10.1007/s40858-020-00382-5},
doi = {10.1007/S40858-020-00382-5/METRICS},
issn = {19832052},
year = {2020},
date = {2020-01-01},
journal = {Tropical Plant Pathology},
volume = {45},
issue = {6},
pages = {646-657},
publisher = {Springer Science and Business Media Deutschland GmbH},
abstract = {Huanglongbing (HLB) is considered the most devastating bacterial disease in citriculture. It is caused by the phloem-limited α-proteobacterium ‘Candidatus Liberibacter asiaticus’ (CLas). All citrus varieties are susceptible to HLB and many studies have been conducted in an attempt to identify potential targets involved in bacterial virulence and pathogenicity mechanisms. One of the most important virulence factors of bacterial pathogens is the effector molecule that can modify host cell structures, facilitate infection, and/or trigger host defense responses. In this study, 14 CLas candidates for secreted hypothetical proteins were predicted by a bioinformatics pipeline, one of which (NSEC01) was predicted to carry a nuclear localization sequence (NLS). Putative effectors HLBSP04 and HLBSP21 were conserved in all genomes evaluated, whereas HLBSP01, HLBSP19, and HLBSP30 were exclusively found in CLas. Expression analysis of candidate effector genes was performed in citrus leaves exhibiting five distinct HLB symptoms (asymptomatic, blotchy mottle, zinc deficiency, yellowed veins, and vein corky) and used as a guide for the selection of effectors for further functional studies. Genes coding for candidate effectors HLBSP13, HLBSP20, HLBSP28, and NSEC01 had high expression levels in blotchy mottle leaves, suggesting they could be used as marker genes that may facilitate detection of the pathogen in trees with this HLB symptom. As NSEC01 was exclusively found in this work, it was selected for transient expression in Nicotiana benthamiana leaves, but further investigations need to be carried out. Also, other candidate effectors predicted in this work still need to be functionally characterized. This study allowed discussing and proposing hypotheses about CLas classification according to virulence and lifestyle, which can help understand the citrus-CLas interaction.},
keywords = {‘Candidatus Liberibacter asiaticus’, Citrus HLB, Gene expression, Secretion, Virulence factors},
pubstate = {published},
tppubtype = {article}
}
Huanglongbing (HLB) is considered the most devastating bacterial disease in citriculture. It is caused by the phloem-limited α-proteobacterium ‘Candidatus Liberibacter asiaticus’ (CLas). All citrus varieties are susceptible to HLB and many studies have been conducted in an attempt to identify potential targets involved in bacterial virulence and pathogenicity mechanisms. One of the most important virulence factors of bacterial pathogens is the effector molecule that can modify host cell structures, facilitate infection, and/or trigger host defense responses. In this study, 14 CLas candidates for secreted hypothetical proteins were predicted by a bioinformatics pipeline, one of which (NSEC01) was predicted to carry a nuclear localization sequence (NLS). Putative effectors HLBSP04 and HLBSP21 were conserved in all genomes evaluated, whereas HLBSP01, HLBSP19, and HLBSP30 were exclusively found in CLas. Expression analysis of candidate effector genes was performed in citrus leaves exhibiting five distinct HLB symptoms (asymptomatic, blotchy mottle, zinc deficiency, yellowed veins, and vein corky) and used as a guide for the selection of effectors for further functional studies. Genes coding for candidate effectors HLBSP13, HLBSP20, HLBSP28, and NSEC01 had high expression levels in blotchy mottle leaves, suggesting they could be used as marker genes that may facilitate detection of the pathogen in trees with this HLB symptom. As NSEC01 was exclusively found in this work, it was selected for transient expression in Nicotiana benthamiana leaves, but further investigations need to be carried out. Also, other candidate effectors predicted in this work still need to be functionally characterized. This study allowed discussing and proposing hypotheses about CLas classification according to virulence and lifestyle, which can help understand the citrus-CLas interaction.
Soratto, Tatiany Aparecida Teixeira; Curtolo, Maiara; Marengo, Samanta; Dezotti, Ana Lúcia; Lima, Rômulo Pedro Macêdo; Gazaffi, Rodrigo; Machado, Marcos Antonio; Cristofani-Yaly, Mariângela
QTL and eQTL mapping associated with host response to Candidatus Liberibacter asiaticus in citrandarins Journal Article
Em: Tropical Plant Pathology, vol. 45, iss. 6, pp. 626-645, 2020, ISSN: 19832052.
Resumo | Links | BibTeX | Tags: citrandarins, Gene expression, huanglongbing, Phenotypic analysis
@article{Soratto2020,
title = {QTL and eQTL mapping associated with host response to Candidatus Liberibacter asiaticus in citrandarins},
author = {Tatiany Aparecida Teixeira Soratto and Maiara Curtolo and Samanta Marengo and Ana Lúcia Dezotti and Rômulo Pedro Macêdo Lima and Rodrigo Gazaffi and Marcos Antonio Machado and Mariângela Cristofani-Yaly},
url = {https://link.springer.com/article/10.1007/s40858-020-00372-7},
doi = {10.1007/S40858-020-00372-7/METRICS},
issn = {19832052},
year = {2020},
date = {2020-01-01},
journal = {Tropical Plant Pathology},
volume = {45},
issue = {6},
pages = {626-645},
publisher = {Springer Science and Business Media Deutschland GmbH},
abstract = {Huanglongbing (HLB) is a severe disease of citrus caused by the bacterium Candidatus Liberibacter. In America, the most common species is Candidatus Liberibacter asiaticus (CLas). In a previous study of the Citrus-HLB pathosystem, our group found differences in CLas multiplication in a population of hybrids obtained by hybridization between Citrus sunki Hort. Ex Tan. and Poncirus trifoliata (L.) Raf. cv. Rubidoux. The bacterial concentration was higher in C. sunki than in P. trifoliata. Thus, this study aims to map phenotypic (QTL) and gene expression (eQTL) data associated with host response to CLas in the linkage groups (LGs) of the previously constructed parental maps of C. sunki and P. trifoliata cv. Rubidoux. For the phenotypic analysis, we used a population of 79 F1 hybrids (termed citrandarins) between C. sunki and P. trifoliata. CLas and starch were quantified in the leaves of the plants two years after pathogen inoculation, allowing the classification of hybrids as resistant, tolerant, and susceptible. The expression of 14 candidate genes was measured in 72 hybrids of the population and used as expression data for the eQTL mapping. We located nine QTL and 52 eQTL in the C. sunki map and 17 QTL and 40 eQTL in the P. trifoliata map. The overlap of eQTL of the majority of genes with QTL from the phenotypic data indicates that the genes are related to the phenotype and are probably related to pathogen infection.},
keywords = {citrandarins, Gene expression, huanglongbing, Phenotypic analysis},
pubstate = {published},
tppubtype = {article}
}
Huanglongbing (HLB) is a severe disease of citrus caused by the bacterium Candidatus Liberibacter. In America, the most common species is Candidatus Liberibacter asiaticus (CLas). In a previous study of the Citrus-HLB pathosystem, our group found differences in CLas multiplication in a population of hybrids obtained by hybridization between Citrus sunki Hort. Ex Tan. and Poncirus trifoliata (L.) Raf. cv. Rubidoux. The bacterial concentration was higher in C. sunki than in P. trifoliata. Thus, this study aims to map phenotypic (QTL) and gene expression (eQTL) data associated with host response to CLas in the linkage groups (LGs) of the previously constructed parental maps of C. sunki and P. trifoliata cv. Rubidoux. For the phenotypic analysis, we used a population of 79 F1 hybrids (termed citrandarins) between C. sunki and P. trifoliata. CLas and starch were quantified in the leaves of the plants two years after pathogen inoculation, allowing the classification of hybrids as resistant, tolerant, and susceptible. The expression of 14 candidate genes was measured in 72 hybrids of the population and used as expression data for the eQTL mapping. We located nine QTL and 52 eQTL in the C. sunki map and 17 QTL and 40 eQTL in the P. trifoliata map. The overlap of eQTL of the majority of genes with QTL from the phenotypic data indicates that the genes are related to the phenotype and are probably related to pathogen infection.
2019
Freitas, Eliane Cristina De; Ucci, Amanda Piovesan; Teixeira, Elaine Cristina; Pedroso, Gisele Audrei; Hilario, Eduardo; Zocca, Vitória Fernanda Bertolazzi; Paiva, Gabriela Barbosa De; Ferreira, Henrique; Pedrolli, Danielle Biscaro; Bertolini, Maria Célia
The copper-inducible copAB operon in Xanthomonas citri subsp. citri is regulated at transcriptional and translational levels Journal Article
Em: Microbiology (United Kingdom), vol. 165, iss. 3, pp. 355-365, 2019, ISSN: 14652080.
Resumo | Links | BibTeX | Tags: Copper resistance, Gene expression, Short upstream ORF, Xanthomonas citri
@article{nokey,
title = {The copper-inducible copAB operon in Xanthomonas citri subsp. citri is regulated at transcriptional and translational levels},
author = {Eliane Cristina De Freitas and Amanda Piovesan Ucci and Elaine Cristina Teixeira and Gisele Audrei Pedroso and Eduardo Hilario and Vitória Fernanda Bertolazzi Zocca and Gabriela Barbosa De Paiva and Henrique Ferreira and Danielle Biscaro Pedrolli and Maria Célia Bertolini},
url = {https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.000767},
doi = {10.1099/MIC.0.000767/CITE/REFWORKS},
issn = {14652080},
year = {2019},
date = {2019-01-01},
journal = {Microbiology (United Kingdom)},
volume = {165},
issue = {3},
pages = {355-365},
publisher = {Microbiology Society},
abstract = {Upstream open reading frames (ORFs) are frequently found in the 5¢-flanking regions of genes and may have a regulatory role in gene expression. A small ORF (named cohL here) was identified upstream from the copAB copper operon in Xanthomonascitri subsp. citri (Xac). We previously demonstrated that copAB expression was induced by copper and that gene inactivation produced a mutant strain that was unable to grow in the presence of copper. Here, we address the role of cohL in copAB expression control. We demonstrate that cohL expression is induced by copper in a copAB-independent manner. Although cohL is transcribed, the CohL protein is either not expressed in vivo or is synthesized at undetectable levels. Inactivation of cohL (X. citri cohL polar mutant strain) leads to an inability to synthesize cohL and copAB transcripts and consequently the inability to grow in the presence of copper. Bioinformatic tools predicted a stem-loop structure for the cohL-copAB intergenic region and revealed that this region may arrange itself in a secondary structure. Using in vitro gene expression, we found out that the structured 5¢-UTR mRNA of copAB is responsible for sequestering the ribosome-binding site that drives the translation of copA. However, copper alone was not able to release the sequence. Based on the results, we speculate that cohL plays a role as a regulatory RNA rather than as a protein-coding gene.},
keywords = {Copper resistance, Gene expression, Short upstream ORF, Xanthomonas citri},
pubstate = {published},
tppubtype = {article}
}
Upstream open reading frames (ORFs) are frequently found in the 5¢-flanking regions of genes and may have a regulatory role in gene expression. A small ORF (named cohL here) was identified upstream from the copAB copper operon in Xanthomonascitri subsp. citri (Xac). We previously demonstrated that copAB expression was induced by copper and that gene inactivation produced a mutant strain that was unable to grow in the presence of copper. Here, we address the role of cohL in copAB expression control. We demonstrate that cohL expression is induced by copper in a copAB-independent manner. Although cohL is transcribed, the CohL protein is either not expressed in vivo or is synthesized at undetectable levels. Inactivation of cohL (X. citri cohL polar mutant strain) leads to an inability to synthesize cohL and copAB transcripts and consequently the inability to grow in the presence of copper. Bioinformatic tools predicted a stem-loop structure for the cohL-copAB intergenic region and revealed that this region may arrange itself in a secondary structure. Using in vitro gene expression, we found out that the structured 5¢-UTR mRNA of copAB is responsible for sequestering the ribosome-binding site that drives the translation of copA. However, copper alone was not able to release the sequence. Based on the results, we speculate that cohL plays a role as a regulatory RNA rather than as a protein-coding gene.
Freitas, Eliane Cristina De; Ucci, Amanda Piovesan; Teixeira, Elaine Cristina; Pedroso, Gisele Audrei; Hilario, Eduardo; Zocca, Vitória Fernanda Bertolazzi; Paiva, Gabriela Barbosa De; Ferreira, Henrique; Pedrolli, Danielle Biscaro; Bertolini, Maria Célia
The copper-inducible copAB operon in Xanthomonas citri subsp. citri is regulated at transcriptional and translational levels Journal Article
Em: Microbiology (United Kingdom), vol. 165, iss. 3, pp. 355-365, 2019, ISSN: 14652080.
Resumo | Links | BibTeX | Tags: Copper resistance, Gene expression, Short upstream ORF, Xanthomonas citri
@article{nokey,
title = {The copper-inducible copAB operon in Xanthomonas citri subsp. citri is regulated at transcriptional and translational levels},
author = {Eliane Cristina De Freitas and Amanda Piovesan Ucci and Elaine Cristina Teixeira and Gisele Audrei Pedroso and Eduardo Hilario and Vitória Fernanda Bertolazzi Zocca and Gabriela Barbosa De Paiva and Henrique Ferreira and Danielle Biscaro Pedrolli and Maria Célia Bertolini},
url = {https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.000767},
doi = {10.1099/MIC.0.000767/CITE/REFWORKS},
issn = {14652080},
year = {2019},
date = {2019-01-01},
journal = {Microbiology (United Kingdom)},
volume = {165},
issue = {3},
pages = {355-365},
publisher = {Microbiology Society},
abstract = {Upstream open reading frames (ORFs) are frequently found in the 5¢-flanking regions of genes and may have a regulatory role in gene expression. A small ORF (named cohL here) was identified upstream from the copAB copper operon in Xanthomonascitri subsp. citri (Xac). We previously demonstrated that copAB expression was induced by copper and that gene inactivation produced a mutant strain that was unable to grow in the presence of copper. Here, we address the role of cohL in copAB expression control. We demonstrate that cohL expression is induced by copper in a copAB-independent manner. Although cohL is transcribed, the CohL protein is either not expressed in vivo or is synthesized at undetectable levels. Inactivation of cohL (X. citri cohL polar mutant strain) leads to an inability to synthesize cohL and copAB transcripts and consequently the inability to grow in the presence of copper. Bioinformatic tools predicted a stem-loop structure for the cohL-copAB intergenic region and revealed that this region may arrange itself in a secondary structure. Using in vitro gene expression, we found out that the structured 5¢-UTR mRNA of copAB is responsible for sequestering the ribosome-binding site that drives the translation of copA. However, copper alone was not able to release the sequence. Based on the results, we speculate that cohL plays a role as a regulatory RNA rather than as a protein-coding gene.},
keywords = {Copper resistance, Gene expression, Short upstream ORF, Xanthomonas citri},
pubstate = {published},
tppubtype = {article}
}
Upstream open reading frames (ORFs) are frequently found in the 5¢-flanking regions of genes and may have a regulatory role in gene expression. A small ORF (named cohL here) was identified upstream from the copAB copper operon in Xanthomonascitri subsp. citri (Xac). We previously demonstrated that copAB expression was induced by copper and that gene inactivation produced a mutant strain that was unable to grow in the presence of copper. Here, we address the role of cohL in copAB expression control. We demonstrate that cohL expression is induced by copper in a copAB-independent manner. Although cohL is transcribed, the CohL protein is either not expressed in vivo or is synthesized at undetectable levels. Inactivation of cohL (X. citri cohL polar mutant strain) leads to an inability to synthesize cohL and copAB transcripts and consequently the inability to grow in the presence of copper. Bioinformatic tools predicted a stem-loop structure for the cohL-copAB intergenic region and revealed that this region may arrange itself in a secondary structure. Using in vitro gene expression, we found out that the structured 5¢-UTR mRNA of copAB is responsible for sequestering the ribosome-binding site that drives the translation of copA. However, copper alone was not able to release the sequence. Based on the results, we speculate that cohL plays a role as a regulatory RNA rather than as a protein-coding gene.
Oliveira, Tiago Silva; Granato, Laís Moreira; Galdeano, Diogo Manzano; Marques, João Paulo Rodrigues; Coerini, Luciane Fender; Machado, Juliana Freitas-Astúa Marcos Antonio; Oliveira, Tiago Silva; Granato, Laís Moreira; Galdeano, Diogo Manzano; Marques, João Paulo Rodrigues; Coerini, Luciane Fender; Machado, Juliana Freitas-Astúa Marcos Antonio
Genetic analysis of salicylic acid-mediated defenses responses and histopathology in the huanglongbing pathosystem Journal Article
Em: Citrus Research & Technology, vol. 40, pp. 1-13, 2019, ISSN: 21775419.
Resumo | Links | BibTeX | Tags: callose deposition, Gene expression, Index terms: SAR, plant-pathogen interaction
@article{Oliveira2019,
title = {Genetic analysis of salicylic acid-mediated defenses responses and histopathology in the huanglongbing pathosystem},
author = {Tiago Silva Oliveira and Laís Moreira Granato and Diogo Manzano Galdeano and João Paulo Rodrigues Marques and Luciane Fender Coerini and Juliana Freitas-Astúa Marcos Antonio Machado and Tiago Silva Oliveira and Laís Moreira Granato and Diogo Manzano Galdeano and João Paulo Rodrigues Marques and Luciane Fender Coerini and Juliana Freitas-Astúa Marcos Antonio Machado},
url = {http://www.citrusrt.periodikos.com.br/article/doi/10.4322/crt.18419},
doi = {10.4322/CRT.18419},
issn = {21775419},
year = {2019},
date = {2019-01-01},
journal = {Citrus Research & Technology},
volume = {40},
pages = {1-13},
publisher = {Centro de Citricultura Sylvio Moreira/Instituto Agronômico},
abstract = {The emergence of citrus huanglongbing (HLB) has been a constraint for worldwide citrus growers. HLB disease is associated to species of the biothrophic bacteria Candidatus Liberibacter spp. (CaL). In this study, we assessed the transcriptional status of salicylic acid (SA) genes and associated defenses between two contrasting citrus genotypes during challenge with Ca. Liberibacter asiaticus or Ca. Liberibacter americanus. Citrus sinensis exhibited the most evident alterations in gene expression of evaluated genes, when compared with Poncirus trifoliata. Upstream pathway SA genes showed a slight upward regulation in C. sinensis. Salicylic acid biosynthesis and accumulation might be impaired as we observed a low expression level of SA biosynthesis related genes. Moreover, genes associated to SA metabolism showed a slight induction. These results may account for the absence of significant downstream defense response related to salicylic acid. Leaf anatomical analysis revealed callose accumulation in both HLB infected, C. sinensis and P. trifoliata sieve tube elements (STE), although only C. sinensis exhibited collapsed STE. Our data corroborate other studies and suggest that the SA biosynthesis and metabolism related genes might be involved in the contrasting response to CaL in different citrus genotypes. Additionally, we suggest that collapsed STE might have a prominent implication in symptomatology of highly susceptible plants.},
keywords = {callose deposition, Gene expression, Index terms: SAR, plant-pathogen interaction},
pubstate = {published},
tppubtype = {article}
}
The emergence of citrus huanglongbing (HLB) has been a constraint for worldwide citrus growers. HLB disease is associated to species of the biothrophic bacteria Candidatus Liberibacter spp. (CaL). In this study, we assessed the transcriptional status of salicylic acid (SA) genes and associated defenses between two contrasting citrus genotypes during challenge with Ca. Liberibacter asiaticus or Ca. Liberibacter americanus. Citrus sinensis exhibited the most evident alterations in gene expression of evaluated genes, when compared with Poncirus trifoliata. Upstream pathway SA genes showed a slight upward regulation in C. sinensis. Salicylic acid biosynthesis and accumulation might be impaired as we observed a low expression level of SA biosynthesis related genes. Moreover, genes associated to SA metabolism showed a slight induction. These results may account for the absence of significant downstream defense response related to salicylic acid. Leaf anatomical analysis revealed callose accumulation in both HLB infected, C. sinensis and P. trifoliata sieve tube elements (STE), although only C. sinensis exhibited collapsed STE. Our data corroborate other studies and suggest that the SA biosynthesis and metabolism related genes might be involved in the contrasting response to CaL in different citrus genotypes. Additionally, we suggest that collapsed STE might have a prominent implication in symptomatology of highly susceptible plants.
Rogerio, Luana Aparecida; Galdeano, Diogo Manzano; Arena, Gabriella Dias; Nunes, Maria Andreia; Machado, Marcos Antonio; Novelli, Valdenice Moreira
Reference genes for gene expression studies by RT-qPCR in Brevipalpus yothersi (Acari: Tenuipalpidae), the mite vector of citrus leprosis virus Journal Article
Em: Scientific Reports 2019 9:1, vol. 9, iss. 1, pp. 1-9, 2019, ISSN: 2045-2322.
Resumo | Links | BibTeX | Tags: Gene expression, Gene expression profiling, RNAi
@article{Rogerio2019,
title = {Reference genes for gene expression studies by RT-qPCR in Brevipalpus yothersi (Acari: Tenuipalpidae), the mite vector of citrus leprosis virus},
author = {Luana Aparecida Rogerio and Diogo Manzano Galdeano and Gabriella Dias Arena and Maria Andreia Nunes and Marcos Antonio Machado and Valdenice Moreira Novelli},
url = {https://www.nature.com/articles/s41598-019-42993-2},
doi = {10.1038/s41598-019-42993-2},
issn = {2045-2322},
year = {2019},
date = {2019-01-01},
journal = {Scientific Reports 2019 9:1},
volume = {9},
issue = {1},
pages = {1-9},
publisher = {Nature Publishing Group},
abstract = {Quantitative reverse transcription PCR (RT-qPCR) is a high-throughput method to analyze the transcriptional expression of genes. Currently, no reference genes have been described for evaluating gene expression in Brevipalpus yothersi, the false spider mite, a polyphagous that act as vector of the citrus leprosis virus C (CiLV-C), an important citrus disease. This study aimed to identify the most stable reference genes in B. yothersi. The RT-qPCR expression data for selected genes were evaluated from three conditions: different developmental stages, plant hosts and acquisition of CiLV-C. To analyze the stability of the candidate reference genes we used ΔCq method, GeNorm, NormFinder, BestKeeper and RefFinder. Ubiq and GAPDH are best suited for normalizing gene expression data in viruliferous and non-viruliferous mites. Ubiq, EF1α and GAPDH are the most stable for different developmental stages. RPL13 and RPL32 are the best reference genes for approaches to B. yothersi in different host plants. Considering all the experimental conditions, Ubiq, EF1α, and GAPDH were the most stable genes. Here we developed an accurate and comprehensive RT-qPCR strategy for use in B. yothersi gene expression analysis. These results will improve the understanding of the biology of the false spider mites and their role as virus vectors.},
keywords = {Gene expression, Gene expression profiling, RNAi},
pubstate = {published},
tppubtype = {article}
}
Quantitative reverse transcription PCR (RT-qPCR) is a high-throughput method to analyze the transcriptional expression of genes. Currently, no reference genes have been described for evaluating gene expression in Brevipalpus yothersi, the false spider mite, a polyphagous that act as vector of the citrus leprosis virus C (CiLV-C), an important citrus disease. This study aimed to identify the most stable reference genes in B. yothersi. The RT-qPCR expression data for selected genes were evaluated from three conditions: different developmental stages, plant hosts and acquisition of CiLV-C. To analyze the stability of the candidate reference genes we used ΔCq method, GeNorm, NormFinder, BestKeeper and RefFinder. Ubiq and GAPDH are best suited for normalizing gene expression data in viruliferous and non-viruliferous mites. Ubiq, EF1α and GAPDH are the most stable for different developmental stages. RPL13 and RPL32 are the best reference genes for approaches to B. yothersi in different host plants. Considering all the experimental conditions, Ubiq, EF1α, and GAPDH were the most stable genes. Here we developed an accurate and comprehensive RT-qPCR strategy for use in B. yothersi gene expression analysis. These results will improve the understanding of the biology of the false spider mites and their role as virus vectors.
Santos, Ivanildes Conceição; Almeida, Alex Alan Furtado; Pirovani, Carlos Priminho; Costa, Marcio Gilberto Cardoso; Conceição, Alessandro Santos; Filho, Walter Santos Soares; Filho, Mauricio Antônio Coelho; Gesteira, Abelmon Silva
Physiological, biochemical and molecular responses to drought conditions in field-grown grafted and ungrafted citrus plants Journal Article
Em: Environmental and Experimental Botany, vol. 162, pp. 406-420, 2019, ISSN: 0098-8472.
Resumo | Links | BibTeX | Tags: Citrus rootstock, Gene expression, Oxidative stress, photosynthesis, water deficit
@article{Santos2019,
title = {Physiological, biochemical and molecular responses to drought conditions in field-grown grafted and ungrafted citrus plants},
author = {Ivanildes Conceição Santos and Alex Alan Furtado Almeida and Carlos Priminho Pirovani and Marcio Gilberto Cardoso Costa and Alessandro Santos Conceição and Walter Santos Soares Filho and Mauricio Antônio Coelho Filho and Abelmon Silva Gesteira},
doi = {10.1016/J.ENVEXPBOT.2019.03.018},
issn = {0098-8472},
year = {2019},
date = {2019-01-01},
journal = {Environmental and Experimental Botany},
volume = {162},
pages = {406-420},
publisher = {Elsevier},
abstract = {The physiological, biochemical and molecular alterations developed by plants under soil water restriction, especially rootstock, is important to elucidate the mechanisms associated to drought tolerance. In order to verify the influence of rootstock and grafting on drought tolerance of citrus plants, we examined water status, photosynthesis, oxidative stress and gene expression in grafted and ungrafted citrus plants subjected to different soil water regimes (well-watered, moderate and severe drought and rehydrated) under field conditions. Analysis were performed in roots and leaves of six plant combinations: ungrafted ‘Rangpur Santa Cruz’ lime (RL) and ‘Sunki Maravilha’ mandarin (SM), RL and SM grafted onto themselves (RL/RL and SM/SM) and ‘Valencia’ sweet orange (VO) grafted onto RL or SM (VO/RL and VO/SM). Drought stress reduced chloroplastid pigment contents and limited the photosynthetic rate, mainly in RL/RL plants. In contrast, the lowest leaf osmotic potential, the best osmotic adjustment and the greater phenotypic plasticity were observed in SM, SM/SMand VO/SM plants. Phenotypic plasticity and the main components of multivariate analysis showed that the physiological variables, such as Fv/F0, F0/Fm and Fv/Fm, were more responsive to soil water deficit. An increase in the superoxide dismutase (SOD) activity and transcript abundance of mitochondrial SOD [Mn-Fe], especially in the scion onto RL, and chloroplastidic SOD [Cu -Zn], inthe scion onto SM, was also observed under water deficit. Besides, an induction of transcription of genes coding for catalase (CAT X2) and ascorbate peroxidase (APX) was also observed, respectively, in leaves and roots of plants subjected to water deficit. However, such transcription increment did not affect the activity of these enzymes, especially in RL/RL plants. RL plants were more effective in soil water extraction than SM plants, evidenced mainly by the maintenance of leaf gas exchange and increase in the leaf relative water content after rehydration. The grafted plants showed alterations in their responses to the stress factor, expressed by a reduction of photosynthetic rates, changes in pigment contents, activity of antioxidant enzymes and expression of genes associated to the antioxidant metabolism, mainly in RL/RL plants. Plants with ‘Valencia’ scion, regardless of the rootstock studied, showed similar physiological, biochemical and molecular responses when exposed to soil water deficit. Multivariate analysis showed that Fv/F0, F0/Fm and Fv/Fm were the most important physiological variables for discriminating citrus plants subjected to control and soil water deficit conditions.},
keywords = {Citrus rootstock, Gene expression, Oxidative stress, photosynthesis, water deficit},
pubstate = {published},
tppubtype = {article}
}
The physiological, biochemical and molecular alterations developed by plants under soil water restriction, especially rootstock, is important to elucidate the mechanisms associated to drought tolerance. In order to verify the influence of rootstock and grafting on drought tolerance of citrus plants, we examined water status, photosynthesis, oxidative stress and gene expression in grafted and ungrafted citrus plants subjected to different soil water regimes (well-watered, moderate and severe drought and rehydrated) under field conditions. Analysis were performed in roots and leaves of six plant combinations: ungrafted ‘Rangpur Santa Cruz’ lime (RL) and ‘Sunki Maravilha’ mandarin (SM), RL and SM grafted onto themselves (RL/RL and SM/SM) and ‘Valencia’ sweet orange (VO) grafted onto RL or SM (VO/RL and VO/SM). Drought stress reduced chloroplastid pigment contents and limited the photosynthetic rate, mainly in RL/RL plants. In contrast, the lowest leaf osmotic potential, the best osmotic adjustment and the greater phenotypic plasticity were observed in SM, SM/SMand VO/SM plants. Phenotypic plasticity and the main components of multivariate analysis showed that the physiological variables, such as Fv/F0, F0/Fm and Fv/Fm, were more responsive to soil water deficit. An increase in the superoxide dismutase (SOD) activity and transcript abundance of mitochondrial SOD [Mn-Fe], especially in the scion onto RL, and chloroplastidic SOD [Cu -Zn], inthe scion onto SM, was also observed under water deficit. Besides, an induction of transcription of genes coding for catalase (CAT X2) and ascorbate peroxidase (APX) was also observed, respectively, in leaves and roots of plants subjected to water deficit. However, such transcription increment did not affect the activity of these enzymes, especially in RL/RL plants. RL plants were more effective in soil water extraction than SM plants, evidenced mainly by the maintenance of leaf gas exchange and increase in the leaf relative water content after rehydration. The grafted plants showed alterations in their responses to the stress factor, expressed by a reduction of photosynthetic rates, changes in pigment contents, activity of antioxidant enzymes and expression of genes associated to the antioxidant metabolism, mainly in RL/RL plants. Plants with ‘Valencia’ scion, regardless of the rootstock studied, showed similar physiological, biochemical and molecular responses when exposed to soil water deficit. Multivariate analysis showed that Fv/F0, F0/Fm and Fv/Fm were the most important physiological variables for discriminating citrus plants subjected to control and soil water deficit conditions.
2018
Pereira, Suzam L. S.; Martins, Cristina P. S.; Sousa, Aurizangela O.; Camillo, Luciana R.; Araújo, Caroline P.; Alcantara, Grazielle M.; Camargo, Danielle S.; Cidade, Luciana C.; Almeida, Alex Alan F. De; Costa, Marcio G. C.
Em: PLOS ONE, vol. 13, iss. 6, pp. e0199187, 2018, ISSN: 1932-6203.
Resumo | Links | BibTeX | Tags: Arabidopsis thaliana, Citrus, Drought adaptation, Gene expression, Genetically modified plants, Leaves, Nicotiana, Oranges
@article{Pereira2018b,
title = {Genome-wide characterization and expression analysis of citrus NUCLEAR FACTOR-Y (NF-Y) transcription factors identified a novel NF-YA gene involved in drought-stress response and tolerance},
author = {Suzam L. S. Pereira and Cristina P. S. Martins and Aurizangela O. Sousa and Luciana R. Camillo and Caroline P. Araújo and Grazielle M. Alcantara and Danielle S. Camargo and Luciana C. Cidade and Alex Alan F. De Almeida and Marcio G. C. Costa},
url = {https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0199187},
doi = {10.1371/JOURNAL.PONE.0199187},
issn = {1932-6203},
year = {2018},
date = {2018-01-01},
journal = {PLOS ONE},
volume = {13},
issue = {6},
pages = {e0199187},
publisher = {Public Library of Science},
abstract = {Nuclear factor Y (NF-Y) is a ubiquitous transcription factor found in eukaryotes. It is composed of three distinct subunits called NF-YA, NF-YB and NF-YC. NF-Ys have been identified as key regulators of multiple pathways in the control of development and tolerance to biotic and abiotic factors. The present study aimed to identify and characterize the complete repertoire of genes coding for NF-Y in citrus, as well as to perform the functional characterization of one of its members, namely CsNFYA5, in transgenic tobacco plants. A total of 22 genes coding for NF-Y were identified in the genomes of sweet orange (Citrus sinensis) and Clementine mandarin (C. clementina), including six CsNF-YAs, 11 CsNF-YBs and five CsNF-YCs. Phylogenetic analyses showed that there is a NF-Y orthologous in the Clementine genome for each sweet orange NF-Y gene; this was not observed when compared to Arabidopsis thaliana. CsNF-Y proteins shared the same conserved domains with their orthologous proteins in other organisms, including mouse. Analysis of gene expression by RNA-seq and EST data demonstrated that CsNF-Ys have a tissue-specific and stress inducible expression profile. qRT-PCR analysis revealed that CsNF-YA5 exhibits differential expression in response to water deficit in leaves and roots of citrus plants. Overexpression of CsNF-YA5 in transgenic tobacco plants contributed to the reduction of H2O2 production under dehydration conditions and increased plant growth and photosynthetic rate under normal conditions and drought stress. These biochemical and physiological responses to drought stress promoted by CsNF-YA5 may confer a productivity advantage in environments with frequent short-term soil water deficit.},
keywords = {Arabidopsis thaliana, Citrus, Drought adaptation, Gene expression, Genetically modified plants, Leaves, Nicotiana, Oranges},
pubstate = {published},
tppubtype = {article}
}
Nuclear factor Y (NF-Y) is a ubiquitous transcription factor found in eukaryotes. It is composed of three distinct subunits called NF-YA, NF-YB and NF-YC. NF-Ys have been identified as key regulators of multiple pathways in the control of development and tolerance to biotic and abiotic factors. The present study aimed to identify and characterize the complete repertoire of genes coding for NF-Y in citrus, as well as to perform the functional characterization of one of its members, namely CsNFYA5, in transgenic tobacco plants. A total of 22 genes coding for NF-Y were identified in the genomes of sweet orange (Citrus sinensis) and Clementine mandarin (C. clementina), including six CsNF-YAs, 11 CsNF-YBs and five CsNF-YCs. Phylogenetic analyses showed that there is a NF-Y orthologous in the Clementine genome for each sweet orange NF-Y gene; this was not observed when compared to Arabidopsis thaliana. CsNF-Y proteins shared the same conserved domains with their orthologous proteins in other organisms, including mouse. Analysis of gene expression by RNA-seq and EST data demonstrated that CsNF-Ys have a tissue-specific and stress inducible expression profile. qRT-PCR analysis revealed that CsNF-YA5 exhibits differential expression in response to water deficit in leaves and roots of citrus plants. Overexpression of CsNF-YA5 in transgenic tobacco plants contributed to the reduction of H2O2 production under dehydration conditions and increased plant growth and photosynthetic rate under normal conditions and drought stress. These biochemical and physiological responses to drought stress promoted by CsNF-YA5 may confer a productivity advantage in environments with frequent short-term soil water deficit.
Erpen, L.; Tavano, E. C. R.; Harakava, R.; Dutt, M.; Grosser, J. W.; Piedade, S. M. S.; Mendes, B. M. J.; Filho, F. A. A. Mourão
Isolation, characterization, and evaluation of three Citrus sinensis-derived constitutive gene promoters Journal Article
Em: Plant Cell Reports, vol. 37, iss. 8, pp. 1113-1125, 2018, ISSN: 07217714.
Resumo | Links | BibTeX | Tags: Cis-regulatory elements, Cultivar improvement, Gene expression, sweet orange
@article{Erpen2018,
title = {Isolation, characterization, and evaluation of three Citrus sinensis-derived constitutive gene promoters},
author = {L. Erpen and E. C. R. Tavano and R. Harakava and M. Dutt and J. W. Grosser and S. M. S. Piedade and B. M. J. Mendes and F. A. A. Mourão Filho},
url = {https://link.springer.com/article/10.1007/s00299-018-2298-1},
doi = {10.1007/S00299-018-2298-1/METRICS},
issn = {07217714},
year = {2018},
date = {2018-01-01},
journal = {Plant Cell Reports},
volume = {37},
issue = {8},
pages = {1113-1125},
publisher = {Springer Verlag},
abstract = {Key message: Regulatory sequences from the citrus constitutive genes cyclophilin (CsCYP), glyceraldehyde-3-phosphate dehydrogenase C2 (CsGAPC2), and elongation factor 1-alpha (CsEF1) were isolated, fused to the uidA gene, and qualitatively and quantitatively evaluated in transgenic sweet orange plants. Abstract: The 5′ upstream region of a gene (the promoter) is the most important component for the initiation and regulation of gene transcription of both native genes and transgenes in plants. The isolation and characterization of gene regulatory sequences are essential to the development of intragenic or cisgenic genetic manipulation strategies, which imply the use of genetic material from the same species or from closely related species. We describe herein the isolation and evaluation of the promoter sequence from three constitutively expressed citrus genes: cyclophilin (CsCYP), glyceraldehyde-3-phosphate dehydrogenase C2 (CsGAPC2), and elongation factor 1-alpha (CsEF1). The functionality of the promoters was confirmed by a histochemical GUS assay in leaves, stems, and roots of stably transformed citrus plants expressing the promoter-uidA construct. Lower uidA mRNA levels were detected when the transgene was under the control of citrus promoters as compared to the expression under the control of the CaMV35S promoter. The association of the uidA gene with the citrus-derived promoters resulted in mRNA levels of up to 60–41.8% of the value obtained with the construct containing CaMV35S driving the uidA gene. Moreover, a lower inter-individual variability in transgene expression was observed amongst the different transgenic lines, where gene constructs containing citrus-derived promoters were used. In silico analysis of the citrus-derived promoter sequences revealed that their activity may be controlled by several putative cis-regulatory elements. These citrus promoters will expand the availability of regulatory sequences for driving gene expression in citrus gene-modification programs.},
keywords = {Cis-regulatory elements, Cultivar improvement, Gene expression, sweet orange},
pubstate = {published},
tppubtype = {article}
}
Key message: Regulatory sequences from the citrus constitutive genes cyclophilin (CsCYP), glyceraldehyde-3-phosphate dehydrogenase C2 (CsGAPC2), and elongation factor 1-alpha (CsEF1) were isolated, fused to the uidA gene, and qualitatively and quantitatively evaluated in transgenic sweet orange plants. Abstract: The 5′ upstream region of a gene (the promoter) is the most important component for the initiation and regulation of gene transcription of both native genes and transgenes in plants. The isolation and characterization of gene regulatory sequences are essential to the development of intragenic or cisgenic genetic manipulation strategies, which imply the use of genetic material from the same species or from closely related species. We describe herein the isolation and evaluation of the promoter sequence from three constitutively expressed citrus genes: cyclophilin (CsCYP), glyceraldehyde-3-phosphate dehydrogenase C2 (CsGAPC2), and elongation factor 1-alpha (CsEF1). The functionality of the promoters was confirmed by a histochemical GUS assay in leaves, stems, and roots of stably transformed citrus plants expressing the promoter-uidA construct. Lower uidA mRNA levels were detected when the transgene was under the control of citrus promoters as compared to the expression under the control of the CaMV35S promoter. The association of the uidA gene with the citrus-derived promoters resulted in mRNA levels of up to 60–41.8% of the value obtained with the construct containing CaMV35S driving the uidA gene. Moreover, a lower inter-individual variability in transgene expression was observed amongst the different transgenic lines, where gene constructs containing citrus-derived promoters were used. In silico analysis of the citrus-derived promoter sequences revealed that their activity may be controlled by several putative cis-regulatory elements. These citrus promoters will expand the availability of regulatory sequences for driving gene expression in citrus gene-modification programs.
Pereira, Suzam L. S.; Martins, Cristina P. S.; Sousa, Aurizangela O.; Camillo, Luciana R.; Araújo, Caroline P.; Alcantara, Grazielle M.; Camargo, Danielle S.; Cidade, Luciana C.; Almeida, Alex Alan F. De; Costa, Marcio G. C.
Em: PLOS ONE, vol. 13, iss. 6, pp. e0199187, 2018, ISSN: 1932-6203.
Resumo | Links | BibTeX | Tags: Arabidopsis thaliana, Citrus, Drought adaptation, Gene expression, Genetically modified plants, Leaves, Nicotiana, Oranges
@article{Pereira2018,
title = {Genome-wide characterization and expression analysis of citrus NUCLEAR FACTOR-Y (NF-Y) transcription factors identified a novel NF-YA gene involved in drought-stress response and tolerance},
author = {Suzam L. S. Pereira and Cristina P. S. Martins and Aurizangela O. Sousa and Luciana R. Camillo and Caroline P. Araújo and Grazielle M. Alcantara and Danielle S. Camargo and Luciana C. Cidade and Alex Alan F. De Almeida and Marcio G. C. Costa},
url = {https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0199187},
doi = {10.1371/JOURNAL.PONE.0199187},
issn = {1932-6203},
year = {2018},
date = {2018-01-01},
journal = {PLOS ONE},
volume = {13},
issue = {6},
pages = {e0199187},
publisher = {Public Library of Science},
abstract = {Nuclear factor Y (NF-Y) is a ubiquitous transcription factor found in eukaryotes. It is composed of three distinct subunits called NF-YA, NF-YB and NF-YC. NF-Ys have been identified as key regulators of multiple pathways in the control of development and tolerance to biotic and abiotic factors. The present study aimed to identify and characterize the complete repertoire of genes coding for NF-Y in citrus, as well as to perform the functional characterization of one of its members, namely CsNFYA5, in transgenic tobacco plants. A total of 22 genes coding for NF-Y were identified in the genomes of sweet orange (Citrus sinensis) and Clementine mandarin (C. clementina), including six CsNF-YAs, 11 CsNF-YBs and five CsNF-YCs. Phylogenetic analyses showed that there is a NF-Y orthologous in the Clementine genome for each sweet orange NF-Y gene; this was not observed when compared to Arabidopsis thaliana. CsNF-Y proteins shared the same conserved domains with their orthologous proteins in other organisms, including mouse. Analysis of gene expression by RNA-seq and EST data demonstrated that CsNF-Ys have a tissue-specific and stress inducible expression profile. qRT-PCR analysis revealed that CsNF-YA5 exhibits differential expression in response to water deficit in leaves and roots of citrus plants. Overexpression of CsNF-YA5 in transgenic tobacco plants contributed to the reduction of H2O2 production under dehydration conditions and increased plant growth and photosynthetic rate under normal conditions and drought stress. These biochemical and physiological responses to drought stress promoted by CsNF-YA5 may confer a productivity advantage in environments with frequent short-term soil water deficit.},
keywords = {Arabidopsis thaliana, Citrus, Drought adaptation, Gene expression, Genetically modified plants, Leaves, Nicotiana, Oranges},
pubstate = {published},
tppubtype = {article}
}
Nuclear factor Y (NF-Y) is a ubiquitous transcription factor found in eukaryotes. It is composed of three distinct subunits called NF-YA, NF-YB and NF-YC. NF-Ys have been identified as key regulators of multiple pathways in the control of development and tolerance to biotic and abiotic factors. The present study aimed to identify and characterize the complete repertoire of genes coding for NF-Y in citrus, as well as to perform the functional characterization of one of its members, namely CsNFYA5, in transgenic tobacco plants. A total of 22 genes coding for NF-Y were identified in the genomes of sweet orange (Citrus sinensis) and Clementine mandarin (C. clementina), including six CsNF-YAs, 11 CsNF-YBs and five CsNF-YCs. Phylogenetic analyses showed that there is a NF-Y orthologous in the Clementine genome for each sweet orange NF-Y gene; this was not observed when compared to Arabidopsis thaliana. CsNF-Y proteins shared the same conserved domains with their orthologous proteins in other organisms, including mouse. Analysis of gene expression by RNA-seq and EST data demonstrated that CsNF-Ys have a tissue-specific and stress inducible expression profile. qRT-PCR analysis revealed that CsNF-YA5 exhibits differential expression in response to water deficit in leaves and roots of citrus plants. Overexpression of CsNF-YA5 in transgenic tobacco plants contributed to the reduction of H2O2 production under dehydration conditions and increased plant growth and photosynthetic rate under normal conditions and drought stress. These biochemical and physiological responses to drought stress promoted by CsNF-YA5 may confer a productivity advantage in environments with frequent short-term soil water deficit.
Tavano, Eveline Carla Rocha; Erpen, Ligia; Aluisi, Bianca; Harakava, Ricardo; Lopes, João Roberto Spotti; Vieira, Maria Lucia Carneiro; Piedade, Sônia Maria De Stefano; Mendes, Beatriz Madalena Januzzi; Filho, Francisco Assis Alves Mourão
Em: https://doi.org/10.1080/14620316.2018.1493361, vol. 94, iss. 2, pp. 210-219, 2018, ISSN: 14620316.
Resumo | Links | BibTeX | Tags: antimicrobial peptide, Bacteria, Citrus sinensis, disease resistance, Gene expression, huanglongbing
@article{Tavano2018,
title = {Sweet orange genetic transformation with the attacin A gene under the control of phloem-specific promoters and inoculation with Candidatus Liberibacter asiaticus},
author = {Eveline Carla Rocha Tavano and Ligia Erpen and Bianca Aluisi and Ricardo Harakava and João Roberto Spotti Lopes and Maria Lucia Carneiro Vieira and Sônia Maria De Stefano Piedade and Beatriz Madalena Januzzi Mendes and Francisco Assis Alves Mourão Filho},
url = {https://www.tandfonline.com/doi/abs/10.1080/14620316.2018.1493361},
doi = {10.1080/14620316.2018.1493361},
issn = {14620316},
year = {2018},
date = {2018-01-01},
journal = {https://doi.org/10.1080/14620316.2018.1493361},
volume = {94},
issue = {2},
pages = {210-219},
publisher = {Taylor & Francis},
abstract = {An alternative approach to control Huanglongbing disease in Citrus is the development of transgenic plants expressing genes that may influence pathogen development. We report herein the production ...},
keywords = {antimicrobial peptide, Bacteria, Citrus sinensis, disease resistance, Gene expression, huanglongbing},
pubstate = {published},
tppubtype = {article}
}
An alternative approach to control Huanglongbing disease in Citrus is the development of transgenic plants expressing genes that may influence pathogen development. We report herein the production ...