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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
In: In Vitro Cellular and Developmental Biology - Plant, vol. 56, iss. 3, pp. 280-289, 2020, ISSN: 14752689.
Abstract | 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.
2019
Pinheiro, Thaísa T.; Peres, Lázaro E. P.; Purgatto, Eduardo; Latado, Rodrigo R.; Maniero, Rodolfo A.; Martins, Mônica M.; Figueira, Antonio
Citrus carotenoid isomerase gene characterization by complementation of the “Micro-Tom” tangerine mutant Journal Article
In: Plant Cell Reports, vol. 38, iss. 5, pp. 623-636, 2019, ISSN: 07217714.
Abstract | Links | BibTeX | Tags: Carotenoid biosynthesis, CRTISO, Functional genomics, sweet orange, Tomato
@article{Pinheiro2019,
title = {Citrus carotenoid isomerase gene characterization by complementation of the “Micro-Tom” tangerine mutant},
author = {Thaísa T. Pinheiro and Lázaro E. P. Peres and Eduardo Purgatto and Rodrigo R. Latado and Rodolfo A. Maniero and Mônica M. Martins and Antonio Figueira},
url = {https://link.springer.com/article/10.1007/s00299-019-02393-2},
doi = {10.1007/S00299-019-02393-2/METRICS},
issn = {07217714},
year = {2019},
date = {2019-01-01},
journal = {Plant Cell Reports},
volume = {38},
issue = {5},
pages = {623-636},
publisher = {Springer Verlag},
abstract = {Key message: Complementation of the “Micro-Tom” tomato tangerine mutant with a Citrus CRTISO allele restores the wild-type fruit carotenoid profile, indicating that the Citrus allele encodes an authentic functional carotenoid isomerase. Abstract: Citrus fruits are rich in carotenoids; the genus offers a large diversity in composition, yet to be fully explored to improve fruit nutritional quality. As perennial tree species, Citrus lack the resources for functional genetic studies, requiring the use of model plant systems. Here, we used the “Micro-Tom” (MT) tomato carrying the tangerine mutation (t), deficient for the carotenoid isomerase (CRTISO) gene, to functionally characterize the homologous C. sinensis genes. We identified four putative loci in the C. sinensis genome, named CsCRTISO, CsCRTISO-Like 1, CsCRTISO-Like 2, and CsCRTISO-Like 2B, with the latter as a presumed duplication of CRTISO-Like 2. In general, all the Citrus paralogs showed less expression specialization than the tomato ones, with CsCRTISO being the most expressed gene in all tissues analyzed. MT-t plants were successfully complemented with the CsCRTISO, and fruits showed a carotenoid profile similar to the control, indicating that the Citrus allele indeed encodes an authentic functional carotenoid isomerase and that the signal peptide is functional in tomato. MT was silenced using an inverted repeat of a fragment from the Citrus CRTISO resulting in a stronger phenotype than MT-t. MT-t and MT silenced for CRTISO presented an overall decrease in transcript accumulation of all genes from the biosynthesis pathway. The expression of the Citrus CRTISO gene is able to restore the biosynthesis of carotenoids with the appropriate regulation in MT-t. The decrease in transcript accumulation in MT-t and MT-CRTISO-suppressed lines reinforces previous suggestions that transcriptional regulation of the carotenoid biosynthesis involves regulatory loops by intermediate products.},
keywords = {Carotenoid biosynthesis, CRTISO, Functional genomics, sweet orange, Tomato},
pubstate = {published},
tppubtype = {article}
}
Key message: Complementation of the “Micro-Tom” tomato tangerine mutant with a Citrus CRTISO allele restores the wild-type fruit carotenoid profile, indicating that the Citrus allele encodes an authentic functional carotenoid isomerase. Abstract: Citrus fruits are rich in carotenoids; the genus offers a large diversity in composition, yet to be fully explored to improve fruit nutritional quality. As perennial tree species, Citrus lack the resources for functional genetic studies, requiring the use of model plant systems. Here, we used the “Micro-Tom” (MT) tomato carrying the tangerine mutation (t), deficient for the carotenoid isomerase (CRTISO) gene, to functionally characterize the homologous C. sinensis genes. We identified four putative loci in the C. sinensis genome, named CsCRTISO, CsCRTISO-Like 1, CsCRTISO-Like 2, and CsCRTISO-Like 2B, with the latter as a presumed duplication of CRTISO-Like 2. In general, all the Citrus paralogs showed less expression specialization than the tomato ones, with CsCRTISO being the most expressed gene in all tissues analyzed. MT-t plants were successfully complemented with the CsCRTISO, and fruits showed a carotenoid profile similar to the control, indicating that the Citrus allele indeed encodes an authentic functional carotenoid isomerase and that the signal peptide is functional in tomato. MT was silenced using an inverted repeat of a fragment from the Citrus CRTISO resulting in a stronger phenotype than MT-t. MT-t and MT silenced for CRTISO presented an overall decrease in transcript accumulation of all genes from the biosynthesis pathway. The expression of the Citrus CRTISO gene is able to restore the biosynthesis of carotenoids with the appropriate regulation in MT-t. The decrease in transcript accumulation in MT-t and MT-CRTISO-suppressed lines reinforces previous suggestions that transcriptional regulation of the carotenoid biosynthesis involves regulatory loops by intermediate products.
2018
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
In: Plant Cell Reports, vol. 37, iss. 8, pp. 1113-1125, 2018, ISSN: 07217714.
Abstract | 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.
2017
Campos, Kelly Aparecida Fernandes; Azevedo, Fernando Alves; Bastianel, Marinês; Cristofani-Yaly, Mariângela
RESISTANCE TO ALTERNARIA BROWN SPOT OF NEW CITRUS HYBRIDS Journal Article
In: Revista Brasileira de Fruticultura, vol. 39, iss. 5, 2017, ISSN: 0100-2945.
Abstract | Links | BibTeX | Tags: Alternaria alternata, genetic breeding, Murcott tangor, Pêra de Abril, sweet orange
@article{nokey,
title = {RESISTANCE TO ALTERNARIA BROWN SPOT OF NEW CITRUS HYBRIDS},
author = {Kelly Aparecida Fernandes Campos and Fernando Alves Azevedo and Marinês Bastianel and Mariângela Cristofani-Yaly},
url = {http://www.scielo.br/j/rbf/a/N8ZSkkBFwbgW5k6rjN6ffjv/?lang=en},
doi = {10.1590/0100-29452017613},
issn = {0100-2945},
year = {2017},
date = {2017-01-01},
journal = {Revista Brasileira de Fruticultura},
volume = {39},
issue = {5},
publisher = {Sociedade Brasileira de Fruticultura},
abstract = {ABSTRACT Alternaria brown spot (ABS) disease is caused by the fungus of Alternaria alternata f. sp. citri, which causes injury in leaves, branches and fruits of citrus. The action of the pathogen is directly related to the presence of toxin receptors in susceptible genotypes. The objective of this study was to characterize a population of citrus hybrids obtained from controlled crosses between Pêra de Abril sweet orange and the hybrid of Murcott tangor x Pêra sweet orange (TM x LP 163) for response to ABS through the in vitro inoculation of fungal spores in young detached leaves. The fungus was isolated from the lesions of Murcott tangor fruits that exhibited ABS symptoms. Two hundred thirty-five hybrids were evaluated, and 70 (30%) showed different levels of disease symptoms on detached leaves after 72 hours of inoculation with the fungus, and 165 (70%) were asymptomatic. The frequency of segregation observed (165R:70S) and high level of heritability (h2g = 0.91) suggest that few genes may be involved in controlling the inheritance of ABS resistance in citrus.},
keywords = {Alternaria alternata, genetic breeding, Murcott tangor, Pêra de Abril, sweet orange},
pubstate = {published},
tppubtype = {article}
}
ABSTRACT Alternaria brown spot (ABS) disease is caused by the fungus of Alternaria alternata f. sp. citri, which causes injury in leaves, branches and fruits of citrus. The action of the pathogen is directly related to the presence of toxin receptors in susceptible genotypes. The objective of this study was to characterize a population of citrus hybrids obtained from controlled crosses between Pêra de Abril sweet orange and the hybrid of Murcott tangor x Pêra sweet orange (TM x LP 163) for response to ABS through the in vitro inoculation of fungal spores in young detached leaves. The fungus was isolated from the lesions of Murcott tangor fruits that exhibited ABS symptoms. Two hundred thirty-five hybrids were evaluated, and 70 (30%) showed different levels of disease symptoms on detached leaves after 72 hours of inoculation with the fungus, and 165 (70%) were asymptomatic. The frequency of segregation observed (165R:70S) and high level of heritability (h2g = 0.91) suggest that few genes may be involved in controlling the inheritance of ABS resistance in citrus.
Campos, Kelly Aparecida Fernandes; Azevedo, Fernando Alves; Bastianel, Marinês; Cristofani-Yaly, Mariângela
RESISTANCE TO ALTERNARIA BROWN SPOT OF NEW CITRUS HYBRIDS Journal Article
In: Revista Brasileira de Fruticultura, vol. 39, iss. 5, 2017, ISSN: 0100-2945.
Abstract | Links | BibTeX | Tags: Alternaria alternata, genetic breeding, Murcott tangor, Pêra de Abril, sweet orange
@article{nokey,
title = {RESISTANCE TO ALTERNARIA BROWN SPOT OF NEW CITRUS HYBRIDS},
author = {Kelly Aparecida Fernandes Campos and Fernando Alves Azevedo and Marinês Bastianel and Mariângela Cristofani-Yaly},
url = {http://www.scielo.br/j/rbf/a/N8ZSkkBFwbgW5k6rjN6ffjv/?lang=en},
doi = {10.1590/0100-29452017613},
issn = {0100-2945},
year = {2017},
date = {2017-01-01},
journal = {Revista Brasileira de Fruticultura},
volume = {39},
issue = {5},
publisher = {Sociedade Brasileira de Fruticultura},
abstract = {ABSTRACT Alternaria brown spot (ABS) disease is caused by the fungus of Alternaria alternata f. sp. citri, which causes injury in leaves, branches and fruits of citrus. The action of the pathogen is directly related to the presence of toxin receptors in susceptible genotypes. The objective of this study was to characterize a population of citrus hybrids obtained from controlled crosses between Pêra de Abril sweet orange and the hybrid of Murcott tangor x Pêra sweet orange (TM x LP 163) for response to ABS through the in vitro inoculation of fungal spores in young detached leaves. The fungus was isolated from the lesions of Murcott tangor fruits that exhibited ABS symptoms. Two hundred thirty-five hybrids were evaluated, and 70 (30%) showed different levels of disease symptoms on detached leaves after 72 hours of inoculation with the fungus, and 165 (70%) were asymptomatic. The frequency of segregation observed (165R:70S) and high level of heritability (h2g = 0.91) suggest that few genes may be involved in controlling the inheritance of ABS resistance in citrus.},
keywords = {Alternaria alternata, genetic breeding, Murcott tangor, Pêra de Abril, sweet orange},
pubstate = {published},
tppubtype = {article}
}
ABSTRACT Alternaria brown spot (ABS) disease is caused by the fungus of Alternaria alternata f. sp. citri, which causes injury in leaves, branches and fruits of citrus. The action of the pathogen is directly related to the presence of toxin receptors in susceptible genotypes. The objective of this study was to characterize a population of citrus hybrids obtained from controlled crosses between Pêra de Abril sweet orange and the hybrid of Murcott tangor x Pêra sweet orange (TM x LP 163) for response to ABS through the in vitro inoculation of fungal spores in young detached leaves. The fungus was isolated from the lesions of Murcott tangor fruits that exhibited ABS symptoms. Two hundred thirty-five hybrids were evaluated, and 70 (30%) showed different levels of disease symptoms on detached leaves after 72 hours of inoculation with the fungus, and 165 (70%) were asymptomatic. The frequency of segregation observed (165R:70S) and high level of heritability (h2g = 0.91) suggest that few genes may be involved in controlling the inheritance of ABS resistance in citrus.
Curtolo, Maiara; Cristofani-Yaly, Mariângela; Gazaffi, Rodrigo; Takita, Marco Aurélio; Figueira, Antonio; Machado, Marcos Antonio
QTL mapping for fruit quality in Citrus using DArTseq markers Journal Article
In: BMC Genomics, vol. 18, iss. 1, pp. 1-16, 2017, ISSN: 14712164.
Abstract | Links | BibTeX | Tags: hybrids, Integrated linkage map, Mandarins, sweet orange, Synteny
@article{Curtolo2017,
title = {QTL mapping for fruit quality in Citrus using DArTseq markers},
author = {Maiara Curtolo and Mariângela Cristofani-Yaly and Rodrigo Gazaffi and Marco Aurélio Takita and Antonio Figueira and Marcos Antonio Machado},
url = {https://bmcgenomics.biomedcentral.com/articles/10.1186/s12864-017-3629-2},
doi = {10.1186/S12864-017-3629-2/TABLES/5},
issn = {14712164},
year = {2017},
date = {2017-01-01},
journal = {BMC Genomics},
volume = {18},
issue = {1},
pages = {1-16},
publisher = {BioMed Central Ltd.},
abstract = {Background: Citrus breeding programs have many limitations associated with the species biology and physiology, requiring the incorporation of new biotechnological tools to provide new breeding possibilities. Diversity Arrays Technology (DArT) markers, combined with next-generation sequencing, have wide applicability in the construction of high-resolution genetic maps and in quantitative trait locus (QTL) mapping. This study aimed to construct an integrated genetic map using full-sib progeny derived from Murcott tangor and Pera sweet orange and DArTseq™ molecular markers and to perform QTL mapping of twelve fruit quality traits. A controlled Murcott x Pera crossing was conducted at the Citrus Germplasm Repository at the Sylvio Moreira Citrus Centre of the Agronomic Institute (IAC) located in Cordeirópolis, SP, in 1997. In 2012, 278F1 individuals out of a family of 312 confirmed hybrid individuals were analyzed for fruit traits and genotyped using the DArTseq markers. Using OneMap software to obtain the integrated genetic map, we considered only the DArT loci that showed no segregation deviation. The likelihood ratio and the genomic information from the available Citrus sinensis L. Osbeck genome were used to determine the linkage groups (LGs). Results: The resulting integrated map contained 661 markers in 13 LGs, with a genomic coverage of 2,774cM and a mean density of 0.23 markers/cM. The groups were assigned to the nine Citrus haploid chromosomes; however, some of the chromosomes were represented by two LGs due the lack of information for a single integration, as in cases where markers segregated in a 3:1 fashion. A total of 19 QTLs were identified through composite interval mapping (CIM) of the 12 analyzed fruit characteristics: fruit diameter (cm), height (cm), height/diameter ratio, weight (g), rind thickness (cm), segments per fruit, total soluble solids (TSS, %), total titratable acidity (TTA, %), juice content (%), number of seeds, TSS/TTA ratio and number of fruits per box. The genomic sequence (pseudochromosomes) of C. sinensis was compared to the genetic map, and synteny was clearly identified. Further analysis of the map regions with the highest LOD scores enabled the identification of putative genes that could be associated with the fruit quality characteristics. Conclusion: An integrated linkage map of Murcott tangor and Pera sweet orange using DArTseq™ molecular markers was established and it was useful to perform QTL mapping of twelve fruit quality traits. The next generation sequences data allowed the comparison between the linkage map and the genomic sequence (pseudochromosomes) of C. sinensis and the identification of genes that may be responsible for phenotypic traits in Citrus. The obtained linkage map was used to assign sequences that had not been previously assigned to a position in the reference genome.},
keywords = {hybrids, Integrated linkage map, Mandarins, sweet orange, Synteny},
pubstate = {published},
tppubtype = {article}
}
Background: Citrus breeding programs have many limitations associated with the species biology and physiology, requiring the incorporation of new biotechnological tools to provide new breeding possibilities. Diversity Arrays Technology (DArT) markers, combined with next-generation sequencing, have wide applicability in the construction of high-resolution genetic maps and in quantitative trait locus (QTL) mapping. This study aimed to construct an integrated genetic map using full-sib progeny derived from Murcott tangor and Pera sweet orange and DArTseq™ molecular markers and to perform QTL mapping of twelve fruit quality traits. A controlled Murcott x Pera crossing was conducted at the Citrus Germplasm Repository at the Sylvio Moreira Citrus Centre of the Agronomic Institute (IAC) located in Cordeirópolis, SP, in 1997. In 2012, 278F1 individuals out of a family of 312 confirmed hybrid individuals were analyzed for fruit traits and genotyped using the DArTseq markers. Using OneMap software to obtain the integrated genetic map, we considered only the DArT loci that showed no segregation deviation. The likelihood ratio and the genomic information from the available Citrus sinensis L. Osbeck genome were used to determine the linkage groups (LGs). Results: The resulting integrated map contained 661 markers in 13 LGs, with a genomic coverage of 2,774cM and a mean density of 0.23 markers/cM. The groups were assigned to the nine Citrus haploid chromosomes; however, some of the chromosomes were represented by two LGs due the lack of information for a single integration, as in cases where markers segregated in a 3:1 fashion. A total of 19 QTLs were identified through composite interval mapping (CIM) of the 12 analyzed fruit characteristics: fruit diameter (cm), height (cm), height/diameter ratio, weight (g), rind thickness (cm), segments per fruit, total soluble solids (TSS, %), total titratable acidity (TTA, %), juice content (%), number of seeds, TSS/TTA ratio and number of fruits per box. The genomic sequence (pseudochromosomes) of C. sinensis was compared to the genetic map, and synteny was clearly identified. Further analysis of the map regions with the highest LOD scores enabled the identification of putative genes that could be associated with the fruit quality characteristics. Conclusion: An integrated linkage map of Murcott tangor and Pera sweet orange using DArTseq™ molecular markers was established and it was useful to perform QTL mapping of twelve fruit quality traits. The next generation sequences data allowed the comparison between the linkage map and the genomic sequence (pseudochromosomes) of C. sinensis and the identification of genes that may be responsible for phenotypic traits in Citrus. The obtained linkage map was used to assign sequences that had not been previously assigned to a position in the reference genome.
Campos, Kelly Aparecida Fernandes; Azevedo, Fernando Alves; Bastianel, Marinês; Cristofani-Yaly, Mariângela
RESISTANCE TO ALTERNARIA BROWN SPOT OF NEW CITRUS HYBRIDS Journal Article
In: Revista Brasileira de Fruticultura, vol. 39, iss. 5, 2017, ISSN: 0100-2945.
Abstract | Links | BibTeX | Tags: Alternaria alternata, genetic breeding, Murcott tangor, Pêra de Abril, sweet orange
@article{nokey,
title = {RESISTANCE TO ALTERNARIA BROWN SPOT OF NEW CITRUS HYBRIDS},
author = {Kelly Aparecida Fernandes Campos and Fernando Alves Azevedo and Marinês Bastianel and Mariângela Cristofani-Yaly},
url = {http://www.scielo.br/j/rbf/a/N8ZSkkBFwbgW5k6rjN6ffjv/?lang=en},
doi = {10.1590/0100-29452017613},
issn = {0100-2945},
year = {2017},
date = {2017-01-01},
journal = {Revista Brasileira de Fruticultura},
volume = {39},
issue = {5},
publisher = {Sociedade Brasileira de Fruticultura},
abstract = {ABSTRACT Alternaria brown spot (ABS) disease is caused by the fungus of Alternaria alternata f. sp. citri, which causes injury in leaves, branches and fruits of citrus. The action of the pathogen is directly related to the presence of toxin receptors in susceptible genotypes. The objective of this study was to characterize a population of citrus hybrids obtained from controlled crosses between Pêra de Abril sweet orange and the hybrid of Murcott tangor x Pêra sweet orange (TM x LP 163) for response to ABS through the in vitro inoculation of fungal spores in young detached leaves. The fungus was isolated from the lesions of Murcott tangor fruits that exhibited ABS symptoms. Two hundred thirty-five hybrids were evaluated, and 70 (30%) showed different levels of disease symptoms on detached leaves after 72 hours of inoculation with the fungus, and 165 (70%) were asymptomatic. The frequency of segregation observed (165R:70S) and high level of heritability (h2g = 0.91) suggest that few genes may be involved in controlling the inheritance of ABS resistance in citrus.},
keywords = {Alternaria alternata, genetic breeding, Murcott tangor, Pêra de Abril, sweet orange},
pubstate = {published},
tppubtype = {article}
}
ABSTRACT Alternaria brown spot (ABS) disease is caused by the fungus of Alternaria alternata f. sp. citri, which causes injury in leaves, branches and fruits of citrus. The action of the pathogen is directly related to the presence of toxin receptors in susceptible genotypes. The objective of this study was to characterize a population of citrus hybrids obtained from controlled crosses between Pêra de Abril sweet orange and the hybrid of Murcott tangor x Pêra sweet orange (TM x LP 163) for response to ABS through the in vitro inoculation of fungal spores in young detached leaves. The fungus was isolated from the lesions of Murcott tangor fruits that exhibited ABS symptoms. Two hundred thirty-five hybrids were evaluated, and 70 (30%) showed different levels of disease symptoms on detached leaves after 72 hours of inoculation with the fungus, and 165 (70%) were asymptomatic. The frequency of segregation observed (165R:70S) and high level of heritability (h2g = 0.91) suggest that few genes may be involved in controlling the inheritance of ABS resistance in citrus.
Sagawa, C. H. D.; Cristofani-Yaly, M.; Novelli, V. M.; Bastianel, M.; Machado, M. A.
Assessing genetic diversity of Citrus by DArT_seq™ genotyping Journal Article
In: https://doi.org/10.1080/11263504.2017.1341438, vol. 152, iss. 4, pp. 593-598, 2017, ISSN: 17245575.
Abstract | Links | BibTeX | Tags: citrus fingerprint, genetic diversity, Molecular markers, Poncirus, sweet orange
@article{Sagawa2017,
title = {Assessing genetic diversity of Citrus by DArT_seq™ genotyping},
author = {C. H. D. Sagawa and M. Cristofani-Yaly and V. M. Novelli and M. Bastianel and M. A. Machado},
url = {https://www.tandfonline.com/doi/abs/10.1080/11263504.2017.1341438},
doi = {10.1080/11263504.2017.1341438},
issn = {17245575},
year = {2017},
date = {2017-01-01},
journal = {https://doi.org/10.1080/11263504.2017.1341438},
volume = {152},
issue = {4},
pages = {593-598},
publisher = {Taylor & Francis},
abstract = {In citrus despite the diversity among cultivated genotypes related to morphological, physiological, and agronomic traits, low polymorphism is detected at the molecular marker level, and the number ...},
keywords = {citrus fingerprint, genetic diversity, Molecular markers, Poncirus, sweet orange},
pubstate = {published},
tppubtype = {article}
}
In citrus despite the diversity among cultivated genotypes related to morphological, physiological, and agronomic traits, low polymorphism is detected at the molecular marker level, and the number ...