Potato Snakin - 1 gene silencing affects cell division, primary metabolism, and cell wall composition
Snakin-1 [SN1] is an antimicrobial cysteine-rich peptide isolated from potato [Solanum tuberosum] that was classified as a member of the Snakin/Gibberellic Acid Stimulated in Arabidopsis protein family. In this work, a transgenic approach was used to study the role of SN1 in planta. Even when overexpressing SN1, potato lines did not show remarkable morphological differences from the wild type; SN1 silencing resulted in reduced height, which was accompanied by an overall reduction in leaf size and severe alterations of leaf shape. Analysis of the adaxial epidermis of mature leaves revealed that silenced lines had 70 percent to 90 percent increases in mean cell size with respect to wild-type leaves. Consequently, the number of epidermal cells was significantly reduced in these lines. Confocal microscopy analysis after agroinfiltration of Nicotiana benthamiana leaves showed that SN1-green fluorescent protein fusion protein was localized in plasma membrane, and bimolecular fluorescence complementation assays revealed that SN1 self-interacted in vivo. We further focused our study on leaf metabolism by applying a combination of gas chromatography coupled to mass spectrometry, Fourier transform infrared spectroscopy, and spectrophotometric techniques. These targeted analyses allowed a detailed examination of the changes occurring in 46 intermediate compounds from primary metabolic pathways and in seven cell wall constituents. We demonstrated that SN1 silencing affects cell division, leaf primary metabolism, and cell wall composition in potato plants, suggesting that SN1 has additional roles in growth and development beyond its previously assigned role in plant defense.
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Subjects: | GREEN FLUORESCENT PROTEIN, SN1 PROTEIN, SOLANUM TUBEROSUM, VEGETABLE PROTEIN, CELL DIVISION, CELL MEMBRANE, CELL WALL, CHEMISTRY, CYTOLOGY, GENE EXPRESSION REGULATION, GENE SILENCING, GENETICS, INFRARED SPECTROSCOPY, MASS FRAGMENTOGRAPHY, METABOLISM, MOLECULAR GENETICS, NUCLEOTIDE SEQUENCE, PHYSIOLOGY, PLANT EPIDERMIS, PLANT LEAF, POTATO, SOLANACEAE, TRANSGENIC PLANT, GAS CHROMATOGRAPHY-MASS SPECTROMETRY, MOLECULAR SEQUENCE DATA, PLANT LEAVES, PLANT PROTEINS, PLANTS, GENETICALLY MODIFIED, SOLANUM TUBEROSUM, SPECTROSCOPY, FOURIER TRANSFORM INFRARED, ARABIDOPSIS, NICOTIANA BENTHAMIANA, , |
Online Access: | http://ceiba.agro.uba.ar/cgi-bin/koha/opac-detail.pl?biblionumber=46536 http://ceiba.agro.uba.ar/cgi-bin/koha/opac-detail.pl?biblionumber= |
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GREEN FLUORESCENT PROTEIN SN1 PROTEIN, SOLANUM TUBEROSUM VEGETABLE PROTEIN CELL DIVISION CELL MEMBRANE CELL WALL CHEMISTRY CYTOLOGY GENE EXPRESSION REGULATION GENE SILENCING GENETICS INFRARED SPECTROSCOPY MASS FRAGMENTOGRAPHY METABOLISM MOLECULAR GENETICS NUCLEOTIDE SEQUENCE PHYSIOLOGY PLANT EPIDERMIS PLANT LEAF POTATO SOLANACEAE TRANSGENIC PLANT GAS CHROMATOGRAPHY-MASS SPECTROMETRY MOLECULAR SEQUENCE DATA PLANT LEAVES PLANT PROTEINS PLANTS, GENETICALLY MODIFIED SOLANUM TUBEROSUM SPECTROSCOPY, FOURIER TRANSFORM INFRARED ARABIDOPSIS NICOTIANA BENTHAMIANA GREEN FLUORESCENT PROTEIN SN1 PROTEIN, SOLANUM TUBEROSUM VEGETABLE PROTEIN CELL DIVISION CELL MEMBRANE CELL WALL CHEMISTRY CYTOLOGY GENE EXPRESSION REGULATION GENE SILENCING GENETICS INFRARED SPECTROSCOPY MASS FRAGMENTOGRAPHY METABOLISM MOLECULAR GENETICS NUCLEOTIDE SEQUENCE PHYSIOLOGY PLANT EPIDERMIS PLANT LEAF POTATO SOLANACEAE TRANSGENIC PLANT GAS CHROMATOGRAPHY-MASS SPECTROMETRY MOLECULAR SEQUENCE DATA PLANT LEAVES PLANT PROTEINS PLANTS, GENETICALLY MODIFIED SOLANUM TUBEROSUM SPECTROSCOPY, FOURIER TRANSFORM INFRARED ARABIDOPSIS NICOTIANA BENTHAMIANA |
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GREEN FLUORESCENT PROTEIN SN1 PROTEIN, SOLANUM TUBEROSUM VEGETABLE PROTEIN CELL DIVISION CELL MEMBRANE CELL WALL CHEMISTRY CYTOLOGY GENE EXPRESSION REGULATION GENE SILENCING GENETICS INFRARED SPECTROSCOPY MASS FRAGMENTOGRAPHY METABOLISM MOLECULAR GENETICS NUCLEOTIDE SEQUENCE PHYSIOLOGY PLANT EPIDERMIS PLANT LEAF POTATO SOLANACEAE TRANSGENIC PLANT GAS CHROMATOGRAPHY-MASS SPECTROMETRY MOLECULAR SEQUENCE DATA PLANT LEAVES PLANT PROTEINS PLANTS, GENETICALLY MODIFIED SOLANUM TUBEROSUM SPECTROSCOPY, FOURIER TRANSFORM INFRARED ARABIDOPSIS NICOTIANA BENTHAMIANA GREEN FLUORESCENT PROTEIN SN1 PROTEIN, SOLANUM TUBEROSUM VEGETABLE PROTEIN CELL DIVISION CELL MEMBRANE CELL WALL CHEMISTRY CYTOLOGY GENE EXPRESSION REGULATION GENE SILENCING GENETICS INFRARED SPECTROSCOPY MASS FRAGMENTOGRAPHY METABOLISM MOLECULAR GENETICS NUCLEOTIDE SEQUENCE PHYSIOLOGY PLANT EPIDERMIS PLANT LEAF POTATO SOLANACEAE TRANSGENIC PLANT GAS CHROMATOGRAPHY-MASS SPECTROMETRY MOLECULAR SEQUENCE DATA PLANT LEAVES PLANT PROTEINS PLANTS, GENETICALLY MODIFIED SOLANUM TUBEROSUM SPECTROSCOPY, FOURIER TRANSFORM INFRARED ARABIDOPSIS NICOTIANA BENTHAMIANA Nahirñak, Vanesa Almasia, Natalia Inés Fernández, Paula Virginia Hopp, Horacio Esteban Estévez, José Manuel Carrari, Fernando Vazquez Rovere, Cecilia Potato Snakin - 1 gene silencing affects cell division, primary metabolism, and cell wall composition |
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Snakin-1 [SN1] is an antimicrobial cysteine-rich peptide isolated from potato [Solanum tuberosum] that was classified as a member of the Snakin/Gibberellic Acid Stimulated in Arabidopsis protein family. In this work, a transgenic approach was used to study the role of SN1 in planta. Even when overexpressing SN1, potato lines did not show remarkable morphological differences from the wild type; SN1 silencing resulted in reduced height, which was accompanied by an overall reduction in leaf size and severe alterations of leaf shape. Analysis of the adaxial epidermis of mature leaves revealed that silenced lines had 70 percent to 90 percent increases in mean cell size with respect to wild-type leaves. Consequently, the number of epidermal cells was significantly reduced in these lines. Confocal microscopy analysis after agroinfiltration of Nicotiana benthamiana leaves showed that SN1-green fluorescent protein fusion protein was localized in plasma membrane, and bimolecular fluorescence complementation assays revealed that SN1 self-interacted in vivo. We further focused our study on leaf metabolism by applying a combination of gas chromatography coupled to mass spectrometry, Fourier transform infrared spectroscopy, and spectrophotometric techniques. These targeted analyses allowed a detailed examination of the changes occurring in 46 intermediate compounds from primary metabolic pathways and in seven cell wall constituents. We demonstrated that SN1 silencing affects cell division, leaf primary metabolism, and cell wall composition in potato plants, suggesting that SN1 has additional roles in growth and development beyond its previously assigned role in plant defense. |
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GREEN FLUORESCENT PROTEIN SN1 PROTEIN, SOLANUM TUBEROSUM VEGETABLE PROTEIN CELL DIVISION CELL MEMBRANE CELL WALL CHEMISTRY CYTOLOGY GENE EXPRESSION REGULATION GENE SILENCING GENETICS INFRARED SPECTROSCOPY MASS FRAGMENTOGRAPHY METABOLISM MOLECULAR GENETICS NUCLEOTIDE SEQUENCE PHYSIOLOGY PLANT EPIDERMIS PLANT LEAF POTATO SOLANACEAE TRANSGENIC PLANT GAS CHROMATOGRAPHY-MASS SPECTROMETRY MOLECULAR SEQUENCE DATA PLANT LEAVES PLANT PROTEINS PLANTS, GENETICALLY MODIFIED SOLANUM TUBEROSUM SPECTROSCOPY, FOURIER TRANSFORM INFRARED ARABIDOPSIS NICOTIANA BENTHAMIANA |
author |
Nahirñak, Vanesa Almasia, Natalia Inés Fernández, Paula Virginia Hopp, Horacio Esteban Estévez, José Manuel Carrari, Fernando Vazquez Rovere, Cecilia |
author_facet |
Nahirñak, Vanesa Almasia, Natalia Inés Fernández, Paula Virginia Hopp, Horacio Esteban Estévez, José Manuel Carrari, Fernando Vazquez Rovere, Cecilia |
author_sort |
Nahirñak, Vanesa |
title |
Potato Snakin - 1 gene silencing affects cell division, primary metabolism, and cell wall composition |
title_short |
Potato Snakin - 1 gene silencing affects cell division, primary metabolism, and cell wall composition |
title_full |
Potato Snakin - 1 gene silencing affects cell division, primary metabolism, and cell wall composition |
title_fullStr |
Potato Snakin - 1 gene silencing affects cell division, primary metabolism, and cell wall composition |
title_full_unstemmed |
Potato Snakin - 1 gene silencing affects cell division, primary metabolism, and cell wall composition |
title_sort |
potato snakin - 1 gene silencing affects cell division, primary metabolism, and cell wall composition |
url |
http://ceiba.agro.uba.ar/cgi-bin/koha/opac-detail.pl?biblionumber=46536 http://ceiba.agro.uba.ar/cgi-bin/koha/opac-detail.pl?biblionumber= |
work_keys_str_mv |
AT nahirnakvanesa potatosnakin1genesilencingaffectscelldivisionprimarymetabolismandcellwallcomposition AT almasianataliaines potatosnakin1genesilencingaffectscelldivisionprimarymetabolismandcellwallcomposition AT fernandezpaulavirginia potatosnakin1genesilencingaffectscelldivisionprimarymetabolismandcellwallcomposition AT hopphoracioesteban potatosnakin1genesilencingaffectscelldivisionprimarymetabolismandcellwallcomposition AT estevezjosemanuel potatosnakin1genesilencingaffectscelldivisionprimarymetabolismandcellwallcomposition AT carrarifernando potatosnakin1genesilencingaffectscelldivisionprimarymetabolismandcellwallcomposition AT vazquezroverececilia potatosnakin1genesilencingaffectscelldivisionprimarymetabolismandcellwallcomposition |
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1774851078146228224 |
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KOHA-OAI-AGRO:465362023-08-17T13:58:37Zhttp://ceiba.agro.uba.ar/cgi-bin/koha/opac-detail.pl?biblionumber=46536http://ceiba.agro.uba.ar/cgi-bin/koha/opac-detail.pl?biblionumber=AAGPotato Snakin - 1 gene silencing affects cell division, primary metabolism, and cell wall compositionNahirñak, VanesaAlmasia, Natalia InésFernández, Paula VirginiaHopp, Horacio EstebanEstévez, José ManuelCarrari, FernandoVazquez Rovere, Cecilia textspaapplication/pdfSnakin-1 [SN1] is an antimicrobial cysteine-rich peptide isolated from potato [Solanum tuberosum] that was classified as a member of the Snakin/Gibberellic Acid Stimulated in Arabidopsis protein family. In this work, a transgenic approach was used to study the role of SN1 in planta. Even when overexpressing SN1, potato lines did not show remarkable morphological differences from the wild type; SN1 silencing resulted in reduced height, which was accompanied by an overall reduction in leaf size and severe alterations of leaf shape. Analysis of the adaxial epidermis of mature leaves revealed that silenced lines had 70 percent to 90 percent increases in mean cell size with respect to wild-type leaves. Consequently, the number of epidermal cells was significantly reduced in these lines. Confocal microscopy analysis after agroinfiltration of Nicotiana benthamiana leaves showed that SN1-green fluorescent protein fusion protein was localized in plasma membrane, and bimolecular fluorescence complementation assays revealed that SN1 self-interacted in vivo. We further focused our study on leaf metabolism by applying a combination of gas chromatography coupled to mass spectrometry, Fourier transform infrared spectroscopy, and spectrophotometric techniques. These targeted analyses allowed a detailed examination of the changes occurring in 46 intermediate compounds from primary metabolic pathways and in seven cell wall constituents. We demonstrated that SN1 silencing affects cell division, leaf primary metabolism, and cell wall composition in potato plants, suggesting that SN1 has additional roles in growth and development beyond its previously assigned role in plant defense.Snakin-1 [SN1] is an antimicrobial cysteine-rich peptide isolated from potato [Solanum tuberosum] that was classified as a member of the Snakin/Gibberellic Acid Stimulated in Arabidopsis protein family. In this work, a transgenic approach was used to study the role of SN1 in planta. Even when overexpressing SN1, potato lines did not show remarkable morphological differences from the wild type; SN1 silencing resulted in reduced height, which was accompanied by an overall reduction in leaf size and severe alterations of leaf shape. Analysis of the adaxial epidermis of mature leaves revealed that silenced lines had 70 percent to 90 percent increases in mean cell size with respect to wild-type leaves. Consequently, the number of epidermal cells was significantly reduced in these lines. Confocal microscopy analysis after agroinfiltration of Nicotiana benthamiana leaves showed that SN1-green fluorescent protein fusion protein was localized in plasma membrane, and bimolecular fluorescence complementation assays revealed that SN1 self-interacted in vivo. We further focused our study on leaf metabolism by applying a combination of gas chromatography coupled to mass spectrometry, Fourier transform infrared spectroscopy, and spectrophotometric techniques. These targeted analyses allowed a detailed examination of the changes occurring in 46 intermediate compounds from primary metabolic pathways and in seven cell wall constituents. We demonstrated that SN1 silencing affects cell division, leaf primary metabolism, and cell wall composition in potato plants, suggesting that SN1 has additional roles in growth and development beyond its previously assigned role in plant defense.GREEN FLUORESCENT PROTEINSN1 PROTEIN, SOLANUM TUBEROSUMVEGETABLE PROTEINCELL DIVISIONCELL MEMBRANECELL WALLCHEMISTRYCYTOLOGYGENE EXPRESSION REGULATIONGENE SILENCINGGENETICSINFRARED SPECTROSCOPYMASS FRAGMENTOGRAPHYMETABOLISMMOLECULAR GENETICSNUCLEOTIDE SEQUENCEPHYSIOLOGYPLANT EPIDERMISPLANT LEAFPOTATOSOLANACEAETRANSGENIC PLANTGAS CHROMATOGRAPHY-MASS SPECTROMETRYMOLECULAR SEQUENCE DATAPLANT LEAVESPLANT PROTEINSPLANTS, GENETICALLY MODIFIEDSOLANUM TUBEROSUMSPECTROSCOPY, FOURIER TRANSFORM INFRAREDARABIDOPSISNICOTIANA BENTHAMIANAPlant Physiology |