Monitoring genetic transformation with RUBY visible reporter in Nicotiana tabaccum L.
Abstract Plant genetic transformation has emerged as a key platform for both fundamental and translational research. The reporter genes are widely used to visualize gene expression patterns, promoter analysis, localization, and stable transformation studies. Marker genes would be expected to exhibit...
Ausführliche Beschreibung
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| Autor*in: |
Jogam, Phanikanth [verfasserIn] Anumula, Vaishnavi [verfasserIn] Sandhya, Dulam [verfasserIn] Manokari, M. [verfasserIn] Venkatapuram, Ajay Kumar [verfasserIn] Achary, V. Mohan Murali [verfasserIn] Shekhawat, Mahipal S. [verfasserIn] Peddaboina, Venkataiah [verfasserIn] Allini, Venkateswar Rao [verfasserIn] |
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| Format: |
E-Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2024 |
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| Schlagwörter: |
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| Anmerkung: |
© The Author(s), under exclusive licence to Springer Nature B.V. 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. |
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| Übergeordnetes Werk: |
Enthalten in: Plant cell, tissue and organ culture - Springer Netherlands, 1981, 157(2024), 1 vom: Apr. |
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| Übergeordnetes Werk: |
volume:157 ; year:2024 ; number:1 ; month:04 |
| Links: |
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| DOI / URN: |
10.1007/s11240-024-02752-2 |
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SPR055528171 |
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| 245 | 1 | 0 | |a Monitoring genetic transformation with RUBY visible reporter in Nicotiana tabaccum L. |
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| 520 | |a Abstract Plant genetic transformation has emerged as a key platform for both fundamental and translational research. The reporter genes are widely used to visualize gene expression patterns, promoter analysis, localization, and stable transformation studies. Marker genes would be expected to exhibit characteristics such as robustness, chemical affordability, visualization without any special equipment, and a non-destructive detection approach. Betalains are tyrosine-derived red-yellow pigments found in the caryophyllale plant group that are capable of being used as a visual reporter marker to identify plant transformation. In this study, we transformed Nicotiana tabacum with a RUBY vector construct consisting of three gene clusters, CYP76AD1, DODA, and 5-GT, along with a hygromycin selective marker for visible reporter study. After three weeks of transformation, the betalain pigments were easily detected in the explant leaf tissue. The RUBY-transformed shoots displayed a reddish color that could easily be distinguished from the non-transgenic green shoots during tissue culture. Besides the reddish color, RUBY transformed shoots displayed phenotypically similar to nontransformed shoots and did not show any negative effect during multiple shoots differentiation, proliferation, regeneration, and in the initial stage of plant development. However, the RUBY-expressed plants exhibited slightly reduced plant height and delayed flowering time compared to the wild-type plant. Transgenic plants were further confirmed by PCR amplification of a 341 bp inter DNA fragment of the gene construct. The betalain pigments were observed during all stages of plant development (stem, root, and leaves), beginning with the in vitro shoot buds of genetically transformed explants. Additionally, microstructural analysis of the leaf clearly indicates that betalain pigments accumulated more in the vascular bundle, lamina, trichome, and especially stomata guard cell. A comparative cross-section of the stem and root clearly revealed the presence of betalain pigment within the cortex, endodermis, epidermis, xylem, and phloem tissues of the RUBY-transformed plant. Our findings provide conclusive evidence for the potential application of the RUBY for the visual screening of genetic transformations. | ||
| 520 | |a Key message The transgenic tobacco (Nicotiana tabacum L.) plants were produced with a non-invasive visual reporter (RUBY) gene. The accumulation of betalin pigment was observed in various plant parts during the different developmental stages. | ||
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| 700 | 1 | |a Anumula, Vaishnavi |e verfasserin |4 aut | |
| 700 | 1 | |a Sandhya, Dulam |e verfasserin |4 aut | |
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| 700 | 1 | |a Achary, V. Mohan Murali |e verfasserin |4 aut | |
| 700 | 1 | |a Shekhawat, Mahipal S. |e verfasserin |4 aut | |
| 700 | 1 | |a Peddaboina, Venkataiah |e verfasserin |4 aut | |
| 700 | 1 | |a Allini, Venkateswar Rao |e verfasserin |4 aut | |
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10.1007/s11240-024-02752-2 doi (DE-627)SPR055528171 (SPR)s11240-024-02752-2-e DE-627 ger DE-627 rakwb eng 630 640 570 540 VZ 42.03 bkl 42.40 bkl 48.03 bkl 48.56 bkl Jogam, Phanikanth verfasserin aut Monitoring genetic transformation with RUBY visible reporter in Nicotiana tabaccum L. 2024 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s), under exclusive licence to Springer Nature B.V. 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. Abstract Plant genetic transformation has emerged as a key platform for both fundamental and translational research. The reporter genes are widely used to visualize gene expression patterns, promoter analysis, localization, and stable transformation studies. Marker genes would be expected to exhibit characteristics such as robustness, chemical affordability, visualization without any special equipment, and a non-destructive detection approach. Betalains are tyrosine-derived red-yellow pigments found in the caryophyllale plant group that are capable of being used as a visual reporter marker to identify plant transformation. In this study, we transformed Nicotiana tabacum with a RUBY vector construct consisting of three gene clusters, CYP76AD1, DODA, and 5-GT, along with a hygromycin selective marker for visible reporter study. After three weeks of transformation, the betalain pigments were easily detected in the explant leaf tissue. The RUBY-transformed shoots displayed a reddish color that could easily be distinguished from the non-transgenic green shoots during tissue culture. Besides the reddish color, RUBY transformed shoots displayed phenotypically similar to nontransformed shoots and did not show any negative effect during multiple shoots differentiation, proliferation, regeneration, and in the initial stage of plant development. However, the RUBY-expressed plants exhibited slightly reduced plant height and delayed flowering time compared to the wild-type plant. Transgenic plants were further confirmed by PCR amplification of a 341 bp inter DNA fragment of the gene construct. The betalain pigments were observed during all stages of plant development (stem, root, and leaves), beginning with the in vitro shoot buds of genetically transformed explants. Additionally, microstructural analysis of the leaf clearly indicates that betalain pigments accumulated more in the vascular bundle, lamina, trichome, and especially stomata guard cell. A comparative cross-section of the stem and root clearly revealed the presence of betalain pigment within the cortex, endodermis, epidermis, xylem, and phloem tissues of the RUBY-transformed plant. Our findings provide conclusive evidence for the potential application of the RUBY for the visual screening of genetic transformations. Key message The transgenic tobacco (Nicotiana tabacum L.) plants were produced with a non-invasive visual reporter (RUBY) gene. The accumulation of betalin pigment was observed in various plant parts during the different developmental stages. Betalain (dpeaa)DE-He213 Tobacco (dpeaa)DE-He213 Visual marker (dpeaa)DE-He213 Microscopy (dpeaa)DE-He213 Anumula, Vaishnavi verfasserin aut Sandhya, Dulam verfasserin aut Manokari, M. verfasserin aut Venkatapuram, Ajay Kumar verfasserin aut Achary, V. Mohan Murali verfasserin aut Shekhawat, Mahipal S. verfasserin aut Peddaboina, Venkataiah verfasserin aut Allini, Venkateswar Rao verfasserin aut Enthalten in Plant cell, tissue and organ culture Springer Netherlands, 1981 157(2024), 1 vom: Apr. (DE-627)27093278X (DE-600)1478391-5 1573-5044 nnns volume:157 year:2024 number:1 month:04 https://dx.doi.org/10.1007/s11240-024-02752-2 X:SPRINGER Resolving-System lizenzpflichtig Volltext SYSFLAG_0 GBV_SPRINGER SSG-OLC-PHA SSG-OPC-FOR GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_211 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_647 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_2548 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4328 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 42.03 VZ 42.40 VZ 48.03 VZ 48.56 VZ AR 157 2024 1 04 |
| spelling |
10.1007/s11240-024-02752-2 doi (DE-627)SPR055528171 (SPR)s11240-024-02752-2-e DE-627 ger DE-627 rakwb eng 630 640 570 540 VZ 42.03 bkl 42.40 bkl 48.03 bkl 48.56 bkl Jogam, Phanikanth verfasserin aut Monitoring genetic transformation with RUBY visible reporter in Nicotiana tabaccum L. 2024 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s), under exclusive licence to Springer Nature B.V. 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. Abstract Plant genetic transformation has emerged as a key platform for both fundamental and translational research. The reporter genes are widely used to visualize gene expression patterns, promoter analysis, localization, and stable transformation studies. Marker genes would be expected to exhibit characteristics such as robustness, chemical affordability, visualization without any special equipment, and a non-destructive detection approach. Betalains are tyrosine-derived red-yellow pigments found in the caryophyllale plant group that are capable of being used as a visual reporter marker to identify plant transformation. In this study, we transformed Nicotiana tabacum with a RUBY vector construct consisting of three gene clusters, CYP76AD1, DODA, and 5-GT, along with a hygromycin selective marker for visible reporter study. After three weeks of transformation, the betalain pigments were easily detected in the explant leaf tissue. The RUBY-transformed shoots displayed a reddish color that could easily be distinguished from the non-transgenic green shoots during tissue culture. Besides the reddish color, RUBY transformed shoots displayed phenotypically similar to nontransformed shoots and did not show any negative effect during multiple shoots differentiation, proliferation, regeneration, and in the initial stage of plant development. However, the RUBY-expressed plants exhibited slightly reduced plant height and delayed flowering time compared to the wild-type plant. Transgenic plants were further confirmed by PCR amplification of a 341 bp inter DNA fragment of the gene construct. The betalain pigments were observed during all stages of plant development (stem, root, and leaves), beginning with the in vitro shoot buds of genetically transformed explants. Additionally, microstructural analysis of the leaf clearly indicates that betalain pigments accumulated more in the vascular bundle, lamina, trichome, and especially stomata guard cell. A comparative cross-section of the stem and root clearly revealed the presence of betalain pigment within the cortex, endodermis, epidermis, xylem, and phloem tissues of the RUBY-transformed plant. Our findings provide conclusive evidence for the potential application of the RUBY for the visual screening of genetic transformations. Key message The transgenic tobacco (Nicotiana tabacum L.) plants were produced with a non-invasive visual reporter (RUBY) gene. The accumulation of betalin pigment was observed in various plant parts during the different developmental stages. Betalain (dpeaa)DE-He213 Tobacco (dpeaa)DE-He213 Visual marker (dpeaa)DE-He213 Microscopy (dpeaa)DE-He213 Anumula, Vaishnavi verfasserin aut Sandhya, Dulam verfasserin aut Manokari, M. verfasserin aut Venkatapuram, Ajay Kumar verfasserin aut Achary, V. Mohan Murali verfasserin aut Shekhawat, Mahipal S. verfasserin aut Peddaboina, Venkataiah verfasserin aut Allini, Venkateswar Rao verfasserin aut Enthalten in Plant cell, tissue and organ culture Springer Netherlands, 1981 157(2024), 1 vom: Apr. (DE-627)27093278X (DE-600)1478391-5 1573-5044 nnns volume:157 year:2024 number:1 month:04 https://dx.doi.org/10.1007/s11240-024-02752-2 X:SPRINGER Resolving-System lizenzpflichtig Volltext SYSFLAG_0 GBV_SPRINGER SSG-OLC-PHA SSG-OPC-FOR GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_211 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_647 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_2548 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4328 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 42.03 VZ 42.40 VZ 48.03 VZ 48.56 VZ AR 157 2024 1 04 |
| allfields_unstemmed |
10.1007/s11240-024-02752-2 doi (DE-627)SPR055528171 (SPR)s11240-024-02752-2-e DE-627 ger DE-627 rakwb eng 630 640 570 540 VZ 42.03 bkl 42.40 bkl 48.03 bkl 48.56 bkl Jogam, Phanikanth verfasserin aut Monitoring genetic transformation with RUBY visible reporter in Nicotiana tabaccum L. 2024 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s), under exclusive licence to Springer Nature B.V. 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. Abstract Plant genetic transformation has emerged as a key platform for both fundamental and translational research. The reporter genes are widely used to visualize gene expression patterns, promoter analysis, localization, and stable transformation studies. Marker genes would be expected to exhibit characteristics such as robustness, chemical affordability, visualization without any special equipment, and a non-destructive detection approach. Betalains are tyrosine-derived red-yellow pigments found in the caryophyllale plant group that are capable of being used as a visual reporter marker to identify plant transformation. In this study, we transformed Nicotiana tabacum with a RUBY vector construct consisting of three gene clusters, CYP76AD1, DODA, and 5-GT, along with a hygromycin selective marker for visible reporter study. After three weeks of transformation, the betalain pigments were easily detected in the explant leaf tissue. The RUBY-transformed shoots displayed a reddish color that could easily be distinguished from the non-transgenic green shoots during tissue culture. Besides the reddish color, RUBY transformed shoots displayed phenotypically similar to nontransformed shoots and did not show any negative effect during multiple shoots differentiation, proliferation, regeneration, and in the initial stage of plant development. However, the RUBY-expressed plants exhibited slightly reduced plant height and delayed flowering time compared to the wild-type plant. Transgenic plants were further confirmed by PCR amplification of a 341 bp inter DNA fragment of the gene construct. The betalain pigments were observed during all stages of plant development (stem, root, and leaves), beginning with the in vitro shoot buds of genetically transformed explants. Additionally, microstructural analysis of the leaf clearly indicates that betalain pigments accumulated more in the vascular bundle, lamina, trichome, and especially stomata guard cell. A comparative cross-section of the stem and root clearly revealed the presence of betalain pigment within the cortex, endodermis, epidermis, xylem, and phloem tissues of the RUBY-transformed plant. Our findings provide conclusive evidence for the potential application of the RUBY for the visual screening of genetic transformations. Key message The transgenic tobacco (Nicotiana tabacum L.) plants were produced with a non-invasive visual reporter (RUBY) gene. The accumulation of betalin pigment was observed in various plant parts during the different developmental stages. Betalain (dpeaa)DE-He213 Tobacco (dpeaa)DE-He213 Visual marker (dpeaa)DE-He213 Microscopy (dpeaa)DE-He213 Anumula, Vaishnavi verfasserin aut Sandhya, Dulam verfasserin aut Manokari, M. verfasserin aut Venkatapuram, Ajay Kumar verfasserin aut Achary, V. Mohan Murali verfasserin aut Shekhawat, Mahipal S. verfasserin aut Peddaboina, Venkataiah verfasserin aut Allini, Venkateswar Rao verfasserin aut Enthalten in Plant cell, tissue and organ culture Springer Netherlands, 1981 157(2024), 1 vom: Apr. (DE-627)27093278X (DE-600)1478391-5 1573-5044 nnns volume:157 year:2024 number:1 month:04 https://dx.doi.org/10.1007/s11240-024-02752-2 X:SPRINGER Resolving-System lizenzpflichtig Volltext SYSFLAG_0 GBV_SPRINGER SSG-OLC-PHA SSG-OPC-FOR GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_211 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_647 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_2548 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4328 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 42.03 VZ 42.40 VZ 48.03 VZ 48.56 VZ AR 157 2024 1 04 |
| allfieldsGer |
10.1007/s11240-024-02752-2 doi (DE-627)SPR055528171 (SPR)s11240-024-02752-2-e DE-627 ger DE-627 rakwb eng 630 640 570 540 VZ 42.03 bkl 42.40 bkl 48.03 bkl 48.56 bkl Jogam, Phanikanth verfasserin aut Monitoring genetic transformation with RUBY visible reporter in Nicotiana tabaccum L. 2024 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s), under exclusive licence to Springer Nature B.V. 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. Abstract Plant genetic transformation has emerged as a key platform for both fundamental and translational research. The reporter genes are widely used to visualize gene expression patterns, promoter analysis, localization, and stable transformation studies. Marker genes would be expected to exhibit characteristics such as robustness, chemical affordability, visualization without any special equipment, and a non-destructive detection approach. Betalains are tyrosine-derived red-yellow pigments found in the caryophyllale plant group that are capable of being used as a visual reporter marker to identify plant transformation. In this study, we transformed Nicotiana tabacum with a RUBY vector construct consisting of three gene clusters, CYP76AD1, DODA, and 5-GT, along with a hygromycin selective marker for visible reporter study. After three weeks of transformation, the betalain pigments were easily detected in the explant leaf tissue. The RUBY-transformed shoots displayed a reddish color that could easily be distinguished from the non-transgenic green shoots during tissue culture. Besides the reddish color, RUBY transformed shoots displayed phenotypically similar to nontransformed shoots and did not show any negative effect during multiple shoots differentiation, proliferation, regeneration, and in the initial stage of plant development. However, the RUBY-expressed plants exhibited slightly reduced plant height and delayed flowering time compared to the wild-type plant. Transgenic plants were further confirmed by PCR amplification of a 341 bp inter DNA fragment of the gene construct. The betalain pigments were observed during all stages of plant development (stem, root, and leaves), beginning with the in vitro shoot buds of genetically transformed explants. Additionally, microstructural analysis of the leaf clearly indicates that betalain pigments accumulated more in the vascular bundle, lamina, trichome, and especially stomata guard cell. A comparative cross-section of the stem and root clearly revealed the presence of betalain pigment within the cortex, endodermis, epidermis, xylem, and phloem tissues of the RUBY-transformed plant. Our findings provide conclusive evidence for the potential application of the RUBY for the visual screening of genetic transformations. Key message The transgenic tobacco (Nicotiana tabacum L.) plants were produced with a non-invasive visual reporter (RUBY) gene. The accumulation of betalin pigment was observed in various plant parts during the different developmental stages. Betalain (dpeaa)DE-He213 Tobacco (dpeaa)DE-He213 Visual marker (dpeaa)DE-He213 Microscopy (dpeaa)DE-He213 Anumula, Vaishnavi verfasserin aut Sandhya, Dulam verfasserin aut Manokari, M. verfasserin aut Venkatapuram, Ajay Kumar verfasserin aut Achary, V. Mohan Murali verfasserin aut Shekhawat, Mahipal S. verfasserin aut Peddaboina, Venkataiah verfasserin aut Allini, Venkateswar Rao verfasserin aut Enthalten in Plant cell, tissue and organ culture Springer Netherlands, 1981 157(2024), 1 vom: Apr. (DE-627)27093278X (DE-600)1478391-5 1573-5044 nnns volume:157 year:2024 number:1 month:04 https://dx.doi.org/10.1007/s11240-024-02752-2 X:SPRINGER Resolving-System lizenzpflichtig Volltext SYSFLAG_0 GBV_SPRINGER SSG-OLC-PHA SSG-OPC-FOR GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_211 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_647 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_2548 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4328 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 42.03 VZ 42.40 VZ 48.03 VZ 48.56 VZ AR 157 2024 1 04 |
| allfieldsSound |
10.1007/s11240-024-02752-2 doi (DE-627)SPR055528171 (SPR)s11240-024-02752-2-e DE-627 ger DE-627 rakwb eng 630 640 570 540 VZ 42.03 bkl 42.40 bkl 48.03 bkl 48.56 bkl Jogam, Phanikanth verfasserin aut Monitoring genetic transformation with RUBY visible reporter in Nicotiana tabaccum L. 2024 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © The Author(s), under exclusive licence to Springer Nature B.V. 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. Abstract Plant genetic transformation has emerged as a key platform for both fundamental and translational research. The reporter genes are widely used to visualize gene expression patterns, promoter analysis, localization, and stable transformation studies. Marker genes would be expected to exhibit characteristics such as robustness, chemical affordability, visualization without any special equipment, and a non-destructive detection approach. Betalains are tyrosine-derived red-yellow pigments found in the caryophyllale plant group that are capable of being used as a visual reporter marker to identify plant transformation. In this study, we transformed Nicotiana tabacum with a RUBY vector construct consisting of three gene clusters, CYP76AD1, DODA, and 5-GT, along with a hygromycin selective marker for visible reporter study. After three weeks of transformation, the betalain pigments were easily detected in the explant leaf tissue. The RUBY-transformed shoots displayed a reddish color that could easily be distinguished from the non-transgenic green shoots during tissue culture. Besides the reddish color, RUBY transformed shoots displayed phenotypically similar to nontransformed shoots and did not show any negative effect during multiple shoots differentiation, proliferation, regeneration, and in the initial stage of plant development. However, the RUBY-expressed plants exhibited slightly reduced plant height and delayed flowering time compared to the wild-type plant. Transgenic plants were further confirmed by PCR amplification of a 341 bp inter DNA fragment of the gene construct. The betalain pigments were observed during all stages of plant development (stem, root, and leaves), beginning with the in vitro shoot buds of genetically transformed explants. Additionally, microstructural analysis of the leaf clearly indicates that betalain pigments accumulated more in the vascular bundle, lamina, trichome, and especially stomata guard cell. A comparative cross-section of the stem and root clearly revealed the presence of betalain pigment within the cortex, endodermis, epidermis, xylem, and phloem tissues of the RUBY-transformed plant. Our findings provide conclusive evidence for the potential application of the RUBY for the visual screening of genetic transformations. Key message The transgenic tobacco (Nicotiana tabacum L.) plants were produced with a non-invasive visual reporter (RUBY) gene. The accumulation of betalin pigment was observed in various plant parts during the different developmental stages. Betalain (dpeaa)DE-He213 Tobacco (dpeaa)DE-He213 Visual marker (dpeaa)DE-He213 Microscopy (dpeaa)DE-He213 Anumula, Vaishnavi verfasserin aut Sandhya, Dulam verfasserin aut Manokari, M. verfasserin aut Venkatapuram, Ajay Kumar verfasserin aut Achary, V. Mohan Murali verfasserin aut Shekhawat, Mahipal S. verfasserin aut Peddaboina, Venkataiah verfasserin aut Allini, Venkateswar Rao verfasserin aut Enthalten in Plant cell, tissue and organ culture Springer Netherlands, 1981 157(2024), 1 vom: Apr. (DE-627)27093278X (DE-600)1478391-5 1573-5044 nnns volume:157 year:2024 number:1 month:04 https://dx.doi.org/10.1007/s11240-024-02752-2 X:SPRINGER Resolving-System lizenzpflichtig Volltext SYSFLAG_0 GBV_SPRINGER SSG-OLC-PHA SSG-OPC-FOR GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_211 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_647 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2118 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_2548 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4328 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 42.03 VZ 42.40 VZ 48.03 VZ 48.56 VZ AR 157 2024 1 04 |
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Enthalten in Plant cell, tissue and organ culture 157(2024), 1 vom: Apr. volume:157 year:2024 number:1 month:04 |
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Betalain Tobacco Visual marker Microscopy |
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Jogam, Phanikanth @@aut@@ Anumula, Vaishnavi @@aut@@ Sandhya, Dulam @@aut@@ Manokari, M. @@aut@@ Venkatapuram, Ajay Kumar @@aut@@ Achary, V. Mohan Murali @@aut@@ Shekhawat, Mahipal S. @@aut@@ Peddaboina, Venkataiah @@aut@@ Allini, Venkateswar Rao @@aut@@ |
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<?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01000caa a22002652 4500</leader><controlfield tag="001">SPR055528171</controlfield><controlfield tag="003">DE-627</controlfield><controlfield tag="005">20240425064720.0</controlfield><controlfield tag="007">cr uuu---uuuuu</controlfield><controlfield tag="008">240416s2024 xx |||||o 00| ||eng c</controlfield><datafield tag="024" ind1="7" ind2=" "><subfield code="a">10.1007/s11240-024-02752-2</subfield><subfield code="2">doi</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-627)SPR055528171</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(SPR)s11240-024-02752-2-e</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-627</subfield><subfield code="b">ger</subfield><subfield code="c">DE-627</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="041" ind1=" " ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="082" ind1="0" ind2="4"><subfield code="a">630</subfield><subfield code="a">640</subfield><subfield code="a">570</subfield><subfield code="a">540</subfield><subfield code="q">VZ</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="a">42.03</subfield><subfield code="2">bkl</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="a">42.40</subfield><subfield code="2">bkl</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="a">48.03</subfield><subfield code="2">bkl</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="a">48.56</subfield><subfield code="2">bkl</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Jogam, Phanikanth</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Monitoring genetic transformation with RUBY visible reporter in Nicotiana tabaccum L.</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">2024</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="a">Text</subfield><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="a">Computermedien</subfield><subfield code="b">c</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="a">Online-Ressource</subfield><subfield code="b">cr</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="500" ind1=" " ind2=" "><subfield code="a">© The Author(s), under exclusive licence to Springer Nature B.V. 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">Abstract Plant genetic transformation has emerged as a key platform for both fundamental and translational research. The reporter genes are widely used to visualize gene expression patterns, promoter analysis, localization, and stable transformation studies. Marker genes would be expected to exhibit characteristics such as robustness, chemical affordability, visualization without any special equipment, and a non-destructive detection approach. Betalains are tyrosine-derived red-yellow pigments found in the caryophyllale plant group that are capable of being used as a visual reporter marker to identify plant transformation. In this study, we transformed Nicotiana tabacum with a RUBY vector construct consisting of three gene clusters, CYP76AD1, DODA, and 5-GT, along with a hygromycin selective marker for visible reporter study. After three weeks of transformation, the betalain pigments were easily detected in the explant leaf tissue. The RUBY-transformed shoots displayed a reddish color that could easily be distinguished from the non-transgenic green shoots during tissue culture. Besides the reddish color, RUBY transformed shoots displayed phenotypically similar to nontransformed shoots and did not show any negative effect during multiple shoots differentiation, proliferation, regeneration, and in the initial stage of plant development. However, the RUBY-expressed plants exhibited slightly reduced plant height and delayed flowering time compared to the wild-type plant. Transgenic plants were further confirmed by PCR amplification of a 341 bp inter DNA fragment of the gene construct. The betalain pigments were observed during all stages of plant development (stem, root, and leaves), beginning with the in vitro shoot buds of genetically transformed explants. Additionally, microstructural analysis of the leaf clearly indicates that betalain pigments accumulated more in the vascular bundle, lamina, trichome, and especially stomata guard cell. A comparative cross-section of the stem and root clearly revealed the presence of betalain pigment within the cortex, endodermis, epidermis, xylem, and phloem tissues of the RUBY-transformed plant. Our findings provide conclusive evidence for the potential application of the RUBY for the visual screening of genetic transformations.</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">Key message The transgenic tobacco (Nicotiana tabacum L.) plants were produced with a non-invasive visual reporter (RUBY) gene. 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|
| author |
Jogam, Phanikanth |
| spellingShingle |
Jogam, Phanikanth ddc 630 bkl 42.03 bkl 42.40 bkl 48.03 bkl 48.56 misc Betalain misc Tobacco misc Visual marker misc Microscopy Monitoring genetic transformation with RUBY visible reporter in Nicotiana tabaccum L. |
| authorStr |
Jogam, Phanikanth |
| ppnlink_with_tag_str_mv |
@@773@@(DE-627)27093278X |
| format |
electronic Article |
| dewey-ones |
630 - Agriculture & related technologies 640 - Home & family management 570 - Life sciences; biology 540 - Chemistry & allied sciences |
| delete_txt_mv |
keep |
| author_role |
aut aut aut aut aut aut aut aut aut |
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springer |
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true |
| illustrated |
Not Illustrated |
| issn |
1573-5044 |
| topic_title |
630 640 570 540 VZ 42.03 bkl 42.40 bkl 48.03 bkl 48.56 bkl Monitoring genetic transformation with RUBY visible reporter in Nicotiana tabaccum L. Betalain (dpeaa)DE-He213 Tobacco (dpeaa)DE-He213 Visual marker (dpeaa)DE-He213 Microscopy (dpeaa)DE-He213 |
| topic |
ddc 630 bkl 42.03 bkl 42.40 bkl 48.03 bkl 48.56 misc Betalain misc Tobacco misc Visual marker misc Microscopy |
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Jogam, Phanikanth Anumula, Vaishnavi Sandhya, Dulam Manokari, M. Venkatapuram, Ajay Kumar Achary, V. Mohan Murali Shekhawat, Mahipal S. Peddaboina, Venkataiah Allini, Venkateswar Rao |
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monitoring genetic transformation with ruby visible reporter in nicotiana tabaccum l. |
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Monitoring genetic transformation with RUBY visible reporter in Nicotiana tabaccum L. |
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Abstract Plant genetic transformation has emerged as a key platform for both fundamental and translational research. The reporter genes are widely used to visualize gene expression patterns, promoter analysis, localization, and stable transformation studies. Marker genes would be expected to exhibit characteristics such as robustness, chemical affordability, visualization without any special equipment, and a non-destructive detection approach. Betalains are tyrosine-derived red-yellow pigments found in the caryophyllale plant group that are capable of being used as a visual reporter marker to identify plant transformation. In this study, we transformed Nicotiana tabacum with a RUBY vector construct consisting of three gene clusters, CYP76AD1, DODA, and 5-GT, along with a hygromycin selective marker for visible reporter study. After three weeks of transformation, the betalain pigments were easily detected in the explant leaf tissue. The RUBY-transformed shoots displayed a reddish color that could easily be distinguished from the non-transgenic green shoots during tissue culture. Besides the reddish color, RUBY transformed shoots displayed phenotypically similar to nontransformed shoots and did not show any negative effect during multiple shoots differentiation, proliferation, regeneration, and in the initial stage of plant development. However, the RUBY-expressed plants exhibited slightly reduced plant height and delayed flowering time compared to the wild-type plant. Transgenic plants were further confirmed by PCR amplification of a 341 bp inter DNA fragment of the gene construct. The betalain pigments were observed during all stages of plant development (stem, root, and leaves), beginning with the in vitro shoot buds of genetically transformed explants. Additionally, microstructural analysis of the leaf clearly indicates that betalain pigments accumulated more in the vascular bundle, lamina, trichome, and especially stomata guard cell. A comparative cross-section of the stem and root clearly revealed the presence of betalain pigment within the cortex, endodermis, epidermis, xylem, and phloem tissues of the RUBY-transformed plant. Our findings provide conclusive evidence for the potential application of the RUBY for the visual screening of genetic transformations. Key message The transgenic tobacco (Nicotiana tabacum L.) plants were produced with a non-invasive visual reporter (RUBY) gene. The accumulation of betalin pigment was observed in various plant parts during the different developmental stages. © The Author(s), under exclusive licence to Springer Nature B.V. 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. |
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Abstract Plant genetic transformation has emerged as a key platform for both fundamental and translational research. The reporter genes are widely used to visualize gene expression patterns, promoter analysis, localization, and stable transformation studies. Marker genes would be expected to exhibit characteristics such as robustness, chemical affordability, visualization without any special equipment, and a non-destructive detection approach. Betalains are tyrosine-derived red-yellow pigments found in the caryophyllale plant group that are capable of being used as a visual reporter marker to identify plant transformation. In this study, we transformed Nicotiana tabacum with a RUBY vector construct consisting of three gene clusters, CYP76AD1, DODA, and 5-GT, along with a hygromycin selective marker for visible reporter study. After three weeks of transformation, the betalain pigments were easily detected in the explant leaf tissue. The RUBY-transformed shoots displayed a reddish color that could easily be distinguished from the non-transgenic green shoots during tissue culture. Besides the reddish color, RUBY transformed shoots displayed phenotypically similar to nontransformed shoots and did not show any negative effect during multiple shoots differentiation, proliferation, regeneration, and in the initial stage of plant development. However, the RUBY-expressed plants exhibited slightly reduced plant height and delayed flowering time compared to the wild-type plant. Transgenic plants were further confirmed by PCR amplification of a 341 bp inter DNA fragment of the gene construct. The betalain pigments were observed during all stages of plant development (stem, root, and leaves), beginning with the in vitro shoot buds of genetically transformed explants. Additionally, microstructural analysis of the leaf clearly indicates that betalain pigments accumulated more in the vascular bundle, lamina, trichome, and especially stomata guard cell. A comparative cross-section of the stem and root clearly revealed the presence of betalain pigment within the cortex, endodermis, epidermis, xylem, and phloem tissues of the RUBY-transformed plant. Our findings provide conclusive evidence for the potential application of the RUBY for the visual screening of genetic transformations. Key message The transgenic tobacco (Nicotiana tabacum L.) plants were produced with a non-invasive visual reporter (RUBY) gene. The accumulation of betalin pigment was observed in various plant parts during the different developmental stages. © The Author(s), under exclusive licence to Springer Nature B.V. 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. |
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Abstract Plant genetic transformation has emerged as a key platform for both fundamental and translational research. The reporter genes are widely used to visualize gene expression patterns, promoter analysis, localization, and stable transformation studies. Marker genes would be expected to exhibit characteristics such as robustness, chemical affordability, visualization without any special equipment, and a non-destructive detection approach. Betalains are tyrosine-derived red-yellow pigments found in the caryophyllale plant group that are capable of being used as a visual reporter marker to identify plant transformation. In this study, we transformed Nicotiana tabacum with a RUBY vector construct consisting of three gene clusters, CYP76AD1, DODA, and 5-GT, along with a hygromycin selective marker for visible reporter study. After three weeks of transformation, the betalain pigments were easily detected in the explant leaf tissue. The RUBY-transformed shoots displayed a reddish color that could easily be distinguished from the non-transgenic green shoots during tissue culture. Besides the reddish color, RUBY transformed shoots displayed phenotypically similar to nontransformed shoots and did not show any negative effect during multiple shoots differentiation, proliferation, regeneration, and in the initial stage of plant development. However, the RUBY-expressed plants exhibited slightly reduced plant height and delayed flowering time compared to the wild-type plant. Transgenic plants were further confirmed by PCR amplification of a 341 bp inter DNA fragment of the gene construct. The betalain pigments were observed during all stages of plant development (stem, root, and leaves), beginning with the in vitro shoot buds of genetically transformed explants. Additionally, microstructural analysis of the leaf clearly indicates that betalain pigments accumulated more in the vascular bundle, lamina, trichome, and especially stomata guard cell. A comparative cross-section of the stem and root clearly revealed the presence of betalain pigment within the cortex, endodermis, epidermis, xylem, and phloem tissues of the RUBY-transformed plant. Our findings provide conclusive evidence for the potential application of the RUBY for the visual screening of genetic transformations. Key message The transgenic tobacco (Nicotiana tabacum L.) plants were produced with a non-invasive visual reporter (RUBY) gene. The accumulation of betalin pigment was observed in various plant parts during the different developmental stages. © The Author(s), under exclusive licence to Springer Nature B.V. 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. |
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| score |
7.401725 |