Overexpression of SrDXS1 and SrKAH enhances steviol glycosides content in transgenic Stevia plants

Abstract Background Stevia rebaudiana produces sweet-tasting steviol glycosides (SGs) in its leaves which can be used as natural sweeteners. Metabolic engineering of Stevia would offer an alternative approach to conventional breeding for enhanced production of SGs. However, an effective protocol for...
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Autor*in:	Junshi Zheng [verfasserIn] Yan Zhuang [verfasserIn] Hui-Zhu Mao [verfasserIn] In-Cheol Jang [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2019

Schlagwörter:	1-deoxy-d-xylulose-5-phosphate synthase 1 Kaurenoic acid hydroxylase Metabolic engineering Stevia transformation Steviol glycosides Transgenic Stevia

Übergeordnetes Werk:	In: BMC Plant Biology - BMC, 2003, 19(2019), 1, Seite 16
Übergeordnetes Werk:	volume:19 ; year:2019 ; number:1 ; pages:16

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.1186/s12870-018-1600-2

Katalog-ID:	DOAJ020000669

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520			\|a Abstract Background Stevia rebaudiana produces sweet-tasting steviol glycosides (SGs) in its leaves which can be used as natural sweeteners. Metabolic engineering of Stevia would offer an alternative approach to conventional breeding for enhanced production of SGs. However, an effective protocol for Stevia transformation is lacking. Results Here, we present an efficient and reproducible method for Agrobacterium-mediated transformation of Stevia. In our attempts to produce transgenic Stevia plants, we found that prolonged dark incubation is critical for increasing shoot regeneration. Etiolated shoots regenerated in the dark also facilitated subsequent visual selection of transformants by green fluorescent protein during Stevia transformation. Using this newly established transformation method, we overexpressed the Stevia 1-deoxy-d-xylulose-5-phosphate synthase 1 (SrDXS1) and kaurenoic acid hydroxylase (SrKAH), both of which are required for SGs biosynthesis. Compared to control plants, the total SGs content in SrDXS1- and SrKAH-overexpressing transgenic lines were enhanced by up to 42–54% and 67–88%, respectively, showing a positive correlation with the expression levels of SrDXS1 and SrKAH. Furthermore, their overexpression did not stunt the growth and development of the transgenic Stevia plants. Conclusion This study represents a successful case of genetic manipulation of SGs biosynthetic pathway in Stevia and also demonstrates the potential of metabolic engineering towards producing Stevia with improved SGs yield.
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allfields	10.1186/s12870-018-1600-2 doi (DE-627)DOAJ020000669 (DE-599)DOAJ6fe0bf2e950b4b96a8750f9c24c39bb5 DE-627 ger DE-627 rakwb eng QK1-989 Junshi Zheng verfasserin aut Overexpression of SrDXS1 and SrKAH enhances steviol glycosides content in transgenic Stevia plants 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Background Stevia rebaudiana produces sweet-tasting steviol glycosides (SGs) in its leaves which can be used as natural sweeteners. Metabolic engineering of Stevia would offer an alternative approach to conventional breeding for enhanced production of SGs. However, an effective protocol for Stevia transformation is lacking. Results Here, we present an efficient and reproducible method for Agrobacterium-mediated transformation of Stevia. In our attempts to produce transgenic Stevia plants, we found that prolonged dark incubation is critical for increasing shoot regeneration. Etiolated shoots regenerated in the dark also facilitated subsequent visual selection of transformants by green fluorescent protein during Stevia transformation. Using this newly established transformation method, we overexpressed the Stevia 1-deoxy-d-xylulose-5-phosphate synthase 1 (SrDXS1) and kaurenoic acid hydroxylase (SrKAH), both of which are required for SGs biosynthesis. Compared to control plants, the total SGs content in SrDXS1- and SrKAH-overexpressing transgenic lines were enhanced by up to 42–54% and 67–88%, respectively, showing a positive correlation with the expression levels of SrDXS1 and SrKAH. Furthermore, their overexpression did not stunt the growth and development of the transgenic Stevia plants. Conclusion This study represents a successful case of genetic manipulation of SGs biosynthetic pathway in Stevia and also demonstrates the potential of metabolic engineering towards producing Stevia with improved SGs yield. 1-deoxy-d-xylulose-5-phosphate synthase 1 Kaurenoic acid hydroxylase Metabolic engineering Stevia transformation Steviol glycosides Transgenic Stevia Botany Yan Zhuang verfasserin aut Hui-Zhu Mao verfasserin aut In-Cheol Jang verfasserin aut In BMC Plant Biology BMC, 2003 19(2019), 1, Seite 16 (DE-627)335489060 (DE-600)2059868-3 14712229 nnns volume:19 year:2019 number:1 pages:16 https://doi.org/10.1186/s12870-018-1600-2 kostenfrei https://doaj.org/article/6fe0bf2e950b4b96a8750f9c24c39bb5 kostenfrei http://link.springer.com/article/10.1186/s12870-018-1600-2 kostenfrei https://doaj.org/toc/1471-2229 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 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_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 19 2019 1 16
spelling	10.1186/s12870-018-1600-2 doi (DE-627)DOAJ020000669 (DE-599)DOAJ6fe0bf2e950b4b96a8750f9c24c39bb5 DE-627 ger DE-627 rakwb eng QK1-989 Junshi Zheng verfasserin aut Overexpression of SrDXS1 and SrKAH enhances steviol glycosides content in transgenic Stevia plants 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Background Stevia rebaudiana produces sweet-tasting steviol glycosides (SGs) in its leaves which can be used as natural sweeteners. Metabolic engineering of Stevia would offer an alternative approach to conventional breeding for enhanced production of SGs. However, an effective protocol for Stevia transformation is lacking. Results Here, we present an efficient and reproducible method for Agrobacterium-mediated transformation of Stevia. In our attempts to produce transgenic Stevia plants, we found that prolonged dark incubation is critical for increasing shoot regeneration. Etiolated shoots regenerated in the dark also facilitated subsequent visual selection of transformants by green fluorescent protein during Stevia transformation. Using this newly established transformation method, we overexpressed the Stevia 1-deoxy-d-xylulose-5-phosphate synthase 1 (SrDXS1) and kaurenoic acid hydroxylase (SrKAH), both of which are required for SGs biosynthesis. Compared to control plants, the total SGs content in SrDXS1- and SrKAH-overexpressing transgenic lines were enhanced by up to 42–54% and 67–88%, respectively, showing a positive correlation with the expression levels of SrDXS1 and SrKAH. Furthermore, their overexpression did not stunt the growth and development of the transgenic Stevia plants. Conclusion This study represents a successful case of genetic manipulation of SGs biosynthetic pathway in Stevia and also demonstrates the potential of metabolic engineering towards producing Stevia with improved SGs yield. 1-deoxy-d-xylulose-5-phosphate synthase 1 Kaurenoic acid hydroxylase Metabolic engineering Stevia transformation Steviol glycosides Transgenic Stevia Botany Yan Zhuang verfasserin aut Hui-Zhu Mao verfasserin aut In-Cheol Jang verfasserin aut In BMC Plant Biology BMC, 2003 19(2019), 1, Seite 16 (DE-627)335489060 (DE-600)2059868-3 14712229 nnns volume:19 year:2019 number:1 pages:16 https://doi.org/10.1186/s12870-018-1600-2 kostenfrei https://doaj.org/article/6fe0bf2e950b4b96a8750f9c24c39bb5 kostenfrei http://link.springer.com/article/10.1186/s12870-018-1600-2 kostenfrei https://doaj.org/toc/1471-2229 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 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_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 19 2019 1 16
allfields_unstemmed	10.1186/s12870-018-1600-2 doi (DE-627)DOAJ020000669 (DE-599)DOAJ6fe0bf2e950b4b96a8750f9c24c39bb5 DE-627 ger DE-627 rakwb eng QK1-989 Junshi Zheng verfasserin aut Overexpression of SrDXS1 and SrKAH enhances steviol glycosides content in transgenic Stevia plants 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Background Stevia rebaudiana produces sweet-tasting steviol glycosides (SGs) in its leaves which can be used as natural sweeteners. Metabolic engineering of Stevia would offer an alternative approach to conventional breeding for enhanced production of SGs. However, an effective protocol for Stevia transformation is lacking. Results Here, we present an efficient and reproducible method for Agrobacterium-mediated transformation of Stevia. In our attempts to produce transgenic Stevia plants, we found that prolonged dark incubation is critical for increasing shoot regeneration. Etiolated shoots regenerated in the dark also facilitated subsequent visual selection of transformants by green fluorescent protein during Stevia transformation. Using this newly established transformation method, we overexpressed the Stevia 1-deoxy-d-xylulose-5-phosphate synthase 1 (SrDXS1) and kaurenoic acid hydroxylase (SrKAH), both of which are required for SGs biosynthesis. Compared to control plants, the total SGs content in SrDXS1- and SrKAH-overexpressing transgenic lines were enhanced by up to 42–54% and 67–88%, respectively, showing a positive correlation with the expression levels of SrDXS1 and SrKAH. Furthermore, their overexpression did not stunt the growth and development of the transgenic Stevia plants. Conclusion This study represents a successful case of genetic manipulation of SGs biosynthetic pathway in Stevia and also demonstrates the potential of metabolic engineering towards producing Stevia with improved SGs yield. 1-deoxy-d-xylulose-5-phosphate synthase 1 Kaurenoic acid hydroxylase Metabolic engineering Stevia transformation Steviol glycosides Transgenic Stevia Botany Yan Zhuang verfasserin aut Hui-Zhu Mao verfasserin aut In-Cheol Jang verfasserin aut In BMC Plant Biology BMC, 2003 19(2019), 1, Seite 16 (DE-627)335489060 (DE-600)2059868-3 14712229 nnns volume:19 year:2019 number:1 pages:16 https://doi.org/10.1186/s12870-018-1600-2 kostenfrei https://doaj.org/article/6fe0bf2e950b4b96a8750f9c24c39bb5 kostenfrei http://link.springer.com/article/10.1186/s12870-018-1600-2 kostenfrei https://doaj.org/toc/1471-2229 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 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_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 19 2019 1 16
allfieldsGer	10.1186/s12870-018-1600-2 doi (DE-627)DOAJ020000669 (DE-599)DOAJ6fe0bf2e950b4b96a8750f9c24c39bb5 DE-627 ger DE-627 rakwb eng QK1-989 Junshi Zheng verfasserin aut Overexpression of SrDXS1 and SrKAH enhances steviol glycosides content in transgenic Stevia plants 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Background Stevia rebaudiana produces sweet-tasting steviol glycosides (SGs) in its leaves which can be used as natural sweeteners. Metabolic engineering of Stevia would offer an alternative approach to conventional breeding for enhanced production of SGs. However, an effective protocol for Stevia transformation is lacking. Results Here, we present an efficient and reproducible method for Agrobacterium-mediated transformation of Stevia. In our attempts to produce transgenic Stevia plants, we found that prolonged dark incubation is critical for increasing shoot regeneration. Etiolated shoots regenerated in the dark also facilitated subsequent visual selection of transformants by green fluorescent protein during Stevia transformation. Using this newly established transformation method, we overexpressed the Stevia 1-deoxy-d-xylulose-5-phosphate synthase 1 (SrDXS1) and kaurenoic acid hydroxylase (SrKAH), both of which are required for SGs biosynthesis. Compared to control plants, the total SGs content in SrDXS1- and SrKAH-overexpressing transgenic lines were enhanced by up to 42–54% and 67–88%, respectively, showing a positive correlation with the expression levels of SrDXS1 and SrKAH. Furthermore, their overexpression did not stunt the growth and development of the transgenic Stevia plants. Conclusion This study represents a successful case of genetic manipulation of SGs biosynthetic pathway in Stevia and also demonstrates the potential of metabolic engineering towards producing Stevia with improved SGs yield. 1-deoxy-d-xylulose-5-phosphate synthase 1 Kaurenoic acid hydroxylase Metabolic engineering Stevia transformation Steviol glycosides Transgenic Stevia Botany Yan Zhuang verfasserin aut Hui-Zhu Mao verfasserin aut In-Cheol Jang verfasserin aut In BMC Plant Biology BMC, 2003 19(2019), 1, Seite 16 (DE-627)335489060 (DE-600)2059868-3 14712229 nnns volume:19 year:2019 number:1 pages:16 https://doi.org/10.1186/s12870-018-1600-2 kostenfrei https://doaj.org/article/6fe0bf2e950b4b96a8750f9c24c39bb5 kostenfrei http://link.springer.com/article/10.1186/s12870-018-1600-2 kostenfrei https://doaj.org/toc/1471-2229 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 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_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 19 2019 1 16
allfieldsSound	10.1186/s12870-018-1600-2 doi (DE-627)DOAJ020000669 (DE-599)DOAJ6fe0bf2e950b4b96a8750f9c24c39bb5 DE-627 ger DE-627 rakwb eng QK1-989 Junshi Zheng verfasserin aut Overexpression of SrDXS1 and SrKAH enhances steviol glycosides content in transgenic Stevia plants 2019 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Background Stevia rebaudiana produces sweet-tasting steviol glycosides (SGs) in its leaves which can be used as natural sweeteners. Metabolic engineering of Stevia would offer an alternative approach to conventional breeding for enhanced production of SGs. However, an effective protocol for Stevia transformation is lacking. Results Here, we present an efficient and reproducible method for Agrobacterium-mediated transformation of Stevia. In our attempts to produce transgenic Stevia plants, we found that prolonged dark incubation is critical for increasing shoot regeneration. Etiolated shoots regenerated in the dark also facilitated subsequent visual selection of transformants by green fluorescent protein during Stevia transformation. Using this newly established transformation method, we overexpressed the Stevia 1-deoxy-d-xylulose-5-phosphate synthase 1 (SrDXS1) and kaurenoic acid hydroxylase (SrKAH), both of which are required for SGs biosynthesis. Compared to control plants, the total SGs content in SrDXS1- and SrKAH-overexpressing transgenic lines were enhanced by up to 42–54% and 67–88%, respectively, showing a positive correlation with the expression levels of SrDXS1 and SrKAH. Furthermore, their overexpression did not stunt the growth and development of the transgenic Stevia plants. Conclusion This study represents a successful case of genetic manipulation of SGs biosynthetic pathway in Stevia and also demonstrates the potential of metabolic engineering towards producing Stevia with improved SGs yield. 1-deoxy-d-xylulose-5-phosphate synthase 1 Kaurenoic acid hydroxylase Metabolic engineering Stevia transformation Steviol glycosides Transgenic Stevia Botany Yan Zhuang verfasserin aut Hui-Zhu Mao verfasserin aut In-Cheol Jang verfasserin aut In BMC Plant Biology BMC, 2003 19(2019), 1, Seite 16 (DE-627)335489060 (DE-600)2059868-3 14712229 nnns volume:19 year:2019 number:1 pages:16 https://doi.org/10.1186/s12870-018-1600-2 kostenfrei https://doaj.org/article/6fe0bf2e950b4b96a8750f9c24c39bb5 kostenfrei http://link.springer.com/article/10.1186/s12870-018-1600-2 kostenfrei https://doaj.org/toc/1471-2229 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2008 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_2031 GBV_ILN_2038 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2056 GBV_ILN_2057 GBV_ILN_2061 GBV_ILN_2111 GBV_ILN_2113 GBV_ILN_2190 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 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_4338 GBV_ILN_4367 GBV_ILN_4700 AR 19 2019 1 16
language	English
source	In BMC Plant Biology 19(2019), 1, Seite 16 volume:19 year:2019 number:1 pages:16
sourceStr	In BMC Plant Biology 19(2019), 1, Seite 16 volume:19 year:2019 number:1 pages:16
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	1-deoxy-d-xylulose-5-phosphate synthase 1 Kaurenoic acid hydroxylase Metabolic engineering Stevia transformation Steviol glycosides Transgenic Stevia Botany
isfreeaccess_bool	true
container_title	BMC Plant Biology
authorswithroles_txt_mv	Junshi Zheng @@aut@@ Yan Zhuang @@aut@@ Hui-Zhu Mao @@aut@@ In-Cheol Jang @@aut@@
publishDateDaySort_date	2019-01-01T00:00:00Z
hierarchy_top_id	335489060
id	DOAJ020000669
language_de	englisch
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Metabolic engineering of Stevia would offer an alternative approach to conventional breeding for enhanced production of SGs. However, an effective protocol for Stevia transformation is lacking. Results Here, we present an efficient and reproducible method for Agrobacterium-mediated transformation of Stevia. In our attempts to produce transgenic Stevia plants, we found that prolonged dark incubation is critical for increasing shoot regeneration. Etiolated shoots regenerated in the dark also facilitated subsequent visual selection of transformants by green fluorescent protein during Stevia transformation. Using this newly established transformation method, we overexpressed the Stevia 1-deoxy-d-xylulose-5-phosphate synthase 1 (SrDXS1) and kaurenoic acid hydroxylase (SrKAH), both of which are required for SGs biosynthesis. Compared to control plants, the total SGs content in SrDXS1- and SrKAH-overexpressing transgenic lines were enhanced by up to 42–54% and 67–88%, respectively, showing a positive correlation with the expression levels of SrDXS1 and SrKAH. Furthermore, their overexpression did not stunt the growth and development of the transgenic Stevia plants. 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callnumber-first	Q - Science
author	Junshi Zheng
spellingShingle	Junshi Zheng misc QK1-989 misc 1-deoxy-d-xylulose-5-phosphate synthase 1 misc Kaurenoic acid hydroxylase misc Metabolic engineering misc Stevia transformation misc Steviol glycosides misc Transgenic Stevia misc Botany Overexpression of SrDXS1 and SrKAH enhances steviol glycosides content in transgenic Stevia plants
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topic_title	QK1-989 Overexpression of SrDXS1 and SrKAH enhances steviol glycosides content in transgenic Stevia plants 1-deoxy-d-xylulose-5-phosphate synthase 1 Kaurenoic acid hydroxylase Metabolic engineering Stevia transformation Steviol glycosides Transgenic Stevia
topic	misc QK1-989 misc 1-deoxy-d-xylulose-5-phosphate synthase 1 misc Kaurenoic acid hydroxylase misc Metabolic engineering misc Stevia transformation misc Steviol glycosides misc Transgenic Stevia misc Botany
topic_unstemmed	misc QK1-989 misc 1-deoxy-d-xylulose-5-phosphate synthase 1 misc Kaurenoic acid hydroxylase misc Metabolic engineering misc Stevia transformation misc Steviol glycosides misc Transgenic Stevia misc Botany
topic_browse	misc QK1-989 misc 1-deoxy-d-xylulose-5-phosphate synthase 1 misc Kaurenoic acid hydroxylase misc Metabolic engineering misc Stevia transformation misc Steviol glycosides misc Transgenic Stevia misc Botany
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title	Overexpression of SrDXS1 and SrKAH enhances steviol glycosides content in transgenic Stevia plants
ctrlnum	(DE-627)DOAJ020000669 (DE-599)DOAJ6fe0bf2e950b4b96a8750f9c24c39bb5
title_full	Overexpression of SrDXS1 and SrKAH enhances steviol glycosides content in transgenic Stevia plants
author_sort	Junshi Zheng
journal	BMC Plant Biology
journalStr	BMC Plant Biology
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lang_code	eng
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author_browse	Junshi Zheng Yan Zhuang Hui-Zhu Mao In-Cheol Jang
container_volume	19
class	QK1-989
format_se	Elektronische Aufsätze
author-letter	Junshi Zheng
doi_str_mv	10.1186/s12870-018-1600-2
author2-role	verfasserin
title_sort	overexpression of srdxs1 and srkah enhances steviol glycosides content in transgenic stevia plants
callnumber	QK1-989
title_auth	Overexpression of SrDXS1 and SrKAH enhances steviol glycosides content in transgenic Stevia plants
abstract	Abstract Background Stevia rebaudiana produces sweet-tasting steviol glycosides (SGs) in its leaves which can be used as natural sweeteners. Metabolic engineering of Stevia would offer an alternative approach to conventional breeding for enhanced production of SGs. However, an effective protocol for Stevia transformation is lacking. Results Here, we present an efficient and reproducible method for Agrobacterium-mediated transformation of Stevia. In our attempts to produce transgenic Stevia plants, we found that prolonged dark incubation is critical for increasing shoot regeneration. Etiolated shoots regenerated in the dark also facilitated subsequent visual selection of transformants by green fluorescent protein during Stevia transformation. Using this newly established transformation method, we overexpressed the Stevia 1-deoxy-d-xylulose-5-phosphate synthase 1 (SrDXS1) and kaurenoic acid hydroxylase (SrKAH), both of which are required for SGs biosynthesis. Compared to control plants, the total SGs content in SrDXS1- and SrKAH-overexpressing transgenic lines were enhanced by up to 42–54% and 67–88%, respectively, showing a positive correlation with the expression levels of SrDXS1 and SrKAH. Furthermore, their overexpression did not stunt the growth and development of the transgenic Stevia plants. Conclusion This study represents a successful case of genetic manipulation of SGs biosynthetic pathway in Stevia and also demonstrates the potential of metabolic engineering towards producing Stevia with improved SGs yield.
abstractGer	Abstract Background Stevia rebaudiana produces sweet-tasting steviol glycosides (SGs) in its leaves which can be used as natural sweeteners. Metabolic engineering of Stevia would offer an alternative approach to conventional breeding for enhanced production of SGs. However, an effective protocol for Stevia transformation is lacking. Results Here, we present an efficient and reproducible method for Agrobacterium-mediated transformation of Stevia. In our attempts to produce transgenic Stevia plants, we found that prolonged dark incubation is critical for increasing shoot regeneration. Etiolated shoots regenerated in the dark also facilitated subsequent visual selection of transformants by green fluorescent protein during Stevia transformation. Using this newly established transformation method, we overexpressed the Stevia 1-deoxy-d-xylulose-5-phosphate synthase 1 (SrDXS1) and kaurenoic acid hydroxylase (SrKAH), both of which are required for SGs biosynthesis. Compared to control plants, the total SGs content in SrDXS1- and SrKAH-overexpressing transgenic lines were enhanced by up to 42–54% and 67–88%, respectively, showing a positive correlation with the expression levels of SrDXS1 and SrKAH. Furthermore, their overexpression did not stunt the growth and development of the transgenic Stevia plants. Conclusion This study represents a successful case of genetic manipulation of SGs biosynthetic pathway in Stevia and also demonstrates the potential of metabolic engineering towards producing Stevia with improved SGs yield.
abstract_unstemmed	Abstract Background Stevia rebaudiana produces sweet-tasting steviol glycosides (SGs) in its leaves which can be used as natural sweeteners. Metabolic engineering of Stevia would offer an alternative approach to conventional breeding for enhanced production of SGs. However, an effective protocol for Stevia transformation is lacking. Results Here, we present an efficient and reproducible method for Agrobacterium-mediated transformation of Stevia. In our attempts to produce transgenic Stevia plants, we found that prolonged dark incubation is critical for increasing shoot regeneration. Etiolated shoots regenerated in the dark also facilitated subsequent visual selection of transformants by green fluorescent protein during Stevia transformation. Using this newly established transformation method, we overexpressed the Stevia 1-deoxy-d-xylulose-5-phosphate synthase 1 (SrDXS1) and kaurenoic acid hydroxylase (SrKAH), both of which are required for SGs biosynthesis. Compared to control plants, the total SGs content in SrDXS1- and SrKAH-overexpressing transgenic lines were enhanced by up to 42–54% and 67–88%, respectively, showing a positive correlation with the expression levels of SrDXS1 and SrKAH. Furthermore, their overexpression did not stunt the growth and development of the transgenic Stevia plants. Conclusion This study represents a successful case of genetic manipulation of SGs biosynthetic pathway in Stevia and also demonstrates the potential of metabolic engineering towards producing Stevia with improved SGs yield.
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title_short	Overexpression of SrDXS1 and SrKAH enhances steviol glycosides content in transgenic Stevia plants
url	https://doi.org/10.1186/s12870-018-1600-2 https://doaj.org/article/6fe0bf2e950b4b96a8750f9c24c39bb5 http://link.springer.com/article/10.1186/s12870-018-1600-2 https://doaj.org/toc/1471-2229
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up_date	2024-07-04T01:44:35.174Z
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Overexpression of SrDXS1 and SrKAH enhances steviol glycosides content in transgenic Stevia plants

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