Factors affecting Agrobacterium -mediated transformation efficiency of kumquat seedling internodal stem segments

Yang, Li; Hu, Wei; Yu-ming, Xie; Li, Yang; Deng, Zhili

doi:10.1016/j.scienta.2016.06.018

Cited by 8 publications

(7 citation statements)

References 27 publications

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“…Earlier studies used both etiolated seedlings and light-grown seedlings for citrus transformation [ 18 , 19 , 20 , 21 , 22 , 23 ]. Our study shows higher transformation efficiency can be achieved using explants derived from light-grown seedlings, consistent with the study in kumquat that a relatively higher transformation efficiency was obtained when light-grown seedlings were used as explant source [ 29 ]. It is known that light reduces auxin levels in plant tissues [ 30 , 31 ].…”

Section: Discussionsupporting

confidence: 91%

An Improved Procedure for Agrobacterium-Mediated Transformation of ‘Carrizo’ Citrange

Tang

Liu

et al. 2022

Plants

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Although several protocols for genetic transformation of citrus have been published, it is highly desirable to further improve its efficiency. Here we report treatments of Agrobacterium cells and citrus explants prior to and during co-cultivation process to enhance transformation efficiency using a commercially used rootstock ‘Carrizo’ citrange [Citrus sinensis (L.) Osb. × Poncirius trifoliata (L.) Raf.] as a model plant. We found explants from light-grown seedlings exhibited higher transformation efficiency than those from etiolated seedlings. We pre-cultured Agrobacterium cells in a 1/10 MS, 0.5 g/L 2-(N-morpholino) ethanesulfonic acid (MES) and 100 µM acetosyringone liquid medium for 6 h at 25 °C before used to infect citrus explants. We incubated epicotyl segments in an MS liquid medium containing 13.2 µM 6-BA, 4.5 µM 2,4-D, 0.5 µM NAA for 3 h at 25 °C prior to Agrobacterium infection. In the co-cultivation medium, we added 30 µM paclobutrazol and 10 µM lipoic acid. Each of these treatments significantly increased the efficiencies of transformation up to 30.4% (treating Agrobacterium with acetosyringone), 31.8% (treating explants with cytokinin and auxin), 34.9% (paclobutrazol) and 38.6% (lipoic acid), respectively. When the three treatments were combined, we observed that the transformation efficiency was enhanced from 11.5% to 52.3%. The improvement of genetic transformation efficiency mediated by these three simple treatments may facilitate more efficient applications of transgenic and gene editing technologies for functional characterization of citrus genes and for genetic improvement of citrus cultivars.

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Section: Discussionsupporting

confidence: 91%

An Improved Procedure for Agrobacterium-Mediated Transformation of ‘Carrizo’ Citrange

Tang

Liu

et al. 2022

Plants

Self Cite

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show abstract

“…Earlier studies used both etiolated seedlings and light-grown seedlings for citrus transformation [18][19][20][21][22][23]. Our study shows higher transformation efficiency can be achieved using explants derived from light-grown seedlings, consistent with the study in kumquat that a relatively higher transformation efficiency was obtained when light-grown seedlings were used as explant source [29]. It is known that light reduces auxin levels in plant tissues [30][31].…”

Section: Discussionsupporting

confidence: 88%

An Improved Procedure for Agrobacterium-Mediated Transformation of Citrus

Li¹,

Tang²,

Liu³

et al. 2022

Preprint

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Although several protocols for genetic transformation of citrus have been published, it is highly desirable to further improve its efficiency. Here we report treatments of Agrobacterium cells and citrus explants prior to and during co-cultivation process to enhance transformation efficiency using a commercially used rootstock 'Carrizo' Citrange as a model plant. We pre-cultured Agrobacterium cells in a 1/10 MS, 0.5 g/L 2-(N-morpholino) ethanesulfonic acid (MES) and 100 µM acetosyringone liquid medium for 6 hours at 25 ℃ before used to infect citrus explants. We incubated epicotyl segments in an MS liquid medium containing 13.2 µM 6-BA, 4.5 µM 2,4-D, 0.5 µM NAA for 3 hours at 25℃ prior to Agrobacterium infection. In the co-cultivation medium, we added 30 µM paclobutrazol and 10 µM lipoic acid. Each of these three treatments significantly increased the efficiencies of transformation. When the three treatments were combined, we observed that the transformation efficiency was enhanced from 11.5% to 52.3%. The improvement of genetic transformation efficiency mediated by these three simple treatments may facilitate more efficient applications of transgenic and gene editing technologies for functional characterization of citrus genes and for genetic improvement of cultivated citrus varieties.

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“…This value is lower than the efficiency reported for other mandarin cultivars (KHAWALE et al, 2006;BACHCHU et al, 2011). Besides genotype dependence and regeneration frequency, the genetic transformation efficiency is influenced by other factors, including the Agrobacterium strain and density and the co-cultivation procedure (DUTT; GROSSER, 2009;YANG et al, 2016). The genetic transformation efficiency may also be influenced by gene constructs.…”

Section: Genetic Transformationmentioning

confidence: 72%

“…However, some species, such as sour orange (GUTIÉRREZ et al 1997;GHORBEL et al, 2000), Rangpur lime , and mandarins are considered recalcitrant to the process. This low genetic transformation efficiency may be influenced by factors related to Agrobacterium inoculation procedure, co-cultivation conditions, and in vitro regeneration efficiency of explants after infection with Agrobacterium (COSTA et al, 2002;RODRIGUEZ et al, 2008;GROSSER, 2009;YANG et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

In vitro organogenesis and genetic transformation of mandarin cultivars

et al. 2019

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The in vitro organogenesis of Fremont (Citrus clementina x ), Citrus reticulataThomas (Citrus reticulata), and Nules (Citrus clementina) mandarins was evaluated aiming to optimize a regeneration protocol that could be applied in genetic transformation. The use of epicotyl-derived explants resulted in higher explant responsiveness and number of shoots developed per explant when compared with the use of internodal-derived explants. The highest efficiency in shoot regeneration was observed in the presence of 1 mg L-1 of BAP, regardless of the explant type and cultivar. The in vitro organogenesis protocol produced transgenic plants from three mandarin cultivars expressing attA gene under the control of phloem-specific promoters.

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Factors affecting Agrobacterium -mediated transformation efficiency of kumquat seedling internodal stem segments

Cited by 8 publications

References 27 publications

An Improved Procedure for Agrobacterium-Mediated Transformation of ‘Carrizo’ Citrange

An Improved Procedure for Agrobacterium-Mediated Transformation of ‘Carrizo’ Citrange

An Improved Procedure for Agrobacterium-Mediated Transformation of Citrus

In vitro organogenesis and genetic transformation of mandarin cultivars

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