The importance of salicylic acid and an improved plant condition in determining success in agrobacterium-mediated transformation

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Abstract

A succeßful Agrobacterium-mediated transformation requires a balanced interaction between the plant and Agrobacterium. We hypothesize that blocking salicylic acid (SA) biosynthesis could increase the plant's susceptibility and gene transfer events. In this study, the influence of the SA biosynthesis on transformation efficiency was investigated. We tested the effect of increasing concentrations of SA inhibitors such as 2-amidoindane-2-phosphonic acid (AIP) and paclobutrazol (PBZ) in Petunia hybrida and Nicotiana benthamiana. As a control, the effect of adding SA was examined. In addition, the effect of lipoic acid (LA) was studied. LA, an antioxidant, is considered to act as a general transformation enhancer by improving the plant's condition. Adding SA inhibitors should lead to improved conditions for Agrobacterium and adding LA should improve conditions for the plant. Explants from fresh leaves of P. hybrida and N. benthamiana were inoculated with A. tumefaciens strain AGL1(pBinGlyRed-Asc1) containing dsRed as a reporter gene. Transient dsRed expreßion was determined at 14 days after co-cultivation by counting red fluorescing spots and transformation efficiency was determined from the number of explants with calli surviving on selection media at 30 days. In P. hybrida, 10 ?M AIP and PBZ resulted in the highest transient and stable transformation. However in N. benthamiana, only 50 ?M AIP showed a significant increase and only in transient expreßion, and no evidence of any stimulatory effect of PBZ was observed. Adding SA always decreased the efficiencies in both plant species. Furthermore, 10 and 1 ?M LA significantly increased transformation efficiency in P. hybrida and N. benthamiana, respectively, with leß browning and necrosis. Our preliminary results suggested that blocking SA biosynthesis promoted gene transfer but responses were variable and not equally succeßful between the two plant species. However, repeating these experiments providing us with more data might clarify the picture further.

Original languageEnglish
Pages (from-to)65-69
JournalActa Horticulturae
Volume1087
DOIs
Publication statusPublished - 2015

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Agrobacterium
salicylic acid
lipoic acid
Petunia hybrida
Nicotiana benthamiana
phosphorous acid
paclobutrazol
biosynthesis
gene transfer
explants
Agrobacterium radiobacter
reporter genes
necrosis
callus
antioxidants
leaves

Keywords

  • 2-amidoindane-2-phosphonic acid
  • Lipoic acid
  • N. benthamiana
  • P. hybrid
  • Paclobutrazol
  • Transformation efficiency

Cite this

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title = "The importance of salicylic acid and an improved plant condition in determining success in agrobacterium-mediated transformation",
abstract = "A succe{\ss}ful Agrobacterium-mediated transformation requires a balanced interaction between the plant and Agrobacterium. We hypothesize that blocking salicylic acid (SA) biosynthesis could increase the plant's susceptibility and gene transfer events. In this study, the influence of the SA biosynthesis on transformation efficiency was investigated. We tested the effect of increasing concentrations of SA inhibitors such as 2-amidoindane-2-phosphonic acid (AIP) and paclobutrazol (PBZ) in Petunia hybrida and Nicotiana benthamiana. As a control, the effect of adding SA was examined. In addition, the effect of lipoic acid (LA) was studied. LA, an antioxidant, is considered to act as a general transformation enhancer by improving the plant's condition. Adding SA inhibitors should lead to improved conditions for Agrobacterium and adding LA should improve conditions for the plant. Explants from fresh leaves of P. hybrida and N. benthamiana were inoculated with A. tumefaciens strain AGL1(pBinGlyRed-Asc1) containing dsRed as a reporter gene. Transient dsRed expre{\ss}ion was determined at 14 days after co-cultivation by counting red fluorescing spots and transformation efficiency was determined from the number of explants with calli surviving on selection media at 30 days. In P. hybrida, 10 ?M AIP and PBZ resulted in the highest transient and stable transformation. However in N. benthamiana, only 50 ?M AIP showed a significant increase and only in transient expre{\ss}ion, and no evidence of any stimulatory effect of PBZ was observed. Adding SA always decreased the efficiencies in both plant species. Furthermore, 10 and 1 ?M LA significantly increased transformation efficiency in P. hybrida and N. benthamiana, respectively, with le{\ss} browning and necrosis. Our preliminary results suggested that blocking SA biosynthesis promoted gene transfer but responses were variable and not equally succe{\ss}ful between the two plant species. However, repeating these experiments providing us with more data might clarify the picture further.",
keywords = "2-amidoindane-2-phosphonic acid, Lipoic acid, N. benthamiana, P. hybrid, Paclobutrazol, Transformation efficiency",
author = "{Fatihah Binti Hasan Nudin}, Nur and {van Kronenburg-van de Ven}, Bernadette and Iris Tinnenbroek-Capel and F. Krens",
year = "2015",
doi = "10.17660/ActaHortic.2015.1087.7",
language = "English",
volume = "1087",
pages = "65--69",
journal = "Acta Horticulturae",
issn = "0567-7572",
publisher = "International Society for Horticultural Science",

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TY - JOUR

T1 - The importance of salicylic acid and an improved plant condition in determining success in agrobacterium-mediated transformation

AU - Fatihah Binti Hasan Nudin, Nur

AU - van Kronenburg-van de Ven, Bernadette

AU - Tinnenbroek-Capel, Iris

AU - Krens, F.

PY - 2015

Y1 - 2015

N2 - A succeßful Agrobacterium-mediated transformation requires a balanced interaction between the plant and Agrobacterium. We hypothesize that blocking salicylic acid (SA) biosynthesis could increase the plant's susceptibility and gene transfer events. In this study, the influence of the SA biosynthesis on transformation efficiency was investigated. We tested the effect of increasing concentrations of SA inhibitors such as 2-amidoindane-2-phosphonic acid (AIP) and paclobutrazol (PBZ) in Petunia hybrida and Nicotiana benthamiana. As a control, the effect of adding SA was examined. In addition, the effect of lipoic acid (LA) was studied. LA, an antioxidant, is considered to act as a general transformation enhancer by improving the plant's condition. Adding SA inhibitors should lead to improved conditions for Agrobacterium and adding LA should improve conditions for the plant. Explants from fresh leaves of P. hybrida and N. benthamiana were inoculated with A. tumefaciens strain AGL1(pBinGlyRed-Asc1) containing dsRed as a reporter gene. Transient dsRed expreßion was determined at 14 days after co-cultivation by counting red fluorescing spots and transformation efficiency was determined from the number of explants with calli surviving on selection media at 30 days. In P. hybrida, 10 ?M AIP and PBZ resulted in the highest transient and stable transformation. However in N. benthamiana, only 50 ?M AIP showed a significant increase and only in transient expreßion, and no evidence of any stimulatory effect of PBZ was observed. Adding SA always decreased the efficiencies in both plant species. Furthermore, 10 and 1 ?M LA significantly increased transformation efficiency in P. hybrida and N. benthamiana, respectively, with leß browning and necrosis. Our preliminary results suggested that blocking SA biosynthesis promoted gene transfer but responses were variable and not equally succeßful between the two plant species. However, repeating these experiments providing us with more data might clarify the picture further.

AB - A succeßful Agrobacterium-mediated transformation requires a balanced interaction between the plant and Agrobacterium. We hypothesize that blocking salicylic acid (SA) biosynthesis could increase the plant's susceptibility and gene transfer events. In this study, the influence of the SA biosynthesis on transformation efficiency was investigated. We tested the effect of increasing concentrations of SA inhibitors such as 2-amidoindane-2-phosphonic acid (AIP) and paclobutrazol (PBZ) in Petunia hybrida and Nicotiana benthamiana. As a control, the effect of adding SA was examined. In addition, the effect of lipoic acid (LA) was studied. LA, an antioxidant, is considered to act as a general transformation enhancer by improving the plant's condition. Adding SA inhibitors should lead to improved conditions for Agrobacterium and adding LA should improve conditions for the plant. Explants from fresh leaves of P. hybrida and N. benthamiana were inoculated with A. tumefaciens strain AGL1(pBinGlyRed-Asc1) containing dsRed as a reporter gene. Transient dsRed expreßion was determined at 14 days after co-cultivation by counting red fluorescing spots and transformation efficiency was determined from the number of explants with calli surviving on selection media at 30 days. In P. hybrida, 10 ?M AIP and PBZ resulted in the highest transient and stable transformation. However in N. benthamiana, only 50 ?M AIP showed a significant increase and only in transient expreßion, and no evidence of any stimulatory effect of PBZ was observed. Adding SA always decreased the efficiencies in both plant species. Furthermore, 10 and 1 ?M LA significantly increased transformation efficiency in P. hybrida and N. benthamiana, respectively, with leß browning and necrosis. Our preliminary results suggested that blocking SA biosynthesis promoted gene transfer but responses were variable and not equally succeßful between the two plant species. However, repeating these experiments providing us with more data might clarify the picture further.

KW - 2-amidoindane-2-phosphonic acid

KW - Lipoic acid

KW - N. benthamiana

KW - P. hybrid

KW - Paclobutrazol

KW - Transformation efficiency

U2 - 10.17660/ActaHortic.2015.1087.7

DO - 10.17660/ActaHortic.2015.1087.7

M3 - Article

VL - 1087

SP - 65

EP - 69

JO - Acta Horticulturae

JF - Acta Horticulturae

SN - 0567-7572

ER -