Starvation Treatment Increases Transient Expression Frequency of a Foreign Gene Introduced into Eustoma grandiflorum Root Tissues by Particle Bombardment.

Takahashi, Misa; Irifune, Kohei; Hasegawa, Masanori; Kuroda, Hiroyuki; Nishizawa, Shuji; Morikawa, Hiromichi

doi:10.5511/plantbiotechnology1984.13.311

Cited by 3 publications

(2 citation statements)

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“…Moreover, in another study on Lilium, MS medium free of NH 4 NO 3 , KNO 3 , and CaCl 2 improved transformation e ciency to 25.4% [20]. In Eustoma grandi orum, 4 to 8 times higher expression frequency was observed when nitrogen-depleted medium was used for cocultivation [31]. Furthermore, Shara et al indicated that removal of NH 4 NO 3, KH 2 PO 4 and KNO 3 signi cantly increased the transformation e ciency of Papaver bracteatum to 34.3% [32].…”

Section: Discussionmentioning

confidence: 91%

Melatonin strongly enhances the Agrobacterium- mediated transformation of carnation in nitrogen-depleted inoculation and co-cultivation media

Aalami

Azadi

Hadizadeh

et al. 2023

Preprint

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With the rising demand for new cultivars of carnation, efficient transformation protocols are needed to enable the bioengineering of new traits. Here, we established a novel and efficient Agrobacterium-mediated transformation system using callus as the target explant for four commercial carnation cultivars. Leaf-derived calli of all cultivars were inoculated with Agrobacterium tumefaciens strain LBA4404 containing the plasmid pCAMBIA 2301 harboring genes for β-glucuronidase (uidA) and neomycin phosphotransferase (nptII). Polymerase chain reaction (PCR) and histochemical assays confirmed the presence of uidA and β-glucuronidase (GUS), respectively in transgenic shoots. The effect on transformation efficiency of medium composition and the presence of antioxidants during inoculation and co-cultivation was investigated. The transformation efficiency was increased to 5% in Murashige and Skoog (MS) medium lacking KNO3 and NH4NO3, compared to 0.6% efficiency in full-strength medium. The omission of other components including the entire macro- and microelements, Fe or vitamins from MS medium was also observed to increase gene transfer. Transformation efficiency was increased dramatically to 24.4% across all carnation cultivars by the addition of 2 mg/l melatonin to nitrogen-depleted MS medium. Shoot regeneration was also doubled in this treatment. The establishment of this efficient and reliable transformation protocol can advance the development of novel carnation cultivars through molecular breeding approaches.

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Section: Discussionmentioning

confidence: 91%

Melatonin strongly enhances the Agrobacterium- mediated transformation of carnation in nitrogen-depleted inoculation and co-cultivation media

Aalami

Azadi

Hadizadeh

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…Moreover, in another study on Lilium, MS medium free of NH 4 NO 3 , KNO 3 , and CaCl 2 improved transformation efficiency to 25.4% [ 25 ]. In Eustoma grandiflorum , 4 to 8 times higher expression frequency was observed when nitrogen-depleted medium was used for co-cultivation [ 42 ]. Furthermore, Sharafi et al indicated that removal of NH 4 NO 3, KH 2 PO 4 and KNO 3 significantly increased the transformation efficiency of Papaver bracteatum to 34.3% [ 43 ].…”

Section: Discussionmentioning

confidence: 99%

Melatonin strongly enhances the Agrobacterium- mediated transformation of carnation in nitrogen-depleted media

et al. 2023

View full text Add to dashboard Cite

With the rising demand for new cultivars of carnation, efficient transformation protocols are needed to enable the bioengineering of new traits. Here, we established a novel and efficient Agrobacterium-mediated transformation system using callus as the target explant for four commercial carnation cultivars. Leaf-derived calli of all cultivars were inoculated with Agrobacterium tumefaciens strain LBA4404 containing the plasmid pCAMBIA 2301 harboring genes for β-glucuronidase (uidA) and neomycin phosphotransferase (nptII). Polymerase chain reaction (PCR) and histochemical assays confirmed the presence of uidA and β-glucuronidase (GUS), respectively in transgenic shoots. The effect on transformation efficiency of medium composition and the presence of antioxidants during inoculation and co-cultivation was investigated. The transformation efficiency was increased in Murashige and Skoog (MS) medium lacking KNO3 and NH4NO3, and also in MS medium lacking macro and micro elements and Fe to 5% and 3.1% respectively, compared to 0.6% in full-strength medium. Transformation efficiency was increased dramatically to 24.4% across all carnation cultivars by the addition of 2 mg/l melatonin to nitrogen-depleted MS medium. Shoot regeneration was also doubled in this treatment. The establishment of this efficient and reliable transformation protocol can advance the development of novel carnation cultivars through molecular breeding approaches.

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Genetic Transformation of Carnation (Dianthus caryophylus L.)

Nontaswatsri

Fukai

2009

Protocols for in Vitro Propagation of Ornamental Plants

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This chapter describes a rapid and efficient protocol for explant preparation and genetic transformation of carnation. Node explants from greenhouse-grown plants and leaf explants from in vitro plants are infected with Agrobacterium tumefaciens AGL0 harboring pKT3 plasmid, consisting of GUS and NPTII genes. Explant preparation is an important factor to obtain the transformed plants. The GUS-staining area was located only on the cut end of explants and only explants with a cut end close to the connecting area between node and leaf, produced transformed shoots. The cocultivation medium is also an important factor for the successful genetic transformation of carnation node and leaf explants. High genetic transformation efficiency of node and leaf explants cocultured with Agrobacterium tumefaciens was achieved when the explants were cocultivated on a filter paper soaked with water or water and acetosyringone mixture (AS).

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Starvation Treatment Increases Transient Expression Frequency of a Foreign Gene Introduced into Eustoma grandiflorum Root Tissues by Particle Bombardment.

Cited by 3 publications

References 1 publication

Melatonin strongly enhances the Agrobacterium- mediated transformation of carnation in nitrogen-depleted inoculation and co-cultivation media

Melatonin strongly enhances the Agrobacterium- mediated transformation of carnation in nitrogen-depleted inoculation and co-cultivation media

Melatonin strongly enhances the Agrobacterium- mediated transformation of carnation in nitrogen-depleted media

Genetic Transformation of Carnation (Dianthus caryophylus L.)

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