Shoot Regeneration and Agrobacterium-Mediated Transformation of Carnation

Altvorst, Anne-Claire van

doi:10.17660/actahortic.1995.420.24

Cited by 5 publications

(6 citation statements)

References 46 publications

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“…Previous studies reported low transformation e ciency in carnation using several approaches [5,6,9]. In the present study, we signi cantly improved the transformation e ciency of four carnation cultivars.…”

Section: Discussionsupporting

confidence: 48%

“…The type of the target explant is one of the determining factors in the success of transformation system since the regeneration competency of cell types, tissues and organs varies considerably following infection with A. tumefaciens [10]. So far, explants including internodes, leaves, and cotyledons [11,12,13], stems [14], and petals [15,5] have been used as target explant for carnation transformation. Callus has not been utilized in carnation transformation, likely due to challenges encountered during callus regeneration [16].…”

Section: Introductionmentioning

confidence: 99%

“…Carnation transformation using Agrobacterium has been practiced in several investigations, most of which reported rather low transformation e ciency (less than 10%) [5,6,7,8,9]. However, a lack of a transformation system with a high rate of cultivar-independent gene transfer has been remained a principal constraint to the further molecular breeding of this species.…”

Section: Introductionmentioning

confidence: 99%

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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: Discussionsupporting

confidence: 48%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

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

“…Carnation transformation using Agrobacterium has been practiced in several investigations, most of which reported rather low transformation efficiency (less than 10%) [ 6 – 10 ]. However, a lack of a transformation system with a high rate of cultivar-independent gene transfer has been remained a principal constraint to the further molecular breeding of this species.…”

Section: Introductionmentioning

confidence: 99%

“…The type of the target explant is one of the determining factors in the success of transformation system since the regeneration competency of cell types, tissues and organs varies considerably following infection with A. tumefaciens [ 11 , 12 ]. So far, explants including internodes, leaves, and cotyledons [ 13 – 15 ], stems [ 16 ], and petals [ 6 , 17 ] have been used as target explant for carnation transformation. Callus has not been utilized in carnation transformation, likely due to challenges encountered during callus regeneration [ 18 ].…”

Section: Introductionmentioning

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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The role of ethylene in the senescence of carnation flowers, a review

Altvorst

Bovy

1995

Plant Growth Regul

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The senescence of flower petals is a highly regulated developmental process which requires active gene expression and protein synthesis. The biochemical changes associated with petal senescence in carnation flowers include an increase in hydrolytic enzymes, degradation of macro-molecules, increased respiratory activity and a climacteric-like increase in ethylene production. It is clear that the gaseous phytohormone ethylene plays a critical role in the regulation and coordination of senescence processes. Many reviews on physiology and mode of action of ethylene are available. Molecular cloning led to the isolation of genes involved in ethylene biosynthesis and action. This review describes the current status of the studies on regulation of ethylene biosynthesis and ethylene response in carnation flowers. An overview is given of studies on senescence-related gene expression and possibilities to improve postharvest longevity by genetic engineering.Abbreviations: ACC = 1 aminocyclopropane-1-carboxylic acid; AIB = o-amino-isobutyric acid; AOA = amino oxyacetic acid; AVG = aminoethoxyvinyl glycine; DACP = diazocyclopentadiene; EFE = ethylene forming enzyme; MACC = malonyl 1 aminocyclopropane-1-carboxylic acid; MTA = S-methylthio-adenosine; NBD = 2,5 norbomadiene; ppb = parts per billion; SAM = S-adenosyl-methionine; STS = silver thiosulphate.

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Shoot Regeneration and Agrobacterium-Mediated Transformation of Carnation

Cited by 5 publications

References 46 publications

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

The role of ethylene in the senescence of carnation flowers, a review

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