Regeneration and Agrobacterium-mediated genetic transformation in Dianthus chinensis

Zhang, Xiaoni; Wu, Quanshu; Lin, Shengnan; Zhang, Zhen; Wang, Zehao; Wang, Qijian; Yan, Xiuli; Bendahmane, Mohammed; Bao, Manzhu; Fu, Xiaopeng

doi:10.1016/j.scienta.2021.110279

Cited by 7 publications

(4 citation statements)

References 39 publications

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“…A suitable concentration of the infection solution is also an important factor for facilitating successful transformation [20,25]. It was found that as the concentration of the infection solution increased, the transformation efficiency first increased and then decreased.…”

Section: Discussionmentioning

confidence: 99%

A Stable and Efficient Genetic Transformation Method for Embryogenic Callus of Fraxinus mandshurica

Dong

Zhao

et al. 2023

Forests

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Fraxinus mandshurica is a widely used greening and ornamental tree species. However, its genetic transformation system has been hampered by problems such as low transformation efficiency, among others, which can hinder research related to molecular breeding and the analysis of functional genes. Thus, in this study, a novel genetic transformation method for efficient transformation of the embryonic callus of Fraxinus mandshurica was investigated. The method was optimized in terms of factors such as antibiotics, infection solution concentrations, co-culture time, and somatic embryo maturation. The results indicated that the optimal antibiotic concentration was 10 mg·L−1 of hygromycin (Hyg). At this point, the callus proliferation multiple was only 0.12. The highest transformation efficiency was found to be 93.93% when the absorbance of the infection solution concentration at OD600 was 0.4. Interestingly, transformation efficiency was found to be highest (77.9%) at 48 h of co-culture, with a GUS staining rate of 88.23%. The medium for somatic embryo maturation of transformed callus was half-strength MS medium (MS 1/2) containing 60 g·L−1 polyethylene glycol, 1 mg·L−1 abscisic acid, 400 mg·L−1 casein enzymatic hydrolysate (CH), 20 g·L−1 sucrose, 1 g·L−1 activated charcoal, and 5 g·L−1 gellan gum. The medium for somatic embryo germination was MS ½, containing 0.2 mg·L−1 of N-(Phenylmethyl)-9H-purin-6-amine(6-BA) and 5.0 mg·L−1 of gibberellin (GA). These results are of significance for the verification of the gene function and future genetic improvement of Fraxinus mandshurica.

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Section: Discussionmentioning

confidence: 99%

A Stable and Efficient Genetic Transformation Method for Embryogenic Callus of Fraxinus mandshurica

Dong

Zhao

et al. 2023

Forests

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“…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

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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 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 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%

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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Regeneration and Agrobacterium-mediated genetic transformation in Dianthus chinensis

Cited by 7 publications

References 39 publications

A Stable and Efficient Genetic Transformation Method for Embryogenic Callus of Fraxinus mandshurica

A Stable and Efficient Genetic Transformation Method for Embryogenic Callus of Fraxinus mandshurica

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

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

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