Antibiotic resistance markers for plant transformation

Angenon, Geert; Dillen, Willy; Montagu, Marc Van

doi:10.1007/978-94-011-0511-8_9

Cited by 17 publications

(12 citation statements)

References 32 publications

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“…Sjahril et al (2006) reported that 25 mg L -1 hygromycin was used in the regeneration medium for selection of putative transgenic Phalaenopsis orchid plantlets. The results are also consistent with those obtained by Angenon et al (1994) and Cosson et al (2015) as who reported that the selection agent should fully inhibit growth of untransformed plant cells, therefore the lowest concentration of the selection agent that suppresses growth of untransformed cells is generally used. They also demonstrated that the sensitivity of plant cells to the selection agent depends upon the genotype, the explant type and the developmental stage.…”

Section: Discussionsupporting

confidence: 94%

Molecular cloning and expression of a vacuolar Na+/H+ antiporter gene (AgNHX1) in fig (Ficus carica L.) under salt stress

Metwali

Soliman

Fuller

et al. 2015

Plant Cell Tiss Organ Cult

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Soil salinity can be a major limiting factor for productivity in agriculture and forestry and in order to fully utilize saline lands productively in plantation forestry for fig production, the genetic modification of tree species for salt tolerance may be required. Na ? /H ? antiporters have been suggested to play important roles in salt tolerance in plants.Here, we isolated AgNHX1 a vacuolar Na ? /H ? antiporter from a halophytic species Atriplex gmelini and introduced it into fig (Ficus carica L.) cv. Black Mission via Agrobacterium-mediated transformation. Leaf discs explants of fig were co-cultivated for 2 days with Agrobacterium tumefaciens strain LBA 4404 harboring the binary vector pBI121 containing the AgNHX1 gene and the hpt selectable marker gene which encodes hygromycin phosphotransferase. Explants were cultured on MS medium containing 30 mg L -1 hygromycin, 3 % sucrose, 0.2 mg L -1 kinetin and 2.0 mg L -1 2,4-dichlorophenoxyacetic acid solidified with 2.5 g L -1 phytagel in darkness for callus formation. The calli were cultured on MS medium containing 2.0 mg L -1 zeatin riboside in combination with 0.4 mg L -1 indole acetic acid in the light for plant regeneration. Putative regenerated transformant shoots were confirmed by polymerase chain reaction (PCR) and Southern hybridization for the AgNHX1 gene. Reverse transcriptase polymerase chain reaction analysis indicated that the gene was highly expressed in transgenic plants, but the degree of this expression varied among transformants. Overexpression of the AgNHX1 gene conferred high tolerance to salt stress and transgenic fig plants overexpressing AgNHX1 developed normally under salinity conditions compared to those of non-transgenic plants. Salt treated transgenic plants contained high proline and K ? but less Na ? compared to non-transgenic control plants.

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Section: Discussionsupporting

confidence: 94%

Molecular cloning and expression of a vacuolar Na+/H+ antiporter gene (AgNHX1) in fig (Ficus carica L.) under salt stress

Metwali

Soliman

Fuller

et al. 2015

Plant Cell Tiss Organ Cult

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“…During plant transformation experiments, positive selection of transgenic plants from a non-transgenic background is prerequisite to the generation of stable transgenic plants (Angenon et al 1994). A large number of positive selectable marker genes have thus been developed, such as antibiotics, antimetabolite and herbicide resistance genes (Miki and McHugh 2004).…”

Section: Introductionmentioning

confidence: 99%

The 35S promoter used in a selectable marker gene of a plant transformation vector affects the expression of the transgene

Yoo

Bomblies

Yoo

et al. 2005

Planta

147

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Positive selection of transgenic plants is essential during plant transformation. Thus, strong promoters are often used in selectable marker genes to ensure successful selection. Many plant transformation vectors, including pPZP family vectors, use the 35S promoter as a regulatory sequence for their selectable marker genes. We found that the 35S promoter used in a selectable marker gene affected the expression pattern of a transgene, possibly leading to a misinterpretation of the result obtained from transgenic plants. It is likely that the 35S enhancer sequence in the 35S promoter is responsible for the interference, as in the activation tagging screen. This affected expression mostly disappeared in transgenic plants generated using vectors without the 35S sequences within their T-DNA region. Therefore, we suggest that caution should be used in selecting a plant transformation vector and in the interpretation of the results obtained from transgenic approaches using vectors carrying the 35S promoter sequences within their T-DNA regions.

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“…Одним из методов повышения устойчивости растений к фитопатогенам может быть трансформация растений генами антимикробных пептидов, обладающих широким спектром антибиотической и фунгицидной активности. Для отбора трансгенных растений традиционно используют селективные гены устойчивости к антибиотикам и гербицидам или гены репортеры [Angenon et al, 1994]. Полученные с помощью селективных маркеров трансгенные растения представляют потенциальную биологическую опасность, связанную с присутствием в их геноме этих генов и с возможностью их неконтролируемого переноса другим растениям и микроорганизмам.…”

Section: получение биобезопасных безмаркерных растенийunclassified

PRODUCING BIOLOGICALLY SAFE MARKER-FREE CAMELINA SATIVA WITH INCREASED RESISTANCE TO PHYTOPATHOGENS

Захарченко

Furs

Dyachenko

et al. 2018

Book of Proceedings of the All-Russian Scientific Conference With International Participation and Schools of Young Scientists

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Abstract. The marker-free Camelina sativa (L.) plants carrying a gene encoding the antimicrobial peptide cecropin P1 were generated. The vector, free of any plant selective genes of resistance to antibiotics or herbicides, was used for transformation. The transformants were screened by detecting cecropin P1 in plant cells according to the antibacterial activity of plant extracts and enzyme immunoassay. The resulting marker-free plants displayed a considerably increased resistance to phytopathogens.

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Antibiotic resistance markers for plant transformation

Cited by 17 publications

References 32 publications

Molecular cloning and expression of a vacuolar Na+/H+ antiporter gene (AgNHX1) in fig (Ficus carica L.) under salt stress

Molecular cloning and expression of a vacuolar Na+/H+ antiporter gene (AgNHX1) in fig (Ficus carica L.) under salt stress

The 35S promoter used in a selectable marker gene of a plant transformation vector affects the expression of the transgene

PRODUCING BIOLOGICALLY SAFE MARKER-FREE CAMELINA SATIVA WITH INCREASED RESISTANCE TO PHYTOPATHOGENS

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