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2012•Journal of Nuclear Agricultural SciencesRequires access

OPTIMIZATION OF Agrobacterium tumefaciens-MEDIATED TRANSFORMATION SYSTEM OF PEANUT

Jingshan Wang

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Abstract

By evaluating GUS transient expression frequency using embryonic leaflet and callus as explants,several factors which may affect genetic transformation mediated by Agrobacterium tumefaciens were studied.The results showed that Agrobacterium suspension supplemented with 150mg/L MES or 0.50ml/30ml tobacco extract could significantly improve the transformation efficiency.Acupuncture treatment significantly increased the GUS transient expression frequency by 29.1% with multi-dot staining compared with CK(12.1%).High-frequency GUS transient expression of multi-dot staining(70.1%) was obtained using callus as the transformation receptor.Resistant plantlets obtained using this transformation system were grafted and transplanted to the field,then the To positive plants were identified by PCR analysis and the target gene was inherited steadily in to some of T1 plants.

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What this paper is about

By evaluating GUS transient expression frequency using embryonic leaflet and callus as explants,several factors which may affect genetic transformation mediated by Agrobacterium tumefaciens were studied.The results showed that Agrobacterium suspension supplemented with 150mg/L MES or 0.50ml/30ml tobacco extract could significantly improve the transformation efficiency.Acupuncture treatment significantly increased the GUS transient expression frequency by 29.1% with multi-dot staining compared with CK(12.1%).High-frequency GUS transient expression of multi-dot staining(70.1%) was obtained using callus as the transformation receptor.Resistant plantlets obtained using this transformation system were grafted and transplanted to the field,then the To positive plants were identified by PCR analysis and the target gene was inherited steadily in to some of T1 plants.

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Available abstract

By evaluating GUS transient expression frequency using embryonic leaflet and callus as explants,several factors which may affect genetic transformation mediated by Agrobacterium tumefaciens were studied.The results showed that Agrobacterium suspension supplemented with 150mg/L MES or 0.50ml/30ml tobacco extract could significantly improve the transformation efficiency.Acupuncture treatment significantly increased the GUS transient expression frequency by 29.1% with multi-dot staining compared with CK(12.1%).High-frequency GUS transient expression of multi-dot staining(70.1%) was obtained using callus as the transformation receptor.Resistant plantlets obtained using this transformation system were grafted and transplanted to the field,then the To positive plants were identified by PCR analysis and the target gene was inherited steadily in to some of T1 plants.

Key concepts: Agrobacterium tumefaciens, Callus, Transformation (genetics), Agrobacterium, Explant culture, Biology, Molecular biology, Botany

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