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United States Department of Agriculture

Agricultural Research Service

Title: Papaya (Carica Papaya L.)

Authors
item Zhu, Yun - HI AG RESEARCH CENTER
item Fitch, Maureen
item Moore, Paul

Submitted to: Book Chapter
Publication Type: Book / Chapter
Publication Acceptance Date: December 20, 2004
Publication Date: March 20, 2006
Citation: Zhu, Y.J., Fitch, M.M., Moore, P.H. 2006. Papaya (Carica papaya L.). Methods of Molecular, vol 344: Agrobacterium Protocols, 2/e, vol. 2:209-217.

Interpretive Summary: Transgenic papaya plants were initially obtained using particle bombardment, a method having a poor efficiency in producing intact, single copy insertion of transgenes. Subsequently, transformation has been improved using Agrobacterium tumefaciens. With rapid progress being made in genome sequencing and gene discovery, there is a need for quicker, more efficient methods of transformation in order to study the function of these genes. We describe a protocol for Agrobacterium-mediated transformation using carborundum-wounded papaya embryogenic calli. This method should lead to high throughput transformation, which on average produced at least one plant that was positive in PCR, histochemical staining, or by Southern blot hybridization from 10 to 20% of the callus clusters that had been co-cultivation with Agrobacterium. Plants regenerated from the callus clusters in 9 to 13 months.

Technical Abstract: Transgenic papaya plants were initially obtained using particle bombardment, a method having a poor efficiency in producing intact, single copy insertion of transgenes. Subsequently, transformation has been improved using Agrobacterium tumefaciens. With rapid progress being made in genome sequencing and gene discovery, there is a need for quicker, more efficient methods of transformation in order to study the function of these genes. We describe a protocol for Agrobacterium-mediated transformation using carborundum-wounded papaya embryogenic calli. This method should lead to high throughput transformation, which on average produced at least one plant that was positive in PCR, histochemical staining, or by Southern blot hybridization from 10 to 20% of the callus clusters that had been co-cultivation with Agrobacterium. Plants regenerated from the callus clusters in 9 to 13 months.

Last Modified: 7/22/2014
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