Papaya (Carica papaya L.).
Transgenic papaya plants were initially obtained using particle bombardment, a method having poor efficiency in producing intact, single-copy insertion of transgenes. Single-copy gene insertion was improved using Agrobacterium tumefaciens. With progress being made in genome sequencing and gene discovery, there is a need for 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 polymerase chain reaction (PCR), histochemical staining, or by Southern blot hybridization from 10 to 20% of the callus clusters that had been co-cultivated with Agrobacterium. Plants regenerated from the callus clusters in 9 to 13 mo.
Zhu
Yun J
YJ
Hawaii Agriculture Research Center, Aiea, HI, USA.
Fitch
Maureen M M
MM
Moore
Paul H
PH
D016960:Agrobacterium tumefaciens / Q000166:cytology / Q000235:genetics*; D017554:Carbon Compounds, Inorganic / Q000494:pharmacology; D029441:Carica / Q000187:drug effects / Q000196:embryology / Q000235:genetics*; D018929:Cell Culture Techniques; D018920:Coculture Techniques; D003470:Culture Media; D018744:DNA, Plant / Q000737:chemistry; D005966:Glucuronidase / Q000032:analysis; D018546:Hypocotyl / Q000196:embryology / Q000254:growth & development / Q000502:physiology; D018515:Plant Leaves / Q000235:genetics / Q000502:physiology; D018517:Plant Roots / Q000235:genetics / Q000254:growth & development; D030821:Plants, Genetically Modified / Q000254:growth & development; D012038:Regeneration; D012825:Silicon / Q000494:pharmacology; D014170:Transformation, Genetic; D019076:Transgenes; D014414:Tungsten / Q000494:pharmacology
D017554:Carbon Compounds, Inorganic; D003470:Culture Media; D018744:DNA, Plant; C517455:carborundum; D005966:Glucuronidase; D014414:Tungsten; D012825:Silicon