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Journal of Forestry Research

, Volume 13, Issue 2, pp 91–97 | Cite as

Genetic transformation ofPinus taeda by particle bombardment

  • Tang Wei
  • Vanessa Samuels
Article

Abstract

A protocol is presented for genetically engineering loblolly pine (Pinus taeda L.) using particle bombardment. This protocol enabled the routine transformation of loblolly pine plants that were previously difficult to transform. Mature zygotic embryos were used to be bombarded and to generate organogenic callus and transgenic regenerated plants. Plasmid pB48.215 DNA contained a synthetic Bacillus thuringiensis (B.t.) crylAc coding sequence flanked by the double cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase (Nos) terminator sequences, and the selectable marker gene, neomycin phosphotransferase II (nptil) controlled by the promoter of the nopaline synthase gene was introduced into loblolly pine tissues by particle bombardment. The transformed tissues were proliferated and selected by kanamycin resistance conferred by the introduced NPTIi gene. Shoot regeneration was induced from the kanamycin-resistant callus, and transgenic plantlets were then produced. The presence of the introduced genes in the transgenic loblolly pine plants was confirmed by polymerase chain reactions (PCR) analysis, by Southern blot analysis, and insect feeding assays. The recovered transgenic plants were acclimatized and then established in soil.

Keywords

Pinus taeda L. Biolistic transformation Bacillus thuringiensis (B.t.) crylAb Insect feeding bioassay 

CLC

Q55 S791.255.04 

Document code

Abbreviations

BA

benzyladenine

B.t.

Bacillus thuringiensis

CaMV

cauliflower mosaic virus

2,4-D

2,4-dichlorophenoxyacetic acid

IBA

indoie-3-butyric acid

Nos

nopaline synthase

nptll

neomycin phosphotransferase II gene

PCR

polymerase chain reactions

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Copyright information

© Northeast Forestry University 2002

Authors and Affiliations

  1. 1.Forest Biotechnology GroupNorth Carolina State UniversityRaleighUSA

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