Stable transformation and regeneration of transgenic plants of Pinus radiata D.!Don

被引:91
作者
Walter, C [1 ]
Grace, LJ
Wagner, A
White, DWR
Walden, AR
Donaldson, SS
Hinton, H
Gardner, RC
Smith, DR
机构
[1] Forest Res Inst Ltd, Sala St Rotorua, New Zealand
[2] AgResearch, Palmerston North, New Zealand
[3] Univ Auckland, Sch Biol Sci, Auckland 1, New Zealand
关键词
Pinus radiata; transformation; biolistic; embryogenic tissue;
D O I
10.1007/s002990050426
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
A biolistic particle delivery system was used to genetically transform embryogenic tissue of Pinus radiata. The introduced DNA contained a uidA reporter gene under the control of either the tandem CaMV 35S or the artificial Emu promoter, and the npt II selectable marker controlled by the CaMV 35S promoter. The average number of stable, geneticin-resistant lines recovered was 0.5 per 200 mg fresh weight bombarded tissue. Expression of the uidA reporter gene was detected histochemically and fluorimetrically in transformed embryogenic tissue and in derived mature somatic embryos and regenerated plants. The integration of uidA and npt II genes into the Pinus radiata genome was demonstrated using PCR amplification of the inserts and Southern hybridisation analysis. The expression of both genes in transformed tissue was confirmed by Northern hybridisation analysis. More than 150 transgenic Pinus radiata plants were produced from 20 independent transformation experiments with four different embryogenic clones.
引用
收藏
页码:460 / 468
页数:9
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