Sweet pepper ferredoxin-like protein ( pflp) gene as a novel selection marker for orchid transformation.
A novel method for selection of transgenic plants utilizing the sweet pepper ( Capsicum annuum L.) ferredoxin-like protein ( pflp) gene as selection marker and Erwinia carotovora as the selection agent has been developed. An expression vector containing a pflp cDNA driven by a cauliflower mosaic virus 35S promoter was successfully transformed into protocorm-like bodies of Oncidium orchid by Agrobacterium tumefaciens and particle bombardment, respectively. Erwinia carotovora was used as a selection agent to screen transformants, thereby obtaining transgenic plants without the use of an antibiotic selection agent. A total of 32 independent transgenic orchid lines were obtained, out of which 9 transgenic lines (beta-glucuronidase positive) were randomly selected and confirmed by Southern and northern blot analyses. The transgenic orchid plants showed enhanced resistance to E. carotovora, even when the entire plant was challenged with the pathogen. Our results suggest the novel use of the pflp gene as a resistance selection marker in plant genetic engineering strategies. In the future, the use of the pflp gene as a selection marker may facilitate the use of smaller gene constructs due to removal of bulky antibiotic selection and reporter genes. These constructs can then be used to incorporate additional genes of choice.
Institute of BioAgricultural Sciences, Academia Sinica, 11529, Taipei, Taiwan, Republic of China.
Su-Juan
SJ
You
Chia-Hui
CH
Liau
Hsiang-En
HE
Huang
Teng-Yung
TY
Feng
Venkatesh
V
Prasad
Hsin-hao
HH
Hsiao
Jian-Cheng
JC
Lu
Ming-Tsair
MT
Chan
D016960:Agrobacterium tumefaciens / Q000235:genetics; D002934:Cinnamates; D005288:Ferredoxins / Q000235:genetics*; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D005819:Genetic Markers / Q000235:genetics; D006921:Hygromycin B / Q000031:analogs & derivatives* / Q000494:pharmacology; D007113:Immunity, Innate / Q000235:genetics; D009154:Mutation; D029595:Orchidaceae / Q000235:genetics* / Q000378:metabolism; D016973:Pectobacterium carotovorum / Q000254:growth & development; D010935:Plant Diseases / Q000382:microbiology; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic / Q000235:genetics*
D002934:Cinnamates; D005288:Ferredoxins; D005819:Genetic Markers; D010940:Plant Proteins; D012333:RNA, Messenger; C474922:plant ferredoxin-like protein, Capsicum annuum; D006921:Hygromycin B; C026273:hygromycin A