A WOUND-INDUCED PROMOTER DRIVING NPT-II EXPRESSION LIMITED TO DEDIFFERENTIATED CELLS AT WOUND SITES IS SUFFICIENT TO ALLOW SELECTION OF TRANSGENIC SHOOTS


FIREK S., OZCAN S., WARNER S., DRAPER J.

PLANT MOLECULAR BIOLOGY, sa.1, ss.129-142, 1993 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Basım Tarihi: 1993
  • Doi Numarası: 10.1007/bf00039001
  • Dergi Adı: PLANT MOLECULAR BIOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.129-142
  • Anahtar Kelimeler: COMPETENT PLANT CELLS, CONTROL OF MARKER GENE EXPRESSION, PLANT GENETIC ENGINEERING, PLANT TRANSFORMATION, TRANSFORMATION MARKER GENE, WOUND-SITE-SPECIFIC PROMOTER, PATHOGENESIS-RELATED PROTEINS, AGROBACTERIUM-TUMEFACIENS, GENE-EXPRESSION, PLANT TRANSFORMATION, TOBACCO PROTOPLASTS, BIOLOGICAL FUNCTION, CHITINASE PROMOTER, INFECTED TOBACCO, MESOPHYLL-CELLS, PR PROTEINS
  • Ankara Üniversitesi Adresli: Evet

Özet

There is much data to indicate that only a small number of cells in plant explants are competent for stable transformation by Agrobacterium. Circumstantial evidence suggests that certain cells reentering cell division at wound sites are competent for transformation by Agrobacterium. We have discovered a member of the intracellular PR gene family from asparagus (AoPR1) which is strongly expressed upon wounding and during the reactivation of the cell cycle in cultured asparagus cells, but which shows very little expression in intact plant tissues. The promoter from the AoPR1 gene was fused to an intron-containing GUS reporter gene and shown to be more strongly expressed than the commonly used CaMV 35S constitutive promoter in target cells for plant transformation. A transcriptional fusion of the AoPR1 promoter with an NPT-II gene was found to be a very efficient marker for the selection of transgenic tobacco callus. Expression of the AoPR1-NPT-II gene allowed efficient shoot formation on transgenic callus and efficient adventitious root formation on transgenic shoots. These latter observations provided firm evidence that transformation selection marker gene expression is most crucial at the early stages of the transformation process, during the establishment of transformed micro-calli.