Zahra Ghorbanzade1, Mohammad Ahmadabadi1. 1. Department of Biotechnology, Faculty of Agriculture, Azarbaijan Shahid Madani University, Tabriz, Iran.
Abstract
BACKGROUND: A highly efficient genetic transformation system is essential for a successful genetic manipulation of the African violet (Saintpaulia ionantha Wendl.). OBJECTIVES: Developing a particle bombardment-based genetic transformation system for the African violet. MATERIALS AND METHODS: A local cultivar of the African violet from Guilan province was used for transformation experiments. The pFF19G and pBin61-Ech42 vectors were used for transient and stable transformation experiments, respectively. The PCR and RT-PCR techniques were used to verify transgene presence and transcript levels in candidate transgenic lines, respectively. RESULTS: Using leaf explants as target tissues, we transferred an endochitinase gene cDNA into African violet. Transgenic plants were regenerated on selection medium at a reasonable frequency (in average, one stable transgenic line per shot). Molecular analysis of transgenic plants by PCR and RT-PCR techniques confirmed successful integration and expression of transgene in several independent transgenic lines. CONCLUSIONS: Our results provide an efficient stable transformation system for genetic transformation of African violet.
BACKGROUND: A highly efficient genetic transformation system is essential for a successful genetic manipulation of the African violet (Saintpaulia ionantha Wendl.). OBJECTIVES: Developing a particle bombardment-based genetic transformation system for the African violet. MATERIALS AND METHODS: A local cultivar of the African violet from Guilan province was used for transformation experiments. The pFF19G and pBin61-Ech42 vectors were used for transient and stable transformation experiments, respectively. The PCR and RT-PCR techniques were used to verify transgene presence and transcript levels in candidate transgenic lines, respectively. RESULTS: Using leaf explants as target tissues, we transferred an endochitinase gene cDNA into African violet. Transgenic plants were regenerated on selection medium at a reasonable frequency (in average, one stable transgenic line per shot). Molecular analysis of transgenic plants by PCR and RT-PCR techniques confirmed successful integration and expression of transgene in several independent transgenic lines. CONCLUSIONS: Our results provide an efficient stable transformation system for genetic transformation of African violet.
Authors: B Schiedlmeier; R Schmitt; W Müller; M M Kirk; H Gruber; W Mages; D L Kirk Journal: Proc Natl Acad Sci U S A Date: 1994-05-24 Impact factor: 11.205