Organogenesis and genetic transformation of Volkamer lemon (Citrus volkameriana)

• Grant number: 06/05454-6
• Support Opportunities: Scholarships in Brazil - Master
• Start date: April 01, 2007
• End date: August 31, 2008
• Field of knowledge: Agronomical Sciences - Agronomy
• Principal Investigator: Beatriz Madalena Januzzi Mendes
• Grantee: Eveline Carla da Rocha Tavano
• Host Institution: Centro de Energia Nuclear na Agricultura (CENA). Universidade de São Paulo (USP). Piracicaba , SP, Brazil

Abstract

Genetic transformation permits the introduction of genes in plant genoma and can be utilized in order to control problems related to plant diseases caused by microorganisms. Citrus sudden death (MSC) is a citrus disease probably caused by a citrus tristeza virus mutant strain and has caused serious problems to the Brazilian citriculture. Genetic transformation can be utilized in order to obtain MSC resistant plants. A genetic transformation program requires a efficient protocol of in vitro plant regeneration. The objective of this work is to study organogenesis and genetic transformation with Citrus volkameriana. The experiments will be performed with internodal segments as explants, collected from plants cultivated in greenhouse. For organogenesis the explants will be cultivated on EME medium supplemented with BAP (0; 0,5; 1,0; 1,5 mg/L) or with the combination of BAP and NAA (0,5 mg/L). The incubation will be in darkness or under 16 h photoperiod for a 30 days period, at 27C. The evaluation will be done determining the number of explants with shoots. For genetic transformation the explants will be inoculated with Agrobacterium tumefaciens, containing pCAMBIA 2201 plasmid with the nptII selection gene, GUS reporter gene and the CTV coat protein gene, in a hairpin construction. The transgenic shoots will be identified by histochemical test GUS and PCR. The positive shoots will be in vitro micrografted and after development the plantlets will be acclimatized to greenhouse conditions. The confirmation of transgene integration and transcription will be performed by Southern analysis.

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

TAVANO, Eveline Carla da Rocha. In vitro organogenesis and genetic transformation of the Volkamer lemon (Citrus volkameriana) and sour orange (Citrus aurantium). 2008. Master's Dissertation - Universidade de São Paulo (USP). Escola Superior de Agricultura Luiz de Queiroz (ESALA/BC) Piracicaba.