Advanced Indica Rice Transformation

TARGET AUDIENCE

The course is targeted at tissue culturists, breeders, physiologists, and agronomists working on varietal improvement using novel methods such as genetic engineering in the public, private, and NGO sectors.

INTRODUCTION

Rice transformation in Indica rice and elite mega varieties offers enormous opportunities in rice improvement by allowing the introduction of novel genes, which may not be possible through conventional breeding. It is an important tool for rice functional genomic studies. Most rice transformation laboratories use the more amenable japonica cultivars in their research; only a few, including the Genetic Transformation Laboratory (GTL) at IRRI, have developed routine protocols for high-efficiency and reproducible transformation of recalcitrant Indica varieties. GTL routinely produces 6,000 events annually with a transformation efficiency of 30–50%, depending on the promoter and gene combination. A routine co-transformation efficiency of 10–15% is achieved for selectable marker-free transformation. A quick, reliable, low cost molecular assay for the presence of the transgene and copy number was established at IRRI, as well as the capacity to select homozygous plants at the T1 generation.

OBJECTIVES

This training course aims to
  • Provide participants with theoretical knowledge on the principles and application of rice transformation
  • Offer hands-on training on the crucial steps involved in high-frequency, reproducible Agrobacterium-mediated transformation for Indica rice
  • Enable trainees to do molecular characterization of transgenic plants, with emphasis on low-cost, quick-step PCR for gene integration detection and copy number determination
  • Teach trainees molecular and non-molecular techniques on homozygosity screening protocol of transgenic plants
  • Create a forum through which experiences on transgenic rice research and development are shared
COURSE CONTENT

1.     Transformation
  • Immature embryo selection, dehulling, sterilization, isolation, and Agrobacterium co-cultivation
  • Embryogenic callus culture selection
  • Pre-regeneration and plant regeneration
  • Rooting for optimum acclimatization
  • Co-transformation
  • Low-cost PCR for copy number determination
  • GUS staining to detect gene expression
  • ELISA and protein product determination
2.      High-frequency transformation of elite indica mega varieties and quality assurance

3.      Molecular analysis of regenerants
  • Low-cost PCR for copy number determination
  • GUS staining to detect gene expression
  • ELISA and protein product determination
4.      Transgene integration site by TAIL PCR and flanking genomic sequence analysis

5.      Designing primers for zygosity and event--specific screening

TRAINING STAFF
 
IRRI staff from GTL, PBGB, CESD, and C4 Rice Center involved in research, product development, and application of genetic transformation will be invited as resource persons and lab trainers.

DATE:   POSTPONED

VENUE
Genetic Transformation Laboratory and Molecular Marker Applications Lecture Room
Plant Breeding, Genetics and Biotechnology Division
International Rice Research Institute (IRRI)
Los Baños, Laguna, Philippines

CONTACT INFORMATION

Dr. Inez Slamet-Loedin
Senior Scientist, Genetic Transformation Laboratory, IRRI

Dr. Noel Magor
Head, Training Center, IRRI
International Rice Research Institute (IRRI)
DAPO Box 7777, Metro Manila, Philippines
Tel no. (63-2) 580-5600
E-mail: IRRITraining@irri.org
Fax: (63-2) 580-5699; 891-1292; 845-0606
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