Grant Information

DEVELOPMENT OF MATURE BUDWOOD TRANSFORMATION TECHNOLOGY

Sponsoring Institution Agricultural Research Service/USDA
Status ACTIVE
Funding Source USDA INHOUSE
Reporting Frequency Annual
Project Director THOMSON J G
Accession Number 428553
Project Number 2030-21000-020-13T
Dates 2014-10-01 - 2016-12-31
Recipient Organization WESTERN REGIONAL RES CENTER

ALBANY,CA 94710
Keywords citrus
hlb
integration
marker
mature
removal
resistance
targeted
tissue
transformation
Research Effort Applied (0%)
Basic (100%)
Developmental (0%)
Classification Parameters
Knowledge AreaSubject of InvestigationField of SciencePercent
201 - Plant Genome, Genetics, and Genetic Mechanisms 1510 - Corn (for sweetcorn use 1480) 1040 - Molecular biology 30%
201 - Plant Genome, Genetics, and Genetic Mechanisms 1530 - Rice 1040 - Molecular biology 20%
201 - Plant Genome, Genetics, and Genetic Mechanisms 2410 - Cross-commodity research--multiple crops 1040 - Molecular biology 20%
201 - Plant Genome, Genetics, and Genetic Mechanisms 1540 - Hard red winter wheat 1040 - Molecular biology 10%
201 - Plant Genome, Genetics, and Genetic Mechanisms 1541 - Hard red spring wheat 1040 - Molecular biology 10%
201 - Plant Genome, Genetics, and Genetic Mechanisms 1542 - Soft red wheat 1040 - Molecular biology 10%
Goals / Objectives
The proposed research, if successful, will produce a transformation protocol for most, and potentially all citrus cultivars. Constructs will be designed to enhance cell health during transformation and provide an early 'readout' of transformation success. These vectors will be complementary with the RMCE Founder lines research project. Finally, potential disease resistance genes will be provided by an ARS scientist and a Texas A&M scientist and introduced to cultivars successfully found transformation competent via the budwood method.
Methods (unparsed)

Objective 1. Develop optimized transformation vectors for transgenic HLB resistant citrus production. Utilization of an anti-apoptotic gene (anti-death) to reduce the rate of cell death during transformation, increasing the rate of cells surviving transformation. Employ a plant optimized DSRed gene for early, non-destructive, detection of transformed tissue. Use novel recombinases, previously developed, for site specific gene stacking and/or marker gene excision for precise genomic modification during transformation. Use of novel plant derived genetic elements, previously identified, will allow creation of transformation enhancing plant specific constructs for precision breeding technology. The resulting citrus trees will contain citrus derived resistance genes and the capacity for recombinase-mediated gene stacking. These vectors will be designed to introduce potential HLB disease resistance genes to citrus during the transformation development stage of the research. We are in collaboration with an ARS scientist and a Texas A&M scientist for their research on resistance genes to the HLB pathogen. Objectives 2. Develop improved transformation protocol via agro-bacterium. Investigate meristematic budwood for transformation and regeneration technique using traditional agro-bacterium and the enhanced transformation vector from Objective 1. Techniques and media for cell culture and selection will be investigated to maximize rates of transformation, selection of transgenic cells and cell survival.