Annual Report: Abiotic Stress Tolerant Bio-engineered Cereals

The objective is to apply transgenic technologies to enhance environmental stress tolerance in cereal crops. The collaboration combines ACPFG's gene systems and technologies with the field evaluation capabilities of Vibha. A series of transgenic wheat and rice lines are targeted to enhanced tolerance to drought and salinity stresses. Key findings from the second project year 2014/2015 include the completion of all transgenic constructs for rice and wheat. ACPFG and Vibha Agrotech Limited were regularly exchanging progress reports, and preparations for sending transgenic rice to India are ongoing. The project has made significant progress in wheat transformation, with all 44 constructs successfully transformed into Agrobacterium cells. In total, 47 stable transformation experiments were performed, resulting in the identification of 468 transgenic plants with between 1-3 copies of the gene of interest. Progress in rice transformation was initially delayed due to technical difficulties, but the quality of donor material in the winter greenhouse has since improved. The project has conducted 33 transformation experiments with a high success rate, and transgenic plants are now being transferred to the glasshouse. A first batch of T1 seed is expected to be ready for transfer to India by early 2016. Promotor studies in rice and wheat have also made significant progress. In wheat, 14 different promoters are included in the project, and 11 have been included in small-scale hydroponics experiments. Three promoters have shown GUSPlus expression in stained root and/or shoot material, indicating their functionality in the wheat background. A further 4 promoters have been present in material with no visible GUSPlus expression but have tested positive for the presence of the gene. The project has achieved several key accomplishments, including the completion of all target genes and promoters, the production of transgenic rice and wheat using Agrobacterium-based technology, and the study of transgene expression levels in T0 transgenic rice. The process to obtain a permit to import transgenic rice seed to India from Australia has also been initiated. Management and supervision of project activities at ACPFG is provided by the project leader, Dr. Stephan M. Haefele. Regular phone calls, emails, and visits have maintained contact and information exchange with Vibha Agrotech Limited. Activities at Vibha Agrotech Limited are now coordinated by the Director of Research, Dr. Bhimsingh Dahiya. The project has also made progress in field testing of transgenic wheat and barley, with a four-year project agreement signed with the Western Australia Agriculture Authority (WAAA). Three operational field sites for GM wheat testing have been established, and trials have commenced. The project aims to include USAID material for field trials on wheat in the 2016 winter season. Technical issues encountered in the project year were addressed through the booking of a growth room with constant temperature/humidity conditions to allow year-round growth of healthy rice plants. Meetings were held with the Indian project partner to report progress and discuss concerns about the unclear situation at Vibha Agrotech. The proposal for reconstitution of the Institutional Biosafety Committee (IBSC) has been submitted to the Department of Biotechnology (DBT). The project's future directions include the completion of the remaining constructs, the harvesting of transgenic plants, and the submission of documents for the GM rice seed import. The project aims to continue making progress in the development of abiotic stress-tolerant bio-engineered cereals.