1. Please state project objectives and what work was done this quarter to address them: Project # 21-025 focuses on developing HLB-resistant and other improved citrus cultivars to mitigate the devastating effects of HLB on the Florida citrus industry. The project utilizes transgenic and non-transgenic approaches, and all experiments are conducted at the secure field site of the USDA/ARS Picos Research Farm. The goal is to provide sustainable solutions to combat HLB, safeguard the citrus industry, and support economic citrus production.This quarter, The test site received routine horticultural care, and trees were exposed to HLB and psyllid vectors to assess their resistance in a real-world environment. All BRS permits were upheld, and the site underwent inspections to ensure compliance with regulatory requirements. In addition Dr. Zhonglin Mou planted a total of 23 transgenic rootstock lines were planted in the farm on July 24, 2024. These lines include 14 NPR3 RNAi rootstock lines, three NPR3 RNAi rootstock lines top-grafted with Hamlin, and six NPR1 rootstock lines. These transgenic lines were generated through mature tissue transformation. There were planted in the field for fruit/seed production. Dr. Mou’s existing transgenics were also assessed. The NPR1 scion lines: 69 trees were planted in May 2019, 98 in May 2021, and 27 in October 2021. Among those planted in 2019, 15 were produced through mature tissue transformation, and 11 survived. These trees have flowered and are producing fruits. Two lines were chosen for propagation. Budsticks from these two lines have been submitted to DPI on July 15, 2024. Leaf and fruit samples were taken from the mature trees on July 26, 2024 for CLas titer assays. One tree from juvenile transformation has flowered this spring and currently has three fruits on the tree. Other juvenile trees have not flowered. 2. Please state what work is anticipated for next quarter:Next quarter, the team will analyze the data collected from the current field trials to evaluate the efficacy of different HLB-resistant candidates. Researchers will continue to have access to the test site for data collection and analysis purposes. Discussions with Soilcea are still continuing as the USDA continues to review contract documents for the planting of CRSPR trees for trial at this site in a true Florida citrus growing region. Upcoming meetings with Dr. Manjul Dut will be held during the end of August to discuss additional plantings at the site and plans for evlautating existing plantings. 3. Please state budget status (underspend or overspend, and why):This quarter, the project is ontrack for spending. We expect that we will be on track for spending for the remainder of the contract period. 4. Please show all potential commercialization products resulting from this research, and the status of each: The trangenic test site is consistently monitiored and researchers are contacted when their population(s) of trees show promising results for commericalization. This research has the potential to yield HLB-resistant transgenic and non-transgenic citrus cultivars suitable for commercial production. Some materials may be immediately suitable for commercial citrus production. The University of Florida has shown increased interest in the commricalization of some transgenic scions planted in the field that show promising single tree data. The data gathered from the field trials will guide recommendations for advancing materials to larger replicated trials, ensuring the development of commercially viable HLB-resistant citrus varieties.