ObjectiveThis project has one major objective for the Juvenile Tissue Citrus Transformation Facility (JTCTF) to stay open and offer the service for production of transgenic citrus plants to scientific community. By serving the researchers involved in work to find solutions against huanglongbing (HLB) and citrus canker, JTCTF remains integral part of the overall effort to assist Citrus Industry to stay profitable in this period when it is challenged by the threat of emerging diseases. Major accomplishments per objectiveThe objective set for this project was only partially met in 2020. Due to the circumstances associated with the COVID19 pandemic, JTCTF was not open at full capacity for the big part of the year. During the first quarter of 2020, JTCTF worked according to normal schedule up to March 13th. The facility was mostly closed for two weeks after that. Throughout the month of April and the beginning of May, the lab was visited on a daily basis to make sure the equipment was in order, to manage plants in the greenhouse and in the lab, and to take care of cultures from previously performed experiments. Upon the approval of the low level (Phase1) re-opening in May, JTCTF resumed some activities. Because of the surface area of the lab where most of the activities of JTCTF take place, being in Phase1 meant that only one employee can be in the lab at the time. Under such work schedule, it was not possible to run new transformation experiments. We proceeded to process the data from experiments that were already done. In the month of July, JTCTF started operating under the rules of Phase3 and it remained in that mode of operation until the end of 2020. Throughout the period between July and December, facility was able to work at about 40% of the capacity. This level of work was higher than needed because of the lack of new orders. One CREC faculty member announced the possibility of multiple orders but retracted that statement in the fall. This group also cancelled two orders and the work on two other orders was halted in the early phase until the transition of the facility into EBA unit is complete and new price list goes into effect. Employees of the facility paid from the USDA grant, continued to do transformation experiments as the time allowed them.Number of orders that JTCTF worked on in 2020 is seven, and they were all associated with the USDA grants where V. Orbovic is a co-PI. The orders were for production of transgenic Duncan grapefruit, Hamlin sweet orange, and Indian curry leaf plant (Murraya koenigii).The JTCTF produced 108 transgenic plants in 2020. Most of these plants belonged to Duncan grapefruit cultivar, there were also 19 Valencia orange plants, two M. koeinigii plants and one Pomelo plant (Table 1). Considering the role of JTCTF in the common effort of fighting citrus tree diseases, all produced plants were potentially resistant/tolerant to HLB or had ability to kill Asian citrus psyllids. None of the plants were sent outside of the state of Florida and will be used to estimate the effect of transgenes on the ability of citrus plants to sustain HLB. Table 1. Plants produced by CTF in 2020Cultivar Number of plants produced Duncan grapefruit 86 Valencia sweet orange 19 Pomelo 1 M. koenigii 2 During the 2020, JTCTF has done 54 experiments and used 88 appropriate bacterial strains for co-incubation of explants. For those experiments we cut about 80,000 explants. There were three experiments where all explants were contaminated and for seven experiments we had partial of full loss of data because they were not processed on time due to COVID19-related closure of CREC. About 25,000 explant were inspected for the presence of GFP fluorescence and about 9,000 for the pale/white phenotype. We also preformed about 700 PCR reactions with different primer pairs in search for JJ7 transgenic Valencia shoots.In an effort to secure sufficient supply of Duncan grapefruit seeds, I purchased 25 half-bushel bags of Duncan fruit in the beginning of 2020. Because of the low demand, most of the fruit rotted and we had to throw it away during the summer. We also got some Duncan fruit from CUPS at CREC. As opposed to 2019, we asked A. Schuman’s crew to harvest and deliver them to us in small installments during the fall and we managed to utilize all of them. For experiments done with the Hamlin material, we got some seeds from the people who ordered those plants while the rest came from fruit we picked from Block 22, a grove that belongs to CREC. M. koeinigii seeds were extracted from fruit harvested from trees grown at CREC.Major shortcomings, unfinished businessThe flux of employees working in the CTF remained high. One full-time employee left JTCTF in the middle of 2020 and she was not replaced. Another employee for whom there was not sufficient funding left facility at the end of 2020. The labor force projected for the demand of transgenic citrus plants in 2021 is at the level of 1.6 FTE which translates into one full time employee and one part-time employee.The transition of JTCTF into an EBA (educational business activity) unit mandated by UF administration will be completed soon. This process has been going on for the last eight months and once complete will fully re-define the operation of the facility. The opportunities going forwardMy efforts to attract business and get more people to order transgenic plants for the last year and-a-half were not successful. Others involved in management of JTCTF on the higher level are also investing efforts for that to happen. Whether these efforts will yield more orders for JTCTF remains to be seen. Disappearance of COVID19 as a threat may increase activities on all fronts and as a consequence lead to more orders. Publications from this project1) Sinn, J., Held, J., Vosburg, C., Klee, S., Orbovic, V., Taylor, E., Gottwald, T., Stover, E., Moore, G., McNellis, T. (2020) Flowering Locus T chimeric protein induces floral precocity in edible citrus. Plant Biotechnology Journal.2) Zhang, F., Rossignol, P., Huang, T., Wang, Y., May, A., Dupont, C., Orbovic, V., Irish, V. (2020) Reprogramming of stem cell activity to convert thorns into branches. Current Biology, 30:2951-2961.