1. Please state project objectives and what work was done this quarter to address them:
Objective 1: Screen breeding populations for new individuals with exceptional HLB tolerance and fruit quality as populations mature and begin to fruit.
We are currently overseeing five trials that have been planted, one trial was planted before the project period, four were recently planted during the project period. Two additional trials are ready to be planted as soon as the land is cleared and prepped, with a planting date in mid-September. Two additional trials are being propagated to replicate the Coca Cola Company trial in two additional locations. By the end of this reporting period, early mandarin cultivars with HLB tolerance will be evaluated for yield and fruit quality for fresh market. Drone flights have been flown to gather more information about HLB tolerance among accessions. There appear to be UF/IFAS and USDA selections with exceptional tolerance at Eagle Lake. Trees going into maturity are being followed and when the fruit are ready they will be evaluated for quality to determine value to the industry.
Objective 2: Plant new and manage ongoing Stage 2 semi-commercial trials of the selections most likely to succeed in the HLB environment versus commercial standard controls.
We will have a total of at least nine Stage 2 trials that will be planted during the course of this project, probably more. As mentioned above, we have one trial that was planted and established before the project period, a sweet orange trial that is now two years old. Some trees in that trial now have fruit which will be phenotyped this coming season. We have recently planted four trials that are in the ground now. One is a fresh market trial at CREC with some selections that are orange-like, another is a sweet orange trial at CREC and a third is a seedless lemon trial versus controls on CREC land. We have also planted a trial on private land in Eagle Lake, FL with sweet orange and other selections. All trials have standard controls and they are in randomized complete block designs. I will provide maps at an upcoming meeting among Rick Dantzler, Dr. Futch and Dr. Mattia. We are also leaving no stone unturned and propagating everything with potential value to the industry from the plant improvement team at CREC to evaluate for HLB tolerance and fruit quality. All trials have either control rootstocks or control combinations, depending on the trial. For the trial to be planted in mid-September, the trees are ready to plant and the nursery will deliver when the land is ready. We will incorporate compost into the soil to aid in increased cation exchange capacity, water holding capacity, nutrient delivery and oganic matter content for the sandy soil that is being used for this CREC trial site.
Objective 3: Utilize bioinformatic tools to identify genes or other variants involved in HLB tolerance and possible resistance for plant transformation and conventional breeding.
As mentioned previously, we have received the genomic, Hi-C, and transcriptomic data for assembly and annotation of the Hamlin 1-4-1 genome so that it can be compared to Hamlin N13-32 to look for genes and other variants associated with HLB tolerance. We have identified the many of the founders of the USDA and IFAS citrus breeding program and these founders will be deep sequenced to assemble, phase and annotate them so that these can be used to look for alleles that are associated with HLB tolerance among the 100 genotyped accessions. Several assembled, phased and annotated genomes will result from this work that should be useful to those at the Crop Transformation Center in Gainesville and others working on the HLB crisis. We have noticed that genomic data that we received from internal core services in Gainesville was not optimal compared to other labs we have worked with, so we are currently working on getting the highest quality data with the lowest possible cost and choosing a different lab accordingly. We are waiting on the most recent drone and phenotypic data from the Coca Cola Company to make decisions about the most tolerant accessions and how it relates to the drone data collected previously with the intention of choosing the most tolerant accessions for sequencing.
2. Please state what work is anticipated for next quarter:
We plan on deep sequencing, annotating, and phasing the founders and short read sequencing the 100 selections from USDA and IFAS breeding programs as soon as we find the best price for the highest quality data possible. We are actively prepping the land for the stage II trial with the Coca Cola Company at CREC and we plan on following ongoing trials to phenotype advanced selections to compare versus the standard controls. We plan to have meetings with the scientists at the Lawrence Berkeley National Laboratory to discuss data and research activities. The documentation with LBNL will be completed by the next report. We will phenotype all accessions that are earliest in the programs.
3. Please state budget status (underspend or overspend, and why):
We are underspent due to working on Lawrence Berkeley National Laboratory documentation and sequencing decisions. We will be more on track by next reporting period.