Biotechnology to Generate Disease Resistant Mature Citrus as a Service

Biotechnology to Generate Disease Resistant Mature Citrus as a Service

Report Date: 09/30/2021
Project: 18-067C   Year: 2021
Category: Plant Improvement
Author: Janice Zale
Sponsor: Citrus Research and Development Foundation

 1. Please state project objectives and what work was done this quarter to address them:         The objectives of this project are to produce disease resistant, commercially & agronomically desirable, mature citrus transgenics, intragenics, & non-GMO gene edited trees using Agrobacterium & biolistics as a service for research & commercialization. The research focus of this project is to improve transformation efficiencies, so that the mature citrus protocols become more productive, decrease prices for scientists, & contribute more to financial self-sufficiency of our lab.        Our facility received an order from California to overexpress a citrus peptide in Hamlin, Kuharske & US942 using biolistic transformation to produce intragenics (all citrus sequences). The plants produced using this technology are not regulated  & can be provided to the growers rapidly after field testing.  We have started co-bombardments in these cultivars using other vectors until we receive the peptide. If this technology is successful, I will contact other scientists who might be interested in plant improvement using this method. For another scientist, 19 intragenics were produced using Agrobacterium transformation of mature EV1 scions.  Twenty-four flowering shoots were produced & micrografted onto rootstock for another scientist. This quarter, a total of 52 transgenic or intragenic shoots were produced in EV1 or Kuharske and micrografted for different scientists.   RNAi was also used to silence genes in another transformation experiment & a manuscript was published. A manuscript describing a protocol to increase transformation efficiency in mature citrus was submitted for review. Another manuscript will be submitted soon about a new intragenic selectable marker.  2. Please state what work is anticipated for next quarter:        Co-bombardments to overexpress the citrus peptide will continue.  We must finish some orders & new orders for Valencia, Early Valencia I, & Kuharske are pending.  We just completed an experiment with Agrobacterium mediated gene editing & the results are pending.    3. Please state budget status (underspend or overspend, and why): CRDF funding is sufficient, but my EBA has been revised to better fund salaries. I am not sure how long it will take to move through the UF system. In the interim, Dr. Rogers funds the rest of our salaries.   


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