CTV-T36 vectors as a tool to induce efficient flowering in citrus seedlings

CTV-T36 vectors as a tool to induce efficient flowering in citrus seedlings

Report Date: 11/13/2023
Project: 21-014   Year: 2023
Category: Other
Author: Choaa El Mohtar
Sponsor: Citrus Research and Development Foundation

1. Please state project objectives and what work was done this quarter to address them: Induce efficient flowering in citrus seedlings by overexpressing FT3 and knocking out negative regulators of flowering. Work done during this quarter:In this quarter, our focus was to amplify virions from agroinfiltration of the constructs replacing the p33 gene of CTV with FT3 gene from citrus and from the double gene constructs which have a silencing suppressor gene cassette in addition to the FT3 gene cassette which should boost the stability of the FT3 gene cassette within the CTV vector. The CTV vector expressing FT3 as replacement of p33 recently went systemic in N. benthamiana. However, the double gene constructs failed to go systemic in N. benthamiana in the first attempt. It prompted us to repeat the agroinfiltration and test replication in the infiltrated leaves. In the second attempt, the double gene construct replicated in the infiltrated leaves  based on ELISA and are waiting for it to go systemic to carry into citrus. The major success this quarter is the flowering of the rooted transgenic FT3 mainly Line 3 after almost 1.5 years of rooting.  2. Please state what work is anticipated for next quarter: In the next quarter, we will inoculate citrus with the newly generated CTV-FT3 vector which has the FT3 replacing the p33 gene cassette and work on carrying the double gene vectors expressing FT3 and silencing suppressors into citrus upon going systemic in N. benthamiana. Despite not being uniform in all branches, transgenic Carrizo lines are starting to flower. Thus, upon hardening of tissue we will generate the combinations of CTV-RNAi strategies targeting negative regulators of flowering with transgenic rootstock expressing FT3 to induce better flowering in citrus scions.   3. Please state budget status (underspend or overspend, and why): On budget   


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