Generation of non-transgenic HLB-resistant sweet orange varieties using CRISPR-Cas technology

Generation of non-transgenic HLB-resistant sweet orange varieties using CRISPR-Cas technology

Report Date: 02/12/2023
Project: 21-028   Year: 2023
Category: Plant Improvement
Author: Nian Wang
Sponsor: Citrus Research and Development Foundation

1. Please state project objectives and what work was done this quarter to address them: The purpose of this project is to generate non-transgenic HLB resistant Valencia and Hamlin sweet orange plants using CRISPR-Cas technology. Objective 1. Generate non-transgenic HLB resistant/tolerant Valencia and Hamlin sweet orange plants by mutation of HLB susceptibility genes. In total, six putative S genes will be edited. Constructs needed for CRISPR genome editing are being made. Multiple edited lines were generated for ACD2 gene. However, further confirmation demonstrated none of them was biallelic/homozygous mutant. We are genenerating more lines for ACD2 and other target genes. Objective 2. Generate cisgenic genome modified Valencia and Hamlin sweet orange plants by knock-in the gene encoding MaSAMP from Microcitrus. We are optimizing the knock-in method using the CRISPR technology. We have made some progress in knock-in methodology using non-transgenic approach. We will use the developed method for knockin wiht MaSAMP.  2. Please state what work is anticipated for next quarter:To generate more genome edited lines for ACD2 and other target genes. Using the developed method for knockin with MaSAMP.     3. Please state budget status (underspend or overspend, and why):On schedule.     


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