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ARS Home » Northeast Area » Kearneysville, West Virginia » Appalachian Fruit Research Laboratory » Innovative Fruit Production, Improvement, and Protection » Research » Publications at this Location » Publication #398140
Research Project: Improving Fruit Crop Traits Through Advanced Genomic, Breeding, and Management Technologies

Location: Innovative Fruit Production, Improvement, and Protection

Title: A novel and efficient Agrobacterium-mediated transient gene expression in citrus epicotyls and mature stem tissues

Author
item LI, YANJUN - University Of Connecticut
item HU, WEI - Jiangxi Agricultural University
item AHMAD, SHOWKAT - University Of London
item Liu, Zongrang
item CHENG, BAOPING - Guangdong Academy Of Agricultural Sciences
item Duan, Hui
item LI, YI - University Of Connecticut

Submitted to: Fruit Research
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/28/2024
Publication Date: N/A
Citation: N/A

Interpretive Summary: The use of gene editing technology can improve agronomical traits in fruit trees, but current gene editing technology also frequently incorporates transgenes into the plant genome, which raises concerns about the impact on environmental and ecological integrity, as well as food safety. This study tested various parameters in citrus tissues in order to enhance transgene-free gene editing efficiency. A number of parameters, including tissue age, chemicals, and hormones, have been tested for enhancing transient gene expression efficiency, and optimal concentrations, tissue ages, and treatment times have been found. In combination with the optimized parameters, transient gene expression was enhanced by sixfold. The result of this study has an implication in improvement of horticulturally important traits in citrus, peach, and apple crops.

Technical Abstract: Agrobacterium-mediated transient gene expression is a powerful technique for rapidly evaluating gene expression in higher plants. This study aimed to improve transient expression levels of T-DNA genes in citrus by investigating the effects of various factors, including seedling age, tissue treatments, Agrobacterium incubation medium and duration, hormone combinations, methylation inhibitors, antioxidants, and more. We tested and optimized these parameters, and observed significant increases in transient gene expression in juvenile epicotyls of 'Carrizo' citrange and mature stem tissues of Pineapple sweet orange, Valencia orange, and Washington navel orange. Our results demonstrated up to a six-fold increase in transient GUS gene expression, highlighting the effectiveness of these simple and inexpensive treatments. The juvenile and mature citrus explants used in this study displayed high levels of transient expression, which may provide a valuable tool for studying phloem-associated diseases such as Huanglongbing (HLB), facilitating rapid analysis of factors involved in HLB pathogenicity. This optimized method may also offer a promising tool for advancing genetic studies and improving the efficiency of Agrobacterium-mediated transgene-free editing in citrus and other economically significant perennial crops.