Antisense Oligonucleotide as a New Technology Application for CsLOB1 Gene Silencing Aiming at Citrus Canker Resistance

Autor(es)

de Lima, Luiz Felipe Franco ; Carvalho, Isis Gabriela Barbosa ; de Souza-Neto, Reinaldo Rodrigues ; dos Santos, Larissa da Silva ; Nascimento, Cesar Augusto ; Takita, Marco Aurelio ; Tavora, Fabiano Touzdjian Pinheiro Kohlrausch ; Mehta, Angela ; de Souza, Alessandra Alves

• Tipo de documento: Artigo Científico
• Fonte: PHYTOPATHOLOGY; v. 114, n. 8, p. 8-pg., 2024-08-14.

Resumo

Citrus canker disease, caused by Xanthomonas citri subsp. citri, poses a significant threat to global citrus production. The control of the disease in the field relies mainly on the use of conventional tools such as copper compounds, which are harmful to the environment and could lead to bacterial resistance. This scenario stresses the need for new and sustainable technologies to control phytopathogens, representing a key challenge in developing studies that translate basic into applied knowledge. During infection, X. citri subsp. citri secretes a transcriptional activator-like effector that enters the nucleus of plant cells, activating the expression of the canker susceptibility gene LATERAL ORGAN BOUNDARIES 1 (LOB1). In this study, we explored the use of antisense oligonucleotides (ASOs) with phosphorothioate modifications to transiently inhibit the gene expression of CsLOB1 in Citrus sinensis. We designed and validated three potential ASO sequences, which led to a significant reduction in disease symptoms compared with the control. The selected ASO3-CsLOB1 significantly decreased the expression level of CsLOB1 when delivered through two distinct delivery methods, and the reduction of the symptoms ranged from approximately 15 to 83%. Notably, plants treated with ASO3 did not exhibit an increase in symptom development over the evaluation period. This study highlights the efficacy of ASO technology, based on short oligonucleotide chemically modified sequences, as a promising tool for controlling phytopathogens without the need for genetic transformation or plant regeneration. Our results demonstrate the potential of ASOs as a biotechnological tool for the management of citrus canker disease. (AU)

• Processo FAPESP: 21/09689-8 - Aplicação de oligos antisense para silenciamento dos genes pds, lob1 e calS7 de Citrus sinensis
• Beneficiário: Larissa da Silva dos Santos
• Modalidade de apoio: Bolsas no Brasil - Programa Capacitação - Treinamento Técnico
• Processo FAPESP: 17/13885-1 - Uso de CRISPR/Cas9 na edição de genes de suscetibilidade de Citrus sinensis a Xanthomonas citri subsp. Citri
• Beneficiário: Reinaldo Rodrigues de Souza Neto
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 20/04795-1 - Transformação de variedades de laranja doce para estudo funcional e/ou edição de genes visando resistência a doenças bacterianas de citros
• Beneficiário: Isis Gabriela Barbosa Carvalho
• Modalidade de apoio: Bolsas no Brasil - Programa Capacitação - Treinamento Técnico
• Processo FAPESP: 19/23217-1 - Desenvolvimento de nanoformulação visando o manejo sustentável de fitopatógenos agrícolas
• Beneficiário: Alessandra Alves de Souza
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 22/04112-7 - Aplicação de oligos antisense para silenciamento dos genes LOB1 e EXPANSINA de Citrus sinensis
• Beneficiário: Cesar Augusto Nascimento
• Modalidade de apoio: Bolsas no Brasil - Programa Capacitação - Treinamento Técnico