Vargas-Salinas Mayela, Medina-Hernández Diana, Arcos-Ortega Guadalupe Fabiola, Luis-Villaseñor Irasema Elizabeth, Holguín-Peña Ramón Jaime
Centro de Investigaciones Biológicas del Noroeste, Instituto Politécnico Nacional 195, Col. Playa Palo de Santa Rita, 23096 La Paz, Baja California Sur Mexico.
Facultad de Ciencias del Mar, Universidad Autónoma de Sinaloa (UAS) Mazatlán, 82000 Sinaloa, Mexico.
3 Biotech. 2021 Mar;11(3):114. doi: 10.1007/s13205-021-02653-7. Epub 2021 Feb 3.
This study compared the transcriptional changes in plants treated with homologous sequences derived from (PepGMV) and heterologous sequences that derived from another begomovirus, (ToChLPV) prior to infection by PepGMV. The results of microarray analyses identified upregulated genes associated with RNAi such as DCL2, DCL4, AGO3, AGO7, AGO10, NRPD2B (Pol IV), DRB3, CMT3, RDR6. The components that participate in different RNAi pathways were identified, including methylation induced by both constructs, as well as the code of these genes in and its counterpart in through different genome assembly. The expression of these genes was validated by quantitative reverse transcription polymerase chain reaction (RT-qPCR), where DCL3, DCL4, AGO1-1, AGO2, RDR6 and PPR1 showed increased expression during plant protection with the heterologous construct compared to those protected with the homologous construct. The results of this study confirmed the activation of the gene silencing mechanism at the transcriptional level with both constructs and established the possibility of their use as a protection system for both homologous and heterologous sequences.
本研究比较了在被番茄潜叶蛾花叶病毒(PepGMV)感染之前,用源自番茄潜叶蛾花叶病毒(PepGMV)的同源序列和源自另一种双生病毒,即番茄智利潜叶蛾病毒(ToChLPV)的异源序列处理的植物中的转录变化。微阵列分析结果确定了与RNA干扰相关的上调基因,如DCL2、DCL4、AGO3、AGO7、AGO10、NRPD2B(聚合酶IV)、DRB3、CMT3、RDR6。确定了参与不同RNA干扰途径的成分,包括两种构建体诱导的甲基化,以及这些基因在番茄潜叶蛾花叶病毒及其通过不同基因组组装的对应物中的编码。通过定量逆转录聚合酶链反应(RT-qPCR)验证了这些基因的表达,与用同源构建体保护的植物相比,在用异源构建体进行植物保护期间,DCL3、DCL4、AGO1-1、AGO2、RDR6和PPR1的表达增加。本研究结果证实了两种构建体在转录水平上激活了基因沉默机制,并确立了将其用作同源和异源序列保护系统的可能性。