Institute of Sugar Beet Research, Göttingen, Germany.
SESVanderHave, Tienen, Belgium.
Plant Biotechnol J. 2024 Aug;22(8):2129-2141. doi: 10.1111/pbi.14333. Epub 2024 Mar 15.
Eukaryotic translation initiation factors (eIFs) are important for mRNA translation but also pivotal for plant-virus interaction. Most of these plant-virus interactions were found between plant eIFs and the viral protein genome-linked (VPg) of potyviruses. In case of lost interaction due to mutation or deletion of eIFs, the viral translation and subsequent replication within its host is negatively affected, resulting in a recessive resistance. Here we report the identification of the Beta vulgaris Bv-eIF(iso)4E as a susceptibility factor towards the VPg-carrying beet chlorosis virus (genus Polerovirus). Using yeast two-hybrid and bimolecular fluorescence complementation assays, the physical interaction between Bv-eIF(iso)4E and the putative BChV-VPg was detected, while the VPg of the closely related beet mild yellowing virus (BMYV) was found to interact with the two isoforms Bv-eIF4E and Bv-eIF(iso)4E. These VPg-eIF interactions within the polerovirus-beet pathosystem were demonstrated to be highly specific, as single mutations within the predicted cap-binding pocket of Bv-eIF(iso)4E resulted in a loss of interaction. To investigate the suitability of eIFs as a resistance resource against beet infecting poleroviruses, B. vulgaris plants were genome edited by CRISPR/Cas9 resulting in knockouts of different eIFs. A simultaneous knockout of the identified BMYV-interaction partners Bv-eIF4E and Bv-eIF(iso)4E was not achieved, but Bv-eIF(iso)4E plants showed a significantly lowered BChV accumulation and decrease in infection rate from 100% to 28.86%, while no influence on BMYV accumulation was observed. Still, these observations support that eIFs are promising candidate genes for polerovirus resistance breeding in sugar beet.
真核翻译起始因子(eIFs)对于 mRNA 翻译非常重要,同时也是植物-病毒相互作用的关键。这些植物-病毒相互作用大多数发生在植物 eIFs 与马铃薯 Y 病毒属(Potyviridae)病毒的衣壳蛋白连接(VPg)之间。如果由于 eIFs 的突变或缺失导致相互作用丧失,病毒的翻译和随后在其宿主中的复制将受到负面影响,导致隐性抗性。在这里,我们报告了鉴定出的β 菜豆 Bv-eIF(iso)4E 是携带 VPg 的甜菜黄化病毒(属 Polerovirus)的易感性因子。使用酵母双杂交和双分子荧光互补测定,检测到 Bv-eIF(iso)4E 与假定的 BChV-VPg 之间的物理相互作用,而密切相关的甜菜轻度黄化病毒(BMYV)的 VPg 被发现与两种同工型 Bv-eIF4E 和 Bv-eIF(iso)4E 相互作用。在 polerovirus-甜菜病理系统中,这些 VPg-eIF 相互作用被证明是高度特异性的,因为 Bv-eIF(iso)4E 中预测的帽子结合口袋内的单个突变导致相互作用丧失。为了研究 eIFs 作为针对侵染甜菜的 poleroviruses 的抗性资源的适宜性,通过 CRISPR/Cas9 对 B. vulgaris 植物进行了基因组编辑,导致不同 eIFs 的敲除。没有实现同时敲除鉴定出的 BMYV 相互作用伙伴 Bv-eIF4E 和 Bv-eIF(iso)4E,但 Bv-eIF(iso)4E 植物的 BChV 积累明显降低,感染率从 100%降至 28.86%,而对 BMYV 积累没有影响。尽管如此,这些观察结果支持 eIFs 是甜菜 polerovirus 抗性育种的有前途的候选基因。