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利用酵母双杂交技术筛选与甘蔗黄叶病毒编码的毒力因子 P0 互作的宿主因子。

Screening and Identification of Host Factors Interacting with the Virulence Factor P0 Encoded by Sugarcane Yellow Leaf Virus by Yeast Two-Hybrid Assay.

机构信息

Guangxi Key Laboratory for Sugarcane Biology, Guangxi University, Nanning 530004, China.

College of Agriculture, Guangxi University, Nanning 530004, China.

出版信息

Genes (Basel). 2023 Jul 3;14(7):1397. doi: 10.3390/genes14071397.

Abstract

Sugarcane yellow leaf virus (SCYLV), a member of the genus in the family , causes severe damage and represents a great threat to sugarcane cultivation and sugar industry development. In this study, inoculation of plants with a potato virus X (PVX)-based vector carrying the SCYLV P0 gene induced typical mosaic, leaf rolling symptoms and was associated with a hypersensitive-like response (HLR) necrosis symptom, which is accompanied with a systemic burst of HO and also leads to higher PVX viral genome accumulation levels. Our results demonstrate that SCYLV P0 is a pathogenicity determinant and plays important roles in disease development. To further explore its function in pathogenic processes, a yeast two-hybrid assay was performed to screen the putative P0-interacting host factors. The recombinant plasmid pGBKT7-P0 was constructed as a bait and transformed into the yeast strain Y2HGold. The ROC22 cultivar (an important parental resource of the main cultivar in China) cDNA prey library was constructed and screened by co-transformation with the P0 bait. We identified 28 potential interacting partners including those involved in the optical signal path, plant growth and development, transcriptional regulation, host defense response, and viral replication. To our knowledge, this is the first time we have reported the host proteins interacting with the P0 virulence factor encoded by sugarcane yellow leaf virus. This study not only provides valuable insights into elucidating the molecular mechanism of the pathogenicity of SCYLV, but also sheds light on revealing the probable new pathogenesis of in the future.

摘要

甘蔗黄叶病毒(SCYLV)是呼肠孤病毒科斐济病毒属的成员,可导致严重的危害,对甘蔗种植和糖业发展构成巨大威胁。本研究中,接种携带 SCYLV P0 基因的马铃薯 X 病毒(PVX)载体的甘蔗植株,诱导出典型的花叶、叶片卷曲症状,并伴有过敏样坏死反应(HLR)坏死症状,伴随着 HO 的系统爆发,同时导致更高水平的 PVX 病毒基因组积累。研究结果表明,SCYLV P0 是一个致病性决定因子,在疾病发展中起重要作用。为了进一步探索其在致病过程中的功能,进行了酵母双杂交筛选来筛选可能与 P0 相互作用的宿主因子。构建了 pGBKT7-P0 重组质粒作为诱饵,并转化到酵母菌株 Y2HGold 中。利用该菌株与 P0 诱饵共转化构建 ROC22 品种(中国主要品种的重要亲本资源)cDNA 诱饵文库,并进行筛选。鉴定到 28 个潜在的互作伙伴,包括涉及光信号途径、植物生长发育、转录调控、宿主防御反应和病毒复制的蛋白。据我们所知,这是首次报道与甘蔗黄叶病毒编码的 P0 毒力因子互作的宿主蛋白。该研究不仅为阐明 SCYLV 致病性的分子机制提供了有价值的见解,而且为揭示未来 可能的新发病机制提供了线索。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/977b/10379860/09407d8242d3/genes-14-01397-g001.jpg

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