Abstract:

To elucidate the expression characteristics of the effector protein Pt6647 from Puccinia triticina and further reveal its role in host pathogenicity,the candidate effector protein Pt6647,which was previously identified as highly expressed during host-pathogen interaction,was selected for analysis.We performed gene cloning via PCR,sequence alignment using the NCBI database,signal peptide prediction and functional validation to assess its secretory activity,subcellular localization to determine its cellular distribution,and Quantitative Real-time PCR(qRT-PCR)to examine its expression profile during infection.The open reading frame(ORF)of Pt6647,765 bp in length,was successfully cloned.The gene was highly conserved across eight different physiological races of P.triticina,with only two amino acid variations observed.The Pt6647 protein consisted of 254 amino acids,with its N-terminal 1—18 amino acids confirmed as a functional signal peptide for secretion.Subcellular localization analysis revealed that the full-length Pt6647 protein localized to the cell membrane,while the signal peptide-truncated variant(Pt6647^Δsp)was distributed in the nucleus,cytoplasm,and cell membrane.Expression profiling demonstrated that the transcription of Pt6647 peaked at 36 hours post-inoculation(hpi).The effector Pt6647 was successfully cloned and characterized.Its specific subcellular localization and early infection expression peak suggest that Pt6647 plays a critical role in the initial stages of pathogen invasion.

Key words: Puccinia triticina, Effector, Gene cloning, Quantitative Real-time PCR, Subcellular localization

CLC Number: 

• S512.1小麦

• S435.1禾谷类作物病虫害