Transcriptional Dynamics of PsZFPK1 in P. sojae

Materials from different developmental stages of P. sojae, including mycelia (MY), sporulating hyphae (SP), zoospores (ZO), cystospores (CY), germinated cystospores (GC), and the process of infection (at 1.5, 3, 6, 12, 24, and 48 h), were collected as described previously [29 (link)]. Total RNA was extracted using the SV Total RNA Isolation kit (Promega, Madison, WI, USA) following the recommended protocol, and RNA quality was tested via agarose gel electrophoresis. First-strand cDNA was synthesized from 1 μg of total RNA using the FastKing Reverse Transcriptase kit (Tiangen, Beijing, China). Quantitative real-time PCR (qRT-PCR) was performed on a Bio-Rad CFX Connect Real-Time PCR System using SuperReal qPCR Mix (Tiangen) following the manufacturer’s instructions. Actin sequence from P. sojae was used as a reference gene, and the primers for this experiment are listed in Table S1. Relative PsZFPK1 transcript levels were calculated using the 2^−ΔΔCT method [30 (link)]. Means and standard deviations were calculated using data from three biological replicates.