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Detection of transcripts of defense response genes related to the immunity marker gene NbHIN1 , salicylic acid signaling marker gene NbPR1 , and jasmonic acid signaling marker gene NbLOX4 by reverse transcription-quantitative PCR (RT-qPCR). The transcripts were detected in 4-week-old Nicotiana benthamiana leaves 2 days after agro-infiltration with the 18 random selective secretome members BAX, VdEG1, and the cotton A08G47475 (thionin protein) used as positive controls to induce the N. benthamiana immunity response. The RT-qPCR experiments were performed three times. Error bars represent standard errors. Asterisks * ( P < 0.05) and ** ( P < 0.01) indicate a significant difference relative to the agro-infiltration GFP control in unpaired Student’s t -tests

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