Fig.1 LAMP primer design of P.parvum(A) and P.versatile(B)

Fig.2 Agarose gel diagram and melting curve of Quantitative Real-time PCR primer specificity verification of P.parvum M.2 000 bp Marker;1—16.P.parvum YT22221,P.versatile FN22511,P.atrosepticum FN20412,P.parmentieri YL22131,P.polaris WW22521,P.punjabense XW22812,P.carotovorum FN22613,P.brasiliensis XW22111,P.polonicum BY21311,Erwinia rhapontici BYQ31,Pantoea agglomerans FN22611, Serratia marcescens GX23711,Psedomonas straminea GX23461,Enterobacter soil GX23381,plant DNA of potato,ddH₂O.The same as Fig.10-A.

Fig.3 Agarose gel diagram and melting curve of Quantitative Real-time PCR primer specificity verification of P.versatile M.2 000 bp Marker;1—16.P.versatile FN22511, P.parvum YT22221,P.atrosepticum FN20412,P.parmentieri YL22131,P.polaris WW22521,P.punjabense XW22812,P.carotovorum FN22613,P.brasiliensis XW22111,P.polonicum BY21311,Erwinia rhapontici BYQ31,Pantoea agglomerans FN22611, Serratia marcescens GX23711,Psedomonas straminea GX23461,Enterobacter soil GX23381,Plant DNA of potato,ddH₂O.The same as Fig.10-B.

Fig.4 Optimization of Quantitative Real-time PCR primer concentration conditions for P.parvum Different lowercase letters indicate significant differences(P<0.05).The same as Fig.5—7.

Fig.5 Optimization of Quantitative Real-time PCR primer concentration conditions for P.versatile

Fig.6 Optimization of annealing temperature conditions for Quantitative Real-time PCR of P.parvum

Fig.7 Optimization of annealing temperature conditions for Quantitative Real-time PCR of P.versatile

Fig.8 Quantitative Real-time PCR primer sensitivity detection of P.parvum A.Primer XX3-F/R pathogen gDNA sensitivity detection.1—7.DNA concentrations were 100,10,1,0.1,1×10^-2,1×10^-3,1×10^-4 ng/μL;8.Control;the same as Fig.9-A,C.B.Primer XX3-F/R sensitivity detection of DNA in artificially inoculated plant samples.1—6.DNA concentrations were 100,10,1,1×10^-2,1×10^-3 ng/μL;7.Control;the same as Fig.9-B,D.

Fig.9 Quantitative Real-time PCR primer sensitivity detection of P.versatile A.Primer DY1-F/R pathogen gDNA sensitivity detection;B.Primer DY1-F/R sensitivity detection of DNA in artificially inoculated plant samples; C.Primer DY2-F/R pathogen gDNA sensitivity detection;D.Primer DY2-F/R sensitivity detection of DNA in artificially inoculated plant samples.

Fig.10 Validation results of specificity of LAMP primers and agarose gel diagram for P.parvum(A) and P.versatile(B)

Fig.11 LAMP reaction conditions optimization results and agarose gel diagram M.2 000 bp Marker.A.Optimization results of LAMP primer concentration conditions for P.parvum;B.Optimization results of LAMP reaction time for P.parvum;C.Optimization results of LAMP reaction temperature of P.parvum;D.Optimization results of LAMP primer concentration conditions for P.versatile;E.Optimization results of LAMP reaction time for P.versatile;F.Optimization results of LAMP reaction temperature of P.versatile;1—4.The final concentrations of primers were 0.4 μmol/L for inner primers(0.1 μmol/L for outer primers),0.8 μmol/L for inner primers(0.2 μmol/L for outer primers),1.2 μmol/L for inner primers(0.3 μmol/L for outer primers),and 1.6 μmol/L for inner primers(0.4 μmol/L for outer primers).

Fig.12 LAMP sensitivity detection results and agarose gel diagram of P.parvum M.2 000 bp Marker.A.Pathogen gDNA sensitivity detection;B.Sensitivity detection of DNA in artificially inoculated plant samples.The same as Fig.13.

Fig.13 LAMP sensitivity detection results and agarose gel diagram of P.versatile

Fig.14 Samples from each stem segment after inoculation 12,24,36 h were detected by Quantitative Real-time PCR

Fig.15 Samples from each tuber segment after inoculation 12,24,48 h were detected by Quantitative Real-time PCR

Fig.16 Quantitative Real-time PCR detection of drosophila samples mixed with different OD₆₀₀ values in feed culture medium

Fig.17 Results and agarose gel diagram of samples from each stem segment after inoculation 12,24,36 h were detected by LAMP Marker.2 000 bp Marker.A.P.parvum LAMP;B.P.versatile LAMP.The same as Fig.18.

Fig.18 Results and agarose gel diagram of samples from each tuber segment after inoculation 12,24,48 h were detected by LAMP

Fig.19 LAMP detection drosophila samples feed medium inoculated with 500 μL of bacterial fluid with different OD₆₀₀ values M.2 000 bp Marker;A.P.parvum LAMP;B.P.versatile LAMP;1.H₂O;2—6.OD₆₀₀ are 0.1,0.2,0.4,0.6,0.8.