To obtain GFP-labeled VdaSm-GFP, the pNEO-olic GFP plasmid (stored in our laboratory) was transferred to VdaSm. G418 was used to select the transformants on PDA medium.
To generate the knockout plasmids, pKOVsmNoxB, pKOVsmPls1 and pKOVsmCrz1, upstream and downstream genomic sequences of these genes were amplified with the corresponding primers (Table S1 ). The paired sequences were inserted into a position flanking the hygromycin resistant cassette of the vector pGKO-HPT with the Exnase MultiS (Vazyme, Nanjing, China), and then the knock-out plasmids were transformed as previously described [35 (link)].
To produce the complemented strains, including VdΔnoxb/VdNoxB and VdΔpls1/VdPls1, GFP-fused VdNoxB and VdPls1 under the native promoter were introduced into VdΔnoxb and VdΔpls1, respectively. RFP-fused VdKar2 under the Tef promoter were introduced into VdΔnoxb/VdNoxB and VdΔpls1/VdPls1, to detect the location of VdNoxB and VdPls1, respectively. The fusion plasmids were constructed as previously described [22 (link)], and the primers are listed inTable S1 .
To generate the knockout plasmids, pKOVsmNoxB, pKOVsmPls1 and pKOVsmCrz1, upstream and downstream genomic sequences of these genes were amplified with the corresponding primers (
To produce the complemented strains, including VdΔnoxb/VdNoxB and VdΔpls1/VdPls1, GFP-fused VdNoxB and VdPls1 under the native promoter were introduced into VdΔnoxb and VdΔpls1, respectively. RFP-fused VdKar2 under the Tef promoter were introduced into VdΔnoxb/VdNoxB and VdΔpls1/VdPls1, to detect the location of VdNoxB and VdPls1, respectively. The fusion plasmids were constructed as previously described [22 (link)], and the primers are listed in
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