Figure 2: Deletion and complementation of FgLAI12 in F. graminearum.
(A) The left border (LB) and right border (RB) were amplified from the wild type (a). ΔFgLAI12 mutant (d) was created by recombination between the knockout vector (b) and FgLAI12 (c). (B) The nucleotide sequence of FgLAI12 was cloned (a), and ligated into the complementation vector (b). The T-DNA region of the complementation vector was recombined into ΔFgLAI12 to create C-FgLAI12 (c). SacI, ApaI, SpeI, HindIII, NcoI and SpeI indicate the restriction enzymes used. (C) PCR verification of ΔFgLAI12 using the primer pairs P5 forward + P5 reverse and P7 forward + P7 reverse. (D) Verification of expression of FgLAI12 using the primer pair RJ-FgLAI12 forward + RJ-FgLAI12 reverse. FgGAPDH were used as a control. (E) Copy numbers of HPH determined by qPCR. Different letters (a and b) above each column indicate a significant difference (P < 0.05; n = 3). (F) Effect of SA and LA on expression of FgLAI12 in hyphae, as determined using the primer pair RJ-FgLAI12 (Table 1). Hyphae were collected on the 4th day after adding SA and LA. FgGAPDH was used as a reference. “F”, forward; “R”, reverse.
