Fig. 1. Details of mutations and ARMS-PCR primers (A: SdhBP225F; B: SdhBH272R). Mutations in genomic sequences are indicated by dotted box. Bold characters in primers are the positions of point mutations. Lower italic letters are the extra mismatch bases. Direction of primers are indicated by arrows.
Fig. 2. Electrophoretogram of tetra-primer ARMS-PCR system for detection of the Sdh-BP225F and SdhBH272R mutation associated with boscalid resistance. (A) The 426 bp and 330 bp fragments are directed to the P225F mutant (B1~B3). The 426 bp and 142 bp fragments on agarose gel are directed to the native Pro225 codon (B4~B9). The mixture of the mutant and the native were shown behind, with three fragments (B1+B4 and B1+ B7). (B) The 540 bp and 192 bp fragments are directed to the H272R mutant (B4~B6). The 540 bp and 394 bp fragments are directed to the native His272 codon (B1~B3, B7~B9). The mixture of the mutant and the native were shown behind, with three fragments (B1+B4 and B4+ B7).
Table 1. Botrytis cinerea strains for ARMS-PCR validation
Table 2. Primers used in this study
Table 3. Boscalid resistance monitoring by tetra-primer ARMS-PCR, conidial germination, and sequencing
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