2.5. MAT Gene Amplification and Mating Type Assignment

To further understand the possible reproductive mode of the population of each Calonectria species identified in this study, the mating type idiomorph of each representative Calonectria isolate of an identified species was identified. Mating type primer pairs Cal_MAT111_F/Cal_MAT111_R and Cal_MAT121_F/ Cal_MAT121_R were used to amplify the MAT1-1-1 and MAT1-2-1 genes in all selected isolates using the protocol described by Li and co-authors [35] (Table 1). For the heterothallic Calonectria species based on MAT gene amplification results, the species with an adequate number of isolates as one population, the possibility that recombination had taken place was tested. A two-tailed exact binomial method [38] was used, and each population that included isolates of both mating types was tested in R version 3.6.1 to evaluate whether the MAT1-1 and MAT1-2 frequency significantly deviated from a 1:1 ratio.

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