As co-infection increased the accumulation of 21- and 22-nt M-vsiRNAs, the hotspots distributed in the (+)- and (−)-sense strands of MCMV genome were both increased in co-infected maize plants, which mainly located in the P32/P50, P31 and CP coding regions and the 3′-UTR region (Fig. 5E). This evidence concerns the gene CP and coinfection.