Molecular monitoring of Metarhizium spp. (Hypocreales: Clavicipitaceae) in sugarcane plantationsThe aim of this study was to characterize the diversity of Metarhizium in a sugarcane plantation and to monitor the efficacy and persistence of the isolate ESALQ 1604 of M. anisopliae after field application, by molecular techniques.Isolates were recovered from samples of soil, root and infected spittlebugs of two areas in Iracemápolis-SP: one with application of the fungus (isolate ESALQ 5310) and mechanical harvesting and another without fungal application and manual harvest preceded by burning.The isolates from insects were recovered from nymphs and adults collected 51, 42 and 1 day (s) before application of the fungus and 7, 30, 60 and 90 days after application.Isolates of soil and root were obtained 90 days pre-application of the fungus and 30 and 90 days post-application.The application of M. anisopliae was carried out in 9/Jan/15 with suspension of 3,72 x 10 6 viable conidia/mL at a volume rate of 150 l/ha.In the molecular analyses, 22 isolates from the same areas collected in 18/12/2012 were also included.The haplotype diversity of Metarhizium was obtained by genotyping 10 microsatellite markers of 213 isolates from soil samples, 22 from roots and 73 from spittlebugs.The specific identification of fungi was obtained by sequencing the gene 5'-TEF of 61 isolated from soil, 6 from root and 13 from spittlebug selected among the different haplotypes generated by analysis of microsatellite markers.Among 310 isolates genotyped, 156 haplotypes of the fungus were obtained, of these, 132 from soil isolates, 17 of root and 20 of spittlebug.The highest haplotype diversity was found in soil h=0.989 and the smallest in spittlebug h = 0.779.The genetic divergence among isolates from insect, soil and root was significant (ФST = 0.303), being the largest proportion of the variation within (69.6%) than among (30.3%)these groups.The mortality of spittlebugs by M. anisopliae before application ranged from 0 to 14.8% for nymphs and 7.3-18.1% for adults.Seven days after application, 50% of nymph mortality and 10.7% of adult mortality was observed.The isolate applied on the field was recovered only in spittlebugs and only 7, 30 and 60 days, post-application, and accounted for 50%, 50% and 70.5% of all insects killed by M. anisopliae, respectively.The pest population reduced after 90 days and no spittlebug infected was observed in this sample date.The species and clades found in isolates from soil were M. robertsii (clades Mrob 1, Mrob 2 and Mrob 4), M. anisopliae (Mani 1 and Mani 2) and M. brunneum.In roots the species found were M. brunneum and M. anisopliae (Mani 2).In adults and nymphs of spittlebug only haplotypes of a single clade (Mani 2) of M. anisopliae were found.The isolate ESALQ 5310 applied in previous years in the area was never recovered.The results obtained in this work revelead the great diversity of haplotype Metarhizium spp.and the impact of local populations of M. anisopliae on population regulation of spittlebugs in sugarcane.