Hessian Fly Population Characterization

Four different Hessian fly populations were obtained from bread wheat from the major wheat producing areas in Morocco: Doukkala, Fes, Chaouia, and Safi (Table 1). The plants containing Hessian fly pupae were collected in winter 2017 and 2018 and they were reared in a growth room under constant conditions (Temperature 25 ± 1 °C, Relative Humidity 51.7%) from which adults were allowed to emerge. The first generation of adults were collected. A laboratory population kept at the International Center of Agricultural Research in Dry Areas (ICARDA) in Rabat, Morocco, was also included in the screening analysis (Table 1). All samples collected were placed separately (one adult per tube) in 100% ethanol and stored at room temperature until use. Before the DNA extraction, samples were rinsed with sterile water then the DNA of the whole fly was isolated using a modified CTAB (Cetyl Trimethyl Ammonium Bromide) method [46 (link)]. The quality and quantity of DNA samples was tested using a Q5000 micro-volume UV Vis spectrophotometer (Quawell Technology, San Jose, CA, USA). DNA samples were stored in Eppendorf tubes at −20 °C until PCR amplification and amplicon sequencing analysis.