Identification of Sitotorga cerealella digestive system predominant symbiotic bacteria

Subject Areas : entomology and othea arthropods

raheleh moradi ¹ , M. Kazazi ² , GH. Khodakaramiyan ³ , E. Sedaghatfar ⁴

1 - Plant protection
2 - عضو هیات علمی گروه حشره شناسی دانشگاه بوعلی سینا همدان
3 - عضو هیات علمی گروه بیماری شناسی گیاهی دانشگاه بوعلی سینا همدان
4 - عضو هیات علمی گروه بیماری شناسی گیاهی دانشگاه آزاد اسلامی واحد اراک

Received: 2022-01-03 Accepted : 2022-02-21 Published : 2022-02-20

Keywords:

Abstract :

In this study,symbiotic bacteria of Sitotorga cerealella were identified. Containers containing barley were stored in a growth chamber at a temperature of 24 ° C and a relative humidity of 65% after contamination. To separate the bacteria of Sitotorga cerealella gastrointestinal tract,10 healthy larvae were selected from the reared population. After disinfection, the larvae gastrointestinal tract was removed with ethanol 70%, sodium hypochlorite and distilled water5%. Then, one milliliter of 0.1 M phosphate buffer with pH 7 was used to homogenize the gastrointestinal tract. The homogeneous and diluted contents were cultured in nutrient agar medium. To perform polymerase chain reactions from 5 μl of Master Mix ,2 μl of extracted DNA, 1.5 μl of RP2 primer and 1.5 μl of FD1 primer and 15 μl Distilled Water that the total volume of these materials should reach 25 microliters. The PCR process was performed using general primers FD1 and RP2 and 16srRNA gene sequence in a thermocycle.The PCR product sequencing was performed in collaboration with South Korea&amps Bioneer and the results were reviewed at the NCBI site. The phenotypic characteristics of the isolated bacteria were determined by standard bacteriological methods. After reviewing all the prepared colonies and comparing the obtained results, the highest frequency in terms of number of colonies was specific to Enterococcus mundtii and Enterobacter hormaechei was less abundant in the gastrointestinal tract of Sitotorga cerealella. Also, according to electrophoresis, the highest bands observed belong to Enterobacter hormaechei and the shortest band belongs to Enterococcus mundtii.