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Establishment and characterization of three embryonic cell lines of beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae)

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Abstract

Three cell lines (QAU-Se-E-1, -2 and -3, or Se-1, -2 and -3 for short) were established from eggs of beet armyworm (Spodoptera exigua) that have been passaged stably for more than 60 times in TNM-FH medium supplemented with 10 % fetal bovine serum. The cell lines consisted of round and spindle-shaped cells. The round cells accounted for 96.82, 84.34 and 83.16 % of the cells in the three cell lines, respectively, with cell diameters of 16.21 ± 0.72, 15.63 ± 0.58 and 13.06 ± 0.44 μm. Random amplified polymorphic DNA and analysis of the CO I gene showed that the three cell lines were all derived from S. exigua. Growth curves at passage 30 were determined and the results showed that the cell population doubling times were 59.03, 49.08 and 49.91 h, respectively. The three cell lines can be infected by S. exigua multiple nucleopolyhedrovirus (SeMNPV). Se-3 was extremely susceptible to the virus with an infection rate of 97.52 % 4 days after the inoculation and produced 2.02 × 106 OBs per mL of culture. Flow cytometry analysis showed that some of Se-1 and Se-2 cells had apoptosis after infection, whereas Se-3 cells did not. Bioassays showed that the virulence of the SeMNPV proliferated from Se-3 was similar to that from the insect with LC50 of 5.55 × 105 and 2.64 × 105 OBs/mL. Therefore, the cell lines can be used to study the SeMNPV-host interactions and mechanisms underlying the interactions.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (31272376), Graduate Innovation Project of Qingdao Agricultural University (QYC201307), and Tai-Shan Scholar Construction Foundation of Shandong Province, China.

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Authors and Affiliations

  1. Key Lab of Integrated Crop Pest Management of Shandong Province, College of Agronomy and Plant Protection, Qingdao Agricultural University, Qingdao, 266109, Shandong, People’s Republic of China

    Rui Su, Gui-Ling Zheng, Fang-Hao Wan & Chang-You Li

  2. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences (CAAS), Beijing, 100193, People’s Republic of China

    Fang-Hao Wan

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  1. Rui Su
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  2. Gui-Ling Zheng
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  3. Fang-Hao Wan
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Correspondence to Chang-You Li.

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Su, R., Zheng, GL., Wan, FH. et al. Establishment and characterization of three embryonic cell lines of beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae). Cytotechnology 68, 1223–1232 (2016). https://doi.org/10.1007/s10616-015-9882-9

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  • DOI: https://doi.org/10.1007/s10616-015-9882-9

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