Korean Journal of Environmental Biology (환경생물)

Korean Society of Environmental Biology (한국환경생물학회)
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Assessment of phosphine susceptibility and genetic analysis of dld and cyt-b5-r expression and mutations in Korean Tribolium castaneum

국내 거짓쌀도둑거저리의 포스핀 감수성 평가 및 포스핀 저항성 관련 유전자인 dld와 cyt-b5-r의 발현량과 점 돌연변이 분석

  • Donghyeon Kim (Department of Applied Biosciences, Kyungpook National University) ;
  • Jinuk Yang (Department of Integrative Biology, Kyungpook National University) ;
  • Junyeong Park (Department of Integrative Biology, Kyungpook National University) ;
  • Bongsu Kim (Plant Quarantine Technology Center, Animal and Plant Quarantine Agency(APQA)) ;
  • Jun-Ran Kim (Plant Quarantine Technology Center, Animal and Plant Quarantine Agency(APQA)) ;
  • Sung-Eun Lee (Department of Applied Biosciences, Kyungpook National University)
  • 김동현 (경북대학교 응용생명과학과) ;
  • 양진욱 (경북대학교 농생명융합공학과) ;
  • 박준영 (경북대학교 농생명융합공학과) ;
  • 김봉수 (농림축산검역본부 식물검역기술개발센터) ;
  • 김준란 (농림축산검역본부 식물검역기술개발센터) ;
  • 이성은 (경북대학교 응용생명과학과)
  • Received : 2024.08.09
  • Accepted : 2024.09.23
  • Published : 2024.09.30
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Abstract

Phosphine (PH3) fumigation has been widely used for controlling storedgrain insect pests, causing the development of resistance of stored-grain insect pests to phosphine. PH3 resistance in Sitophilus oryzae has been reported in Korea. However, PH3 resistance in Tribolium castaneum has not been reported yet. This study was conducted to determine susceptibilities of T. castaneum collected from five different domestic locations to PH3. The susceptibility to PH3 was investigated using the FAO fumigation method. All domestic T. castaneum individuals were controlled by PH3 at 0.04 g m-3 . At 0.01 g m-3, T. castaneum collected from two domestic locations did not exhibit 100% mortality. A P45S point mutation in dihydrolipoamide dehydrogenase (dld) gene was found in a PH3-resistant strain of T. castaneum (Aus07), but not in five domestic stains or a PH3-susceptible strain (Aus10). No significant difference was found in dld or cyt-b5-r gene expression across all tested strains. However, the Gyeongju-collected strain of T. castaneum showed more than a 1.7-fold increase in cyt-b5-r expression compared to the Aus07 strain. cDNA sequence analysis revealed that P45S (C133T) in the dld gene was only present in Aus07. A characteristic single nucleotide polymorphism in the cyt-b5-r gene sequence was identified in the five domestic strains. This study suggests that it is necessary to continuously monitor PH3-susceptibility of T. castaneum in Korea to quickly identify resistant individuals and prevent the spread of PH3 resistance through rapid control.

포스핀(PH3) 훈증은 저곡해충을 방제하기 위해 널리 사용되어 왔으며, 이러한 높은 의존도에 의해 저곡해충에서 포스핀에 대한 저항성이 유발되었다. 국내에서는 쌀바구미에서 포스핀 저항성이 보고된 바 있으나, 거짓쌀도둑거저리에서는 포스핀 저항성이 보고된 바 없다. 본 연구는 국내 다섯 지역에서 채집한 거짓쌀도둑거저리를 이용하여 포스핀에 대한 감수성을 평가하기 위해 수행되었다. 포스핀에 대한 감수성은 FAO 훈증법을 통해 조사하였으며, 실험에 사용한 모든 국내 거짓쌀도둑거저리 개체는 0.04 g m-3 농도의 포스핀에서 사멸되었다. 0.01 g m-3 농도의 포스핀에서는 국내 두 지역에서 채집한 거짓쌀도둑거저리에서 100%의 사멸이 이루어지지 않았다. dihydrolipoamide dehydrogenase (dld) 유전자의 P45S 점돌연변이는 포스핀 저항성 거짓쌀도둑거저리 계통(Aus07)에서 확인되었으나, 포스핀 감수성 거짓쌀도둑거저리 계통(Aus10)과 국내 다섯 지역에서 채집한 거짓쌀도둑거저리에서는 확인되지 않았다. dld 유전자와 사이토크롬 b5 환원효소(cyt-b5-r) 유전자의 발현량을 비교한 결과, 모든 시험 종에서 유의미한 차이가 발견되지 않았다. 그러나 cyt-b5-r 유전자의 경우에, Aus07에서 발현량이 감소한 것에 비해서 경주 채집 종에서 1.7배 이상 발현량이 높은 것을 확인하였다. 또한, cDNA 염기서열을 분석한 결과, dld 유전자 염기서열의 P45S (C133T)는 Aus07에서만 확인되었으며, 국내 5개 채집 지역의 거짓쌀도둑거저리 종에서는 cyt-b5-r 유전자 염기서열에서 특징적인 염기서열다형성이 확인되었다. 이번 연구는 국내 거짓쌀도둑거저리의 포스핀 감수성을 지속적으로 모니터링하여 저항성 개체를 신속하게 확인하고, 신속한 방제를 통해 포스핀 저항성 확산을 방지할 필요가 있음을 시사한다.

Keywords

Acknowledgement

본 연구는 농림축산검역본부 학술연구용역과제(과제번호 Z-1543086-2023-25-02) 연구비를 지원받아 수행되었습니다.

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