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Rapid Diagnosis of Resistance to Glufosinate-ammonium in Transgenic Sweet Potato

형질전환 고구마에 대한 Glufosinate-ammonium 저항성 간이진단법

  • Kuk, Yong-In (Dept. of Development in Resources Plants, College of Life Science and Natural Resources, Sunchon National University)
  • 국용인 (순천대학교 생명산업과학대학 자원식물개발학과)
  • Received : 2010.08.27
  • Accepted : 2010.11.02
  • Published : 2010.12.30

Abstract

Transgenic herbicide-resistant sweet potato plants [Ipomoea batatas (L.) Lam.] produced through a biolistic transformation were used in this study. The objective of this research was to find out a rapid and reliable assay method for confirming glufosinate-ammonium resistance. The techniques tested include whole-plant bioassay, one leaf bioassay, and leaf disk bioassay. Parameters investigated in this study were leaf injury and ammonium accumulation at 1 and 5 days after treatment of glufosinate-ammonium. In the leaf disk bioassay, leaf injury of the transgenic line 7171 was 1.9-fold less affected by glufosinate-ammonium than the wild type. The leaf injury of 7171 in one leaf and whole-plant bioassays was 59- and 92-fold less affected by glufosinate-ammonium, respectively, compared with that of the wild type. Leaf disk, one leaf, and whole-plant bioassays showed that ammonium accumulation of the 7171 was 2 to 20-, 4 to 43-, and 6 to 115-fold less affected by 0.5-5 mM glufosinate-ammonium than that of the wild type. All three bioassays successfully distinguished the resistance from the transgenic lines, but one leaf bioassay is the simplest and quickest. Leaf injury and ammonium accumulation were the same in leaves 1, 3, 5, 7, and 10 of 3 mM glufosinate-ammonium treated plants or nontreated plants. The one leaf bioassay was chosen as the standard procedure for future confirmation of resistance in transgenic sweet potato because it is a rapid and reliable assay.

본 연구는 유전자총기법에 의해 생산된 형질전환 고구마를 사용하여 glufosinate-ammonium에 대한 저항성 유무를 빠르고, 경제적이며, 신뢰할 수 있는 간이분석법을 찾고자 수행되었다. 저항성 진단법은 whole-plant 진단법, one leaf 진단법 및 leaf disk 진단법을 사용하였고, 이들 진단법에 의해 glufosinateammonium을 처리하고 잎의 피해율과 암모늄 축적량을 조사하였다. Leaf disk 진단법에서 glufosinateammonium 처리 후 형질전환 라인 7171의 잎에 대한 약해는 wild type에 비해 1.9배 낮았다. One leaf 진단법과 whole plant 진단법의 경우 glufosinateammonium 처리 후 7171의 잎에 대한 약해는 wild type에 비해 각각 59배와 92배 적었다. Leaf disk, one leaf와 whole plant 진단법에서 0.5-5mM glufosinateammonium 처리 후 암모늄 축적은 wild type이 7171에 비해 각각 2-20, 4-43 및 6-115배 더 많이 축적되었다. 이들 3가지 방법 모두 형질전환 고구마에 대한 glufosinate-ammonium 저항성 유무를 판단하는데 가능한 방법이나 one leaf 진단법이 가장 간편하면서 빠른 진단방법이었다. 그러나 이 방법은 엽령별 내성차이가 존재할 수 있어 고구마 엽이 10개 전개되었을 때 1, 3, 5, 7, 9엽을 각각 채취하여 3mM glufosinateammonium을 처리시 잎의 피해율과 암모늄 축적을 엽령별로 조사한 결과 유의적인 차이를 보이지 않았다. 따라서 분석에 소요되는 기간, 시료량, 정확성 등을 고려할 때 형질전환 고구마의 저항성을 판별하는데 one leaf 진단법이 가장 적합한 것으로 판명 되었다.

Keywords

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