| 摘要 |
PURPOSE:To obtain the subject DNA probe capable of accurately judging whether mycelia of mycorrhizal fungus are taking roots of a host plant such as pine tree and multiplied or not and of rapidly and simply detecting a mycorrhizal fungus, containing a DNA specifically bondable complementary with nucleic acid of mycorrhizal fungus. CONSTITUTION:Preservation strain IFO 6,918 of Armillaria matsudake and separated strain Ls. No1 of Lyophyllum aggregatum are subjected to liquid culture, mycelia are grown on the whole area of a medium, collected and frozen with liquid nitrogen. The mycelia are ground by a homogenizer in liquid nitrogen and extracted with 10mM tris hydrochloric acid buffer solution (pH 8.00) containing 0.1M NaCl. The extracted solution is treated with phenol and a DNA is precipitated by adding an alcohol to the extracted substance and separated. RNA is decomposed and removed. The DNA is amplified with two kinds of primers composed of 18 SrRNA derived from a mycorrhizal fungus by PCR reaction and the amplified DNA is confirmed to be specifically hybridized with nucleic acid of mycorrhizal fungus to give the objective DNA probe for detecting a mycorrhizal fungus, specific to nucleic acid derived from a mycorrhizal fungus such as Armillaria matsudake or Lyophyllum aggregatum. |
