1999 Fiscal Year Final Research Report Summary
Detection of powdery mikiew fungus from winter buds by the polymerase chai* reaction using species-specifrc prinerset.
| Project/Area Number |
09660055
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
植物保護
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| Research Institution | College of Technology, Toyama Prefectural University |
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Principal Investigator |
SATO Yukio College of Technology, Associate Prof., 助教授 (20104979)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAMATSU Susumu Bioresources Mie Uriv, Prof., 生物資源学部, 教授 (20260599)
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| Project Period (FY) |
1997 – 1999
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| Keywords | Powdery Mildew / Crape myrtle powdery mildew / species-specific primer set / Detection / PCR |
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| Research Abstract |
Powdery mildew fungi generally overwinter by cleistothecla. On the other hand, the powdery mildew fungus of crape myrtle (Lagerstroemia indica) is known to overwinter in winter buds of the plant. If the fungus latently infecting the winter buds can be detected, we can obtain the information as to when the winter buds are infected with infection frequency of the winter buds. This may clarify the life cycle of this fungus, which leads to the possibility of disease forecasting. We thus tried to develop the detection method of powdery mildew from the winter buds of crape myrtle by the polymerase chain reaction using species-specific primer set. The results are as follows.
1)DNA sequences of the ITS regions including 5.8S rDNA were determined for various powdery mildew fungi Based on the sequences, four primers specific to the powdery mildew fungus (Erysiphe australiana) of crape myrtle were constructed 2) Only the DNA of the powdery mildew of crape myrtle was amplified by the specific primer set, which indicates the usefulness of the primer set 3) Some faint bands were detected when fungal isolates from leaves of crape myrtle were subjected to the PCR amplification using the specific primer set. Some of these bands could not be removed even if the annealing temperature was changed 4)PCR-RFLP using the restriction enzyme Xhol I was useful to differentiate foe crape myrtle powdery mildew from the fungal isolates. 5) The minimum limit of detection the primer set SA2/SA3 was 0.0025pg DNA for the crape myrtle fungus, 6) Danas successfully extracted from the leaves of crape myrtle by using Dneasy Plant Mini- Kit combined with cleaning by Hepese.
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