DNA barcoding of selected plant pathogenic fungi in Singapore
Molecular techniques, like Polymerase Chain Reaction (PCR) and DNA sequencing, have been recognized as an important tool in fungal species identification. However, the current available protocols for fungi have many drawbacks, including exposure to harmful chemicals such as phenol and chloroform, lo...
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| Format: | Final Year Project |
| Language: | English |
| Published: |
2013
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| Online Access: | http://hdl.handle.net/10356/53821 |
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| Institution: | Nanyang Technological University |
| Language: | English |
| Summary: | Molecular techniques, like Polymerase Chain Reaction (PCR) and DNA sequencing, have been recognized as an important tool in fungal species identification. However, the current available protocols for fungi have many drawbacks, including exposure to harmful chemicals such as phenol and chloroform, low DNA yield and purity and also complex and time-consuming steps. In this experiment, pure fungal cultures were first isolated and then used for the testing of a protocol involving either of two different DNA extraction methods. Method 1 involved the physical disruption of the fungal cell wall by a hand-held drill and liquid nitrogen whereas Method 2 utilizes microwave irradiation. From the experiment, the most common plant pathogenic fungi isolated were Colletotrichum sp., Fusarium sp., Macrophoma sp., Pestalotiopsis sp. and Phomopsis sp. and the protocol, depending on the Methods 1 and 2 used, worked successfully on only certain fungi types. Method 1 worked best for Macrophoma sp. while Method 2 for Pestalotiopsis sp.. For Colletotrichum sp., both methods produced varying results while for Fusarium sp. and Phomopsis sp., results, although varying, were produced only with Method 2. |
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