I have some mango fruit samples in our lab. Which is the best Anthracnose extraction method from mango fruit?
Take an early stage lesion of typical anthracnose. Surface disinfect the fruit with 10% chlorox bleach for 4 minutes. Let the surface disinfected fruit dry off under the sterile hood.
Flame the scalpel and excise the anthracnose lesion. Continue to make small pieces from the margin of the lesion and then plant these in general media such as potato dextrose agar. When you can identify the colonies with characteristic acervuli then purifify and multiply.
Antracnose fungi can be co mingled with bacteria acidified PDA helps but ideally to get to a single spore isolation your will eliminate and co contaiminant.
While PDA can work for sporulation increase oatmeal can be more productive.
Once you have purified source colonies the preservation on slants under sterile mineral oil works well.
For large quantities of spores grow on sterilized millet or and sorghum in Erlenmeyer flasks under 12 lighting.
Thanks Paul.
Can 70% ethanol replace 10% chlorox? What about use of sodium hypochlorite?
Chlorox is sodium hypochlorite. Generally the Clorox bottle has 5.25% sodium hypochlorite which when dliluted 9 to 1 or the 10% which is 0.525% sodium hypochlorite that is pretty standard. Check your bottle and brand for the concentration it claims.
The time of submerging in 10% Chlorox or 0.525 sodium hypochlorite is 4 minutes. Some people use 70% ethanol for 1 minute and then give 1 minute rinse in sterile destilled water as a standard procedure.
Ethanol at times can promote bacteria. The drying period in the sterile hood prevents excess water from interfering with the growth of the fungus if the material is too wet it favors bacterial contamination.
Some people use flaming of lesioned tissues and plate but this is a little standardized. Flaming works well for internal wilf fungi from twigs of plants suffering vascular wilt diseases.
If you use old lesions for isolation you tend to get a lot of secondary organisms. Secondary organisms will take over the older central part of the lesion.
Good hunting.
The Mango anthranose organism is usually identified as Colletotrichum gloeosporioides and that fungus can show a lot of isolate variations and forms.
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