Genomic DNA Isolation and analysis of Botryodiplodia theobromae, a tropical fruits pathogenic fungus

Noor Hasniza, Md Zin (2003) Genomic DNA Isolation and analysis of Botryodiplodia theobromae, a tropical fruits pathogenic fungus. [Final Year Project Report] (Unpublished)

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Abstract

Botryodiplodia theobromae, the plane pathogenic fungus that cause rot disease to the plant was grown on natural media. This study compare the effect of four factors to the enzymes activity and gene analysis from genomic DNA extracted. The factors are media, range of time for fungal growth, ratio of fruit to water (in media) and shaking condition of cultures during fungal growth. The natural media are citrus and banana; two types of tropical fruits that have different water content. Enzymes extracted were analyzed for amylase, cellulase and xylanase activity by using enzyme assay and also for protein activity. Protein activity was studied for the additional information. The results indicate that xylanase has the highest activity among the three enzymes studied. Water ratio at 1: 1 is also shows the highest activity. Banana media shows the faster growth of the fungus compared to citrus media due to the fruit's nutrient composition. Shaking factor affecting the fungal growth further more promotes the enzyme activity. In gene analysis, APAI gene and ApA2 gene that responsible to code the amylase enzyme activity was detected at 250bp. The gene was cloned into Escherichia coli JM 109. The enzyme activity data suggest that xylanase play the most important role as degrading enzyme for fruit degradation compared to other enzymes. Data from genes analysis give us an idea to exploit the amylase potential as degrading enzyme in many industrial fields

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