Molecular Identification of the Local Isolated Fungi Aureobasidium pullulans

Abstract

Twenty fungal isolates of Aureobasidium were isolated from many healthy leaves ofdifferent citrus trees. Fungal isolates understudy were identified by using morphological andcultural characters. The results revealed that the isolates are belonged to the species A .pullulans.Molecular identification of the isolated fungi were carried out using the specific-PCR reaction. Theresults showed 17 bands of purified genomic DNA from local fungal isolates of similar size ranging(500-600)bp. These bands have the same size to that obtained from the standard strain A. pullulansNRRL 58560. On the other hand, it did not show any band of the DNA as a result of the Specific-PCR reaction specialize of isolates (Ap15 and Ap17 and Ap18) in the agarose gel.The sequence of the nitrogenous bases for the Specific PCR products were determined forthree chosen local isolates. The results were analyzed using DNA Blast / NCBI program revealedthat there is a similarity higher than 95% between these sequences and those of the standardstrains of A. pullulans already recorded in the Gene Bank. Additionally, the point mutationswhether they are substitution or addition or deletion were observed in some positions of thenitrogenous bases after comparing the sequences of the tested local isolates and the standard.These mutations may occur spontaneously.This analysis results sent to GenBank using the website of the NCBI and introduced inGenBank record as standard isolates and given a cod number in the GenBank as follows: Ap1.sqnAp1 KX964610, Ap7.sqn Ap7: KX964611, Ap14.sqn Ap14 KX964612 This confirms the validityof the results obtained in this study and diagnosis of the validity of the local isolates under study.