A RAPID AND EFFICIENT METHOD FOR THE ISOLATION OF MITOCHONDRIAL DNA FROM SMALL AMOUNTS OF SOME FRUIT TREES

Abstract

the plant mitochondrial genome is a source of potentially useful markers for studies of phylogeny and population genetics. The isolation of high quality mtDNA from plants containing a high content of polyphenolics has been a difficult problem because the presence of metabolites that interfere with DNA isolation procedures and downstream applications such as PCR and DNA restriction. The present study describes a rapid and easy protocol that provides mtDNA of sufficient purity from small amounts (milligrams to 1gram) leaves tissue from different species of fruit trees such as pear (Pyrus betulaefolia), apple (Anna), olive (Kalamata), mango (Sukkary), and banana (Grand Nain), containing a high content of polyphenolics. The purity of mtDNA was confirmed by PCR amplification of genomic, mitochondrial gene and RAPD profiles. The mitochondrial COXIII gene of 400 bp was amplified while the genomic β-actin gene was not amplified. The technique is fast, inexpensive, reproducible, and suitable for PCR-based markers.