Specific and Rapid Detection of Camellia oleifera Anthracnose Pathogen by Nested-PCR

Abstract

Camellia oleifera is an economical important plant in southern China for edible oil production. Anthracnose is a serious disease that limited its development. The internal transcribed spacer (ITS) regions and the 5.8S rRNA gene of strain C1 of the pathogenic fungus Colletetrichum gloeosporioides were sequenced in order to design specific PCR primers for pathogen detection. Alignment of the sequence data of strain C1 and the other Colletetrichum species obtained from the Genbank were made using CLUSTAL W. Based on the aligned ITS sequences, specific primers for C. gloeosporioides were developed (YT1 and YT2). The infecting pathogens were successfully detected with our specific primer set and showed high specificity. The result showed that the nested-PCR reaction was at least 10,000- fold more specific than that of the simple PCR method. This new method provides a useful technique to further study disease cycle and for early prediction of anthracnose of Camellia oleifera.