Abstract:

Being able to rapidly and reliably detect Botryosphaeria dothidea will enable better disease monitoring and control efforts. In the present study, we developed conventional PCR, nested PCR, and LAMP assays in order to specifically differentiate between B. dothidea and other fungal species. Then, the sensitivity of the three detection methods was compared and analyzed. Finally, these assays were used to detect diseased samples collected from fields. The results showed that the conventional PCR, nested PCR and LAMP assays were able to detect and diagnose fruit diseases caused by B. dothidea. Sensitivity analyses revealed that the detection limit for conventional PCR assay was 10^-1 ng/μL of B. dothidea DNA, whereas the nested PCR assay had a 10^-4 ng/μL detection limit, and the LAMP assay was sensitive to as little as 10^-5 ng/μL of fungal DNA. The nested PCR and LAMP assays can be reliably utilized to diagnose fruit diseased samples from field caused by B. dothidea. The positive rates of the LAMP detection, nested PCR, conventional PCR and traditional identification method were 65.0%, 62.5%, 58.8% and 27.5% respectively.

Key words: fruit diseases, Botryosphaeria dothidea, PCR, LAMP, detection