Isolation and identification of the pathogen causing leaf spot disease on a rare and endan-gered plant species Rhododendron huadingense
Rhododendron huadingense is a rare and endangered plant species endemic to Zhejiang Province,and leaf spot is an important disease of this plant.R.huadingense leaves infected with leaf spot disease were used as materi-als.Based on pathogen isolation,purification,and pathogenicity identification,both morphological and molecular bi-ology methods were applied to determine the fungal pathogen.The results showed that one fungal strain designated HDDJ was isolated from the diseased leaves.Using Koch's postulates,the fungus caused leaf spot disease,and the characteristics of the lesions were consistent with those observed in the field.The colonies of HDDJ were white,then gradually turned gray,and finally became black with fluffy aerial mycelium.The hyphal characteristics of HDDJ were identical to those of Botryosphaeria dothidea.Fragments of actin and tubulin genes were cloned by using PCR,and phylogenetic analysis results revealed that both were clustered with B.dothidea.According to the morphological and molecular identification results,it was confirmed that the fungal pathogen causing R.huadingense leaf spot disease was B.dothidea.Isolation and identification of R.huadingense leaf spot disease laid a foundation for this disease control in the future.
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