Cloning and characterization of a Nep1-like gene from anthracnose fungus Colletotrichum gloeosporioides
Date Issued
2011
Date
2011
Author(s)
Kan, Chia-Cheng
Abstract
Colletotrichum gloeosporioides (Penz.) Penz.& Sacc., which causes anthracnose on a wide range of crops, is regarded as a hemibiotrophic pathogen. In the biotrophic stage of infection, no obvious necrosis symptom appears on the host; however, in the necrotrophic stage, C. gloeosporioides causes rapid and wide-spread cell death of plant cells, which constitutes the lesion symptom characteristic of anthracnose. Nep1-like proteins (NLPs), which were named after the necrosis and ethylene-inducing protein (NEP1) of Fusarium oxysporum f. sp. erythroxyli, comprise a large family of secreted proteins that cause rapid cell death and general stress responses on dicotyledonous plants. In this study, we found that C. gloeosporioides contains a single copy gene encoding NLP and named it CgNlp1. Analysis by reverse transcriptase-polymerase chain reaction indicated that the CgNlp1 message was detectable in mycelia harvested from different cultures and inoculation of the mango fruit. Moreover, its expression was enhanced by alkalization and as the infection proceeding. To characterize the function, we expressed the gene by agroinfiltration, and found that the protein of CgNlp1 caused necrosis on the tobacco plants 36 hours post infiltration. These results indicated that CgNlp1 expression is related with environmental factor and infection, also its gene product is able to cause cell death on plants. We prospect to develop a PEG-mediated transformation system for functional analysis. Whether CgNlp1 plays an essential role in the pathogenesis of C. gloeosporioides awaits further investigation.
Subjects
Colletotrichum gloeosporioides
secreted protein
Nep1-like gene
necrosis
PEG-mediated transformation
Type
thesis
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