Keywords: Fungi; Colletotrichum gloeosporiodes; Pathogen; Endophyte; Protein profiling
Class – Sordariomycetes
Family - Glomerallaceae
The genus Colletotrichum comprises ~ 600 species . Exist in two stages – telomorph (sexual stage) called as Glomerella cingulata and anamorph (asexual) Colletotrichum gloeosporiodes. The generic name Colletotrichum was introduced by Corda for C.lineola, a species found associated with a member of the Apiaceae in the Czech Republic . It attacks over 3,200 species of monocot and dicot plants. Distribution wise Colletotricum has Cosmopolitan nature. Almost all of the plants are susceptible to one or more species of Colletotricum.
Eg: Taxomyces andreanae which produces taxol.
Proteomics complements other functional genomics approaches including transcriptomics . Colletotrichum exhibit different forms in their life cycle even within a single species, like biotrophy, necrotrophy, endophytic and latency. Each stage brings about different interaction and during these stages many proteins are released due to differential gene expression.
Biotrophy ⇨ Necrotrophy ⇨ Endophytic ⇨ Latency
Biotrophic fungi have variations in their tubular cells, the hyphae. Haustorium is the specialized hyphae.
Necrotrophic fungi secrete toxins and kill host cells as soon as they enter. They take the nutrients from plants and kill them. Endophytic fungi doesn’t sporulate at all. They can be seen inside the cell. They live in mutualism. Latency period is that between sporulation and germination. It remains in an inactive stage and there will be no growth.
Protein profiling can give an overall picture about the conversion of a pathogen to an endophyte. There are reports for the presence of plant like S8A – subtilisin in the genome of Colletotrichum. Evidence of molecular mimicry for the production of subtilisin by horizontal gene transfer from the plants has been reported . These proteins show evidence of lateral gene transfer from plants to a Colletotrichum ancestor. It’s possible that Colletotrichum acquire and use plant like subtilisins to manipulate host metabolism . The increase of plant-pathogen interaction analysis, focused on the plant–fungus association is a topic of rising interest in the last five years; even when the limitation in sequence availability in public databases is also challenging . Hence it’s a unique opportunity for comparative proteomic analysis of their association with plants.
2. Differentially expressed spots are subjected to in gel trypsin digestion .
3. After crystallization by standard procedures, the peptides are analysed by MALDI-TOF MSor MALDI-TOF/TOF MS/MS using Proteomics analyser mass spectrometer . Peptides are identified by MASCOT search engine.
Grinyer, et al. used a combination of 2D-PAGE and mass spectrometry for the identification of proteins from filamentous fungi . Hernández-Macedo, et al. carried out fungal protein vizualisation by 2DPAGE . A filamentous fungi used as a model organism is Aspergillus sp. The majority of the Aspergillus proteome research is still represented by quantitative 2D studies and less than 10% of the predicted whole proteome of Aspergillus sp [42-44]. Has been identified and quantified until now. Some of the fungal proteins which are studied are shown in Table 1.
Author & Year
Mycellial intracellular proteome
Carberry, et al. 
Intracellular proteome of dormant conidia
Teutschbein, et al. 
Cytosolic proteome reference map
Lu, et al. 
Intracellular proteome reference map
Jami, et al. 
One of the constrain of proteomic analysis of filamentous fungi is that only less than 10% of the predicted whole proteome are identifiable. Under laboratory conditions, only a fraction of the total genome gives rise to proteins. Expression of other proteins will be under different nutritional and environmental conditions in nature.
2. Develop systematic understanding of virulence factors in pathogenic fungi. Fernandez-Acero, et al. .
3.Providing insight related to systematic metabolic flux changes
Endophytic fungi are an important bio resource which are not utilised properly due to lack of understanding about the physiological and biochemical aspects. If utilised properly, these organism can have lot of application in medicine, industry and agriculture.
Proteomic analysis is a powerful tool capable of resolving thousands of proteins and thereby allows a systematic understanding of the molecular events that occur within an organism’s various physiological states, protein expression level of specific genes, and through cellular fractionation can provide spatially specific information on protein expression. For C. gloeosporiodes, post genomic studies (especially in the area of proteomics) are just beginning to take place and considering the importance of the organism as both a pathogen and endophyte, it is surprising that proteome profiling is yet to be done. Our current studies are focussed on the same.
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