Gene expression and proteome analysis during the Musa acuminata-Mycosphaerella musicola interaction

R.N.G. Miller, M.A.N. Passos, F.L. Emediato, V.O. Cruz, O. Silva Jr., O.L. Franco, N.G. Oliveira-Junior
Sigatoka leaf spot disease in banana (Musa), caused by the fungal pathogen Mycosphaerella musicola, can cause significant reduction in functional leaf area and premature fruit ripening. In order to develop a functional genomics resource for this crop which offers insights into molecular mechanisms involved in host responses to this biotic stress, an analysis of differential gene expression was conducted in 'Calcutta 4' (AA genome, Musa acuminata ssp. burmannicoides) and 'Grande Naine' (AAA genome, Cavendish subgroup), contrasting in resistance to the fungal pathogen. Total cDNA was prepared from whole plant leaf materials, both uninfected and artificially challenged with pathogen conidiospores. From a subset of defense-related genes identified in silico based upon available 454 GS-FLX Titanium-derived sequence transcripts, gene expression was further examined by RT-qPCR. Data were analyzed using programs Rest and Miner to calculate primer efficiency and differential gene expression. Transcripts displaying a significant change in expression between infected and non-infected treatments included, amongst others, chitinases, thaumatin, glutathione transferases, a putative avrRpt2-induced AIG2 protein, a Germin/oxalate oxidase, a regulator of pathogen resistance responses of RPS2 and RPM1 genes, and an IAA-amino acid hydrolase. Simultaneously, the proteomic profile for this pathosystem was conducted from the inoculated and non-inoculated leaf materials. Isoelectric focusing of proteins, followed by second-dimension separation using 12% SDS polyacrylamide gels, enabled differential protein spots to be identified for characterization by MALDI-TOF-TOF. Analysis of gene and protein expression in this pathosystem is important for furthering our understanding of defense mechanisms in Musa and development of disease control strategies.
Miller, R.N.G., Passos, M.A.N., Emediato, F.L., Cruz, V.O., Silva Jr., O., Franco, O.L. and Oliveira-Junior, N.G. (2016). Gene expression and proteome analysis during the Musa acuminata-Mycosphaerella musicola interaction. Acta Hortic. 1114, 149-152
DOI: 10.17660/ActaHortic.2016.1114.21
proteomics, pyrosequencing, Sigatoka leaf spot, transcriptomics

Acta Horticulturae