Comparative analysis of expressed genes from cacao meristems infected by Moniliophthora perniciosa

Abstract

Background and aims: Witches' broom disease is caused by the hemibiotrophic basidiomycete Moniliophthora perniciosa, and is one of the most important diseases of cacao in the western hemisphere. Because very little is known about the global process of such disease development, expressed sequence tags (ESTs) were used to identify genes expressed during the Theobroma cacao-Moniliophthora perniciosa interaction.

Methods: Two cDNA libraries corresponding to the resistant (RT) and susceptible (SP) cacao-M. perniciosa interactions were constructed from total RNA, using the DB SMART Creator cDNA library kit (Clontech). Clones were randomly selected, sequenced from the 5' end and analysed using bioinformatics tools including in silico analysis of the differential gene expression.

Key results: A total of 6884 ESTs were generated from the RT and SP cDNA libraries. These ESTs were composed of 2585 singlets and 341 contigs for a total of 2926 non-redundant sequences. The redundancy of the libraries was low and their specificity high when compared with the few other cacao libraries already published. Sequence analysis allowed the assignment of a putative functional category for 54 % of sequences, whereas approx. 22 % of sequences corresponded to unknown function and approx. 24 % of sequences did not show any significant similarity with other proteins present in the database. Despite the similar overall distribution of the sequences in functional categories between the two libraries, qualitative differences were observed. Genes involved during the defence response to pathogen infection or in programmed cell death were identified, such as pathogenesis related-proteins, trypsin inhibitor or oxalate oxidase, and some of them showed an in silico differential expression between the resistant and the susceptible interactions.

Conclusions: As far as is known this is the first EST resource from the cacao-M. perniciosa interaction and it is believed that it will provide a significant contribution to the understanding of the molecular mechanisms of the resistance and susceptibility of cacao to M. perniciosa, to develop strategies to control witches' broom, and as a source of polymorphism for molecular marker development and marker-assisted selection.

Fig. 1.

Histogram showing the distribution of ESTs by contigs. The contig size is the number of EST/contig.

Fig. 2.

Distribution of the 1719 RT and 1207 SP unigene sequences into functional classes. The 16 broad categories that were used for classification during the semi-automatic annotation are indicated, as well as the number of corresponding sequences. Only one class was assigned to each sequence.

Fig. 3.

Venn diagram showing the distribution of the sequences present in cacao libraries. (A) From Verica et al. (2004). The percentage values were based on the total sequence number (4465). (B) From Jones et al. (2002). The percentage values were based on the total sequence number (4004).