Electrophoretic karyotype analysis of Crinipellis perniciosa, the causal agent of witches' broom disease of Theobroma cacao
ABSTRACT Pulse-field gel electrophoresis (PFGE) was used to determine the genome size and characterize karyotypic differences in isolates of the cacao biotype of Crinipellis perniciosa (C-biotype). The karyotype analysis of four isolates from Brazil revealed that this biotype could be divided into two genotypes: one presenting six chromosomal bands and the other presenting eight. The size of the chromosomes ranged from 2.7 to 5.3 Mb. The different genotypes correlate with telomere-based PCR analysis. The isolates with six chromosomal bands had two that appeared to be doublets, as shown by densitometric analysis, indicating that the haploid chromosome number for this biotype is eight. The size of the haploid genomes was estimated at approximately 30 Mb by both PFGE and Feulgen-image analysis. DNA hybridization revealed that the rDNA sequences are clustered on a single chromosome and these sequences were located on different chromosomes in an isolate dependent manner. This is the first report of genome size and chromosomal polymorphism for the C-biotype of C. perniciosa.
SourceAvailable from: Mateus Ferreira Santana[Show abstract] [Hide abstract]
ABSTRACT: Transposase sequence analysis is an important technique used to detect the presence of transposable elements in a genome. Putative transposase sequence was analyzed in the genome of the phytopathogenic fungus Moniliophthora perniciosa, the causal agent of witches' broom disease of cocoa. Sequence comparisons of the predicted transposase peptide indicate a close relationship with the transposases from the elements of the Tc1-Mariner superfamily. The analysis of the distribution of transposase sequence was done by means of PCR and Southern blot techniques in different isolates of the fungus belonging to C-, L-, and S-biotypes and collected from various geographical areas. The distribution profile of the putative transposase sequence suggests the presence of polymorphic copies among the isolates from C-biotypes. The total DNA hybridization profile of each isolate was used to calculate genetic distance and group by the UPGMA method. C-biotype isolates colleted from of the Bahia showed two hybridization profiles for the transposase sequence. Thus the two different fingerprinting profiles for transposase sequence reported here by Southern analysis could also be correlated to the presence of two different genotypes in Bahia, Brazil.Tropical Plant Pathology 10/2011; 36(5):276-286. DOI:10.1590/S1982-56762011000500002 · 0.55 Impact Factor
[Show abstract] [Hide abstract]
ABSTRACT: Moniliophthora perniciosa is a fungus that causes witches’ broom disease (WBD) in the cacao tree (Theobroma cacao). The M. perniciosa genome contains different transposable elements; this prompted an evaluation of the use of its retrotransposons as molecular markers for population studies. The inter-retrotransposon amplified polymorphism (IRAP) and retrotransposon-microsatellite amplified polymorphism (REMAP) techniques were used to study the variability of 70 M. perniciosa isolates from different geographic origins and biotypes. A total of 43 loci was amplified. Cluster analysis of different geographical regions of C biotype revealed two large groups in the state of Bahia, Brazil. Techniques using retrotransposon-based molecular markers showed advantages over previously used molecular techniques for the study of genetic variability in M. perniciosa.European Journal of Plant Pathology 11/2012; 134(3). DOI:10.1007/s10658-012-0031-4 · 1.71 Impact Factor
[Show abstract] [Hide abstract]
ABSTRACT: The production of polyclonal sera against plant pathogens allows the development of immunoassays to quickly identify and characterize the pathogens at low cost. Polyclonal sera were produced against Crinipellis perniciosa, the causal agent of witches' broom disease of cacao (Theobromae cacao) using hyphae extracts from isolates collected in Pará and Bahia states, to identify and distinguish isolates from various regions and hosts. Serum 1 was produced from isolate ESJOH-1 (collected in Pará) and serum 2, from isolate CP-85 (from Bahia). There was no cross reaction between the two sera with mycelia extracts from Alternaria solani, Marasmius sp., Oudemansiella canarii and Verticillium fungicola. The sera presented high affinity cross reaction with Moniliophthora roreri and medium- to low cross-reaction against Marasmius cladophyllus. The sera were evaluated against isolates from biotype-C, collected in the states of Amazonas, Bahia, Mato Grosso, Rondônia and Pará. The serological reaction of both sera did not allow to distinguish the isolates' site of origin (Pará and Bahia), nor did they differentiate isolates of biotype C. The antigen binding sites were different for serum 1 and 2, because both exhibited different response to the same strains. However, these differences in affinity did not allow for differentiating the origin of collection, host or biotype, but allowed for the identification of C. perniciosa belonging to biotypes C and S.Fitopatologia Brasileira 10/2005; 30(5):482-488. DOI:10.1590/S0100-41582005000500004