Evolution of GHF5 endoglucanase gene structure in plant-parasitic nematodes: no evidence for an early domain shuffling event.
BACKGROUND
Endo-1,4-beta-glucanases or cellulases from the glycosyl hydrolase family 5 (GHF5) have been found in numerous bacteria and fungi, and recently also in higher eukaryotes, particularly in plant-parasitic nematodes (PPN). The origin of these genes has been attributed to horizontal gene transfer from bacteria, although there still is a lot of uncertainty about the origin and structure of the ancestral GHF5 PPN endoglucanase. It is not clear whether this ancestral endoglucanase consisted of the whole gene cassette, containing a catalytic domain and a carbohydrate-binding module (CBM, type 2 in PPN and bacteria) or only of the catalytic domain while the CBM2 was retrieved by domain shuffling later in evolution. Previous studies on the evolution of these genes have focused primarily on data of sedentary nematodes, while in this study, extra data from migratory nematodes were included.


RESULTS
Two new endoglucanases from the migratory nematodes Pratylenchus coffeae and Ditylenchus africanus were included in this study. The latter one is the first gene isolated from a PPN of a different superfamily (Sphaerularioidea); all previously known nematode endoglucanases belong to the superfamily Tylenchoidea (order Rhabditida). Phylogenetic analyses were conducted with the PPN GHF5 endoglucanases and homologous endoglucanases from bacterial and other eukaryotic lineages such as beetles, fungi and plants. No statistical incongruence between the phylogenetic trees deduced from the catalytic domain and the CBM2 was found, which could suggest that both domains have evolved together. Furthermore, based on gene structure data, we inferred a model for the evolution of the GHF5 endoglucanase gene structure in plant-parasitic nematodes. Our data confirm a close relationship between Pratylenchus spp. and the root knot nematodes, while some Radopholus similis endoglucanases are more similar to cyst nematode genes.


CONCLUSIONS
We conclude that the ancestral PPN GHF5 endoglucanase gene most probably consisted of the whole gene cassette, i.e. the GHF5 catalytic domain and the CBM2, rather than that it evolved by domain shuffling. Our evolutionary model for the gene structure in PPN GHF5 endoglucanases implies the occurrence of an early duplication event, and more recent gene duplications at genus or species level.
Kyndt
Tina
T
Department of Molecular Biotechnology, Ghent University, Ghent, Belgium. tina.kyndt@ugent.be
Haegeman
Annelies
A
Gheysen
Godelieve
G
D000818:Animals; D001483:Base Sequence; D002480:Cellulase / Q000737:chemistry / Q000235:genetics*; D019143:Evolution, Molecular; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics*; D007438:Introns; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009348:Nematoda / Q000737:chemistry / Q000145:classification / Q000201:enzymology* / Q000235:genetics*; D010802:Phylogeny; D010944:Plants / Q000469:parasitology*; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment
D015801:Helminth Proteins; D002480:Cellulase