Genomic and expression analysis of glycosyl hydrolase family 35 genes from rice (Oryza sativa L.).
BACKGROUND
Many plant beta-galactosidases (Bgals) have been well characterized and their deduced biological functions mainly involve degradation of structural pectins, xyloglucans or arabinogalactoproteins in plant cell walls. However, gene multiplicity in glycosyl hydrolase family 35 (GH35), to which these proteins belong, implies diverse functions. In this study, the gene multiplicity, apparent evolutionary relationships and transcript expression of rice Bgal genes were examined, in order to predict their biological functions.


RESULTS
Fifteen rice Bgal genes were identified in the plant genome, one of which encodes a protein similar to animal Bgals (OsBgal9), and the remaining 14 fall in a nearly plant-specific subfamily of Bgals. The presence of both classes of Bgals in bryophytes, as well as vascular plants, suggests both gene lineages were present early in plant evolution. All 15 proteins were predicted to contain secretory signal sequences, suggesting they have secretory pathway or external roles. RT-PCR and database analysis found two distinct lineages to be expressed nearly exclusively in reproductive tissues and to be closely related to Arabidopsis Bgals expressed most highly in flower and pollen. On the other hand, OsBgal6 is expressed primarily in young vegetative tissues, and alternative splicing in panicle prevents its production of full-length protein in this reproductive tissue. OsBgal11 also showed alternative splicing to produce different length proteins. OsBgal13 produced by recombinant expression in Escherichia coli hydrolyzed alpha-L-arabinoside in addition to beta-D-galactoside and beta-(1-->3)-, beta-(1-->4)- and beta-(1-->6)- linked galacto-oligosaccharides.


CONCLUSIONS
Rice GH35 contains fifteen genes with a diversity of protein sequences, predicted locations and expression and splicing patterns that suggest that OsBgals enzymes may play a variety of roles in metabolism of cell wall polysaccharides, glycoproteins and glycolipids.
Tanthanuch
Waraporn
W
National Synchrotron Research Center, 111 University Avenue, Nakhon Ratchasima 30000, Thailand. waraporn@nsrc.or.th
Chantarangsee
Mallika
M
Maneesan
Janjira
J
Ketudat-Cairns
James
J
D017398:Alternative Splicing / Q000235:genetics; D032461:Chromosomes, Plant / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D005091:Exons / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D006027:Glycosides / Q000378:metabolism; D006868:Hydrolysis; D007438:Introns / Q000235:genetics; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D009699:N-Glycosyl Hydrolases / Q000235:genetics*; D009844:Oligosaccharides / Q000378:metabolism; D012275:Oryza / Q000201:enzymology* / Q000235:genetics*; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018626:Retroelements / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D018076:DNA, Complementary; D006027:Glycosides; D007527:Isoenzymes; D009844:Oligosaccharides; D010940:Plant Proteins; D018626:Retroelements; D001616:beta-Galactosidase; D009699:N-Glycosyl Hydrolases