Glycoside-specific glycosyltransferases catalyze regio-selective sequential glucosylations for a sesame lignan, sesaminol triglucoside.

Sesame (Sesamum indicum) seeds contain a large number of lignans, phenylpropanoid-related plant specialized metabolites. (+)-Sesamin and (+)-sesamolin are major hydrophobic lignans, whereas (+)-sesaminol primarily accumulates as a water-soluble sesaminol triglucoside (STG) with a sugar chain branched via ß1→2 and ß1→6-O-glucosidic linkages [i.e. (+)-sesaminol 2-O-ß-d-glucosyl-(1→2)-O-ß-d-glucoside-(1→6)-O-ß-d-glucoside]. We previously reported that the 2-O-glucosylation of (+)-sesaminol aglycon and ß1→6-O-glucosylation of (+)-sesaminol 2-O-ß-d-glucoside (SMG) are mediated by UDP-sugar-dependent glucosyltransferases (UGT), UGT71A9 and UGT94D1, respectively. Here we identified a distinct UGT, UGT94AG1, that specifically catalyzes the ß1→2-O-glucosylation of SMG and (+)-sesaminol 2-O-ß-d-glucosyl-(1→6)-O-ß-d-glucoside [termed SDG(ß1→6)]. UGT94AG1 was phylogenetically related to glycoside-specific glycosyltransferases (GGTs) and co-ordinately expressed with UGT71A9 and UGT94D1 in the seeds. The role of UGT94AG1 in STG biosynthesis was further confirmed by identification of a STG-deficient sesame mutant that predominantly accumulates SDG(ß1→6) due to a destructive insertion in the coding sequence of UGT94AG1. We also identified UGT94AA2 as an alternative UGT potentially involved in sugar-sugar ß1→6-O-glucosylation, in addition to UGT94D1, during STG biosynthesis. Yeast two-hybrid assays showed that UGT71A9, UGT94AG1, and UGT94AA2 were found to interact with a membrane-associated P450 enzyme, CYP81Q1 (piperitol/sesamin synthase), suggesting that these UGTs are components of a membrane-bound metabolon for STG biosynthesis. A comparison of kinetic parameters of these UGTs further suggested that the main ß-O-glucosylation sequence of STG biosynthesis is ß1→2-O-glucosylation of SMG by UGT94AG1 followed by UGT94AA2-mediated ß1→6-O-glucosylation. These findings together establish the complete biosynthetic pathway of STG and shed light on the evolvability of regio-selectivity of sequential glucosylations catalyzed by GGTs.

Comparative effects of sesame seeds differing in lignan contents and composition on fatty acid oxidation in rat liver.

We compared the physiological activities of sesame seeds rich in lignans from three varieties (Gomazou, Maruhime and Maruemon), and those from a conventional cultivar (Masekin) in rats. The sum of the values of fat-soluble lignans (sesamin and sesamolin) in seeds of Gomazou, Maruhime and Maruemon varieties was approximately double the value in Masekin. Seeds from Maruemon contained fat-soluble lignan most exclusively as sesamin while other varieties contained sesamin and sesamolin at about a 2:1 ratio. After a 16 d experiment, sesame seeds, added at 200 g/kg to the experimental diets, increased the activity and mRNA levels of fatty acid oxidation enzymes. Increases were stronger with seeds rich in lignans than with seeds from Masekin. In contrast, sesame seeds lowered the activity and mRNA levels of lipogenic enzymes. However, sesame seeds from all the varieties were comparable in affecting these parameters. Serum triacylglycerol concentrations were lower in rats fed diets containing sesame seeds rich in lignans than in those fed a diet free of sesame seeds or a diet containing seeds from the Masekin variety. Serum malondialdehyde (a marker of lipid peroxidation) was lower in rats fed diets containing sesame seeds rich in lignans than in those fed a sesame seed-free diet or Masekin diet. It is apparent that sesame seeds rich in lignans, irrespective of lignan composition, more profoundly affect hepatic fatty acid oxidation and serum triacylglycerol levels and possibly attenuate oxidative stress. Therefore, consumption of sesame seeds rich in lignans hopefully results in physiological activity to promote health.

Quantitative trait locus responsible for resistance to Aphanomyces root rot (black root) caused by Aphanomyces cochlioides Drechs. in sugar beet.

Aphanomyces root rot, caused by Aphanomyces cochlioides Drechs., is one of the most serious diseases of sugar beet (Beta vulgaris L.). Identification and characterization of resistance genes is a major task in sugar beet breeding. To ensure the effectiveness of marker-assisted screening for Aphanomyces root rot resistance, genetic analysis of mature plants' phenotypic and molecular markers' segregation was carried out. At a highly infested field site, some 187 F(2) and 66 F(3) individuals, derived from a cross between lines 'NK-310mm-O' (highly resistant) and 'NK-184mm-O' (susceptible), were tested, over two seasons, for their level of resistance to Aphanomyces root rot. This resistance was classified into six categories according to the extent and intensity of whole plant symptoms. Simultaneously, two selected RAPD and 159 'NK-310mm-O'-coupled AFLP were used in the construction of a linkage map of 695.7 cM. Each of nine resultant linkage groups was successfully anchored to one of nine sugar beet chromosomes by incorporating 16 STS markers. Combining data for phenotype and molecular marker segregation, a single QTL was identified on chromosome III. This QTL explained 20% of the variance in F(2) population (in the year 2002) and 65% in F(3) lines (2003), indicating that this QTL plays a major role in the Aphanomyces root rot resistance. This is the first report of the genetic mapping of resistance to Aphanomyces-caused diseases in sugar beet.