Identification and quantitative analysis of polyphenolic compounds from the indigo plant (Polygonum tinctorium Lour).

The indigo plant (Polygonum tinctorium Lour) has been used traditionally as a medicinal plant with a variety of biological effects. Of these, polyphenolic ingredients are postulated to contribute to these activities. However, the identification and quantification of polyphenolic compounds in indigo plants have not been conducted comprehensively until now. This study was undertaken to identify the related ingredients by combined instrumental analyses using ultra-performance liquid chromatography electrospray-ionisation mass spectrometry and gas chromatography-mass spectrometry after the extracts of plant tissues were fractionated by absorption column chromatography. These analyses allowed the identification of kaempferol, quercetin-3-O-glucuronide, quercetin, kaempferol-3-O-glucopyranoside, caffeic acid, chlorogenic acid and tentative 3,5,4'-trihydroxy-6,7-methylenedioxyflavone. Furthermore, predominant polyphenolic compounds were quantified by reverse-phase high-performance liquid chromatography and capillary gas chromatography, revealing the higher proportions of kaempferol, quercetin-3-O-glucuronide and quercetin among them. The results indicate that the indigo plant is a promising source for flavonoids and the related compounds with beneficial medicinal effects.

Biochemical mechanism on GABA accumulation during fruit development in tomato.

A large amount of gamma-aminobutyric acid (GABA) was found to accumulate in tomato (Solanum lycopersicum) fruits before the breaker stage. Shortly thereafter, GABA was rapidly catabolized after the breaker stage. We screened the GABA-rich tomato cultivar 'DG03-9' which did not show rapid GABA catabolism after the breaker stage. Although GABA hyperaccumulation and rapid catabolism in fruits is well known, the mechanisms are not clearly understood. In order to clarify these mechanisms, we performed comparative studies of 'Micro-Tom' and 'DG03-9' fruits for the analysis of gene expression levels, protein levels and enzymatic activity levels of GABA biosynthesis- and catabolism-related enzymes. During GABA accumulation, we found positive correlations among GABA contents and expression levels of SlGAD2 and SlGAD3. Both of these genes encode glutamate decarboxylase (GAD) which is a key enzyme of GABA biosynthesis. During GABA catabolism, we found a strong correlation between GABA contents and enzyme activity of alpha-ketoglutarate-dependent GABA transaminase (GABA-TK). The contents of glutamate and aspartate, which are synthesized from GABA and glutamate, respectively, increased with elevation of GABA-TK enzymatic activity. GABA-TK is the major GABA transaminase form in animals and appears to be a minor form in plants. In 'DG03-9' fruits, GAD enzymatic activity was prolonged until the ripening stage, and GABA-TK activity was significantly low. Taken together, our results suggest that GAD and GABA-TK play crucial roles in GABA accumulation and catabolism, respectively, in tomato fruits.

Genome wide cDNA-AFLP analysis of genes rapidly induced by combined sucrose and ABA treatment in rice cultured cells.

We identified 27 genes induced by combined sucrose and ABA treatment from rice cultured cells with cDNA-AFLP. Thirteen of these up-regulated genes were induced 30 min after the co-treatment. This suite of genes includes starch biosynthesis related genes. Type A genes were expressed only in the presence of both sucrose and ABA. Type B genes were expressed in the presence of sucrose or ABA and the expression was dramatically enhanced by the co-treatment of sucrose and ABA. These results indicate that multiple steps of starch biosynthesis and other processes may be regulated by at least two different pathways.