Differences In Flavonoid Contents Research Articles

Germplasm Resources and Metabolite Marker Screening of High-Flavonoid Tartary Buckwheat (Fagopyrum tataricum).

Tartary buckwheat is an annual minor cereal crop with a variety of secondary metabolites, endowing it with a high nutritional and medicinal value. Flavonoids constitute the primary compounds of Tartary buckwheat. Recently, metabolomics, as an adjunct breeding method, has been increasingly employed in crop research. This study explores the correlation between the total flavonoid content (TFC) and antioxidant capacity in 167 Tartary buckwheat varieties. Ten Tartary buckwheat varieties with significant differences in flavonoid content and antioxidant capacity were selected by cluster analysis. With the use of liquid chromatography-mass spectrometry, 58 flavonoid compounds were identified, namely, 42 flavonols, 10 flavanols, 3 flavanones, 1 isoflavone, 1 anthocyanidin, and 1 proanthocyanidin. Different samples were clearly separated by employing principal component analysis and partial least-squares discriminant analysis. Eight differential flavonoid compounds were further selected through volcano plots and variable importance in projection. Differential metabolites were highly correlated with TFC and antioxidant capacity. Finally, metabolic markers of kaempferol-3-O-hexoside, kaempferol-7-O-glucoside, and naringenin-O-hexoside were determined by the random forest model. The findings provide a basis for the selection and identification of Tartary buckwheat varieties with high flavonoid content and strong antioxidant activity.

Bioactive Contents, BSLT Toxicity and antioxidants from Apis dorsata Binghami and Apis mellifera nest extracts

Beenest is rich in secondary metabolites because honeycombs, among others, are formed from plant resins (propolis). This study aimed to analyze differences in flavonoid content and in vitro antioxidant activity of Apis dorsata Binghami and Apis mellifera nest extracts. The samples used were A. dorsata from North Sulawesi and A. mellifera from South Sulawesi. The honeycomb was extracted using 95% ethanol solvent. Honeycomb extract was analyzed for its flavonoid content by the HPLC method, toxicity was tested by the BSLT method, and in vitro antioxidant activity was tested by the DPPH method. The results of the analysis of flavonoids showed that A. dorsata nest extract produced 21 types of compounds while A. mellifera produced 26 types of compounds. The toxicity test results showed that the A. dorsata nest extract had a better LC50 of 245,691 mg/l than the A. mellifera nest extract with an LC50 of 443,701 mg/l. The in vitro antioxidant test results of A. dorsata nest extract were more robust, namely IC 50 1.161 mg/l, compared to A. mellifera nest extract IC 50 2.404 mg/l. However, both were included in the category of powerful antioxidants. In vitro, anticancer test results on MCF-7 cells, A. mellifera nest extract was active with IC 50 100.02mg/l. Compared to A. dorsata extract, it was active with IC 50 102.217mg/l, but the two were not significantly different. Based on the analysis of flavonoid content, toxicity test, and antioxidant test, A.dorsata and A.mellifera honeycomb extracts have potential as in vitro antioxidants

Ticktrefoil (Desmodium) species as a nutraceutical forage resource for animals

AbstractTicktrefoil (Desmodium) species from the USDA, ARS, Plant Genetic Resources Conservation Unit (PGRCU) germplasm collection in Griffin, GA, were evaluated for their potential livestock nutraceutical (nutritional + pharmaceutical) value in a field small plot study at the Fort Valley State University (FVSU) Agricultural Research Station, Fort Valley, GA. Horse marmalade (Desmodium discolor Vogel), greenleaf ticktrefoil (Desmodium intortum [Mill.] Urb.), panicledleaf ticktrefoil (Desmodium paniculatum [L.] DC.), Hawaii ticktrefoil (Desmodium sandwicense E. May) and twisted ticktrefoil (Desmodium tortuosum [Sw.] DC.) were established in a greenhouse and transplanted to field plots, with plant material from each species harvested, freeze‐dried, ground and analyzed for nutrient content and bioactivity. Crude protein (CP) levels in D. discolor and D. tortuosum averaged 233 g/kg, whereas CP in D. intortum, D. paniculatum and D. sandwicense ranged from 99 to 157 g/kg. The Desmodium species had in vitro true digestibility values of 755 to 878 g/kg. There were no detectable condensed tannins in D. discolor and D. tortuosum, whereas D. intortum, D. paniculatum and D. sandwicense averaged 88, 89 and 73 g/kg total condensed tannin, respectively. There were species differences in flavonoid content (p < .01), with the highest delphinidin and cyanidin concentrations in D. sandwicense and D. paniculatum, respectively. There were no detectable levels of delphinidin or cyanidin for D. discolor or D. tortuosum. Quercitin and kaempferol concentrations were highest in D. intortum and D. paniculatum, respectively, whereas D. discolor and D. tortuosum had the lowest values. The highest protein precipitable phenolics and total phenolic concentrations were for D. paniculatum. Desmodium species have potential as nutraceutical forage for livestock and warrant further investigation.

Transcriptome and metabolome analysis of flavonol synthesis in apricot fruits.

Apricot fruits are edible and serve as a source of medicinal compounds. Flavonols are important plant secondary metabolites that have antioxidant and antitumor effects and may promote cardiovascular health. The flavonoid content in three stages of the 'Kuijin' and the 'Katy' was observed, followed by the combination of metabolome and transcriptome analysis to explore the metabolic basis of flavonol synthesis. The differences in the metabolite contents between stages (of the same cultivar) and between cultivars (at the same stage) revealed decreases in the flavonoid content as fruits developed (i.e., from 0.28 mg/g to 0.12 mg/g in 'Kuijin' and from 0.23 mg/g to 0.05 mg/g in 'Katy'). To decipher the regulation of flavonol synthesis in apricot (Prunus armeniaca L.), the metabolomes and transcriptomes of fruit pulp at three developmental stages of 'Kuijin' and the 'Katy' were analyzed. A total of 572 metabolites were detected in 'Kuijin' and the 'Katy' pulp, including 111 flavonoids. The higher flavonol content young 'Kuijin' fruits at 42 days after full bloom is mainly due to 10 types of flavonols. Three pairs of significant differences in flavonol content were identified. From these three comparison groups, three structural genes were strongly correlated with the levels of 10 types of flavonols (Pearson correlation coefficients > 0.8, p value < 0.05), including PARG09190, PARG15135, and PARG17939. The weighted gene co-expression network analysis showed that the turquoise module genes were highly correlated with flavonol contents (P < 0.01). There were 4897 genes in this module. Out of 4897 genes, 28 transcription factors are associated with 3 structural genes based on weight value. Two of the transcription factors are not only associated with PARG09190 but also with PARG15135, indicating their critical importance in the flavonols biosynthesis. The two TFs are PARG27864 and PARG10875. These findings provide new insights into the biosynthesis of flavonols and may explain the significant differences in flavonoid content between the 'Kuijin' and the 'Katy' cultivars. Moreover, it will aid in genetic improvement to enhance the nutritional and health value of apricots.

Transcriptome and proteome associated analysis of flavonoid metabolism in haploid Ginkgo biloba

Having different number if genome copies affect transcription and metabolite production of plants. This may be due to different gene transcription and protein expression, but the reasons for this remains poorly known. Here we measured flavonoid content in leaves of three haploid and diploid grafted plants of Ginkgo biloba, a model gymnosperm important economically for its flavonoid content. We reported the first combined transcriptomic and proteomic analysis of the difference in flavonoid content in three haploid ginkgos to investigate the effect of haploidy. Haploids had always smaller leaves and flavonoid content than the diploids. The selected haploid had also generally lower gene dosage than the selected diploid, with 1149 up-regulated (46.8 %) and 1309 down-regulated (53.2 %) among 2452 differentially expressed genes (DEGs). Of 686 differentially expressed proteins (DEPs) detected, 289 proteins (42.1 %) were upregulated, and 397 proteins (57.9 %) were downregulated in haploids. A particular attention deserves the downregulation of PAL, PAM, FLS, OMT1 hub genes involved in flavonoid biosynthesis regulation. Our study confirms the trend of haploids to have lower metabolic contents and points that lower flavonoid content in ginkgo monoploids could be due to reduced dosage of the corresponding regulatory genes and downregulation of genes involved in flavonoid synthesis.

Identification of the key flavonoid and lipid synthesis proteins in the pulp of two sea buckthorn cultivars at different developmental stages

BackgroundSea buckthorn is an economically important woody plant for desertification control and water soil conservation. Its berry pulp is rich in flavonoids and unsaturated fatty acids. Cultivars containing high oil and flavonoid contents have higher economic value and will increase in the planting area. However, the cause of the differences in oil and flavonoid contents among cultivars is still unclear. The influence of key enzymes in the lipid and flavonoid synthesis pathways on their content needs to be explored and clarified.ResultsThe flavonoid content in XE (Xin’e 3) was 54% higher than that in SJ (Suiji 1). Rutin was the main flavonoid in sea buckthorn pulp, and the differences in the rutin content could cause flavonoid differences between the two cultivars. The oil content of XE was 31.58% higher than that of SJ, and the difference in oil content was highest at 50–70 DAF. High-throughput proteomics was used to quantify key enzymes of flavonoid and lipid synthesis pathways in two cultivars at three developmental stages. By functional annotation and KEGG analysis, 41 key enzymes related to phenylpropanoid biosynthesis, flavonoid biosynthesis, flavone and flavonol biosynthesis, fatty acid biosynthesis and TAG biosynthesis were quantified. CHS, F3H, ANS, fabD, FATA, FAB2, LPIN and plcC showed significant differences between the two cultivars. In addition, we quantified 6 oleosins. With the exception of a 16 kDa oleosin, the other oleosins in the two cultivars were positively correlated with oil content.ConclusionsIn the flavonoid synthesis pathway, CHS and F3H were the main enzymes responsible for the difference in flavonoid content between the two cultivars. In the lipid synthesis pathway, LPIN, plcC and MGD were the main enzymes with different contents in the middle to late stages. Higher contents of LPIN and plcC in XE than in SJ could cause DAG to generate TAG from PC, since the difference in DGAT between the two cultivars was not significant. Investigating the causes of flavonoid and oil content differences among different cultivars from the perspective of proteomics, could provide a basis for understanding the regulatory mechanism of flavonoids and lipid synthesis in sea buckthorn pulp.

Comparison of flavonoid content and antioxidant activity in calyces of two roselle varieties (Hibiscus sabdariffa L.)

Free radicals can cause various degenerative diseases such as cancer and cardiovascular disease. Flavonoids have antioxidant abilities that inhibit damage from free radicals. The roselle antioxidant mechanism is associated with high flavonoid content. Red and purple roselles are two varieties of roselle (Hibiscus sabdariffa L.) cultivated in Indonesia. The aim of this research was to analyze the differences in flavonoid content and antioxidant activity in the calyces of two varieties of roselle (Hibiscus sabdariffa L.), red and purple. Total flavonoid analysis and antioxidant activity testing were done by spectrophotometry in the visible region. Total flavonoid content was analyzed by pretreatment with aluminium chloride and sodium acetate. Antioxidant activity was tested with 2,2-diphenyl-1-picrylhydrazyl. The total flavonoids for both red and purple roselle calyces ethanolic extract were 27.55 and 39.18 mg quercetin equivalent per gram. The half maximal inhibitory concentration values obtained for both red and purple roselle calyces ethanolic extract a were 51.59 and 42.17 μg/mL. The total flavonoid content and antioxidant activity of purple roselle calyces ethanolic extract was higher and stronger than red roselle calyces ethanolic extract.

Molecular mechanisms of the irradiation-induced accumulation of polyphenols in star anise (Illicium verum Hook. f.)

Irradiation technology has been shown to be a possible preservation technology used to prolong the shelf life and provide microbial decontamination of spices. In this study, the molecular mechanisms of the irradiation-induced accumulation of polyphenols in star anise were explored using metabolomics combined with ultra-high-performance liquid chromatography quadrupole-Orbitrap (UHPLC-Q-Orbitrap) high-resolution mass spectrometry. Fifty-six star anise samples from different areas processed with 2–10 kGy doses of irradiation were compared. The effect of post irradiation storage times ranging from 0 to 70 days was also evaluated. A total of 161 metabolites with variable importance on projection greater than 1 were characterized, and significant differences in flavonoid content in star anise were observed after irradiation. Absolute quantification was used for the quantification of distinctive compounds, and the limit of quantitation of the components was 1.90–6.53 μg/kg. The flavonoid content was stable on the 28th day of storage. The average content of flavonoids increased by 2–9 times due to the breaking of glycoside bonds in star anise by irradiation. C15H11O7+m/z 303.13045 and C3H3O2+m/z 71.01276 were defined as the characteristic fragments of flavonoids according to the mechanism of fragmentation. Checking the extraction ion chromatogram (EIC) characteristic fragments allows straightforward screening of untargeted flavonoids.

Varietal differences in flavonoid and antioxidant activity in Japanese soybean accessions.

Soybean seeds contain many antioxidants, including flavonoids and other phytochemicals. Isoflavone is a phytoestrogen that mimics estrogenic effects on target tissues and also exerts antioxidant activity by sequestering free radicals. Despite many cultivars developed to date, varietal differences in flavonoid content and antioxidant activity in Japanese soybean accessions remain less well characterized. Here, we evaluated the seed content of isoflavones, total flavonoids, and total phenolics in 26 soybean accessions. Next, the antioxidant activity of each accession was examined using antioxidant responsive element (ARE) linked to a luciferase reporter in human HepG2 stable cells. The relative ARE luciferase activity rate of all soybean accessions varied up to 4-fold which ranged from 1.00 to 4.02; and 22 accessions exhibited significant antioxidant activities. Correlation analysis indicated that the level of total isoflavone moderately correlated with antioxidant activity.

Identification and characterization of long non-coding RNAs regulating flavonoid biosynthesis in Ginkgo biloba leaves

Flavonoids are the main secondary metabolites in Ginkgo biloba leaves. Most previous studies focused on the molecular mechanisms of structural genes and transcription factors regulating flavonoid synthesis. Few reports have examined the long non-coding RNAs (lncRNAs) regulating flavonoid synthesis in G. biloba. Here, we performed high-throughput sequencing of three stages of G. biloba leaves with significant differences in flavonoid contents. We identified 2124 lncRNAs, including 1032 intergenic lncRNAs and 1092 antisense lncRNAs. Enrichment analysis revealed that cis-regulated target genes of upregulated lncRNAs were significantly enriched in the flavonoid biosynthesis pathway. Among these, 28 differentially expressed structural genes in this pathway and 14 key transcription factors may be regulated by multiple lncRNAs. In addition, we found that lncRNAs may act as precursors and endogenous target mimics of miRNAs to indirectly regulate protein-coding genes (PCgenes). More importantly, we found that multiple lncRNAs may serve as the targets of miR156a, miR172a, miR396a, miR828a, and miR858a and participate in the synthesis of ginkgo flavonoids by forming lncRNA (target)–miRNA–PCgene networks. Taken together, our results characterize the differences in the structure and expression levels of G. biloba lncRNAs and protein-coding genes, and demonstrating that lncRNAs may participate in the regulation of biosynthesis of G. biloba flavonoids in several ways.

Identification and Quantification of Antioxidant Compounds in Clarified Cashew Apple Juice ‘Cajuína’

Background:Cajuína is obtained through clarification of cashew apple juice (Anacardium occidentale L.), a non-diluted and non-fermented beverage, which is prepared from the edible part of the pseudo-fruit.Objective:This study sought to identify and quantify phenolic compounds, determine vitamin C content, total flavonoid content, and the antioxidant activity of two commercial brands of clarified cashew apple juice (cajuína) produced in Piauí, Brazil.Methods:Spectrophotometric and high performance liquid chromatography were used.Results:A high vitamin C content was found mainly in brand A. A high total phenolic content was observed in brand A, and there was no significant difference in flavonoid content between the two brands. Cajuína had high antioxidant activity, which was confirmed by two methods. The highest value was found for brand A by the DPPH method.Conclusion:We concluded that this clarified cashew apple juice has a high antioxidant compound content, and it is a functional food.

Carotenoid Accumulation and Its Contribution to Flower Coloration of Osmanthus fragrans.

Among naturally occurring pigments, carotenoids are importantly involved in the photosynthesis of plants and responsible for the coloration of petals and fruits. Osmanthus fragrans Lour., a famous ornamental plant, has many cultivars with different flower color. Petal coloration in O. fragrans mainly depends on the kinds of carotenoids and their contents. To investigate the mechanism of flower coloration in different cultivars, an analysis of phenotypic classification, phytochemistry, as well as the expression of carotenoid metabolism genes based on different groups was performed in the present study. Two main clusters including the orange-red cluster containing Aurantiacus cultivars and the yellowish-white cluster containing the other three cultivar groups were classified using the CIEL∗a∗b∗ system. No significant differences in flavonoid contents were observed between these two clusters. However, carotenoids, especially α-carotene and β-carotene, were found to have crucial roles in the diversity of floral coloration among the different cultivars. Carotenoid compositions in the petals of cultivars from both clusters consisted of α-carotene, β-carotene, α-cryptoxanthin, β-cryptoxanthin, lutein, and zeaxanthin, but carotenoid accumulation patterns during the flowering process were different. The petals of the yellowish-white cultivars exhibited high contents of β-carotene, lutein and α-carotene, whereas the petals of the orange-red cultivars mainly contained β-carotene and α-carotene. The profound diversity in the total carotenoid concentrations in the two clusters was determined by the transcript levels of OfCCD4. Furthermore, the accumulation of upstream products with orange color in orange-red cultivars was partially due to the low expression of OfCHYB, whereas the relatively higher OfCHYB expression in the petals of the yellowish-white cultivars led to higher proportions of lutein, which is yellow. We also found that downregulation of OfLCYE, which encodes 𝜀-ring cyclase, indicated that the carotenoid flux of most cultivars mainly resulted in more β, β-branched products. Additionally, carotenoid biosynthesis in green tissues and petals was compared, revealing the tissue specificity of carotenoid accumulation in O. fragrans. Therefore, the effects of multiple genes on carotenoid accumulation give rise to the colorful O. fragrans.

Polyphenolic compounds and essential oil analysis of selected species of the genus Thymus / Analýza fenolových zložiek a analýza silice vybraných druhov rodu Thymus

Abstract The content of phenolic compounds (total phenolic compounds, tannins, flavonoids and anthocyanidins) of three species of thyme (Thymus pulegioides, Thymus pannonicus, Thymus praecox) of different origin (Bohemian-Moravian highlands, Křivoklat and Považsky Inovec) was determined using spectrophotometric methods of European Pharmacopoeia 8th edition. Furthermore, the determination of the content of essential oil and analysis of its constituents was realised. The amount of total phenolics was determined by a spectrophotometric method using the Folin-Ciocalteu reagent. Their content ranged from 3.87 to 8.86%. The content of tannins was established on a preliminary determination of the total phenolics, followed by adsorption of tannins on hide powder. The amount of tannins in our samples varied from 1.96 to 5.65%. The content of total flavonoids was determined by a spectrophotometric method using aluminium chloride. Quantitative analysis of flavonoids has shown that content in our samples ranged from 0.59 to 1.52% expressed as luteolin-7-O-glucoside (λ = 392 nm) and from 0.41 to 1.12% expressed as rutin (λ = 420 nm). Anthocyanins represent a small amount of total polyphenol content in Thymus species. In our samples, the content of anthocyanins expressed as cyanidin-3-O-glucoside varied from 0.02 to 0.1%. The determination of essential oil was realised by method of European Pharmacopoeia 8th edition. Its contents in our samples ranged from 0.2 to 0.75%. Gas chromatography-mass spectrometry was used for essential oil analysis. According to the presence of main monoterpene in Thymus essential oil, we can distinguish three chemotypes: thymol, carvacrol and linalool. Due to variations of chemical constituents of thyme essential oil, it was possible to observe a relationship between occurrence of certain chemotype and origin of sample. Differences in content of flavonoids have not shown a significant relationship to locality of origin.

Influence of modified atmosphere packaging on 'Star Ruby' grapefruit phytochemicals.

Modified atmosphere packaging (MAP) can extend the shelf life of salads, vegetables, and fruits by generating a storage environment with low O2, high CO2, and high humidity. The current study investigates the effect of modified atmosphere and humidity generated by two plastic films, microperforated bags (MIPBs) and macroperforated bags (MAPBs), on the levels of phytochemicals present in 'Star Ruby' grapefruits (Citrus paradisi, Macf.) stored for 16 weeks at 10 °C. Control fruits were stored without any packaging film. Juice samples were analyzed every 4 weeks for ascorbic acid, carotenoids, limonoids, flavonoids, and furocoumarins and assessed for quality parameters. MAP significantly reduced weight loss compared to control grapefruits. Control fruits had more β-carotene, lycopene, and furocoumarin compared with the fruits in MAP. Flavonoid content was highest in fruits stored in MAPB (P < 0.05), while fruits stored in MIPB showed no significant difference in flavonoid content compared to control (P > 0.05). The MAP treatments did not significantly affect ascorbic acid, limonoids, or fruit quality parameters, including total soluble solids, acidity, ripening ratio, decay and disorders, fruit taste, and off-flavors after 16 weeks of storage. These results suggest that MAP can be used to maintain the quality of 'Star Ruby' grapefruit with no detrimental effect on health-promoting phytochemicals.

Flavonoid and Leaf Gas Exchange Responses of Centella asiatica to Acute Gamma Irradiation and Carbon Dioxide Enrichment under Controlled Environment Conditions

The study was couducted to investigate the effects of gamma irradiation and CO2 on flavonoid content and leaf gas exchange in C.asiatica. For flavonoid determination, the design was a split split plot based on Randomized Complete Block Design (RCBD). For other parameters, the designs were split plots. Statistical tests revealed significant differences in flavonoid contents of Centella asiatica leaves between different growth stages and various CO2 treatments. CO2 400, G20 (400 = ambient CO2; G20 = Plants exposed to 20 Gy) showed 82.90% higher total flavonoid content (TFC) in the 5th week than CO2 400 as control at its best harvest time (4th week). Increasing the concentration of CO2 from 400 to 800 μmol/mol had significant effects on TFC and harvesting time. In fact, 800 μmol/mol resulted in 171.1% and 66.62% increases in TFC for control and irradiated plants, respectively. Moreover, increasing CO2 concentration reduced the harvesting time to three and four weeks for control and irradiated plants, respectively. Enhancing CO2 to 800 µmol/mol resulted in a 193.30% (CO2 800) increase in leaf biomass compared to 400 µmol/mol and 226.34% enhancement in irradiated plants (CO2 800, G20) [800 = Ambient CO2; G20 = Plants exposed to 20 Gy] than CO2 400, G20. In addition, the CO2 800, G20 had the highest amount of flavonoid*biomass in the 4th week. The results of this study indicated that all elevated CO2 treatments had higher PN than the ambient ones. The findings showed that when CO2 level increased from 400 to 800 µmol/mol, stomatal conductance, leaf intercellular CO2 and transpiration rate had the tendency to decrease. However, water use efficiency increased in response to elevated CO2 concentration. Returning to the findings of this study, it is now possible to state that the proposed method (combined CO2 and gamma irradiation) has the potential to increase the product value by reducing the time to harvest, increasing the yield per unit area via boosting photosynthesis capacity, as well as increasing biochemicals (flavonoids) per gram DM.

Teor e rendimento de flavonóides em calêndula (Calendula officinalis L.) cultivada com diferentes lâminas de irrigação

A Calendula officinalis L. apresenta uso ornamental, culinário e terapêutico. A inflorescência é usada com ação cicatrizante e antiinflamatória. O experimento analisou o efeito de quatro lâminas de irrigação em condições de campo, na produção de flavonóides em calêndula. As lâminas baseadas na evaporação do Tanque Classe A (ECA) foram: 0, 50, 100 e 150% ECA; sendo aplicadas desde o início do florescimento até o final do ciclo da cultura. O teor de flavonóides não apresentou diferenças estatísticas. Os maiores valores de matéria seca de flores e rendimento de flavonóides foram obtidos no tratamento sem irrigação. O teor de prolina foi maior para o tratamento com estresse por excesso hídrico. A deficiência hídrica acumulada para o tratamento sem irrigação sinalizou uma possível morte por deficiência hídrica. Com a ocorrência da precipitação no período de pré-florescimento, a planta, originária do Mediterrâneo, respondeu com florescimento abundante em massa e em número de flores. Assim, para Presidente Prudente, caso não ocorra a reposição hídrica do solo por precipitação, é necessário o uso de irrigação complementar com 200 mm apenas no período de pré-florescimento, para induzir ao máximo florescimento e máximo rendimento de flavonóides por hectare.

Transcriptional and biochemical signatures of divergence in natural populations of two species of New Zealand alpinePachycladon

New Zealand is diverse in alpine and subalpine environments, a consequence of Late Tertiary tectonic and climatic change. However, few studies have sought to evaluate the importance of these environments as abiotic drivers in the diversification of plant species. Of particular interest is the Late Tertiary radiation of Pachycladon, an endemic New Zealand genus of alpine cress. Here we report observations on genome-wide levels of differential expression measured in the habitats of two closely related species of Pachycladon with distinct altitudinal preferences. Using Arabidopsis microarrays, we have identified 310 predominantly hormone- and stress-response genes up-regulated in Pachycladon fastigiata and 324 genes up-regulated in Pachycladon enysii. Expression patterns for glucosinolate biosynthesis and hydrolysis genes (MAM1, MAM-I, MAM-D, AOP2, ESP, ESM1) as well as flavonoid biosynthesis genes (F3'H, FLS, FAH1) were found to be species specific. Predicted differences in flavonoid contents were partly confirmed by high performance liquid chromatography analysis. Differences in glucosinolate profiles and glucosinolate hydrolysis products obtained by high performance liquid chromatography and gas chromatography-mass spectrometry analysis, respectively, also supported inferences from expression analyses. Five glucosinolate chemotypes were matched to known Arabidopsis ecotypes, and the potential adaptive significance of these chemotypes has been discussed. Our findings, in contrast to expectations for evolution of the New Zealand flora, suggest that biotic drivers, such as plant-herbivore interactions, are likely to be as important as abiotic drivers in the diversification of Pachycladon.

The Aqueous Extracts ofPassiflora alataandPassiflora edulisReduce Anxiety-Related Behaviors Without Affecting Memory Process in Rats

Several species of Passiflora have been employed widely as a folk medicine because of sedative and tranquillizer activities. In this study, we evaluate the effects on anxiety and memory process of two popularly used Passiflora species. To this aim, male Wistar rats (weighing 250-300 g) were intraperitoneally injected with the aqueous extract of Passiflora alata or Passiflora edulis (25, 50, 100, or 150 mg/kg; single injection) 30 minutes prior to the elevated plus-maze test, inhibitory avoidance test, or habituation to an open-field apparatus. The effects of both species of Passiflora were compared with that of diazepam (1 mg/kg), a standard anxiolytic drug. Our findings revealed that, similar to diazepam, the treatment with P. alata (100 and 150 mg/kg) and P. edulis (50, 100, and 150 mg/kg) induced anxiolytic-like effects in rats. Memory was not affected by the treatment with any dose of P. alata or P. edulis, but diazepam disrupted memory process in rats. Phytochemical analysis showed that the content of flavonoids of the aqueous extract of P. edulis is almost twice that of P. alata. These differences in contents of flavonoids could explain the lower active doses of the aqueous extract of P. edulis in inducing anxiolytic-like effects compared to P. alata. In conclusion, our findings suggest that, distinct from diazepam, the aqueous extract of both species of Passiflora induced anxiolytic-like effects in rats without disrupting memory process.

Organ-specific distribution of phenolic compounds in bilberry (Vaccinium myrtillus) and ‘northblue’ blueberry (Vaccinium corymbosum x V. angustifolium)

Blueberries and bilberries are recognized as some of the best sources of flavonoids, especially anthocyanins. The contents of flavonoids (anthocyanins, proanthocyanidins, flavonols) and hydroxycinnamic acids in the flower, fruit skin and pulp, leaf and rhizome of bilberry and the blueberry cultivar ‘Northblue’ were analyzed using high-performance liquid chromatography combined with diode-array detection. The most striking difference in the fruits was the predominance of hydroxycinnamic acids in blueberry, whereas in bilberry the anthocyanin content was much higher, particularly in the pulp. Differences in flavonoid contents of fruits were already apparent at the flower stage. Bilberry and blueberry leaves both contained high amounts of proanthocyanidins, flavonols and hydroxycinnamic acids. Blueberry rhizomes accumulated high amounts of hydroxycinnamic acids. All plant parts of bilberry and blueberry are potential sources of phenolic compounds for use either as dietary botanicals or by the pharmaceutical industry.

Hypericum Perforatum L., H. Maculatum Crantz., H. Calycinum L. and H. pulchrum L.: Phytochemical and Morphological Studies

Four species of Hypericum growing in Italy were characterized morphologically and chemically: Hypericum perforatum L., H. maculatum Crantz., H. calycinum L. and H. pulchrum L. The composition of secondary metabolites (phloroglucinols, naphthodianthrones, flavonoids) in the aerial parts of plants collected in different habitats was analysed. The four species show different compositions of phloroglucinols and naphthodianthrones, but there was no qualitative difference in flavonoid content of the species analysed. Study of main-constituent variation during the ontogenetic cycle showed that hypericin decreases and hyperforin increases during the reproductive phase. In St. John's Wort, hypericin and hyperforin are thought to be localised in black nodules. Our investigation shows no clear correlation between either the presence or absence of nodules and hypericin or hyperforin content.