Flavone C-glycosides from Coronilla varia

One new and 5 known flavone C-glycosides were isolated from leaves and stems of Coronilla varia. The new compound was shown to be isoorientin 2″-O-rhamnoside. The known compounds were isovitexin, isoorientin, isovitexin 4′-O-glucoside, isoorientin 4′-O-glucoside, and isoorientin 7-O-glucoside.     

On the localization of enzymes related to flavonoid metabolism in sections and tissues of oat primary leaves

During growth of the primary leaves of Avena sativa L., the distribution of extractable L-phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) and chalcone-flavanone isomerase (CFI, EC 5.5.1.6) activities in distinct leaf sections (top section, medium section and meristematic basal section) and in the epidermal and mesophyll tissues were investigated in relation to C-glycosylflavone accumulation. Characteristic changes have been observed in the levels of PAL and CFI activities within the three leaf sections, depending upon their stage of development. An increase in both enzyme activities accompanies a strong flavone accumulation in the section of the leaf that derives from the basal meristem. Highest specific PAL activity is localized in the meristem itself, which is poor in both flavones and CFI activity. Total flavone accumulation was found to be nearly the same in all three leaf tissues, lower and upper epidermis and mesophyll. Similarly, PAL activity is distributed about equally in these tissues in young leaves; in older ones, activity is relatively higher in the lower leaf epidermis. In contrast, CFI is found to be localized almost entirely in the mesophyll and not in the epiderms. Therefore the question arises whether CFI is involved at all in flavone metabolism and whether it may represent, as a marker enzyme, the localization of other specific C₁₅-enzymes of the flavonoid biosynthetic pathway in oat primary leaves.Abbreviations PAL L-phenylalamine ammonia lyase - CFI chalcone-flavanone isomerase     

Two isovitexin 2″-O-glycosides from primary leaves of Secale cereale

Two isovitexin 2″-O-glycosides have been isolated from primary leaves of rye and identified as isovitexin 2″-O-arabinoside and isovitexin 2     

Flavonoids and xanthones from the leaves of Amorphophallus titanum (Araceae)

The flavonoids and xanthones in the leaves of Amorphophallus titanum, which has the largest inflorescence among all Araceous species, were surveyed. Eight C-glycosylflavones, five flavonols, one flavone O-glycoside and two xanthones were isolated and characterized as vitexin, isovitexin, orientin, isoorientin, schaftoside, isoschaftoside, vicenin-2 and lucenin-2 (C-glycosylflavones), kaempferol 3-O-robinobioside, 3-O-rutinoside and 3-O-rhamnosylarabinoside, and quercetin 3-O-robinobioside and 3-O-rutinoside (flavonols), luteolin 7-O-glucoside (flavone), and mangiferin and isomangiferin (xanthones). Although the inflorescence of this species has been surveyed for flavonoids, those of the leaves were reported for the first time.     

The pH dependent substrate specificity of udp-glucose: isovitexin 2″-O-glucosyltransferase in Silene alba

In Silene alba plants the dominant allele of gene Fg controls an enzyme which catalyses the formation of isovitexin 2″-O-glucoside both in petals and green parts. Both isovitexin and isoorientin can act as substrate. K_mvalues for the isovitexin glucosylation are 0.09 mM for isovitexin and 0.3 mM for UDP-glucose, V_max 0.17 nmol min^?1 mg protein^?1. For the isoorientin glucosylation K_m values of 0.45 mM for isoorientin, of 0.75 mM for UDP-glucose and V_max of 0.27 nmol min^?1 mg protein^?1 are found. The pH optima for both substrates differ markedly. For the substrate with one hydroxyl in the B-ring, isovitexin, the pH optimum is pH 8.5. For isoorientin, which has two hydroxyls in the B-ring, a pH optimum of 7.5 is found. These results suggest that the B-ring hydroxylation pattern influences the pH at which the substrate has optimal affinity for the enzyme. The location of the carbon-carbon bound glucose on a the flavonoid skeleton is of importance for enzyme activity as well. Vitexin, which has glucose at the 8-position, was not a substrate. The glucosylation of vitexin could, however, be demonstrated in enzyme extracts of petals of plants, grown from seed collected in Armenia; in these petals apart from isovitexin glycosides, vitexin glycosides are found as well.     

Differential tissue distribution of metabolites in Jacobaea vulgaris,Jacobaea aquatica and their crosses

Plants are attacked by many different herbivores. Some will consume whole leaves or roots, while others will attack specific types of tissue. Thus, insight into the metabolite profiles of different types of leaf tissues is necessary to understand plant resistance against herbivores. Jacobaea vulgaris, J. aquatica and three genotypes of their crossings were used to study the variation in metabolomic profiles between epidermis and mesophyll tissues. Extracts of epidermis and mesophyll tissues were obtained using carborundum abrasion (CA). Subsequently, ¹H nuclear magnetic resonance (NMR) spectroscopy and multivariate data analyses were applied to compare the metabolome profiles. Orthogonal partial least-squares-discriminant analysis (OPLS-DA) resulted in a clear separation of epidermis and mesophyll extracts. The epidermis contained significantly higher amounts of jacaranone and phenylpropanoids, specifically chlorogenic (5-O-CQA) and feruloyl quinic (FQA) acids compared to the mesophyll. In contrast, the mesophyll showed significantly higher concentrations of pyrrolizidine alkaloids (PAs), specifically jacobine and jaconine. The tissue specific distribution of these compounds was constant over all genotypes tested. Phenylpropanoids, 5-O-CQA and FQA, as well as PAs are known for their inhibitory effect on herbivores, especially against thrips. Thrips feeding commences with the penetration of the epidermis, followed by ingestion of sub-epidermal or mesophyll. Thrips thus may have to encounter phenylpropanoids in the epidermis as the first line of defence, before encountering the PAs as the ultimate defence in the mesophyll. The finding of tissue specific defense may have a major impact on studies of plant resistance. We cannot judge resistance using analyses of a whole roots, leafs or flowers. In such a whole-organism approach, the levels of potential defense compounds are far below the real ones encountered in tissues involved in the first line of defense. Instead, it is of great importance to study the defence compounds in the specific tissue to which the herbivore is confined.     

Polyphenols of mature plant,seedling and tissue cultures of Theobroma cacao

The major flavone in mature cocoa leaves is isovitexin, with smaller amounts of vitexin and 7-O-glucosides of apigenin, luteolin and chrysoeriol. F     

C-Glycosylflavones from Avena sativa

Avena sativa leaves, stems and inflorescences contain a range of new C-glycosylflavone 2″-O-glycosides, including vitexin and isoswertisin 2″-rhamnosides, isovitexin and isoorientin 2″-arabinosides. The structure of ‘vitexin 4′-rhamnoside’ from Crataegus oxyacantha is revised in vitexin 2″-rhamnoside.     

UV-microscopic studies on the vacuolar changes caused by the flavone “aglycone” isovitexin inSilene pratensis plants

Summary UV-microscopic and chromatographic studies have been performed on the variation in contents and configuration of the flavones present in epidermal cells of the petals, stem leaves, rosette leaves and cotyledons ofSilene pratensis plants. Most of the flavone contents is located in the vacuole of the upper epidermis cells, the concentration depending on the light intensity at which the plants were grown. In plants able to glycosylate isovitexin in the petals (genotypegG/. gl/gl fg/fg, accumulating isovitexin 7-O-glucoside) the vacuole is completely filled with the UV absorbing flavone. In plants which are unable to glycosylate isovitexin in their petals (genotypeg/g gl/gl fg/fg, accumulating only isovitexin) the upper epidermal cells of stem leaves and petals contain droplet like structures in their vacuoles. At high light intensities these structures increase in mass and become detectable in the visible light. These denser structures often condense to structures with radiating threads.As compared with the accumulation of isovitexin in upper epidermal cells of stem leaves and petals in genotypeg/g gl/gl fg/fg, the cotyledons and the rosette leaves contain two isovitexin glycosides. In the latter organs the upper epidermal cells are very similar to the upper epidermal cells fromgG/. gl/gl fg/fg plants, having a vacuole filled with UV absorbing material. It appears therefore that isovitexin itself causes the formation of the structurés in the cells. It was shown by varying the light intensity that a relative high concentration of isovitexin is necessary for the droplet like structures to appear. Still higher concentrations are needed for the formation of the structures with radiating threads. It is hypothesized that isovitexin interferes with the energy supply of the cells, which therefore are not able to maintain their turgor.     

Characteristics of leaf structure and photosynthetic apparatus within the crown of systematically shadedQuercus petraea andNothofagus procera seedlings

Four-year-old seedlings ofQuercus petraea (Matt.) Liebl. andNothofagus procera (Poepp. et Endl.) Querst were grown outdoors in pots while subjected to full, medium and low irradiances. Shading and decrease in height of leaf attachment generally increased specific leaf area, the diameters of chloroplasts and of palisade and spongy mesophyll cells, but decreased leaf thickness, number of palisade cell layers, length of palisade and spongy mesophyll cells, number of chloroplasts per mesophyll cell and epidermal cell and cuticle thickness, stomata and hair densities per unit leaf area, hair length, maximum hair breath and cell wall thickness in the two species. However, inN. procera grown under full irradiance, leaves at the upper and middle positions had hairs on both upper and lower epidermes, whereas those in other treatments and all leaves in all treatments inQ. petraea, had theirs only on the upper epidermis.     

新疆野苹果不同地理分布与树龄的叶片解剖结构特征

以新疆野苹果为材料,对不同居群及不同树龄叶片解剖结构特征进行了分析研究。结果表明：新疆野苹果叶片属于异面叶,由上表皮、栅栏组织、叶脉、海绵组织和下表皮组成;不同居群新疆野苹果叶片厚度及细胞排列方式有差异,巩留、额敏及托里的野苹果栅栏组织增厚,且栅栏组织/海绵组织比值增大;新源的野苹果叶片及栅栏组织较薄,栅栏组织/海绵组织比值偏低;不同树龄新疆野苹果叶片厚度随着树龄的增大而呈变薄的趋势,多年生野苹果及野苹果“树王”栅栏组织及海绵组织层数最少,厚度较薄。这些结果暗示新疆野苹果通过各部分结构的协调生长来保证光合作用效率,以适应不同的生长环境以及处于不同发育时期的生长状况。本研究为新疆野苹果原位保护、保育提供了实验依据。     

Flavonoid variation in the genus briza

During a survey of 6 Eurasian and 10 South American Briza species for leaf flavonoids, 27 components were found. Twelve of these were identified: tricin 5-glucoside, tricin 7-glucoside, quercetin 3-glucoside, kaempferol 3-glucoside, vitexin, isovitexin, orientin, iso-orientin, and the 4′-O-glucoside of all 4 glycoflavones, 3 of which are reported for the first time. The Eurasian species, with the exception of Briza maxima, are remarkably uniform in their flavonoid pattern, accumulating mainly vitexin and isovitexin; whereas the South American species are characterized by the presence of orientin, iso-orientin and 9 unidentified flavonoids. In Briza media and the South American species, ploidy level is shown to play a large part in flavonoid variation. Examination of 12 diploid and 8 autotetraploid plants of B. media revealed that diploids accumulate vitexin and isovitexin, whereas tetraploids accumulate orientin and iso-orientin, autotetraploidy having apparently upset regulatory genes in the formation of the flavone C-glycosides. Mild alkaline treatment of both isovitexin and iso-orientin was found to give 100% conversion to the corresponding 8-C-glucoside.     

风毛菊属3种植物叶的解剖结构比较

采用石蜡切片法对分布于祁连山海拔5 000 m左右流石滩上菊科风毛菊属水母雪兔子（Saussurea medusa Maxim）、鼠曲雪兔子（Saussurea gnaphalodes (Royle) Sch.）、红叶雪兔子（Saussurea paxiana Diels.）3种植物叶片的解剖结构进行了比较研究,结果表明：叶片表皮细胞均为单层,上下表皮都有气孔分布,气孔不下陷;角质层较厚,叶表面均被单列细胞的表皮毛。3种植物均为异面叶;叶肉栅栏组织较发达,通常由2～3层细胞组成,但栅栏组织细胞排列较疏松;海绵组织存在大量的细胞间隙;叶肉中通气组织发达,且均有不规则裂生性气腔。叶脉维管束中韧皮部都具有异细胞存在。这些共同特征是3种植物对高山地区缺氧、低温、强辐射等自然条件长期适应的结果。但是,3种植物在叶片的外部形态特征、叶肉栅栏组织细胞的特点、维管束发育程度、内分泌结构、不规则裂生气腔等方面又存在明显的不同,表现出3种植物对环境的适应也是存在差异的。     

The effect of light quality on leaf production and development of in vitro-cultured plants of Alternanthera brasiliana Kuntze

We investigated the influence of light quality on the leaf development of Alternanthera brasiliana Kuntze (Amaranthaceae) grown in vitro. Growth parameters including specific leaf mass, thickness, and leaf density were lowest in plants grown under red light. Blue light induced the largest number of leaves/plant, and the largest thickness and area of the leafblade. Green and red lights induced the smallest leaf areas. The thickness of the abaxial-face epidermis and spongy parenchyma of the plants was significantly reduced in plants grown under red light. The thickness of the palisade parenchyma and upper epidermis were significantly increased in plants grown under blue light, compared to the other fluorescent-light treatments. The specific spectral band also influenced the differentiation of mesophyll cells. In the dark and under red light, the mesophyll was homogenous; and in the dark and under green light, the leaves were more compact. Under blue light, the cells displayed the characteristic palisade morphology. The results showed that the increase of a specific parenchyma type was related to a specific spectral band. All spectral-quality treatments reduced the numbers of stomata and trichomes. The results for green light were in some respects similar to those for red light, and in other respects similar to those for blue light, probably because phytochromes and cryptochromes are green-light receptors. This study indicated that Alternanthera plants have strong morphological plasticity induced by light. The results suggest that high-quality Alternanthera can be achieved by culturing the plants in vitro under a combination of blue and red light.     

Palisade mesophyll cell expansion during leaf development in Zinnia elegans (Asteraceae)

Temporal and spatial patterns of palisade mesophyll cell expansion in Zinnia elegans were characterized as a basis for developing a suspension culture model for mesophyll cell expansion. Our objectives were to 1) identify the leaf regions from which cells in various stages of expansion could be selectively isolated for culture, and 2) develop a basis for comparison of rate and extent of mesophyll cell expansion in culture with that in the leaf. Palisade mesophyll cells were isolated from expanding leaves by gentle physical maceration without the use of enzymes. Isolated cells from leaves in different stages of expansion were then measured by computer image analysis. Analysis of size frequency distributions showed that unexpanded cells can be isolated from the entire blade of small leaves or the basal regions of partially expanded leaves. Fully expanded cells can be obtained from the apical and middle regions of partially expanded leaves. Within the leaf, Zinnia mesophyll cells expanded from about 400 μm² to about 2.300 μm² at an estimated rate of 160 μm² d^-1. The percent increase in cell length exceeded the percent increase in cell width. Expansion of mesophyll cells continued for 6–8 d after epidermal expansion ceased. This difference in the timing of cell expansion in epidermal and mesophyll cells indicates that different regulatory factors may be operating in these adjacent tissues and underscores the importance of investigating the regulation of mesophyll cell expansion at the cellular level.     

Flavonoids in the leaves of <Emphasis Type="Italic">Oxalis corniculata </Emphasis>and sequestration of the flavonoids in the wing scales of the pale grass blue butterfly, <Emphasis Type="Italic">Pseudozizeeria </Emphasis><Emphasis Type="Italic">maha</Emphasis>

Three C-glycosylflavones in the leaves of Oxalis corniculata, the host plant of the lycaenid butterfly pale grass blue (Pseudozizeeria maha), were identified as 6-C-glucosylluteolin (isoorientin), 6-C-glucosylapigenin (isovitexin) and isovitexin 7-methyl ether (swertisin). Comparative spectral and HPLC analyses between the leaf extract of the host plants and the wings of P. maha showed selective uptake of the host-plant flavonoid isovitexin to the wings of the butterfly.     

On the Resistance to Transpiration of the Sites of Evaporation within the Leaf

The rates of transpiration from the upper and lower surfaces of leaves of Gossypium hirsutum, Xanthium strumarium, and Zea mays were compared with the rates at which helium diffused across those leaves. There was no evidence for effects of CO₂ concentration or rate of evaporation on the resistance to water loss from the evaporating surface (“resistance of the mesophyll wall to transpiration”) and no evidence for any significant wall resistance in turgid tissues. The possible existence of a wall resistance was also tested in leaves of Commelina communis and Tulipa gesneriana whose epidermis could be easily peeled. Only when an epidermis was removed from a leaf, evaporation from the mesophyll tissue declined. We conclude that under conditions relevant to studies of stomatal behavior, the water vapor pressure at the sites of evaporation is equal to the saturation vapor pressure.     

C-glycosylflavones from parkinsonia aculeata

The only C-glycosylflavones isolated from Parkinsonia aculeata leaves were identified as orientin (instead of epiorientin), isoorientin (instead of parkinsonin-A), vitexin and isovitexin (instead of parkinsonin-B). Orientin was definitely present in the laevorotatory form.     

Tissue-distribution of secondary phenolic biosynthesis in developing primary leaves of Avena sativa L.

Primary leaves of oats (Avena sativa L.) have been used to study the integration of secondary phenolic metabolism into organ differentiation and development. In particular, the tissue-specific distribution of products and enzymes involved in their biosynthesis has been investigated. C-Glucosylflavones along with minor amounts of hydroxycinnamic-acid esters constitute the soluble phenolic compounds in these leaves. In addition, considerable amounts of insoluble products such as lignin and wall-bound ferulic-acid esters are formed. The tissue-specific activities of seven enzymes were determined in different stages of leaf growth. The rate-limiting enzyme of flavonoid biosynthesis in this system, chalcone synthase, together with chalcone isomerase (EC 5.5.1.6) and the terminal enzymes of the vitexin and isovitexin branches of the pathway (a flavonoid O-methyltransferase and an isovitexin arabinosyltransferase) are located in the leaf mesophyll. Since the flavonoids accumulate predominantly (up to 70%) in both epidermal layers, an intercellular transport of products is postulated. In contrast to the flavonoid enzymes, L-phenylalanine ammonia-lyase (EC 4.3.1.5), 4-coumarate: CoA ligase (EC 6.2.1.12), and S-adenosyl-L-methionine: caffeate 3-O-methyltransferase (EC 2.1.1.-), all involved in general phenylpropanoid metabolism, showed highest activities in the basal leaf region as well as in the epidermis and the vascular bundles. We suggest that these latter enzymes participate mainly in the biosynthesis of non-flavonoid phenolic products, such as lignin in the xylem tissue and wall-bound hydroxycinnamic acid-esters in epidermal, phloem, and sclerenchyma tissues.Abbreviations CHI chalcone isomerase - CHS chalcone synthase - 4CL 4-coumarate: CoA ligase - CMT S-adenosyl-L-methionine:caffeate 3-O-methyltransferase - FMT S-adenosyl-L-methionine:vitexin 2-O-rhamnoside 7-O-methyltransferase - HPLC high-performance liquid chromatography - IAT uridine 5-diphosphate L-arabinose:isovitexin 2-O-arabinosyltransferase - PAL L-phenylalanine ammonia-lyase     

冠层部位对金叶女贞叶片结构的影响

金叶女贞是一种在园林绿化上广泛应用的彩叶植物,上层叶金黄色,下层叶深绿色。本文利用常规植物制片技术,研究了叶绿素缺少的金叶女贞上层金叶和下层绿叶的解剖结构。结果表明:(1)金叶女贞上层金叶和下层绿叶的上表皮无显著差异,但是下层绿叶的下表皮细胞较小,气孔密度较低;(2)下层绿叶的叶肉组织较薄,细胞较大,排列疏松;(3)在下层绿叶中叶绿体类囊体片层排列密集,叶肉组织内单位体积叶绿体数量较少;(4)冠层部位对光合色素的积累有明显影响。叶绿素缺少的金叶女贞叶片结构显著受到冠层部位的影响,光合色素的变化幅度较一般植物更大,不同冠层部位叶片叶色有明显差异,可以作为研究植物冠层部位对叶形态、结构和生理生化影响的好材料。