Abscisic Acid Biosynthesis in Leaves and Roots of Xanthium strumarium

Research on the biosynthesis of abscisic acid (ABA) has focused primarily on two pathways: (a) the direct pathway from farnesyl pyrophosphate, and (b) the indirect pathway involving a carotenoid precursor. We have investigated which biosynthetic pathway is operating in turgid and stressed Xanthium leaves, and in stressed Xanthium roots using long-term incubations in ¹⁸O₂. It was found that in stressed leaves three atoms of ¹⁸O from ¹⁸O₂ are incorporated into the ABA molecule, and that the amount of ¹⁸O incorporated increases with time. One ¹⁸O atom is incorporated rapidly into the carboxyl group of ABA, whereas the other two atoms are very slowly incorporated into the ring oxygens. The fourth oxygen atom in the carboxyl group of ABA is derived from water. ABA from stressed roots of Xanthium incubated in ¹⁸O₂ shows a labeling pattern similar to that of ABA in stressed leaves, but with incorporation of more ¹⁸O into the tertiary hydroxyl group at C-1′ after 6 and 12 hours than found in ABA from stressed leaves. It is proposed that the precursors to stress-induced ABA are xanthophylls, and that a xanthophyll lacking an oxygen function at C-6 (carotenoid numbering scheme) plays a crucial role in ABA biosynthesis in Xanthium roots. In turgid Xanthium leaves, ¹⁸O is incorporated into ABA to a much lesser extent than it is in stressed leaves, whereas exogenously applied ¹⁴C-ABA is completely catabolized within 48 hours. This suggests that ABA in turgid leaves is either (a) made via a biosynthetic pathway which is different from the one in stressed leaves, or (b) has a half-life on the order of days as compared with a half-life of 15.5 hours in water-stressed Xanthium leaves. Phaseic acid showed a labeling pattern similar to that of ABA, but with an additional ¹⁸O incorporated during 8′-hydroxylation of ABA to phaseic acid.     

Identifying Three Ecological Chemotypes of Xanthium strumarium Glandular Trichomes Using a Combined NMR and LC-MS Method

Xanthanolides, as the sesquiterpene lactones, are reportedly the major components for the pharmacological properties of X. strumarium L. species. Phytochemical studies indicated that the glandular structures on the surface of plant tissues would form the primary sites for the accumulation of this class of the compounds. As the interface between plants and their natural enemies, glandular trichomes may vary with respect to which of their chemicals are sequestered against different herbivores in different ecologies. However, to date, no data are available on the chemical characterisation of X. strumarium glandular cells. In this study, the trichome secretions of the X. strumarium species originating from nineteen unique areas across eleven provinces in China, were analysed by HPLC, LC-ESI-MS and NMR. For the first time three distinct chemotypes of X. strumarium glandular trichomes were discovered along with the qualitative and quantitative evaluations of their presence of xanthanolides; these were designated glandular cell Types I, II, and III, respectively. The main xanthanolides in Type I cells were 8-epi-xanthatin and xanthumin while no xanthatin was detected. Xanthatin, 8-epi-xanthatin, and xanthumin dominated in Type II cells with comparable levels of each being present. For Type III cells, significantly higher concentrations of 8-epi-xanthatin or xanthinosin (relative to xanthatin) were detected with xanthinosin only being observed in this type. Further research will focus on understanding the ecological and molecular mechanism causing these chemotype differences in X. strumarium glandular structures.     

Ethylene Release from Leaves of Xanthium strumarium L. and Zea mays L.

The release of ethylene into sealed Erlenmeyer flasks by intactleaves and leaf discs of Xanthium strumarium L. a C₃ plant andZea mays L. a C₄ plant were compared both in white light andin darkness. The effects of the presence or absence of addedCO₂ (in the form of sodium bicarbonate) the photosynthetic inhibitor3-[3,4-dichlorophenyl]-l, l-dimethyl urea (DCMU) and 1-aminocyclopropane-1-carboxylicacid (ACC), the precursor of ethylene in higher plants, werealso investigated. The rate of ethylene release from leaf tissue of Xanthium inthe absence of added CO₂ was markedly reduced in the light (i.e.at the CO₂ compensation point). Treatments that would enhancethe CO₂ availability to the tissue (i.e. added bicarbonate,darkness, treatment with DCMU) allowed higher levels of ethylenerelease. Incubation of the tissue with ACC considerably enhancedthe release of ethylene compared to that from the correspondingcontrol tissue without ACC. However, the pattern of ethylenerelease induced by the various treatments was similar with orwithout added ACC. When tissue, in the absence of added CO₂, was transferred fromlight to darkness, and back to light for 90 min periods, theethylene release rates Increased during the interposed darkperiod but resumed the lower rate during the final light period.The addition of CO₂ in the light resulted in a similar rateof ethylene release to that found in the dark. The overall pattern of ethylene release from Zea leaf tissuesubjected to light and dark in the presence or absence of addedCO₂ was similar to that of Xanthium. However, two or three timesmore ethylene was released from maize leaves in the light whenCO² was added compared to that generated in the dark. This isin marked contrast to Xanthium, where, under the light conditionsused, the ethylene release rate in the dark equalled or exceededthat occurring in the light, even in the presence of high levelsof CO₂. A very low rate of ethylene release was observed atthe CO₂ compensation point of maize. A speculative model is presented to explain how photosyntheticactivity might act as a key factor in regulating ethylene evolutionfrom leaf tissue in these experiments. It invokes the conceptof an inhibition by CO₂ of ethylene retention or breakdown thuspermitting more ethylene to be released from the leaves.     

Photosynthesis and Ribulose 1,5-Bisphosphate Concentrations in Intact Leaves of Xanthium strumarium L

The interacting effects of the rate of ribulose 1,5-bisphosphate (RuBP) regeneration and the rate of RuBP utilization as influenced by the amount and activation of RuBP carboxylase on photosynthesis and RuBP concentrations were resolved in experiments which examined the kinetics of the response of photosynthesis and RuBP concentrations after step changes from a rate-saturating to a rate-limiting light intensity in Xanthium strumarium. Because RuBP carboxylase requires several minutes to deactivate in vivo, it was possible to observe the effect of reducing the rate of RuBP regeneration on the RuBP concentration at constant enzyme activation state by sampling very soon after reducing the light intensity. Samples taken over longer time periods showed the effect of changes in enzyme activation at constant RuBP regeneration rate on RuBP concentration and photosynthetic rate. Within 15 s of lowering the light intensity from 1500 to 600 microEinsteins per square meter per second the RuBP concentration in the leaves dropped below the enzyme active site concentration, indicating that RuBP regeneration rate was limiting for photosynthesis. After longer intervals of time, the RuBP concentration in the leaf increased as the RuBP carboxylase assumed a new steady state activation level. No change in the rate of photosynthesis was observed during the interval that RuBP concentration increased. It is concluded that the rate of photosynthesis at the lower light intensity was limited by the rate of RuBP regeneration and that parallel changes in the activation of RuBP carboxylase occurred such that concentrations of RuBP at steady state were not altered by changes in light intensity.     

Effect of Light Quality on Stomatal Opening in Leaves of Xanthium strumarium L

Flux response curves were determined at 16 wavelengths of light for the conductance for water vapor of the lower epidermis of detached leaves of Xanthium strumarium L. An action spectrum of stomatal opening resulted in which blue light (wavelengths between 430 and 460 nanometers) was nearly ten times more effective than red light (wavelengths between 630 and 680 nanometers) in producing a conductance of 15 centimoles per square meter per second. Stomata responded only slightly to green light. An action spectrum of stomatal responses to red light corresponded to that of CO₂ assimilation; the inhibitors of photosynthetic electron transport, cyanazine (2-chloro-4[1-cyano-1-methylethylamino]-6-ethylamino-s-triazine) and 3-(3,4-dichlorophenyl)-1,1-dimethylurea, eliminated the response to red light. This indicates that light absorption by chlorophyll is the cause of stomatal sensitivity to red light. Determination of flux response curves on leaves in the normal position (upper epidermis facing the light) or in the inverted position (lower epidermis facing the light) led to the conclusion that the photoreceptors for blue as well as for red light are located on or near the surfaces of the leaves; presumably they are in the guard cells themselves.     

Structural and Histochemical Investigation of the Glandular Trichomes of Salvia aurea L. Leaves, and Chemical Analysis of the Essential Oil

Anatomical and histological investigations of the secretoryhairs ofSalvia aurea leaves, and identification of the maincomponents of the essential oil were carried out. Two typesof glandular trichome were found: peltate glands, characterizedby a short stalk and a large six to eight-celled head, and capitatetrichomes which were further subdivided into two kinds, thefirst with a short monocellular stalk and two-cellular head(type I), and the second with a multicellular stalk, a neckcell and a small globose unicellular head (type II). Whereaspeltate glands and type I capitate trichomes were always present,type II capitate glands were not found in all leaf samples.The histochemical study suggested an ‘endodermal’role for the stalk cell (peltate and capitate type I) as wellas for the neck cell (capitate type II), preventing the lossof essential oil. Histological reactions also revealed the complexnature of the material secreted by all types ofS. aurea trichome,including polysaccharides, polyphenols and proteins, in additionto the essential oil. Qualitative and quantitative GC-MS analysisof the essential oil revealed camphor to be the main constituent.The findings are discussed in relation to studies of trichomesfrom other members of the Lamiaceae. Salvia aurea L.; glandular trichomes; histochemistry; essential oil     

Xanthatin and xanthinosin from the burs of Xanthium strumarium L. as potential anticancer agents

Xanthatin and xanthinosin, 2 sesquiterpene lactones isolated from the burs of Xanthiun strumarium L. (cocklebur), showed moderate to high in vitro cytotoxic activity in the human cancer cell lines WiDr ATCC (colon), MDA-MB-231 ATCC (breast), and NCI-417 (lung). Xanthatin and xanthinosin were purified as the result of a multi-screening bioassay-guided study of wild plant species of the family Asteraceae, collected from various sites in Saskatchewan, Canada. Seventy-five extracts at a single concentration of 100 microg/mL were evaluated for in vitro cytotoxicity to the human cancer cell lines used. The chloroform extract of Carduus nutans L. (nodding thistle) aerial parts (IC50, 9.3 microg/mL) and the hexane extract of Echinacea angustifolia DC. (narrow-leaved purple coneflower) root (IC50, 4.0 microg/mL) were moderately to highly cytotoxic to the lung cancer cell line. The chloroform extracts of X. strumarium L. burs and Tanacetum vulgare L. (tansy) aerial parts exhibited the highest cytotoxicity for all cell lines tested; their IC50 values, obtained from multidose testing, ranged from 0.1 to 6.2 microg/mL (X. strumarium) and from 2.4 to 9.1 microg/mL (T. vulgare). Further purification of the chloroform fraction of X. strumarium yielded xanthatin and xanthinosin in high yields. This is the first time that these compounds have been reported in the burs of X. strumarium. Their IC50 values are also reported herein.     

Anti-allergic rhinitis effect of caffeoylxanthiazonoside isolated from fruits of Xanthium strumarium L. in rodent animals

The fruits of Xanthium strumarium L. (Asteraceae) have been used extensively in China for treatment of various diseases such as allergic rhinitis (AR), tympanitis, urticaria and arthritis or ozena. This study was designed to systemically investigate the effects of the caffeoylxanthiazonoside (CXT) isolated from fruits of X. strumarium on AR in rodent animals. Animals were orally administered with CXT. Anti-allergic activity of CXT was evaluated by passive cutaneous anaphylaxis test (PCA); acetic acid-induced writhing tests were used to evaluate the analgesic effects of CXT; acetic acid-induced vascular permeability tests were performed to evaluate anti-inflammatory effect of CXT. Then, the model AR in rats was established to evaluate the effects of CXT on AR with the following tests: the sneezing and nasal scratching frequencies, IgE level in serum, and histopathological examinations. Our results demonstrated that CXT had favorable anti-allergic, anti-inflammatory and analgesic effects. Additionally, we found that CXT was helpful to ameliorate the nasal symptoms and to down-regulate IgE levels in AR rats. Thus, we suggested that CXT can be treated as a candidate for treating AR.     

Cytokinins in Xanthium strumarium L.: Distribution in the Plant and Production in the Root System

Cytokinins from leaf laminae, buds, petioles, stems, roots,and root exudate of mature vegetative plants of Xanthium strumariumL. were extracted, fractionated, and partially characterizedby means of column chromatography with Sephadex LH20. Two peaksof cytokinin activity with elution volumes corresponding tozeatin and zeatin riboside were detected, in varying concentrations,in all plant parts. A third cytokinin, detected only in petiolesand in expanding and mature leaves, eluted off the Sephadexcolumn before zeatin riboside. This cytokinin (peak ‘a’)was converted to zeatin or to a zeatin-like cytokinin followingboth acid hydrolysis and treatment with ß-glucosidase.Peak ‘a’ was not detected in buds or in the youngestdeveloping leaves but was the predominant cytokinin presentin half-expanded and more mature leaves. By contrast, the zeatinriboside-like peak (peak ‘b’) constituted the majorcytokinin in root exudate, apical buds, and the youngest developingleaves, while not greatly contributing to the cytokinin contentof mature leaves. The detopped root system was shown to be capable of cytokininproduction. The distribution of cyrtokinins in the plant isdiscussed in relation to their probable origin in the root system.     

Abscisic Acid Content and Stomatal Sensitivity to CO(2) in Leaves of Xanthium strumarium L. after Pretreatments in Warm and Cold Growth Chambers

The degree of stomatal sensitivity to CO₂ was positively correlated with the content of abscisic acid of leaves of Xanthium strumarium grown in a greenhouse and then transferred for 24 hours or more to a cold (5/10 C, night/day) or a warm growth chamber (20/23 C). This correlation did not exist in plants kept in the greehouse continuously (high abscisic acid, no CO₂ sensitivity), nor in plants transferred from the cold to the warm chamber (low abscisic acid, high CO₂ sensitivity). The abscisic acid content of leaves was correlated with water content only within narrow limits, if at all. At equal water contents, prechilled leaves contained more abscisic acid than leaves of plants pretreated in the warm chamber. There appear to be at least two compartments for abscisic acid in the leaf.     

Biochemical and Histochemical Localization of Monoterpene Biosynthesis in the Glandular Trichomes of Spearmint (Mentha spicata)

The primary monoterpene accumulated in the glandular trichomes of spearmint (Mentha spicata) is the ketone (−)-carvone which is formed by cyclization of the C₁₀ isoprenoid intermediate geranyl pyrophosphate to the olefin (−)-limonene, hydroxylation to (−)-trans-carveol and subsequent dehydrogenation. Selective extraction of the contents of the glandular trichomes indicated that essentially all of the cyclase and hydroxylase activities resided in these structures, whereas only about 30% of the carveol dehydrogenase was located here with the remainder located in the rest of the leaf. This distribution of carveol dehydrogenase activity was confirmed by histochemical methods. Electrophoretic analysis of the partially purified carveol dehydrogenase from extracts of both the glands and the leaves following gland removal indicated the presence of a unique carveol dehydrogenase species in the glandular trichomes, suggesting that the other dehydrogenase found throughout the leaf probably utilizes carveol only as an adventitious substrate. These results demonstrate that carvone biosynthesis takes place exclusively in the glandular trichomes in which this natural product accumulates.     

Biosynthesis of Resin Terpenes in Leaves and Glandular Hairs of Newcastelia viscida

Label from ¹⁴CO₂ and [2-¹⁴C]mevalonic acid was incorporatedinto resin components of Newcastelia viscida. About 25% of thelabel from [2-¹⁴C]MVA was incorporated into resin componentsafter 4 h. Approximately 80% of the label in the resin was ina TLC band coinciding with the major terpenoid components, with60% associated with the major tricyclic diterpene, a pimaradienewhich contributes 15% to the leaf resin dry weight. Experimentswith detached trichomes showed about 0?01% incorporation oflabel from [2-¹⁴C]MVA into pimaradiene, with most of the [abelbeing associated with ß-sitosterol. It is proposedthat the glandular hairs are the sites of terpene resin synthesis,as well as secretion, in this species.     

Different effects of variation in Xanthium strumarium L. (Compositae) on two insect seed predators

Summary To determine the relative importance of variation in several plant characters on susceptibility to herbivores, we examined patterns of seed predation by two monophagous insect species and patterns of variation in ten populations of the cocklebur, Xanthium strumarium. Multiple regression analysis disclosed that one seed predator was most influenced by plant chemical variation, the other was significantly influenced by both chemical and morphological variation, but variation in yet another character, general burr size, was most important in conferring resistance to both insects simultaneously. The plant populations differed most in this character. Although many of the plant characters were correlated with each other, those important in determining susceptibility to each insect species were uncorrelated and independent of those conferring resistance to both insects simultaneously.These results imply that ecological similar herbivores may be influenced by different aspects of plant variation, and that predictions of evolutionary responses of local plant populations to herbivory may require knowledge of the structure of local herbivore communities and the dynamics of their establishment.     

Kinetics Studies on the Photoinduction of Xanthium strumarium L. and Isolation of a Flower Primordia Growth-Regulating Fractionl

The kinetics of flower primordia formation have been re-examinedin photoinduced Xanthium strumarium L. to determine the numberof short-day treatments that promote optimum primordia initiationand development. Plants given 4 short-day cycles showed thehighest rate of primordia development. Attempts were made toisolate flower-promoting substance(s) from the young leavesof such optimally induced plants by methanol extraction followedby ether fractionation. A Xanthium bioassay was developed fortesting the flower-promoting activity of aqueous concentratesobtained from the acidic, neutral-basic and total ether extracts.No flower-primordia inducing activity was found in any of thefractions, but growth-accelerating activity on young primordiawas shown by substance(s) present in the combined neutral-basicfraction isolated from photoinduced leaves. ¹C.R.D.A. Contribution No. 170. (Received June 14, 1989; Accepted September 20, 1989)     

Silicon Nanoparticles Mediated Increase in Glandular Trichomes and Regulation of Photosynthetic and Quality Attributes in Mentha piperita L.

Journal of Plant Growth Regulation - Interest in the use of the nanoparticles as plant growth elicitors mushroomed within the last decade and the field is quite intriguing to meet the growing needs...     

Glandular Trichomes of Fagonia L. (Zygophyllaceae) Species: Structure, Development and Secreted Materials

The glandular trichomes ofFagoniaconsist of one secretory celland a multicellular stalk, which develops by division, elongationand elevation of epidermal cells. The latter become seperatedfrom the mesophyll and a subepidermal chamber is formed. Thelength of the stalk, which differs in the various species orvarieties is determined by the number of cell divisions and/or the extent of cell elongation. Although the basic morphologyand development of the trichomes of the species and varietiesexamined are similar, two types of mature trichomes can be distinguished:one occurs in the two examined varieties ofF. mollisand thesecond inF. glutinosaandF. arabica. The secretory cells of thesecond type possess a very thick wall and bear a porous cupuleon their top. Histochemical tests revealed that the sticky substancesecreted by the secretory cells contains mainly polysaccharidesand lipophilic compounds. The secreted material exhibits autofluorescence.InF. mollisvar.hispidachanges in the amount and shape of thefluorescent material inside the secretory cell, during the courseof development, have been observed. The contribution of theglandular trichomes inFagoniaspecies to survival in hot desertconditions is discussed. Fagonia; glandular trichomes; subepidermal chamber; secreted material; adaptation to desert conditions; stalk; fluorescence