Isolation of Nematicidal Polyacetylenes from Carthamus tinctorius L.

Two nematicidal polyacetylenes, 3-cis,11-trans- and 3-trans,11-trans-trideca-1,3,11-triene-5,7,9-triyne, were isolated from flowers of Carthamus tinctorius L., by column chromatography and high speed liquid chromatography under the dark condition.

The nematicidal activities of 3-cis,11-trans- and 3-trans,11-trans-isomer to Aphelenchoides besseyi were 80% at 10 ppm and 95% at 1 ppm, respectively.     

Quantitative profiling of polyacetylenes in tissue cultures and plant parts of three species of the Asteraceae

Polyacetylenes are a group of fatty-acid derived specialized metabolites with several C–C-triple bonds and derived compounds which are widely distributed in the plant kingdom, but are especially abundant and structurally diverse in the Asteraceae family. Despite their interesting structural and biological properties, the biosynthesis of polyacetylenes is only poorly understood. We have used three species of the Asteraceae (Carthamus tinctorius, Tagetes patula, and Arctium lappa) to compare their suitability for studies of polyacetylene biosynthesis when used after cultivation on soil or as tissue culture. The polyacetylene profiles detected in different plant parts together with information from the literature indicate that C. tinctorius seedlings and flowers as well as T. patula roots and flower buds are major sites of polyacetylene biosynthesis. Highest levels of polyacetylenes were detected in T. patula [about 30 µmol/g dry weight (d.w.) thiophenes in roots] while A. lappa contained less than 1 µmol/g d.w.. Methyljasmonate (MeJ)-induced T. patula hairy root cultures proved to be an excellent source of butenynyl-bithiophene (200 µmol/g d.w., 43 mg/g d.w.) while T. patula flower buds could serve as a source of pentenynyl-bithiophene and α-terthienyl (5–10 µmol/g d.w.) and C. tinctorius flowers or seedlings as a source of polyacetylenic C₁₃ hydrocarbons, the biosynthetic precursors of thiophenes (5–10 µmol/g d.w.). Upon addition of elicitors to tissue cultures, highest elicitation factors (between four and seven) were reached for 1,11-tridecadiene-3,5,7,9-tetrayne in C. tinctorius cell suspension cultures with 40 µM MeJ and α-terthienyl in T. patula hairy root cultures with 100 µM MeJ.     

Production of Cephalosporin Antibiotics by Strains Belonging to the Genera Arachnomyces,Anixiopsis and Spiroidium

The absolute configuration of a phytoalexin “Safynol” (3E, 11E)-3, 11-tridecadiene-5, 7, 9-triyne-l, 2-diol (I), separated from Phytophthora drechsleri-infected safflower (Carthamus tinctorius L.) was confirmed. (R) and (S)-safynol were synthesized from 2, 3-O-isopropyli- dene-(S)- and (R)-glyceraldehyde respectively, and the (R) configuration was assigned to the natural product. The inhibitory effects of (R)- and (S)-safynol on mycelial growth of five fungi were almost the same and their ED₅₀ values ranged from 6 to 70 ppm.     

Flavonoids and polyacetylenes in Dahlia tenuicaulis

5,7,4′-Trimethoxyflavanone, 5-hydroxy-7,4′-dimethoxyflavanone, 2′-hydroxy-4,4′,6′-trimeth-oxychalcone, and a new naturally occurring compound, 5,7,4′-trimethoxyflavan-4-ol have been isolated from the leaves of Dahlia tenuicaulis Sorensen. Two chalcones, 4,2′,4′-trihydroxychalcone and 3,2′,4′-trihydroxy-4-methoxychalcone, and 5-hydroxy-7,4′-dimethoxyflavanone have been isolated from the flower heads. Minute amounts of the polyacetylene 1,3-diacetoxy-tetradeca-4,6-diene-8,10,12-triyne have been found in both leaves and flower heads, whereas 1-acetoxy-tetradeca-4,6-diene-8,10,12-triyne was present only in the flower heads.     

New polyacetylene glucosides from the florets of Carthamus tinctorius and their weak anti-inflammatory activities

Eight new linear polyacetylene glucosides (1–8), containing two C₁₀-, one C₁₃- and five C₁₄-acetylenes, together with three known polyacetylenes (9–11) were isolated from the florets of Carthamus tinctorius L. Their structures were elucidated by means of spectroscopic methods and chemical evidence. The absolute configurations of compounds 3–9 were confirmed by Snatzke and Gerards’s method, observing the induced circular dichroism after addition of dirhodium tetrakis (trifluoroacetate) [Rh₂(OCOCF₃)₄] in CHCl₃. All the isolated compounds (1–11) were also tested for inhibitory activities against LPS-induced NO production in murine macrophages and just showed weak activities at concentrations of 1 × 10⁻⁵ M.     

Polyacetylenes of Dahlia

Two Dahlia sherffii strains and three Dahlia pinnata strains and Dahlia rudis Sorensen have been investigated for polyacetylenes. A great number of acetylenic compounds, characterized earlier from other Dahlia species was determined. The two Dahlia sherffii strains and the three Dahlia pinnata strains contain 1-phenylhept-5-ene-1,3-diyne and 1-phenyl-hepta-1,3,5-triyne and derivatives which have previously been found in one Dahlia coccinea strain and in a horticultural form. A comparison between this group of dahlias with others previously investigated, show characteristic differences in the chemical pattern. Dahlia rudis is thus closer to the D. imperialis and the D. tenuicaulis and three of the Dahlia coccinea strains. Dahlia merckii exhibits a quite unique pattern of polyacetylenes.     

白花鬼针草中多烯炔类成分的分离及其生物活性研究

为了解白花鬼针草(Bidens pilosa var.radiata)的化学成分,采用多种色谱技术从其提取物中分离多烯炔类成分,并对其生物活性进行研究.结果表明,从白花鬼针草乙酸乙酯提取部位中分离鉴定出4个多烯炔类化合物,分别为5-acetoxy-2-phenyl-ethinyl-thiophene(1)、1-phen...     

Serotobenine,a Novel Phenolic Amide from Safflower Seeds (Carthamus tinctorius L.)

The structure of a new indole derivative, serotobenine (1), from sfflower meal (Carthamus tinctorius> L.) is proposed based on ¹H- and ¹³C-NMR spectral data and X-ray crystallography. The known compounds N-feruloyltryptamine (2) and N-(p-coumaroyl)tryptamine (3) were also isolated and identified.     

Root herbivory in vitro: Interactions between roots and aphids grown in aseptic coculture

Summary An in vitro coculture system has been established to study interactions between roots and aphids. Eight aphid species (Aphis spiraecola P., Trama rara M., Macrosiphum euphorbiae S., Rhopalosiphum padi L., Sitobion avenae F., Rhopalosiphum maidis F., Metopolophium dirhodum W., and Pemphigus populivenae F.) were reared on six species of hairy root cultures, Carthamus tinctorius L. cv N10, Tagetes patula L., Trichosanthes cucumerina L. var anguina, Hyoscyamus muticus L., Nicotiana tabacum L., and Beta vulgaris L. subsp. vulgaris. All species of aphids survived on root cultures for at least 2 d. Three cocultures have been maintained aseptically for periods ranging from 2 mo. to over 2 yr. The coculture of R. padi on C. tinctorius cv N10 (N10-Rp) was used to study morphological and biochemical responses of roots under aphid herbivory. Aphid herbivory caused browning of cultures, reduced root vegetative growth, and increased production of polyacetylenes in C. tinctorius cv N10 roots. Our results suggest that this coculture system may improve our understanding of interactions between aphids and plant roots.     

Relationship between Floret Elongation and Pigment Synthesis in the Flowers of Carthamus tinctorius

Floret elongation and levels of precarthamin were investigatedin freshly collected flowers of Carthamus tinctorius. Accumulationof precarthamin was found to be induced at the early stagesof floret elongation. [U-¹⁴C]Acetate and [U-¹⁴C]phenylalaninewere incorporated into precarthamin in the detached floretsfrom the flower bud. The results suggest that precarthamin issynthesized via the acetate-shikimate pathway. Carthamus tinctorius L, floret elongation, pigment synthesis, precarthamin     

Effect of salinity on plant growth and biological activities of Carthamus tinctorius L. extracts at two flowering stages

In the present study, we were interested in the effect of salt stress on phenolic and carotenoid contents, antioxidant and antimicrobial activity of two varieties of Carthamus tinctorius (Jawhara and 104) flowers. For this purpose, C. tinctorius flowers from plants grown under four saline treatments (0, 5, 10 and 15 g/L NaCl) were collected at two development stages. As salinity increased up to 10 g/L, results showed that total phenols, flavonoids, condensed tannins and carotenoid contents increased with salinity. Such variability might be of great importance in terms of valorizing this plant as a source of naturally secondary metabolites. Furthermore, our results showed an enhancement of antioxidant activity which was evaluated by four different test systems (DPPH, β-carotene–linoleic acid, chelating and reducing power assays) with increasing stress severity. Obtained results showed that, for the two varieties, salt effect was more pronounced at post flowering stage than full flowering one. The sensitivity test of the methanolic extracts of the harvested flowers was applied against seven human pathogenic bacteria and three yeast strains. Salinity reduced significantly the antimicrobial activity of flower extracts.     

Physical and genetic mapping of safflower chloroplast DNA

Summary A physical map of safflower (Carthamus tinctorius L.) chloroplast DNA has been generated using SalI, PstI, KpnI and HindIII restriction endonucleases. The circular plastid genome (151 kbp) has the usual inverted repeat. Heterologous probes containing psbA, rbcL, atpA or rrnA structural genes mapped colinearly with spinach and other chloroplast genomes.     

Bottlenecks in erucic acid accumulation in genetically engineered ultrahigh erucic acid Crambe abyssinica

Erucic acid is a valuable industrial fatty acid with many applications. The main producers of this acid are today high erucic rapeseed (Brassica napus) and mustard (Brassica juncea), which have 45%–50% of erucic acid in their seed oils. Crambe abyssinica is an alternative promising producer of this acid as it has 55%–60% of erucic acid in its oil. Through genetic modification (GM) of three genes, we have previously increased the level of erucic acid to 71% (68 mol%) in Crambe seed oil. In this study, we further investigated different aspects of oil biosynthesis in the developing GM Crambe seeds in comparison with wild‐type (Wt) Crambe, rapeseed and safflower (Carthamus tinctorius). We show that Crambe seeds have very low phosphatidylcholine‐diacylglycerol interconversion, suggesting it to be the main reason why erucic acid is limited in the membrane lipids during oil biosynthesis. We further show that GM Crambe seeds have slower seed development than Wt, accompanied by slower oil accumulation during the first 20 days after flowering (DAF). Despite low accumulation of erucic acid during early stages of GM seed development, nearly 86 mol% of all fatty acids accumulated between 27 and 50 DAF was erucic acid, when 40% of the total oil is laid down. Likely bottlenecks in the accumulation of erucic acid during early stages of GM Crambe seed development are discussed.     

Genetic variation of Carthamus tinctorius L. and related species revealed by SRAP analysis

Genetic diversity of 23 populations of Carthamus tinctorius L. and two populations of Carthamus lanatus L. in China was investigated using Sequence-related Amplified Polymorphism (SRAP). All populations could be uniquely distinguished by 30 primer combinations with 483 bands and 274 polymorphic bands which generated 57% of polymorphic ratio. Unweighed pair-group method of with arithmetical averages (UPGMA) cluster analysis enabled construction of a dendrogram for estimating genetic distances among different populations. The extreme variation was observed when No. 4 cultivated and No. 13 wild population of C. lanatus were grouped at GS = 0.58, and separated from 23 populations of C. tinctorius at GS = 0.10. The result suggested that the cultivated and wild populations of C. lanatus had close relationship with each other and far relationship with C. tinctorius. Dendrogram also revealed a large genetic variation in 23 C. tinctorius populations; different primer combinations allowed them distinctly distinguished one from others with relatively low genetic similarity. Furthermore, five typical representative fragments in C. lanatus were obtained by four most informative primer combinations, which provided a possibility to distinguish C. lanatus from the C. tinctorius evidently.     

Optimizing Seed Water Content: Relevance to Storage Stability and Molecular Mobility

This research was conducted to determine the optimum moisture content (MC) that gave maximum longevity to seeds. Three species were used to represent seeds with different dry matter reserves, which gives them different sorption properties: maize (Zea mays L.), elm (Ulmus pumila L.) and safflower (Carthamus tinctorius L.). The seeds of elm, safflower, and maize embryos with MC ranging from 0.00–0.15 g H₂O/g dry weight (DW) were stored at 35 °C for different periods of time. The results showed that the optimum MC for seed and embryo storage varied between species (0.057 g H₂O/g DW for maize embryos, 0.045 g H₂O/g DW for elm, and 0.02 g H₂O/g DW for safflower). Drying below this optimum MC increased the aging rate and there were detrimental effects of drying. The relative humidity corresponding to optimum MC in embryos of maize, elms and safflower was about 15%, 12% and 7% respectively, according to the lipid composition of the embryos. The data provided confirmatory evidence that molecular mobility (ΔAzz) in elms, maize and safflower embryos was compatible with the optimum moisture content.     

Construction and mapping of safflower chloroplast DNA recombinants and location of selected gene markers

Summary DNA was isolated and purified from chloroplasts of safflower (Carthamus tinctorius L.), digested with HindIII restriction endonuclease, and ligated into the HindIII site of the plasmid pUC9. Recombinant DNAs were isolated from ampicillin resistant white colonies which grew in the presence of the appropriate indicator, digested with HindIII, and then identified by comparison of agarose gel electrophoretic mobilities. HindIII digests of chloroplast DNA were used as a standard. Such recombinants were radiolabeled and hybridized with Southern blots of PstI, SalI, KpnI, and HindIII single and double digests of safflower chloroplast DNA. A physical map was subsequently generated showing the location of each recombinant on the circular plastid genome. Recombinants containing heterologous chloroplast gene markers from spinach or Euglena were also radiolabeled and mapped. The relative mapping positions of these genes are in good agreement with those which have previously been published for spinach and several other higher plants.     

Introgression potential between safflower (Carthamus tinctorius) and wild relatives of the genus Carthamus

Background  

Safflower, Carthamus tinctorius, is a thistle that is grown commercially for the production of oil and birdseed and recently, as a host for the production of transgenic pharmaceutical proteins. C. tinctorius can cross with a number of its wild relatives, creating the possibility of gene flow from safflower to weedy species. In this study we looked at the introgression potential between different members of the genus Carthamus, measured the fitness of the parents versus the F1 hybrids, followed the segregation of a specific transgene in the progeny and tried to identify traits important for adaptation to different environments.     

Geographical and seasonal variation in water hemlock (Cicuta maculata) toxins

Water hemlock (Cicuta spp.) plants are typically found in wet areas and are toxic to all species of livestock. The toxic components in water hemlock are C₁₇ polyacetylenes, with cicutoxin being the most studied. The objective of this study was to evaluate the variation in cicutoxin and total C₁₇ polyacetylene compounds in water hemlock populations across western North America. Cicutoxin and total C₁₇ polyacetylene concentrations varied among the six collection locations and among plant parts. Tubers contained the highest cicutoxin and total C₁₇ polyacetylene concentrations of all plant parts. Green seeds contained the second most abundant total C₁₇ polyacetylene concentrations. Total C₁₇ polyacetylene and cicutoxin concentrations were also compared in different plant parts at several different phenological stages over the growing season. Cicutoxin and total C₁₇ polyacetylene concentrations in the tubers increased until the green seed stage whereupon the concentrations decreased as the seeds matured and the plant began to senesce. Concentrations of secondary compounds in the stems were consistent with the optimal defense theory in which secondary compounds were higher in stems in earlier development stages compared to later stages and concentrations in seeds were higher than other above ground parts. The toxic compounds are found in all plant parts, with tubers posing the most significant risk of livestock poisoning. Results presented in this study suggest that the toxic risk to livestock likely does not differ between water hemlock (Cicuta maculata) populations across western North America under similar circumstances of ingestion.     

Sesquiterpene and polyacetylene profile of the Bidens pilosa complex (Asteraceae: Heliantheae) from Southeast of Brazil

The pantropical weed Bidens pilosa (Asteraceae) is a species with several taxonomic problems. Recently it has been shown to be a complex of different species. To shed light on this problem in Brazil, dichloromethane extract of leaves of several populations corresponding to three proposed species for the complex in southeastern Brazil (B. pilosa, Bidens alba and Bidens subalternans) were analyzed by GC–MS. Twenty-four substances were detected, of which four resemble polyacetylenes, the others sesquiterpenes. Five sesquiterpenes tentatively identified as E-caryophyllene, α-humulene, germacrene-D, bicyclogermacrene and α-muurolene were found in all three Bidens species. The polyacetylene phenylhepta-1,3,5-triyne was identified only in B. alba. Multivariate analysis (cluster and principal component analyses) separated the three entities, suggesting that these compounds could represent a useful tool to distinguish species in the B. pilosa complex.     

Effect of Different Osmotic Potentials of Media on Germination of Three Safflower Varieties

The effect of osmotic potentials, 0 to ?13.5 atm (?1.37 M Pa), on the germination of three safflower varieties (Carthamus tinctorius L.) was studied at a controlled temperature (30±2°C) using a non-electrolyte substance, polyethelene glycol 6000. The varieties were cv. Ute, cv. Iranian local 3151, and cv. Iranian local 2811. The percent germination of the three varieties averaged over all osmotic potentials were: 86, 77 and 61 for local 3151, local 2811 and “Ute”, respectively. The Ute variety had the highest, and the local 3151 variety had the lowest rate of germination reduction. The variations among the varieties in the percent and rate of germination were due to osmotic effects of the media.