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PubMed1980

Interaction of Gibberellic Acid and Indole-3-acetic Acid in the Growth of Excised Cuscuta Shoot Tips in Vitro.

Gibberellic acid (GA(3)) induced a marked elongation of 2.5-centimeter shoot tips of Cuscuta chinensis Lamk. cultured in vitro. In terms of the absolute amount of elongation, this growth may be the largest reported for an isolated plant system. The response to hormone was dependent on an exogenous carbohydrate supply. The hormone-stimulated growth was due to both cell division and cell elongation. The growth response progressively decreased if GA(3) was given at increasingly later times after culturing, but the decreased growth response could be restored by the application of indole-3-acetic acid (IAA) to the apex. Explants deprived of GA(3) gradually lost their ability to transport IAA basipetally, but this ability was also restored by auxin application. The observations are explained on the basis that: (a) the growth of Cuscuta shoot tip in vitro requires, at least, both an auxin and a gibberellin; and (b) in the absence of gibberellin the cultured shoot tip explants lose the ability to produce and/or transport auxin.

Journal Article↗
PubMed1981

Trehalose Toxicity in Cuscuta reflexa: CORRELATION WITH LOW TREHALASE ACTIVITY.

A toxic effect of alpha,alpha-trehalose in an angiospermic plant, Cuscuta reflexa (dodder), is described. This disaccharide and its analogs, 2-aminotrehalose and 4-aminotrehalose, induced a rapid blackening of the terminal region of the vine which is involved in elongation growth. From the results of in vitro growth of several angiospermic plants and determination of trehalase activity in them, it is concluded that the toxic effect of trehalose in Cuscuta is because of the very low trehalase activity in the vine. As a result, trehalose accumulates in the vine and interferes with some process closely associated with growth. The growth potential of Lemna (a duckweed) in a medium containing trehalose as the carbon source was irreversibly lost upon addition of trehalosamine, an inhibitor of trehalase activity. It is concluded that, if allowed to accumulate within the tissue, trehalose may be potentially toxic or inhibitory to higher plants in general. The presence of trehalase activity in plants, where its substrate has not been found to occur, is envisaged to relieve the plant from the toxic effects of trehalose which it may encounter in soil or during association with fungi or insects.

Journal Article↗
PubMed1982

Trehalose Toxicity in Cuscuta reflexa: SUCROSE CONTENT DECREASES IN SHOOT TIPS UPON TREHALOSE FEEDING.

Trehalose, an alpha,alpha-diglucoside, induced a rapid blackening and death of shoot tips of Cuscuta reflexa (dodder) cultured in vitro. The onset of toxic symptom was delayed if any of the several sugars which support the in vitro growth of Cuscuta was supplied with trehalose. The rate of trehalose uptake or its accumulation in the tissue was not affected by sugar cofeeding. The levels of total and reducing sugars declined appreciably in the trehalose-fed shoot tip explants compared to control tissue cultured in absence of a carbon source. This was not due to an increased rate of respiration of the trehalose-treated tissue. In shoot tips cultured in presence of both trehalose and sucrose, the decline in total and reducing sugars was curtailed. There was a marked fall in the level of sucrose; and invertase activity was higher in trehalose-fed shoot tips. The incorporation of label from [(14)C]glucose into sucrose in the shoot tip explant was reduced as early as 12 h of trehalose feeding. The results suggest that increased utilization of sucrose as well as an inhibition of its synthesis contribute to the drastic fall in the sucrose content upon trehalose feeding.

Journal Article↗
PubMed1982

Trehalose Toxicity in Cuscuta reflexa: Cell Wall Synthesis Is Inhibited upon Trehalose Feeding.

alpha,alpha-Trehalose induced a rapid blackening of the terminal 2.5-centimeter region of excised Cuscuta reflexa Roxb. vine. The incorporation of radioactivity from [(14)C]glucose into alkali-insoluble fraction of shoot tip was markedly inhibited by 12 hours of trehalose feeding to an excised vine. This inhibition was confined to the apical segment of the vine in which cell elongation occurred. The rate of blackening of shoot tip explants was hastened by the addition of gibberellic acid A(3), which promoted elongation growth of isolated Cuscuta shoot tips. The symptom of trehalose toxicity was duplicated by 2-deoxyglucose, which has been shown to be a potent inhibitor of cell wall synthesis in yeast. The observations suggest that trehalose interferes with the synthesis of cell wall polysaccharides, the chief component of which was presumed to be cellulose.

Journal Article↗
PubMed1997

[Chemical constituents of Cuscuta austrolis R.Br].

Australiside A(1), a new diterpenoid glucoside, was isolated together with thymidine(2), caffeic acid(3), p-coumaric acid(4) and caffeic-beta-D-glucoside(5) from Cuscuta australis. Based on spectral analysis and physico-chemical properties the structures of the above-cited five compounds were identified. The structure of compound 1 was deduced as 4-oic acid-7-oxo-kaurene-6 alpha-O-beta-D-glucoside. Compounds 2, 3, 4 and 5 were obtained from Cuscuta for the first time.

Caffeic Acids↗
PubMed2000

Effects of flavonoids from Semen Cuscutae on the reproductive system in male rats.

AIM: To evaluate the effects of the flavonoids extracted from the Semen Cuscutae (FSC) on the reproductive and endocrine functions in male rats. METHODS: (1) FSC were obtained from the semen of Cuscuta sinensis Lam through solvent extraction and polyamide columnar chromatography; (2) Effect of FSC on the reproductive organs was assessed in immature rats. Rats were administered FSC through gastric gavage at a dose of 300 mg/kg per day for 7 days and the weights of testis, epididymis, seminal vesicle and pituitary gland were then observed; (3) To observe the effect of FSC on the reproductive endocrine function: same dose level of FSC was given to male rats of different age groups for 7 days; on day 8, the plasma testosterone (T), estradiol (E2) and LH were determined by RIA, the specific binding of LH was estimated and the testes were weighed. (4) Effect of FSC on LH secretion was assessed in vitro on cultured adenohypophysis. (5) Effect of FSC on T secretion was assessed in vitro on Leydig cell culture. RESULTS: FSC increased the weights of testis, epididymis and pituitary gland, and stimulated T and LH secretion both in vitro and in immature rats. CONCLUSION: FSC invigorates the reproductive system and reproductive endocrine function in male rats.

Animals↗
PubMedDate not supplied

Analysis of flavonoids in Semen Cuscutae by micellar electrokinetic chromatography and high-performance liquid chromatography.

Five different flavonoids in Semen Cuscutae were determined for the first time by micellar electrokinetic chromatography (MEKC) and reversed-phase high-performance liquid chromatography (RP-HPLC). MEKC was carried out on a 68.5-cm fused-silica capillary using 0.035 mol/L sodium dodecyl sulfate and 0.02 mol/L borate-phosphate (pH 7.0) containing 10% methanol as the running buffer. RP-HPLC was performed on a 12.5-cm Hypersil C18 (Agilent Technologies, Palo Alto, CA, U.S.A.) column with a 0.025-mol/L phosphate acid (A) - methanol (B) solvent gradient. Both MEKC and HPLC are rapid, simple, and efficient. The results obtained using these techniques are compared. MEKC and HPLC can be complementary to each other for the quality control of Semen Cuscutae.

Calibration↗
PubMed1990

[Effects of alcohol extract from Polygonatum odoratum (Mill.) Druce and Cuscuta australis R. Br. on immunological function of mice injured by burns].

This paper shows that the alcoholic extract from Polygonatum odoratum and Cuscuta australis increases the percentage of phagocytosis and the index of phagocytosis of macrophages as well as hemolysin and proliferation of lymphocytes up to normal level in mice with II+ burn injury covering 5% body surface. It follows that both Polygonatum odoratum and Cuscuta australis may serve as a kind of immunopotentiator for mammals.

Adjuvants, Immunologic↗
PubMed1997

Physico-chemical and functional characterization of a high molecular weight carboxymethylcellulase from Cuscuta reflexa.

Carboxymethylcellulase (CMCase) was extracted and purified from an angiosperm parasite Cuscuta reflexa free from beta-glucosidase and other enzyme activities. The molecular mass and Stokes' radius of the purified enzyme are 144 kDa and 44 A, respectively. The diffusion coefficient and frictional ratio of the enzyme were 5.15 x 10(-7) cm2/sec and 1.27. The SDS-PAGE revealed homotetrameric nature of the enzyme with a subunit molecular mass of 35 +/- 1 kDa. Titration against DTNB and NBS revealed 19 sulfhydryl groups and 8 tryptophan groups, respectively, per mole of the enzyme. A sharp pH optimum at 5.0 was obtained. Cuscuta CMCase activity is unique amongst plant endoglucanases in being stimulated by Mg2+ and Mn2+ ions and by various thiols. Reaction product analysis, mode of enzyme action and substrate specificity test suggest the endo- nature of the purified CMCase. The enzyme showed K(m) value of 26 +/- 1 mg/ml for carboxymethylcellulose (sodium salt).

Cellulase↗
PubMed2004

Glyphosate inhibits the translocation of green fluorescent protein and sucrose from a transgenic tobacco host to Cuscuta campestris Yunk.

The parasitic plant Cuscuta campestris is dependent on its host for water, assimilates and amino acids. It can be controlled by the herbicide glyphosate, which inhibits 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), resulting in shikimate accumulation. In this study, C. campestris was parasitic on transgenic tobacco plants expressing green fluorescent protein (GFP) in the phloem. Changes in [14C]sucrose and GFP accumulation in the parasite were used as indicators of the herbicide's effect on translocation between the host and parasite. Host plants were treated with glyphosate 22 days after sowing. Shikimate accumulation in the parasite 1 day after glyphosate treatment (DAGT) confirmed EPSPS inhibition in C. campestris. No damage was visible in the host plants for the first 3 DAGT, while during that same time, a significant reduction in [14C]sucrose and GFP accumulation was observed in the parasite. Thus, we propose that the parallel reduction in GFP and sucrose accumulation in C. campestris is a result of a glyphosate effect on the parasite's ability to withdraw assimilates from the host.

Cuscuta↗
PubMed2005

C3 binding glycoprotein from Cuscuta europea induced different cytokine profiles from human PBMC compared to other plant and bacterial immunomodulators.

The immunomodulatory properties of bioactive agents include the ability to induce cytokine production by the activated target cell. The effect of immunomodulatory C3 binding glycoprotein isolated from Cuscuta europea on the induction of human PBMC cytokine synthesis and the cell viability was investigated. Isolated PBMC from healthy donors were cultured for 24 h with C3bgp. We also studied the influence of C3bgp on the cytokine production in LPS, PHA, PWM and Dex treated PBMC. The quantitative determination of TNF-alpha, IL-12, IL-6 and IL-10 was performed in culture supernatant by ELISA. Results obtained demonstrated that C3bgp induced proinflammatory and immunoregulatory cytokine production, in the highest degree IL-12, followed by IL-6 and in lower degree TNF-alpha. IL-12 quantity was significantly increased in C3bgp stimulated cultures in comparison with LPS, PHA and PWM stimulated PBMC. C3bgp also increased IL-12 in PHA or PWM stimulated cultures, but not in LPS stimulated culture. C3bgp significantly increased IL-6 production compared to the PHA and PWM but not to LPS stimulation. On the other side, C3bgp inhibited IL-10 production after LPS, PHA and PWM stimulation. Cell viability in C3bgp stimulated cultures retained on the same level from 72 to 120 h of culturing, in contrast to LPS and PHA stimulated cultures. Based on the results presented, we conclude that the C3bgp exhibited immunomodulatory properties on the human PBMC. The ability of PDTC and Dex to down-regulate the effect of C3bgp on the proinflammatory cytokine production suggests that a part of the mechanism of action of C3bgp is mediated through NF-kB signal transduction pathway.

Adjuvants, Immunologic↗
PubMed2005

Adjuvant effect of ethanol extract of Semen Cuscutae on the immune responses to ovalbumin in mice.

An ethanol extract of Semen Cuscutae (EESC) was evaluated for its adjuvant potentials on the cellular and humoral immune responses of ICR mice against ovalbumin (OVA). ICR mice were immunized subcutaneously with OVA 100 microg alone or with OVA 100 microg dissolved in saline containing aluminum hydroxide gel (Alum) (200 microg), Quil A (10 or 50 microg) or EESC (100, 200 or 400 microg) on Days 1 and 15. Two weeks later (Day 28), concanavalin A (Con A)-, pokeweed (PWM)- and OVA-stimulated splenocyte proliferation and OVA-specific antibody in serum were investigated. EESC significantly enhanced the Con A-, PWM-, and OVA-induced splenocyte proliferation in OVA-immunized mice at a dose of 200 microg (P<0.05 or P<0.025). OVA-specific IgG, IgG1 and IgG2b antibody levels in serum were significantly enhanced by EESC compared with OVA control group (P<0.025). Moreover, enhancing effects of EESC on these OVA-specific antibody responses to OVA in mice were more significant than those of Alum and Quil A (P<0.025). In conclusion, the results suggest that EESC is effective on Th1 and Th2 cell functions, and could be safely used as adjuvant.

Adjuvants, Immunologic↗
PubMed2003

Effect of Cuscuta chinensis glycoside on the neuronal differentiation of rat pheochromocytoma PC12 cells.

Exposure of rat pheochromocytoma PC12 cells to Cuscuta chinensis glycoside induced neuronal differentiation with resulting outgrowth of neurites and increase of acetylcholinesterase activity. A specific inhibitor of mitogen-activated protein kinase (MAPK) kinase, PD98059, prevented this effect of C. chinensis on PC12 cells. These results suggested that C. chinensis glycoside induced neuronal differentiation in PC12 cells linked to the mitogen-activated protein kinase signaling cascade.

Acetylcholinesterase↗
PubMed2004

Isolation of a new 15-membered macrocyclic glycolipid lactone, Cuscutic Resinoside a from the seeds of Cuscuta chinensis: a stimulator of breast cancer cell proliferation.

While searching for new estrogenic compounds from the plant kingdom, we investigated an extract of the seeds of Cuscuta chinensis (Convolvulaceae) which showed potency for stimulating MCF-7 cell proliferation. A novel resin glycoside, cuscutic resinoside A ( 6) was isolated along with five known compounds from the extract. The structure was deduced from its spectral data as (11 S)-hydroxyhexadecanoic acid 11- O-alpha- L-(4- O-2 R,3 R-nilylrhamnopyranosyl)-(1-->2)- O-alpha- L-rhamnopyranosyl-(1,2-lactone) forming a unique 15-membered macrocyclic lactone. The compound significantly stimulated not only MCF-7 cell proliferation but also T47D human breast cancer cells at a concentration of 10 microM. Along with cuscutamine ( 1) and kaempferol ( 4), 6 was tested in the transient luciferase reporter assay and was found to have different luciferase inducing activity characteristics from the other compounds. These results suggest that 6 stimulated cancer cell proliferation by a different mechanism from 1 and 4.

Breast Neoplasms↗
PubMed2003

Carotenoid specificity of light-harvesting complex II binding sites. Occurrence of 9-cis-violaxanthin in the neoxanthin-binding site in the parasitic angiosperm Cuscuta reflexa.

The parasitic angiosperm Cuscuta reflexa has a highly unusual carotenoid composition in that it does not contain neoxanthin, an otherwise ubiquitous component of the major light-harvesting complex protein (LHCIIb) in all other higher plant species studied to date. Combined HPLC and mass spectrometric analysis has enabled us to detect in tissues of C. reflexa two new types of xanthophylls: lutein-5,6-epoxide and 9-cis-violaxanthin. We have isolated the LHCIIb complex from thylakoids and analyzed chlorophyll and carotenoid composition. The data show that the 9-cis-violaxanthin is present in amounts similar to that of neoxanthin in most plants. On the other hand, lutein-5,6-epoxide was found to be in substoichiometric quantities, suggesting a peripheral location similar to the loosely-associated all-trans-violaxanthin and also enabling suitable accessibility for the de-epoxidase (VDE). Absorption spectroscopy revealed close similarities of the excited state energies of neoxanthin and 9-cis-violaxanthin in vitro and in intact LHCIIb complex. Resonance Raman analysis clearly indicates a cis conformation of violaxanthin in the complex, confirming the pigment analysis data and proving that not only does violaxanthin replace neoxanthin as an intrinsic component of LHCIIb in C. reflexa but it also adopts the same 9-cis conformation of neoxanthin. These results suggest that the N1 binding site of LHCIIb preferentially binds 9-cis-5,6-epoxy carotenoids, which has implications for the features of this binding site and its role in the photosystem II antenna assembly and stability.

Binding Sites↗
PubMed2004

Synthesis of phytochelatins and modulation of antioxidants in response to cadmium stress in Cuscuta reflexa--an angiospermic parasite.

Effect of cadmium on growth, antioxidative enzymes namely catalase, peroxidase, glutathione reductase, level of glutathione and phytochelatin synthesis was investigated in callus and seedlings of Cuscuta reflexa. A time, concentration and tissue dependent response of Cd was observed. Cd inhibited the growth of callus and seedlings by 50% at 300 and 500 micromol/L concentrations, respectively. Shorter exposure of low concentration of Cd led to augmentation of antioxidant activity, both in callus and seedlings, while longer exposure and high concentration of Cd led to a concentration dependent decrease in callus. Analysis of phytochelatin (PC) synthesis in callus and seedlings of C. reflexa revealed both quantitative and qualitative changes. Cd at low concentrations led to synthesis of predominantly PC4, while at higher concentrations, PC3 was the major form being synthesized. Amelioration of antioxidative systems of C. reflexa in response to Cd stress might be playing a protective role, alleviating the damaging effects of ROS, generated during Cd stress. Concomitantly, chelation and sequestering of toxic Cd ions in this parasite was mediated by synthesis of PC. The response to Cd stress shown by this holoparasitic plant was found to be similar to those of non-parasitic plants (hosts).

Antioxidants↗
PubMed2004

Relatedness affects competitive performance of a parasitic plant (Cuscuta europaea) in multiple infections.

Theoretical models predict that parasite relatedness affects the outcome of competition between parasites, and the evolution of parasite virulence. We examined whether parasite relatedness affects competition between parasitic plants (Cuscuta europaea) that share common host plants (Urtica dioica). We infected hosts with two parasitic plants that were either half-siblings or nonrelated. Relative size asymmetry between the competing parasites was significantly higher in the nonrelated infections compared to infections with siblings. This higher asymmetry was caused by the fact that the performance of some parasite genotypes decreased and that of others increased when grown in multiple infections with nonrelated parasites. This result agrees with the predictions of theories on the evolution of parasite virulence: to enhance parasite transmission, selection may favour reduced competition with genetically related parasites in hosts infected by several genotypes. However, in contrast to the most common predictions, nonrelated infections were not more virulent than the sibling infections.

Analysis of Variance↗