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The role of indole and other shikimic acid derived maize volatiles in the attraction of two parasitic wasps.

After herbivore attack, plants release a plethora of different volatile organic compounds (VOCs), which results in odor blends that are attractive to predators and parasitoids of these herbivores. VOCs in the odor blends emitted by maize plants (Zea mays) infested by lepidopteran larvae are well characterized. They are derived from at least three different biochemical pathways, but the relative importance of each pathway for the production of VOCs that attract parasitic wasps is unknown. Here, we studied the importance of shikimic acid derived VOCs for the attraction of females of the parasitoids Cotesia marginiventris and Microplitis rufiventris. By incubating caterpillar-infested maize plants in glyphosate, an inhibitor of the 5-enolpyruvylshikimate-3-phospate (EPSP) synthase, we obtained induced odor blends with only minute amounts of shikimic acid derived VOCs. In olfactometer bioassays, the inhibited plants were as attractive to naive C. marginiventris females as control plants that released normal amounts of shikimic acid derived VOCs, whereas naive M. rufiventris females preferred inhibited plants to control plants. By adding back synthetic indole, the quantitatively most important shikimic acid derived VOC in induced maize odors, to inhibited plants, we showed that indole had no effect on the attraction of C. marginiventris and that M. rufiventris preferred blends without synthetic indole. Exposing C. marginiventris females either to odor blends of inhibited or control plants during oviposition experiences shifted their preference in subsequent olfactometer tests in favor of the experienced odor. Further learning experiments with synthetic indole showed that C. marginiventris can learn to respond to this compound, but that this does not affect its choices between natural induced blends with or without indole. We hypothesize that for naïve wasps the attractiveness of an herbivore-induced odor blend is reduced due to masking by nonattractive compounds, and that during oviposition experiences in the presence of complex odor blends, parasitoids strongly associate some compounds, whereas others are largely ignored.

Animals↗

[Determination of shikimic acid in fruit of Illiciaceae plants by HPLC with diode-array detection].

OBJECTIVE: To establish the determination method of Shikimic acid. METHOD: Using HPLC method as the determination method. The separation was performed in a SiO2-NH2 column with a mobile phase of Acetonitrile-2% H3PO4 water solution (95:5); The sample wavelength was 213 nm, reference wavelength 300 nm. RESULT: The average collection was 98.5%, RSD 1.67% (n = 5). CONCLUSION: This method is suitable for the determination of Shkimic acid in herb medicines and preparation containing shikimic acid.

Chromatography, High Pressure Liquid↗

Comparison of shikimic acid determination by capillary zone electrophoresis with direct and indirect detection with liquid chromatography for varietal differentiation of red wines.

Two capillary zone electrophoretic (CZE) methods for determination of shikimic acid in Chilean red wine were developed and compared with a HPLC method. Both electrophoretic methods were carried out by using a reversed electroosmotic flow induced by trimethyl(tetradecyl)ammoniumbromide (TTAB) with indirect detection at 260 nm using p-aminobenzoic acid as a UV-absorbing co-ion or by direct detection at 213 nm. In both cases, the separation was carried out in a 50 microm I.D. uncoated capillary with an effective length of 48 cm, a negative power supply of 30 kV, using a buffer based on bis[2-hydroxyethyl]imino-tris[hydroxymethyl]methane (Bis-Tris), pH 7.0 or 7.5 and hydrodynamic injection. The chromatographic separations were carried out on a C-18 reversed phase column followed by a sulfonyl-styrene-divinylbenzene (S-DVB) ion exclusion column at 70 degrees C with H2SO4 0.02 M as isocratic mobile phase and a flow rate of 0.5 mL min(-1). The three methods allowed the quantification of shikimic acid with quantification limits between 1.0 and 12.0 mg L(-1) and precision between 7.3 and 10.1%, however, only the concentrations obtained by CZE with direct detection were statistically similar to those of HPLC. This parameter was evaluated as analytical tool to verify varietal authenticity of red wines. In all cases, the Cabernet Sauvignon wines presented higher concentrations of shikimic acid, compared with Merlot or Carmenère wines.

Chromatography, High Pressure Liquid↗

Gene controlling the uptake of shikimic acid by Escherichia coli.

Pittard, James (University of Melbourne, Melbourne, Australia), and B. J. Wallace. Gene controlling the uptake of shikimic acid by Escherichia coli. J. Bacteriol. 92:1070-1075. 1966.-Mutants possessing an impaired ability to take up exogenously supplied shikimic acid were isolated. Mutants showing either a complete or partial loss of this activity were found. The mutations have been mapped on the Escherichia coli chromosome and occur in a single gene, situated near to the cluster of genes concerned with histidine biosynthesis. The uptake of dehydroshikimic acid was also affected by these mutations, suggesting a common mechanism for the uptake of these two related intermediates of aromatic biosynthesis.

Carbon Isotopes↗

Expression of a hypovirulence-causing double-stranded RNA is associated with up-regulation of quinic acid pathway and down-regulation of shikimic acid pathway in Rhizoctonia solani.

We reported previously that a 3.6-kb double-stranded RNA, designated as M2, is associated with hypovirulence in the Rhizoctonia solani isolate Rhs 1A1 and proposed that the M2-encoded putative polypeptide A (pA) might interfere with the regulation of the quinate and shikimate pathways. In this study, Western blot analysis showed that a protein band of the predicted size (83 kDa) binds antibodies specific to a pA epitope and is detectable in M2-containing but not in M2-lacking cultures. A mRNA, associated with Rhs 1A1 polysomes immunoprecipitated with anti-pA antibodies, has a sequence basically identical to that of the sense-strand of M2. The normally inducible quinate pathway was constitutively expressed, whereas the shikimate pathway was down-regulated in the M2-containing, hypovirulent Rhs 1A1. Finally, the relative concentration of phenylalanine, precursor of the virulence determinant phenylacetic acid, was correlated with the degree of pathogenicity in the virulent Rhs 1AP but not in the hypovirulent Rhs 1A1.

Gene Expression Regulation, Fungal↗

[Experimental studies on the anti-thrombosis effect of 3,4-oxo-isopropylidene-shikimic acid].

AIM: To study the effect of 3,4-oxo-isopropylidene-shikimic acid (ISA) against arteriovenous shunt and middle cerebral artery thrombosis (MCAT) in rats. METHODS: Arteriovenous shunt model was adopted to measure thrombus weight; The neurologic deficit (ND) and the infarct size (IS) of the middle cerebral thrombosis (MCAT) model induced by FeCl3 were observed; The effect of ISA on platelet aggregation rate of rat and rabbit by giving ISA in vivo and in vitro was studied. RESULTS: ISA 25, 50, 100 and 200 mg.kg-1 ig was shown to reduced the weight of thrombus in arteriovenous shunt model; ISA 50, 100 and 200 mg.kg-1 ig for 2 times in 24 hours, attenuated the ND of rats subjected to MCAT; ISA 100 and 200 mg.kg-1 reduced IS of rats after MCAT by 27.8% and 31.6%, respectively; ISA 50, 100 and 200 mg.kg-1 ig inhibited platelet aggregation of normal rats; ISA 10(-3)-10(-5) mol.L-1, inhibited rabbit platelet aggregation in vitro. CONCLUSION: ISA inhibited thrombosis by anti-platelet-aggregation.

Animals↗

[Protective effect of 3,4-oxo-isopropylidene-shikimic acid on vascular endothelial cell injured by hydrogen peroxide].

AIM: To study the effect of 3,4-oxo-isopropylidene-shikimic acid (ISA) on H2O2 (200 mol.L-1, 4 h) injured human umbilical vein endothelial cells (HUVEC). METHODS: Morphological change was observed under microscop. Cell viability was assessed by MTT assay. The release of intracellular lactate dehydrogenase (LDH) and NO was assessed by colorimetry. Radioimmunoassay was used to assess 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha). RESULTS: Pretreatment with ISA for 6 h alleviated the morphological damage of H2O2 induced HUVECs. At the concentration of 1-100 mumol.L-1, ISA prevented the inhibitory effect on cell viability induced by H2O2 in dose-dependent manner, but increased the ratio of cell viability from 60.4% to 84.3%. ISA reduced LDH release and increased the level of NO and 6-keto-PGF1 alpha in H2O2 induced HUVECs. CONCLUSION: ISA exerted protective effect on H2O2 injured HUVEC.

6-Ketoprostaglandin F1 alpha↗

Antagonistic effects of shikimic acid against focal cerebral ischemia injury in rats subjected to middle cerebral artery thrombosis.

AIM: To study the effects of shikimic acid (SA) on focal cerebral ischemic injury after middle cerebral artery thrombosis (MCAT). METHODS: Thrombosis was induced by FeCl3 in middle cerebral artery of rats. The influences of SA on neurologic deficit (ND), infarct size (IS), brain edema, and cerebral blood flow (CBF) in ischemic region were observed. RESULTS: SA 25 and 50 mg.kg-1 i.p. for 3 d before MCAT attenuated ND, and reduced IS by 51% and 42%; and decreased brain water content from 80.7% to 79.8% and 79.9%; and increased CBF after ischemia from 50.2% of the preischemic level to 75.5% and 73.3%, respectively. In pathologic examination, there was much less thrombosis in MCA in the rat with the pretreatment by SA 25 mg.kg-1. The extent of brain ischemia was much less than that of control. CONCLUSIONS: SA reduced focal cerebral ischemic injury induced by middle cerebral artery thrombosis.

Animals↗

Identification of the gene (aroK) encoding shikimic acid kinase I of Escherichia coli.

DNA sequence analysis has revealed that an unidentified open reading frame (ufr1) is present immediately preceding the aroB gene of Escherichia coli. The predicted protein product of urf1 contains a consensus ATP-binding-site sequence and shows 34% amino acid homology to shikimate kinase II in a 97-amino-acid region. Inactivation of urf1 by insertion of an antibiotic resistance gene had a polar effect on aroB, indicating that these two genes constitute a transcriptional unit. The auxotrophic requirements of a strain mutant for both urf1 and aroL (encoding shikimate kinase II) are consistent with shikimate kinase deficiency. We propose that urf1 encodes shikimate kinase I and that it be designated aroK.

Amino Acid Sequence↗