Effects of temperature, multiple allelochemicals and larval age on the performance of a specialist caterpillar

To understand the mechanisms underlying plant-insect herbivore interactions, it is necessary to examine the simultaneous effects of temperature, food quality and larval age. We examined the simultaneous effects of three allelochemicals (tomatine, rutin and chlorogenic acid) on the performance of first and second instar Manduca sexta larvae under two representative thermal regimes 21 : 10°C and 26 : 15°C for spring and summer, respectively. Thermal regime and allelochemicals interacted to influence the time from egg hatch to ecdysis to the third instar. On average, it took about half as much time to reach the third instar at 26 : 15°C as it did at 21 : 10°C. Separately, tomatine and rutin had a negative effect on developmental time from egg hatch to the third instar, but their simutaneous effects were not additive. Chlorogenic acid significantly reduced the negative effect of tomatine. The magnitude of the allelochemical effect was larger at the cooler thermal regime compared to the warmer regime. For instance, chlorogenic acid by itself had no effect at the 26 : 15°C regime, but at the 21 : 10°C regime it significantly shortened total developmental time. The effect of chlorogenic acid on stadium duration was distinctly different for the two instars. Chlorogenic acid shortened stadium duration of first instar larvae. However, depending on thermal regime and the presence of tomatine, chlorogenic acid had a negative, positive or neutral effect on stadium duration of second instar larvae. Molting duration of second instar larvae was shortened by a half day at the warmer thermal regime but was not affected by the allelochemicals. Final larval weight was influenced by rutin and chlorogenic acid. Caterpillars fed diets containing 20 moles of rutin were on average 10% lighter than those fed plain diet, whereas those fed diets containing 20 moles of chlorogenic adic were on average 7% heavier. However, the effect of chlorogenic acid depended on thermal regime. Overall, our results indicated that: 1) temperature and food quality can interact to influence insect performance and 2) these effects are influenced by larval age.     

Phenyllactic acid but not tropic acid is an intermediate in the biosynthesis of tropane alkaloids in Datura and Brugmansia transformed root cultures

(S)-(-)-Tropic acid is the acidic moiety of the tropane ester alkaloids, hyoscyamine and scopolamine (hyoscine). When tropic acid is fed to transformed root cultures of Datura stramonium L. or a Brugmansia (Datura) Candida x B. aurea hybrid, the formation of these alkaloids is inhibited. Phenyllactic acid, from which the tropoyl moiety is derived, is considerably less inhibitory. Label from (RS)-phenyl[1,3-¹³C₂]lactic acid is incorporated at high levels into apoatropine, littorine, aposcopolamine, hyoscyamine, 7-hydroxyapoatropine, scopolamine and 7-hydroxyhyoscyamine when fed to these cultures. The presence of an excess concentration of unlabelled tropic acid has little influence on the specific incorporation into these products. It is concluded that free tropic acid is not an intermediate in hyoscyamine biosynthesis but rather that the rearrangement of phenyllactic acid occurs subsequent to its esterification.Abbreviations FM fresh mass - NMR nuclear magnetic resonance spectroscopy We are grateful to Drs. N.J. Walton, A.J. Parr, M.J.C. Rhodes (Institue of Food Research, Norwich) and B. Dräger (Münster, Germany) for helpful and critical discussions. We also wish to thank Dr. P. Bachmann (Braunschweig, Germany) for suggesting the use of the DB-17 column to separate littorine from hyoscyamine and for the modified Excel programme used to calculate the specific incorporations, Yannick Ford (AFRC Co-operative Award Studentship, University of Oxford) and Abigael Peerless for their able assistance, Dr. I. Colquhoun for assistance with some of the NMR spectroscopy and Drs. K. Shimomura (Tsukuba, Japan) and T. Hashimoto (Kyoto, Japan) for pure samples of 7-hydroxyhyoscyamine. J.G.W, gratefully acknowledges support from the Nuffield Foundation under the Small Grants Scheme to promote collaborative experimentation and M.A. is grateful to the Ministry of Education, Iran for a research scholarship.     

崖椒茎的化学成分

从崖椒（Zanthoxglum schinifolutm Sieb.et Zucc.)茎的石油醚、二氯甲烷提取物中分离得到8个化合物。经物理常数测定及光谱（UV,IR,MS,NMR)分析鉴定其为（1）白鲜碱（dictamning),（2）茵芋碱（skimmianine),(3)滨蒿内酯（scoparone),(4)崖椒内酯（schinifolin),(5)莨菪亭（scopoletin),（6）7-羟基-8-甲氧基香豆素（7-hydroxy-8-methoxycoumarin),（7）N-甲基弗林辛（N-methylflindersine),（8）β-谷甾醇（β-sitosterol),其中化合物（5）、（6）和（7）为首次从该植物中分离。     

Production of monoclonal antibodies and development of an ELISA for solamargine

The ratio of hapten and bovine serum albumin in an antigen conjugate was determined by matrix-assisted laser desorption/ionization mass spectrometry. A hybridoma secreting monoclonal antibodies against solamargine was produced by fusing splenocytes immunized with a solamargine-bovine serum albumin conjugate with HAT-sensitive mouse myeloma cell line, P3-X63-Ag8-653. Extensive cross-reaction of anti-solamargine antibodies against solasonine appeared. Aglycone of solamargine, solasodine cross-reacted with anti-solamargine antibodies resulting in a 43.8% cross-reaction. Insignificant cross-reaction appeared with tomatine (2.06%). The full measuring range of the assay extends from 57.5 pmol ml^–1 to 11.5 nmol ml^–1 of solamargine.     

Turnover and transport of quinolizidine alkaloids. Diurnal fluctuations of lupanine in the phloem sap,leaves and fruits of Lupinus albus L.

Quinolizidine alkaloids formed in the leaves of Lupinus albus L. are translocated via the phloem to the other plant organs, especially the maturing fruits. Compared with amino-acid transport in the phloem, the alkaloids contribute about 8% to the overall nitrogen being exported from the leaf. Since it is likely that the alkaloids are subsequently degraded in the target tissues a minor role of quinolizidine alkaloids might be nitrogen transport. A marked diurnal fluctuation of alkaloids was observed in the leaves, the phloem sap, the roots and the fruits with an increase during the day and an amplitude of several hundred percent thus providing evidence for a rapid turnover of endogenous alkaloids.Abbreviations QA quinolizidine alkaloids - GLC gas-liquid chromatography     

Alcaloides du Strychnos tchibangensis

The structure of a new bisindole alkaloid has been elucidated by chemical correlation and analysis of ¹³C NMR spectra.     

(+)-5,17-dehydromatrine N-oxide,a new alkaloid in Euchresta japonica

A new lupin alkaloid, (+)-5,17-dehydromatrine N-oxide, was isolated from the fresh aerial parts of Euchresta japonica. Its structure was confirmed by spectrometric data and by direct comparison with a synthetic sample, prepared from (+)-sophoranol ((+)-5-hydroxymatrine). It was also concluded that (+)-5,17-dehydromatrine N-oxide and (+)-matrine N-oxide possess the same configuration with respect to the asymmetric nitrogen by NMR spectra.     

Precursors of ricinine in the castor bean plant

Isotopic tracer experiments confirmed that glycerol and succinic acid are good precursors of the pyridine ring of ricinine in castor bean plants. Tritium from C-2 was lost from tritiated glycerol while tritium from C-1 was retained. Thus a derivative of dihydroxyacetone is likely to be intermediate. By simultaneous feeding of glycerol-1-(3)-[³H] and succinic acid-2(3)-[¹⁴C], it was hoped to find precursors of ricinine containing both labels, but none could be found. There was no evidence for the appearance of labeled quinolinic acid, which is presumed to be a precursor of ricinine.     

Melovinone,an open chain analogue of melochinone from Melochia tomentosa

Melovinone, a new alkaloid isolated from the roots of Melochia tomentosa has been characterized as 3,7,8-trimethoxy-2-methyl-5(5′-phenylpentyl)-4-quinolinone.     

Levels of tropinone-reductase activities influence the spectrum of tropane esters found in transformed root cultures of Datura stramonium L.

The nortropane sulphur analogues 8-thiabicyclo[3.2.1] octan-3-one, 8-thiabicyclo[3.2.1]octan-3a-ol and 8-thiabicyclo[3.2.1]octan-3-ol have been found to have differential effects in vitro on the activities of tropinone reductase I and tropinone reductase II from Datura stramonium L. It has been demonstrated that only tropinone reductase I is able to metabolise 8-thiabicyclo[3.2.1]octan-3-one and that only this enzyme is inhibited by 8-thiabicyclo[3.2.1]octan-3-ol and 8-thiabicyclo[3.2.1]octan-3-ol. A K _m of 0.035 mM was determined for 8-thiabicyclo[3.2.1]octan-3-one and I₅₀ values of 0.081 mM and 0.021 mM for 8-thiabicyclo[3.2.1]octan-3-ol and 8-thiabicyclo[3.2.1]octan-3-ol, respectively. The influence that these differential interactions might have on metabolism was investigated in transformed root cultures of D. stramonium. It was found that when these cultures were grown in the presence of either 8-thiabicyclo[3.2.1]octan-3-one or 8-thiabicyclo[3.2.1]octan-3-ol the spectrum of alkaloids that accumulated was altered from that found in control roots in the manner predicted from the observed effects of these inhibitors on the isolated reductases. The effect could be mimicked by feeding pseudotropine, the product of tropinone reductase II. It is concluded that the relative levels of activity of the two tropinone reductases might play an important role in regulating the balance of tropan-3-ols to tropan-3-ols seen in the spectrum of tropane-alkaloid-producing plants.Abbreviations GC/MS gas chromatography/mass spectrometry; - I₅₀ concentration of inhibitor required to reduce the rate of reaction to half the maximal value; - -TBOL 8-thiabicyclo[3.2.1]octan-3-ol; - -TBOL 8-thiabicyclo[3.2.1]octan-3-ol; - TBON 8-thiabicyclo[3.2.1]octan-3-one; - TR tropinone reductase We are most grateful to J. Eagles (I.F.R., Norwich) for GC/MS analysis, to colleagues at I.P.B.P. and I.F.R. for helpful discussions, to the technical staff (Chemistry, Glasgow) and to W. Millar (Chemistry, Glasgow) for assistance with the reduction of TBON. This work was, in part, supported by a grant to B Dräger from the Deutsche Forschungsgemeinschaft (Dr227/I-I). The research reported here was supported by an Academic Research Collaboration Cooperative Award (project No. 215) from the British Council and the Deutscher Akademischer Austauschdienst to R.J. Robins and B. Dräger.