Leaf variation in iridoid glycoside content ofPlantago lanceolata (Plantaginaceae) and oviposition of the buckeye,Junonia coenia (Nymphalidae)

Summary Chemical analysis of each individual leaf of fivePlantago lanceolata (Plantaginaceae) plants showed that iridoid glycoside content increased from undetectable in the oldest photosynthetic leaves to over 9% dry weight in the youngest leaves. The relative proportion of the two iridoid glycosides inP. lanceolata also changed with leaf age: older leaves had significantly more aucubin, whereas the youngest leaves had primarily or solely catalpol. Oviposition tests with femaleJunonia coenia (Nymphalidae) butterflies, showed that they laid most of their eggs on new leaves.     

粗壮女贞中的新单萜配糖体(英)

从粗壮女贞（Ｌｉｇｕｓｔｒｕｍ ｒｏｂｕｓｔｕｍ）叶子中分离鉴定了４ 个新的单萜配糖体, 命名为粗壮女贞甙Ｄ、Ｇ、Ｈ和Ｌ． 通过波谱及化学方法,它们分别鉴定为香叶醇３′ＯαＬ吡喃鼠李糖基４′顺式香豆酰基βＤ吡喃葡萄糖甙（１） ,６羟基３,７二甲基２Ｅ,７葵二烯醇３′ＯαＬ吡喃鼠李糖基４′反式香豆酰基βＤ吡喃葡萄糖甙（２） ,６羟基３,７二甲基２Ｅ,７葵二烯醇３′ＯαＬ吡喃鼠李糖基４′顺式香豆酰基βＤ吡喃葡萄糖甙（３） 以及６,７二羟基３,７二甲基２Ｅ葵烯醇３′ＯαＬ吡喃鼠李糖基４′顺式香豆酰基βＤ吡喃葡萄糖甙（４） ． 此外,还从该植物得到了７ 个已知化合物,芹菜素（５）、ｃｏｓｍｏｓｉｉｎ（６） 、ｒｈｏｉｆｏｌｉｎ（７） 、山梨糖醇（８）、阿克苷（９） 、β谷甾醇（１０） 和齐墩果酸（１１） ．     

Survival of first instar larvae ofDanaus plexippus (Lepidoptera: Danainae) in relation to cardiac glycoside and latex content ofAsclepias humistrata (Asclepiadaceae)

Summary Our paper addresses field survivorship of first instar monarch butterfly larvae (Danaus plexippus L., Lep.: Danainae) in relation to the dual cardenolide and latex chemical defenses of the sand hill milkweed plant,Asclepias humistrata (Asclepiadaceae) growing naturally in north central Florida. Survival of first instar larvae in the field was 11.5% in the first experiment (15–20 April 1990), and dropped to 3.4% in the second experiment (20–30 April). About 30% of the larvae were found glued to the leaf surface by the milkweed latex. Predator exclusion of non-flying inverte-brates by applying tanglefoot to the plant stems suggested that the balance of the mortality was due to volant inverte-brates, or to falling and/or moving off the plants. Regression analyses to isolate some of the other variables affecting survivorship indicated that first instar mortality was correlated with (1) increasing cardiac glycoside concentration of the leaves, (2) increasing age of the plants, and (3) the temporal increase in concentration of cardiac glycosides in the leaves. The study also provided confirmatory data of previous studies that wild monarch females tend to oviposit onA. humistrata plants containing intermediate concentrations of cardiac glycosides. Cardiac glycoside concentration in the leaves was not correlated with that in the latex. The concentration of cardenolide in the latex is extremely high, constituting an average of 1.2 and 9.5% of the mass of the wet and dry latex, respectively. The data suggest that an increase in water content of the latex is compensated for by an influx of cardenolide with the result that the cardenolide concentration remains constant in the latex systems of plants that are growing naturally. We also observed first instar larvae taking their first bite of milkweed leaves in the field. In addition to confirming other workers findings that monarch larvae possess elaborate sabotaging behaviour of the milkweed's latex system, we discovered that several larvae on their first bite involuntarily imbided a small globule of latex and instantly became cataleptic. This catalepsis, lasting up to 10 min, may have been in response to the high concentration of cardenolide present in the latex ofA. humistrata, more than 10 times that in the leaves. The results of the present study suggest that more attention should be directed to plant chemical defenses upon initial attack by first instar insect larvae, rather than attempting correlations of plant chemistry with older larvae that have already passed the early instar gauntlet. The first bite of neonate insects may be the most critical moment for coping with the chemical defenses of many plants and may play a much more important role in the evolution of insect herbivory than has previously been recognized.     

The iridoid glycoside,catalpol, as a deterrent to the predatorCamponotus floridanus (Formicidae)

Summary We investigated the role of the iridoid glycoside, catalpol, as a deterrent to the predator,Camponotus floridanus. Four laboratory colonies of this ant were offered buckeye caterpillars (Junonia coenia: Nymphalidae) raised on diets with and without catalpol. The same colonies were offered sugar-water solutions containing varying concentrations of catalpol, in both no-choice and choice tests. Regardless of diet, buckeye caterpillars appeared to be morphologically protected from predation by the ants, possibly because of their large spines or tough cuticle. However, buckeyes raised on diets with catalpol had high concentrations of catalpol in their hemolymph; extracts of this high-catalpol hemolymph proved to be an effective deterrent to the ants. When starved ants were not given the choice of food items, they were more likely to consume sucrose solutions that contained 5 mg catalpol/ml or 10 mg catalpol/ml than they were to consume solutions with 20 mg catalpol/ml. When they were given a choice of sugar solution or a sugar solution containing catalpol, the ants avoided solutions with catalpol at any of these concentrations. Ant colony responses to catalpol in sucrose solutions varied considerably over time and among colonies.     

Two herbivore-deterrent iridoid glycosides reduce the in-vitro growth of a specialist but not of a generalist pathogenic fungus of Plantago lanceolata L

Summary. Many secondary plant compounds are involved in defense against both insect herbivores and pathogens. Two secondary plant compounds of Plantago lanceolata, the iridoid glycosides catalpol and its precursor aucubin, are well known for their deterrent effects on generalist and non-adapted specialist insect herbivores. We tested the effects of these compounds on the in-vitro growth of a specialist and generalist fungal pathogen of this host species. Two chemical forms of these iridoids were tested. The glycosides and their aglycones, the products of enzymatic conversion by specific $/Beta$-glucosidase enzymes. The glycosides enhanced growth of both the specialist fungus Diaporthe adunca and the generalist fungus Fusarium moniliforme var. subglutinans. The positive effect of these glycosides on the generalist fungus is in sharp contrast with the generally negative effects of these glysosides on generalist insect herbivores. The aglycones of aucubin and catalpol reduced the growth of the specialist fungus D. adunca, but, contrary to expectation, enhanced the growth of the generalist fungus F. moniliforme var. subglutinans. Effects of aucubin on D. adunca were stronger than effects of catalpol. This was true both for the growth stimulating effects of the glycosides and for the fungitoxic effects of the aglycones. We therefore expect that the effects of these iridoids in P. lanceolata on the specialist fungus will strongly depend on the ratio between catalpol and its precursor aucubin and the chemical form (glycoside or aglycone) in which these compounds are encountered by the fungus during growth. Our results suggest that iridoid glycosides in P. lanceolata can be used as defense against both herbivores and pathogens, but that their effects are highly specific with respect to the natural enemy species that is encountered. Received 11 April 2002; accepted 9 August 2002     

Responses of three mouse species to deterrent chemicals in the monarch butterfly. II. Taste tests using intact monarchs

Summary Peromyscus melanotis is the only one of three mouse species that eats monarch butterflies at their overwintering sites in Mexico. I tested two hypotheses: 1)P. aztecus avoids monarchs because of a bitter taste aversion to cardiac glycosides (CGs) and an inability to reject CG-rich body parts; 2)Reithrodontomys sumichrasti avoids monarchs principally because of a bitter taste aversion to the CGs. None of the species are sensitive to the toxic effects of ingested CGs. Feeding responses of laboratory-reared mice of each species to monarchs with low, medium and high CG concentrations were compared. BothP. aztecus andR. sumichrasti ate significantly fewer of all three types of monarchs thanP. melanotis. ForP. aztecus andR. sumichrasti, the number of monarchs eaten decreased with increasing CG concentration, whereas forP. melanotis, the number remained constant.Peromyscus melanotis andR. sumichrasti developed a feeding technique for rejecting the CG-laden cuticular material, which reduced the bitterness of ingested monarch material. However,R. sumichrasti displayed the technique significantly less often thanP. melanotis; andP. aztecus never developed it. I conclude that high taste sensitivity to CGs and less versatile food handling preventP. aztecus andR. sumichrasti from overcoming the monarch's chemical defenses.     

京尼平对丝素蛋白交联的染色作用

京尼平是一种环烯醚萜类化合物,能够与氨基酸反应生成蓝色素.制备的无色丝素蛋白溶液与京尼平溶液混合后,溶液显蓝紫色.利用紫外可见光光谱和荧光光谱,分析了整个反应过程,考察了温度、酸碱度等影响因素,并对颜色产物对光照的稳定性进行了分析,初步探讨了京尼平对丝素蛋白交联染色作用的机制.结果显示:生成的颜色产物在590 nm处具有特征光吸收,高温和碱性条件有利于反应的进行;交联产物的颜色与反应时pH相关,酸性条件下偏蓝色,碱性条件下偏紫色;生成的颜色产物具有较高的稳定性,于自然光强下放置一个月或5 000 lx光强照射6 h.590 nm吸收值减小不显著.丝素蛋白的内源荧光显著降低,表明反应导致丝素蛋白构象的剧烈变化.京尼平可能与丝素蛋白中某些氨基酸侧链基团反应形成稳定的颜色产物.研究结果对环烯醚萜类化合物用于蚕丝染色具有一定的指导意义.图5表1参18     

粗壮女贞中的新单萜配糖体

从粗壮女贞（Ｌｉｇｕｓｔｒｕｍ ｒｏｂｕｓｔｕｍ）叶子中分离鉴定了４个新的单萜配糖体,命名为粗壮女贞甙Ｄ、Ｇ、Ｈ和Ｌ．通过滤谱及化学方法,它们分别鉴定为香叶醇３‘－Ｏ－μ－Ｌ－南鼠李糖基－４’－顺式香豆酰基－β－Ｄ－吡喃葡萄糖甙（１）,６－羟基－３,７－二甲基－２Ｅ,７葵二烯醇３‘－Ｏ－α－Ｌ－吡喃鼠李糖基－４’反式香豆酰基－β－Ｄ－吡喃葡萄糖甙（２）,６－羟基－３,７－二甲基－２Ｅ,７－葵二烯醇     

Metabolism of the14C-labeled herbicide clodinafop-propargyl in plant cell cultures of wheat and tobacco

The metabolism of ¹⁴C-clodinafop-propargyl (CfP) was examined in cell cultures of wheat (Triticum aestivum L. cv. ‘Heines Koga II’) and tobacco (Nicotiana tabacum L.). Besides the non-transgenic tobacco culture, cultures transformed separately with cDNA of human cytochrome P450-monooxygenases (P450s) CYP1A1, CYP1A2, CYP3A4, CYP2B6 and CYP2C19 were examined. Experiments with wheat were executed in the presence and absence of safener cloquintocet-mexyl (CqM). After 48 h of incubation, only about 10% of applied ¹⁴C was found in media (both tobacco and wheat). Non-extractable residues of ¹⁴C-CfP in wheat cells were 16.54% (without CqM) and 30.87% (with CqM). In all tobacco cultures, 82.41–92.46% of applied radioactivity was recovered in cell extracts. In contrast to wheat, non-extractable residues amounted only to 1.50–2.82%. As determined by radio-thin layer chromatography (TLC) and -high-performance liquid chromatography (HPLC), the parent CfP was not found in the cell extracts of wheat; in tobacco cell extracts, only traces of CfP were detected. After a hydrolysis of assumed carbohydrate conjugates of CfP derived polar ¹⁴C-labeled compounds, TLC and HPLC analysis showed that in wheat, a more complex pattern of metabolites of CfP were observed as compared to all tobacco cultures. In hydrolysates resulting from wheat, the identity of three primary products was confirmed by means of GC-EI-MS: free acid clodinafop (Cf), hydroxy-Cf hydroxylated at the pyridinyl moiety, and 4-(5-chloro-3-fluoropyridin-2-yloxy)phenol. In hydrolysates derived from all tobacco cultures, main metabolite was Cf besides only traces of further unidentified products. Differences among the different tobacco cultures (non-transgenic, transgenic) did not emerge. According to kinetics of disappearance of primary metabolite Cf as well as formation of polar soluble products and non-extractable residues, metabolization of CfP proceeded at a noticeably higher rate in wheat cells treated with safener CqM than in cells without CqM treatment. Thus, these results indicated a stimulation of CfP's metabolism by CqM, although metabolic profiles observed in CqM treated and non-treated cells (after hydrolysis) were qualitatively similar. The findings obtained from all tobacco cultures suggested that with the exception of ester cleavage to Cf, CfP cannot be metabolized by tobacco itself or by the human P450s examined.     

Molecular studies on the ouabain binding site of the Na+, K+-ATPase in milkweed butterflies

Summary. The Na⁺, K⁺-ATPase of the Monarch butterfly (Danaus plexippus) is insensitive to the inhibition by cardiac glycosides due to an amino acid replacement: histidine instead of asparagine at position 122 of the α-subunit representing the ouabain binding site. By PCR amplification of the DNA sequence of this site, a PCR product of 270 bp was obtained from DNA extracted from Danainae species (Danaus plexippus, D. chrysippus, D. gillipus, D. philene, D. genutia, Tirumala hamata, Euploea spp., Parantica weiskei, P. melusine), Sphingidae (Daphnis nerii) and mimics of milkweed butterflies (Hypolimnas missipus, Limenitis archippus and L. arthemis, Nymphalidae). Analysis of the nucleotide sequences revealed that the single point mutation in the ouabain binding domain (AAC-Asn for CAC-His) was present only in Danaus plexippus, but not in the other species investigated. Since these milkweed butterflies also store cardenolides, other structural modifications of the Na⁺, K⁺-ATPase may have occurred or other strategies of cardenolide tolerance have been developed. Received 15 May 2000; accepted 29 June 2000