Effect of iridoid glycoside content on oviposition host plant choice and parasitism in a specialist herbivore

The Glanville fritillary butterfly Melitaea cinxia feeds upon two host plant species in Å land, Finland, Plantago lanceolataand Veronica spicata, both of which produce iridoid glycosides. Iridoids are known to deter feeding or decrease the growth rate of many generalist insect herbivores, but they often act as oviposition cues to specialist butterflies and are feeding stimulants to their larvae. In this study, two iridoid glycosides (aucubin and catalpol) were analyzed by micellar electrokinetic capillary chromatography. We measured the spatial and temporal variation of iridoid glycosides in natural populations of the host plants of M. cinxia. We also analyzed the aucubin and catalpol content in plants in relation to their use by ovipositing females, and in relation to the incidence of parasitism of M. cinxia larvae in natural populations. The mean concentrations of aucubin and catalpol were higher in P. lanceolata than in V. spicata, and catalpol concentrations were higher than aucubin concentrations in both host species. Plantago lanceolata individuals that were used for oviposition by M. cinxia had higher aucubin concentrations than random plants and neighboring plants. Additionally, oviposition and random plants had higher catalpol concentrations than neighboring plants, indicating that ovipositing females select for high iridoid glycoside plants or that oviposition induces iridoid glycoside production in P. lanceolata. Parasitism by the specialist parasitoid wasp Cotesia melitaearum occurred most frequently in larval groups that were feeding on plants with low concentrations of catalpol, irrespective of year, population, and host plant species. Therefore, parasitoids appear to avoid or perform poorly in host larvae with high catalpol content.     

Iridoid glycoside metabolism and sequestration byPoladryas minuta (Lepidoptera: Nymphalidae) feeding onPenstemon virgatus (Scrophulariaceae)

A bivoltine checkerspot butterfly,Poladryas minuta, is aPenstemon specialist, not known to utilize any other plant genus for oviposition and larval feeding. At several intermontane plains sites of central Colorado, the butterfly utilizesPenstemon virgatus as its sole host plant. Analysis of the host plant showed it to contain three cinnamyl-type catalpol esters (scutellarioside-II, globularin, globularicisin) and catalpol. The host plant contained an average of 10% dry weight iridoids, but some variation among individual plants and leaves within plants was noted. Field-collected butterflies contained 2.1–8.7% dry weight catalpol, but no other iridoids. Adults from larvae fedP. virgatus in the lab contained 4.2–9.0% dry weight catalpol and excreted large amounts of catalpol in the meconium. No catalpol was found in the larval frass. Larvae did not consume three alternate iridoid-containing host-plant species, and most eventually died rather than feed on the alternate plants. Larvae did consume small amounts of artificial diets containing the alternate species andP. virgatus, but most went into diapause and some died. Survival was good on artificial diet containing 10% dry weight of the iridoid esters fromP. virgatus. Only catalpol was found in pupae and adults, but it was absent from the larval frass. The cinnamic-type acids expected from larval hydrolysis of the esters were not found in larval frass, pupae, or adults. These results are contrasted with those found for another checkerspot,Euphydryas anicia, which consumes a different host-plant species but was present at one of the same sites withPoladryas minuta.Paper 15 in the series Chemistry of the Scrophulariaceae. Paper 14 Boros, C.M., Stermitz, F.R., and Harris, G.H. 1990.J. Nat. Prod. 5372–80.     

Patterns of Iridoid Glycoside Production and Induction in Plantago lanceolata and the Importance of Plant Age

Induction of allelochemicals is one way that plants efficiently deploy defenses against herbivory. In two separate experiments we investigated the time course of this inductive response and the importance of the timing of herbivory for Plantago lanceolata (Plantaginaceae). We found a localized induced response of catalpol and the ratio of catalpol to total iridoid glycosides in damaged leaves that was evident at d 6 after caterpillars of the specialist Junonia coenia were put onto the plants. On the whole plant level, we detected small, but significant changes in the iridoid glycoside metabolism of P. lanceolata on several different days following herbivory. We also found considerable change in the amounts of allelochemicals produced during P. lanceolata's ontogeny. This ontogenetic effect might help to explain some of the reasons why induction may be difficult to detect in P. lanceolata. We also investigated the importance of the timing of herbivory on P. lanceolata's inductive response, but neither herbivory after 5 wk of growth nor after 6 or 7 wk of growth induced an increase in aucubin or catalpol.     

Euphydryas anicia (Lepidoptera: Nymphalidae) utilization of iridoid glycosides fromCastilleja andBesseya (Scrophulariaceae) host plants

Iridoid glycosides were found to be sequestered by natural populations ofEuphydryas anicia after ingestion from the host plantsBesseya alpina, B. plantaginea, andCastilleja integra. Both major iridoids ofB. alpina, cataipol and aucubin, were found in butterfly populations where this was the only host plant. The catalpol-aucubin ratio was higher in the butterflies than in the host plant. AnE. anicia population which uses bothB. plantaginea andC. integra as host plants was found to sequester cataipol as well as another iridoid, macfadienoside. Macfadienoside was the major iridoid ofC. integra, while catalpol esters were the major iridoids ofB. plantaginea. Although it was a major sequestered iridoid, catalpol was a minor constituent in both host plants. The macfadienoside-catalpol ratio in the butterflies from this population was highly variable, and there appeared to be both sex and individual variation in host plant and/or iridoid glucoside utilization byE. anicia. Although other iridoids were present in the host plants, none was sequestered in more than trace amounts.This work was supported by grant CHE-8213714 to FRS, in part by grant DEB-06961 to PRE from the National Science Foundation, and a grant from the Koret Foundation of San Francisco to PRE. Paper 5 in the series Chemistry of the Scrophulariaceae. Paper 4: Roby, M.R. and Stermitz, F.R. 1984.J. Nat. Prod. 47:854–857.     

Hostplant Suitability and Defensive Chemistry of the Catalpa Sphinx, Ceratomia catalpae

The growth and survival of the Catalpa sphinx, Ceratomia catalpae (Sphingidae), were measured on five different species of Catalpa: C. bignonioides, C. bungeii, C. fargeseii, C. ovata, and C. speciosa. Larval growth varied significantly among these host plant species; however, survival did not differ. Quantification of the iridoid glycoside content of larvae, pupae, adults, larval frass, and leaves of the larval host plant, C. bignonioides, by gas chromatography showed that leaves contained both catalpol and catalposide; larvae, pupae, and frass contained only catalpol; and the adults contained no detectable iridoid glycosides. Amounts were highest in the larvae and declined in the pupal stage. Very small amounts of catalpol were detected in adults of the parasitoid, Cotesia congregata, and in the silken cocoons. The hemolymph in which the parasitoid larvae grew contained over 50% dry weight catalpol. Larvae of C. catalpae often regurgitate when disturbed. This may serve as a defense against predators. A comparison of the growth of larvae pinched with forceps to induce regurgitation with those that were not so treated showed that larvae that were pinched, and usually regurgitated, grew significantly more slowly than those that were not.     

Iridoid glycosides as oviposition stimulants for the buckeye butterfly,Junonia coenia (Nymphalidae)

Females ofJunonia coenia (Nymphalidae), a specialist on plants that contain iridoid glycosides, were found to use aucubin and catalpol, iridoid glycosides typical of a host plant,Plantago lanceolata (Plantaginaceae), as oviposition cues. Incorporating dried ground leaf material or pure iridoid glycosides into agar disks proved to be a very effective method of testing. In no-choice tests and choice tests, females laid more eggs on disks withP. lanceolata leaf material or iridoid glycosides, compared to agar controls. There was variation among individual females in preference for disks withP. lanceolata leaf material versus disks with iridoid glycosides. Females given a choice of three different concentrations of iridoid glycoside (0.2, 0.5, 1.0%) in the agar disks and a control laid more eggs on the disk with the highest concentration of iridoid glycoside.     

响应面分析法优化怀地黄中梓醇提取工艺

目的：优化回流辅助提取怀地黄中的梓醇工艺。方法：在单因素试验基础上,采用响应面法,以梓醇的提取得率为响应值,通过回归分析各工艺参数与响应值之间的关系,并由此预测最佳的工艺条件。结果：回流提取的最佳条件为甲醇体积分数72.94%、提取温度54.8℃、料液比(g/mL)1:18.7、提取时间3.0h。该条件下提取2次,梓醇的提取得率达到3.513%。结论：响应面法优化回流辅助提取怀地黄中梓醇的预测准确、方便,所得的最佳提取工艺条件高效、可行。     

Iridoid glycosides in the nectar ofCatalpa speciosa are unpalatable to nectar thieves

The floral nectar ofCatalpa speciosa has a chemical mechanism that limits thievery. A bioassay employing sphingid larvae,Ceratomia catalpae, shows that catalpa iridoid glycosides are present in the floral nectar. When potential nectar thieves are fed nectar, a sucrose solution of identical concentration, or a sucrose solution plus 0.4% catalpol and 0.4% catalposide (catalpa iridoids), the thieves drink significantly more of the pure sucrose solution than either of the other two sugar sources. Those thieves that drink either the nectar or the sucrose solution plus catalpa iridoids develop behavioral abnormalities including regurgitation and loss of locomotion. The response of the potential nectar thieves to nectar or the sucrose solution plus catalpa iridoids cannot be distinguished by the amount consumed or by their behavior. The legitimate diurnal pollinators ofC. speciosa are not affected by the iridoid glycosides.     

地黄中梓醇的超声提取-树脂分离纯化工艺

研究了地黄中梓醇的超声提取-树脂分离纯化工艺。结果表明,超声提取的最佳工艺条件为：提取溶剂采用68%（体积）的乙醇,液料体积(ml)质量(g)比5:1,超声时间36 min。考察了10种大孔树脂对地黄粗提液的分离纯化效果,其中以H103对梓醇的纯化效果最佳。通过实验进一步考察了H103的动态吸附性能,得到分离纯化的最佳工艺条件为：上柱液浓度为6.15 mg·ml^-1,上柱液流速为1.0 ml·min^-1,洗脱剂为75%乙醇,洗脱流速为0.5 ml·min^-1,洗脱体积为3.33 BV（bed volume）。提取、纯化两步操作后梓醇纯度达到62.39％。     

HPLC测定地黄叶与生地、熟地中梓醇含量

中药地黄中主要的化学成分是环烯醚萜类,其中梓醇含量较高,是地黄中主要活性成分,具有抗癌、利尿、缓泻、降血糖等作用。传统用药部位是其新鲜或干燥块根。本实验选取制药时废弃不用的地黄叶子为原材料,经实验确定以95%乙醇为提取溶剂,超声20 min提取梓醇。并同时从新鲜地黄叶、干地黄叶、生地、熟地中提取梓醇,采用HPLC分别测得提取物中梓醇的含量。结果表明,鲜地黄叶,干地黄叶,生地和熟地中的梓醇含量分别为1.07,9.93,14.24,12.54 mg/g。从地黄叶片中提取梓醇具有一定的可行性。