青橄榄浸膏的提取及其抗氧化活性研究

为优化青橄榄浸膏提取工艺,并探讨其抗氧化性。以茂名盛产的青橄榄为原料,采用超声波辅助乙醇提取法,以总黄酮和总多酚得率为评价指标,考察各因素对青橄榄浸膏提取效果的影响。采用邻苯三酚自氧化法、结晶紫法和DPPH清除能力评价青橄榄浸膏的抗氧化活性。结果显示,浸膏的最佳提取工艺为:乙醇体积分数70%,料液比1∶18 (g∶mL),超声提取温度50℃,时间6 min(超声提取阶段);单纯有机溶剂提取温度60℃,时间45 min(有机溶剂浸提阶段);此条件下总黄酮得率为1. 76%,总多酚得率为15. 53%。终产物浸膏在0. 3 mg/mL浓度下对超氧阴离子自由基抑制率为22. 74%,相当于同等质量浓度的抗坏血酸抑制效果的23. 47%;在0. 02 mg/mL浓度下对羟基自由基的清除率为67. 32%,相当于同等质量浓度的抗坏血酸清除效果的112. 58%;在0. 2 mmol/mL的DPPH溶液体系中,0. 15 mg/mL的浸膏对DPPH的清除率为95. 40%,相当于同等质量浓度的抗坏血酸清除效果的140. 83%;总体来讲,浸膏具有良好的抗氧化能力,虽然对超氧阴离子自由基抑制率弱于抗坏血酸,但羟基自由基的清除率及DPPH清除率均优于抗坏血酸。     

Improved podophyllotoxin production by transformed cultures of Linum album

Various cell and hairy root cultures of L. album were developed and analyzed for podophyllotoxin content. Transformed callus and hairy root cultures developed from infection of stem portions of in vitro-germinated L. album plant with Agrobacterium rhizogenes NCIM 5140 strain were selected on the basis of high podophyllotoxin content and growth. Based on the integration of Ri T(L)-DNA and T(R)-DNA, integration of only the ags and not the rol gene in transformed cell culture indicated fragmented integration pattern. The effect of different cultivation media and carbon source on growth and podophyllotoxin production were studied in shake-flask suspension cultures. Detailed batch growth and production kinetics with sugar consumption profile were also established. Maximum volumetric productivity of 4.40 and 2.75 mg/L per day was obtained in cell suspension and hairy root cultures, respectively.     

橄榄(Canarium album)种子形态结构与数量性状分析

以福建主栽的橄榄（Canarium album（Lour.）Rauesch.）品种‘长营’种子为材料,观察其成熟种子及不同发育时期种子的形态结构,对数量性状进行测量与统计分析,并运用概率分级建立形态指标的正态分布分级标准。结果显示,橄榄种子表型性状和形态结构变异较大,各数量性状变异系数为6.77%~19.04%,具有丰富的遗传多样性;橄榄种子心室类型有12种,其中三心室占比最高（97.80%）;胚数量0~4个不等;经K-S检验,各数量性状符合正态分布,依据各性状可划分为5级分级标准。本研究为橄榄种质的系统学研究提供了种子形态方面的新内容,可为橄榄资源描述的标准化及遗传育种研究提供一定的参考。     

Assays for three enzymes involved in mevalonic acid metabolism

Enzyme assays have been developed for mevalonate (MVA) kinase, mevalonate phosphate (MVAP) kinase and mevalonate pyrophosphate (MVAPP) anhydrodecar-boxylase. The procedures involve radioactively labelled substrates and separation of the reaction products by anion exchange chromatography. The separation on Dowex 1-X2 in self-packed microcolumns is simple, inexpensive and results in good separation of the MVA derivatives from each other. Because separation of MVAPP from isopenteny] pyrophosphate (IPP) was not possible directly, samples or column fractions containing MVAPP and IPP simultaneously were dephosphorylated by alkaline phosphatase. The resulting MVA and isopentenol are then easily separated in the same system. The assays for all three enzymes not onlv allows the determination of activities in crude enzyme preparations but is also applicable to the in vitro determination of intermediate pools in the reaction sequence from MVA to IPP after using ¹⁴C-MVA as substrate. The major advantage is accuracy and ease of use. The utility of the methods was demonstrated for enzyme extracts from the higher plants Chenopodium and spinach as well as for the fungus Phycomyces .     

A Structure‐Activity Relationship between the Veratrum Alkaloids on the Antihypertension and DNA Damage Activity in Mice

Veratrum plant contains a family of compounds called steroidal alkaloids which have been previously reported to cause DNA damage and blood pressure decrease in vivo. In this study, the antihypertensive effects and DNA damage in brain cells of 12 steroidal alkaloids separated from Veratrum plant were all evaluated to develop a relationship among chemical structure, antihypertensive activity and neurotoxicity by utilization of chemical principal component analysis (PCA) and hierarchical cluster analysis (HCA). Twelve steroidal alkaloids markedly reduced high blood pressure of hypertensive mice and also similarly induced varying degrees of DNA single‐strand breaks in mouse cerebellum and cerebral cortex after oral administration. On the basis of the PCA and HCA results, it was suggested that the 3‐carboxylic esters and benzene group play a core role in the DNA damage of brain cells, while more hydroxy groups in the A‐ring and B‐ring structure of jervine‐type alkaloid led to stronger antihypertensive activity. The primary structure, activity and neurotoxicity relationship were discussed briefly.     

Elucidating steroid alkaloid biosynthesis in Veratrum californicum: production of verazine in Sf9 cells

Steroid alkaloids have been shown to elicit a wide range of pharmacological effects that include anticancer and antifungal activities. Understanding the biosynthesis of these molecules is essential to bioengineering for sustainable production. Herein, we investigate the biosynthetic pathway to cyclopamine, a steroid alkaloid that shows promising antineoplastic activities. Supply of cyclopamine is limited, as the current source is solely derived from wild collection of the plant Veratrum californicum. To elucidate the early stages of the pathway to cyclopamine, we interrogated a V. californicum RNA‐seq dataset using the cyclopamine accumulation profile as a predefined model for gene expression with the pattern‐matching algorithm Haystack. Refactoring candidate genes in Sf9 insect cells led to discovery of four enzymes that catalyze the first six steps in steroid alkaloid biosynthesis to produce verazine, a predicted precursor to cyclopamine. Three of the enzymes are cytochromes P450 while the fourth is a γ‐aminobutyrate transaminase; together they produce verazine from cholesterol.     

Enhancement of tyrosinase inhibition of the extract of Veratrum patulum using cellulase

Inhibitors of melanin biosynthesis were screened by using three different methods. The extract of Veratrum patulum contains hydroxystilbene compounds that are potent tyrosinase inhibitors. We evaluated the enzyme inhibitory property on the mushroom tyrosinase of hydroxystilbene compounds including resveratrol, oxyresveratrol, and their analogs. Biotransformation using cellulase of the whole extract brought about an increase in the inhibitory activity of the products on mushroom tyrosinase. The enhancement of tyrosinase inhibition is supposed to increase the concentration of aglycon, which has superior inhibitory activity to its glycoside. Eventually, melanin biosynthesis was inhibited by the enhanced tyrosinase inhibitory activity of the extract. This result indicated that deglycosylation of stilbene compounds has exerted more effective inhibition on the enzyme than that of the unprocessed plant extract.     

Ultrastructural alterations to chloroplasts in triazine-resistant weed biotypes

Ultrastructural, morphometric and physiological techniques were used to determine the consistent chloroplast differences between triazine-resistant (R) and triazine-susceptible (S) biotypes of Amaranthus hybridus L., Chenopodium album L., and Brassica campestris L. All R biotypes had a larger proportion of the chloroplast volume as grana lamellae and a lower proportion of starch and stroma lamellae than S biotypes. In the R biotypes, a greater percentage of grana contain larger numbers of thylakoids per granum. A greater proportion of chlorophyll associated with the light-harvesting chlorophyll alb protein and a lower chlorophyll alb ratio, traits associated with an increase in grana lamellae, were noted in R biotypes. Chloroplasts of S biotypes could be modified to ultrastructural phenocopies of those in R biotypes by treatment with sublethal levels of the PSII inhibiting herbicides, bentazon, diuron, atrazine and prometon. Despite the structural similarities to R biotypes, the modified S biotypes were not resistant to atrazine as determined by fluorescence measurements. Thus, the structural alterations observed are apparently secondary effects of impaired photosynthetic electron transport in R biotypes, and are not the cause of triazine resistance.     

Photosynthetic activity and chloroplast structural characteristics in triazine-resistant biotypes of three weed species

Triazine-resistant (R) and susceptible (S) biotypes of Chenopodium album, Conyza bonaeriensis and Setaria glauca were compared in terms of photosynthetic activity, chloroplast ultrastructure, and polar-lipid composition of thylakoid membranes. Concerning photosynthetic activity, R relative to S biotypes showed lower rates in photosystem II electron transport (R/S ratios in the 0.3-0.5 range), which were correlated with increases in the ratio of I to P levels of the fluorescence emission curve, but similar rates in both whole-chain electron transport by thylakoids and photosynthetic oxygen evolution at different light intensities and at temperatures of 20 and 30°C by leaf discs. Concerning chloroplast structural characteristics, R and S biotypes did not show differences in degree of thylakoid appression and chlorophyll a/b ratio. However, thylakoid polar-lipids of R biotypes exhibited a higher degree of unsaturation, specially, in its monogalactosyl diglyceride fraction; they were richer in trans-hexadecenoic acid in its phospholipid fraction and they had higher contents in monoglactosyl diglyceride. The above results are discussed in relation to possible differences in photosynthetic performance between R and S plants.     

No effect of larval experience on adult host preferences in Polygonia c‐album (Lepidoptera: Nymphalidae): on the persistence of Hopkins’ host selection principle

Abstract 1. The possible effect of juvenile imprinting or ‘chemical legacy’ on the subsequent oviposition – often called the ‘Hopkins’ host selection principle’– has been a controversial but recurrent theme in the literature on host‐plant preference. While it appears possible in principle, experimental support for the hypothesis is equivocal. The present study points out that it is also important to consider its theoretical implications, and asks under what circumstances, if any, it should be favoured by natural selection. 2. Following this reasoning, it is predicted that host preference in the polyphagous butterfly Polygonia c‐album L. (Lepidoptera, Nymphalidae) should not be influenced by larval environment. This was tested by rearing larvae on three natural host plants: the high‐ranked Urtica dioica and the medium‐ranked Salix cinerea and Ribes uva‐crispa, and exposing the naive females to oviposition choices involving the same set of plants. 3. It was found that larval host plant had no effect on oviposition decisions of the adult female. Hence, the Hopkins’ host selection principle does not seem to be applicable in this species. 4. Based on recent insights on how accuracy of environmental versus genetic information should affect the control of developmental switches, the conditions that could favour the use of juvenile cues in oviposition decisions are discussed. Although the Hopkins’ host selection hypothesis cannot be completely ruled out, we argue that the circumstances required for it to be adaptive are so specific that it should not be invoked as a general hypothesis for host selection in plant‐feeding insects.