Evaluation of the yeast-extract signaling pathway leading to silymarin biosynthesis in milk thistle hairy root culture

The biosynthesis of silymarin, a potent antihepatotoxic compound, from the dried fruits of Silybum marianum L. Gaertn in hairy root cultures can be stimulated by a yeast extract elicitor. These results correlated with culture time, and the biosynthesis reached a maximum of 0.47 mg g^?1 DW by 72 h after culture (2-fold higher than the control). Lipoxygenase activity and linoleic acid content were stimulated by this treatment, suggesting that the jasmonate pathway may mediate the elicitor-induced accumulation of silymarin. The H₂O₂ content increased 24 h after elicitation and did not have marked changes between 48 and 72 h. In addition, the tocopherol content (especially α- and δ-tocopherols) increased 72 h after elicitation in comparison with non-treated cultures. Ascorbate had trace changes during feeding time and was lower than the control. The antioxidant activity was assayed by the 1-1-diphenyl-2-picrylhydrazyl stable free radical method and results were calculated base on an IC₅₀ that increased upon treatment, especially 24 h after treatment, with changes related to H₂O₂ content. These observations suggested that reactive oxygen species may mediate elicitor signals to the jasmonate pathway that lead to the production of silymarin.     

Study of different Trichoderma strains on growth characteristics and silymarin accumulation of milk thistle plant

Abstract

Biological control involves the use of beneficial organisms such as Trichoderma that promote positive responses by the plant. Trichoderma species produce and/or release a variety of compounds, including cell wall degrading enzymes and secondary metabolites, which enhance root development, crop productivity and resistance to biotic and abiotic stresses. This study examines the effects of the five Trichoderma strains (M7, KHB, G124-1, G46-3 and G46-7) on milk thistle including morphological characteristics and silymarin accumulation. This research indicated that treating milk thistle by M7 strain, significantly increased the highest branch length of plants, while strain KHB induced the production of silychristin, isosilybin and silymarin. Finally, strain G46-7 dramatically increased silydianin content in milk thistle plant. The findings suggested that certain Trichoderma strains are positive elicitors for promoting growth characteristics and silymarin accumulation in Silybum marianum (L.) Gaertn.     

Interactive effects of pests increase seed yield

Loss in seed yield and therefore decrease in plant fitness due to simultaneous attacks by multiple herbivores is not necessarily additive, as demonstrated in evolutionary studies on wild plants. However, it is not clear how this transfers to crop plants that grow in very different conditions compared to wild plants. Nevertheless, loss in crop seed yield caused by any single pest is most often studied in isolation although crop plants are attacked by many pests that can cause substantial yield losses. This is especially important for crops able to compensate and even overcompensate for the damage. We investigated the interactive impacts on crop yield of four insect pests attacking different plant parts at different times during the cropping season. In 15 oilseed rape fields in Sweden, we estimated the damage caused by seed and stem weevils, pollen beetles, and pod midges. Pest pressure varied drastically among fields with very low correlation among pests, allowing us to explore interactive impacts on yield from attacks by multiple species. The plant damage caused by each pest species individually had, as expected, either no, or a negative impact on seed yield and the strongest negative effect was caused by pollen beetles. However, seed yield increased when plant damage caused by both seed and stem weevils was high, presumably due to the joint plant compensatory reaction to insect attack leading to overcompensation. Hence, attacks by several pests can change the impact on yield of individual pest species. Economic thresholds based on single species, on which pest management decisions currently rely, may therefore result in economically suboptimal choices being made and unnecessary excessive use of insecticides.     

Protective effects of silymarin, a milk thistle (Silybium marianum) derivative on ethanol-induced oxidative stress in liver

The production of reactive oxygen species (ROS) is considered to be a major factor in oxidative cell injury. The antioxidant activity or the inhibition of the generation of free radicals is important in providing protection against such hepatic damage. Silymarin, derived from the milk thistle plant, Silybium marianum, has been used in traditional medicine as a remedy for diseases of the liver and biliary tract. In the present study, the effect of hepatoprotective drug silymarin on body weight and biochemical parameters, particularly, antioxidant status of ethanol-exposed rats was studied and its efficacy was compared with the potent antioxidant, ascorbic acid as well as capacity of hepatic regeneration during abstention. Ethanol, at a dose of 1.6 g/kg body wt/day for 4 wks affected body weight in 16-18 week-old male albino rats (Wistar strain weighing 200-220 g). Thiobarbituric acid reactive substance (TBARS) level, superoxide dismutase (SOD), and glutathione-s-transferase (GST) activities were significantly increased, whereas GSH content, and catalase, glutathione reductase (GR) and GPx (glutathione peroxidase) activities significantly reduced, on ethanol exposure. These changes were reversed by silybin and ascorbic acid treatment. It was also observed that abstinence from ethanol might help in hepatic regeneration. Silybin showed a significant hepatoprotective activity, but activity was less than that of ascorbic acid. Furthermore, preventive measures were more effective than curative treatment.     

乙酸乙酯对果蝇(Drosophila melanogaster)的麻醉效应研究

麻醉是果蝇实验中最基本的操作,乙醚是最常用的麻醉剂。但因为乙醚是二类易制毒化学品而被国家控制使用。报道一种容易获得的试剂——乙酸乙酯对果蝇的麻醉效果。实验采用的麻醉室大小为125cm3,每处理20~30只果蝇,乙酸乙酯剂量为40、80、120μL,以同等剂量的乙醚为对照,每个实验重复4次,用所有果蝇完全麻醉后20min及120min时的未苏醒率为指标评估麻醉效果及安全性。结果表明:乙酸乙酯对果蝇具有麻醉作用;麻醉时乙酸乙酯开始起效应的时间略晚于同等剂量的乙醚,但使果蝇完全麻醉的时间却比同等剂量的乙醚略短或相接近;麻醉持续的时间则长于同等剂量的乙醚。乙酸乙酯麻醉的果蝇,90%以上的果蝇均在120min内苏醒,表明在这些剂量范围内是安全的。乙酸乙酯完全可以替代乙醚用于果蝇的麻醉。     

Establishment and spread of founding populations of an invasive thistle: the role of competition and seed limitation

Successful plant invasions require both the founding and local spread of new populations. High plant densities occur only when founding plants are able to disperse their seeds well locally to quickly colonize and fill the new patch. We test this ability in a 7-year field experiment with Carduus acanthoides, an invasive weed in several North American ecosystems. Founder plants were planted in the center of 64 m² plots and we monitored the recruitment, distribution pattern, mortality, and seed production of the seedlings that originated from these founding plants. Competing vegetation was clipped not at all, once, or twice each year to evaluate the importance of interspecific competition. More seedlings recruited in the intermediate once-clipped plots, and these seedlings also survived better. The control plots had fewer microsites for seedling recruitment; clipping a second time in September stimulated grasses to fill up the gaps. The number of C. acanthoides recruits and their median distances from the founder plants were also explained by the initial seed production of the founding plants. Overall, the experiment shows that the success of founder plants can fluctuate strongly, as 55% of the plots were empty by the sixth year. Our study suggests that the local invasion speed following initial establishment depends strongly on both the propagule pressure and availability of suitable microsites for seedling recruitment and growth.     

Larvicidal and chemosterilant activity of the acetone fraction of petroleum ether extract from Argemone mexicana L seed

The acetone fraction of the petroleum ether extract of seeds from Argemone mexicana L. exhibited larvicidal and growth inhibiting activity against the second instar larvae of Aedes aegypti (Linn). This activity occurred at higher concentrations (200, 100, 50 and 25 ppm). Chemosterilant activity, including reduction in blood meal utilization (27.70%), reduction in fecundity (19.00%), formation of larval-pupal intermediates, formation of pupal-adult intermediates, adult mortality and sterility of first generation eggs (100%), occurred at low concentration (10 ppm).     

Pretreatment of cellulosic wastes to increase enzyme reactivity

The enzymatic hydrolysis of cellulose to glucose is generally a slow reaction. Different pretreatments, such as ball milling to a ?200 mesh or swelling in 1–2% NaOH are reported to increase the reactivity considerably. In this work a fiber fraction from cattle manure was treated in an autoclave for 5–30 min at temperatures ranging from 130–200°C. The reactivity of the cellulose, measured by incubating samples with a commercial cellulase preparation for one hour at 50°C and pH 4.8, was increased by a factor of 4–6 compared to NaOH treatment and 10–12 compared to untreated fiber. The increased reaction rate is probably mostly due to an increase in cellulose availability to enzymatic attack, as structural hemicellulose is hydrolyzed and removed during the treatment. Sugars, produced by hemicellulosis, hydrolysis, will react further to give caramelization products. These side reactions were shown to be suppressed by short treatment times. The treated fiber could support growth of a mixed culture of Trichoderma viride and Candida utilis only after washing, indicating the formation of water soluble inhibitory products during treatment. The treatment with high-temperature steam can probably be used also with other cellulosic materials to increase reactivity. This may be an attractive way to prepare low-valued wastes such as manure fibers, straw, stalks, or corn cobs for fermentation processes to increase the protein content or for use directly as ruminant animal feed.     

Pretreatment of beet molasses to increase pullulan production

Pretreatment of beet molasses with cation exchange resin, sulphuric acid, tricalcium phosphate, potassium ferrocyanide, and ethylenediaminetetraacetic acid and disodium salt (EDTA) to increase the production of pullulan was investigated. Among the above techniques used for the removal of heavy metals, sulphuric acid treatment gave better results regarding polysaccharide concentration, polysaccharide yield, and sugar utilization. Aureobasidium pullulans grown on beet molasses produced a mixture of pullulan and other polysaccharides. The pullulan content of the crude polysaccharide was 30–35%. The addition of nutrients improved the production of polysaccharide. A maximum polysaccharide concentration (32·0±1·0 g litre⁻¹) was achieved in molasses solution (70 g litre ¹ initial sugar concentration, pH 6·5–7·5) treated with sulphuric acid and supplemented with K₂HPO₄ 0·5%, -glutamic acid 1%, olive oil 2·5% and Tween 80 0·5%. In this case, the highest values of biomass dry weight (33·8±1·0 g litre⁻¹), polysaccharide yield (63·5±2·5%), and sugar utilization (97·5±1·5%) were obtained at pH 6·5, 3·5, and 4·5–7·5, respectively.     

Using prairie restoration to curtail invasion of Canada thistle: the importance of limiting similarity and seed mix richness

Theory has predicted, and many experimental studies have confirmed, that resident plant species richness is inversely related to invisibility. Likewise, potential invaders that are functionally similar to resident plant species are less likely to invade than are those from different functional groups. Neither of these ideas has been tested in the context of an operational prairie restoration. Here, we tested the hypotheses that within tallgrass prairie restorations (1) as seed mix species richness increased, cover of the invasive perennial forb, Canada thistle (Cirsium arvense) would decline; and (2) guilds (both planted and arising from the seedbank) most similar to Canada thistle would have a larger negative effect on it than less similar guilds. Each hypothesis was tested on six former agricultural fields restored to tallgrass prairie in 2005; all were within the tallgrass prairie biome in Minnesota, USA. A mixed-model with repeated measures (years) in a randomized block (fields) design indicated that seed mix richness had no effect on cover of Canada thistle. Structural equation models assessing effects of cover of each planted and non-planted guild on cover of Canada thistle in 2006, 2007, and 2010 revealed that planted Asteraceae never had a negative effect on Canada thistle. In contrast, planted cool-season grasses and non-Asteraceae forbs, and many non-planted guilds had negative effects on Canada thistle cover. We conclude that early, robust establishment of native species, regardless of guild, is of greater importance in resistance to Canada thistle than is similarity of guilds in new prairie restorations.     

Narrow rows reduce biomass and seed production of weeds and increase maize yield

Smallholder farmers in southern African countries rely primarily on cultural control and hoe weeding to combat weeds, but often times, they are unable to keep up with the weeding requirements of the crop because of its laboriousness, causing them to incur major yield losses. Optimisation of crop planting pattern could help to increase yield and suppress weeds and to reduce the critical period of weed control and the weeding requirements to attain maximum yield. Experiments were carried out in Zimbabwe during two growing seasons to assess the effect of maize density and spatial arrangement on crop yield, growth and seed production of weeds and to determine the critical period for weeding. Planting maize at 60 cm row distance achieved higher yields and better weed suppression than planting at 75 or 90 cm row distance. Increasing crop densities beyond the customary three to four plants m⁻² gave modest reductions in weed biomass but also diminished crop yields, probably because of increased competition for water and nutrient resources. Maize planted in narrow rows (60 cm) intercepted more radiation and suffered less yield reduction from delaying hoe weeding than those planted in wider rows (75 or 90 cm), and the duration of the weed-free period required to attain maximum grain yield was 3 weeks shorter in the narrow spacing than that in the 75- and 90-cm row spacings. Weeding was more effective in curtailing weed seed production in the narrow row spatial arrangements than in the wide row planting. The results of these studies show that narrow row spacings may reduce weeding requirements and increase yields.     

A shrunken-2 transgene increases maize yield by acting in maternal tissues to increase the frequency of seed development

The maize (Zea mays) shrunken-2 (Sh2) gene encodes the large subunit of the rate-limiting starch biosynthetic enzyme, ADP-glucose pyrophosphorylase. Expression of a transgenic form of the enzyme with enhanced heat stability and reduced phosphate inhibition increased maize yield up to 64%. The extent of the yield increase is dependent on temperatures during the first 4 d post pollination, and yield is increased if average daily high temperatures exceed 33 °C. As found in wheat (Triticum aestivum) and rice (Oryza sativa), this transgene increases maize yield by increasing seed number. This result was surprising, since an entire series of historic observations at the whole-plant, enzyme, gene, and physiological levels pointed to Sh2 playing an important role only in the endosperm. Here, we present several lines of evidence that lead to the conclusion that the Sh2 transgene functions in maternal tissue to increase seed number and, in turn, yield. Furthermore, the transgene does not increase ovary number; rather, it increases the probability that a seed will develop. Surprisingly, the number of fully developed seeds is only ～50% of the number of ovaries in wild-type maize. This suggests that increasing the frequency of seed development is a feasible agricultural target, especially under conditions of elevated temperatures.     

Warming increases the spread of an invasive thistle

Background

Global warming and shifted precipitation regimes increasingly affect species abundances and distributions worldwide. Despite a large literature on species'' physiological, phenological, growth, and reproductive responses to such climate change, dispersal is rarely examined. Our study aims to test whether the dispersal ability of a non-native, wind-dispersed plant species is affected by climate change, and to quantify the ramifications for future invasion spread rates.

Methodology/Principal Findings

We experimentally increased temperature and precipitation in a two-cohort, factorial field study (n = 80). We found an overwhelming warming effect on plant life history: warming not only improved emergence, survival, and reproduction of the thistle Carduus nutans, but also elevated plant height, which increased seed dispersal distances. Using spatial population models, we demonstrate that these empirical warming effects on demographic vital rates, and dispersal parameters, greatly exacerbate spatial spread. Predicted levels of elevated winter precipitation decreased seed production per capitulum, but this only slightly offset the warming effect on spread. Using a spread rate decomposition technique (c*-LTRE), we also found that plant height-mediated changes in dispersal contribute most to increased spread rate under climate change.

Conclusions/Significance

We found that both dispersal and spread of this wind-dispersed plant species were strongly impacted by climate change. Dispersal responses to climate change can improve, or diminish, a species'' ability to track climate change spatially, and should not be overlooked. Methods that combine both demographic and dispersal responses thus will be an invaluable complement to projections of suitable habitat under climate change.     

Crop transformation and the challenge to increase yield potential

Molecular transformation is commonly offered as a hope to overcome the apparent stagnation in crop yield potential. A basic understanding of the resource limits imposed on crops and the yield hierarchy going from gene expression to harvestable yield leads to a rather negative view that transformations of a few, or even of a complex of genes will result directly in major yield increases. Forty years of biochemical and physiological research illustrate the great difficulty in translating research at the basic level into improvements in crop yield. However, there are a few cases where physiological research has led to improved crop cultivars with increased yield. These successes are instructive in highlighting key elements required to achieve success in developing crop cultivars for increased yield.     

新古尼异虫草石油醚提取物的成分分析

【背景】新古尼异虫草具有多种药理作用,是一种具有开发潜力的新资源。但该虫草仍有很多活性物质值得探寻,目前对该虫草活性物质的研究多集中于大分子或极性物质,对小分子或弱极性物质的研究关注甚少。【目的】研究新古尼异虫草石油醚提取物中非极性/弱极性小分子化合物,以期完善该虫草中活性物质的化学指纹图谱库。【方法】利用石油醚对新古尼异虫草进行索氏提取,借助傅里叶变换红外光谱(Fourier transform infrared spectroscopy,FTIR)和气相色谱-质谱(GC-MS)联用技术鉴定分析石油醚萃取物中的物质组成。【结果】FTIR表明该虫草石油醚萃取物中含有C-H、C=O、C-O和C=C等官能团,经GC-MS进一步分析鉴定出109种化合物,主要包括烷烃、芳烃、烯烃、酸、酯、醇、胺等化合物,且首次检出甾类、芳烃类、烷烃类、酰胺类、烯烃类和酚类非极性/弱极性的小分子化合物,其中亚油酸及其同分异构体的相对含量最高(38.33%)。【结论】从新古尼异虫草中提取得到多种小分子活性成分,补充了该虫草中的物质组成,为其高附加值利用提供数据支撑。     

Genetic control of soybean seed oil: II. QTL and genes that increase oil concentration without decreasing protein or with increased seed yield

Soybean [Glycine max (L.) Merrill] seed oil is the primary global source of edible oil and a major renewable and sustainable feedstock for biodiesel production. Therefore, increasing the relative oil concentration in soybean is desirable; however, that goal is complex due to the quantitative nature of the oil concentration trait and possible effects on major agronomic traits such as seed yield or protein concentration. The objectives of the present study were to study the relationship between seed oil concentration and important agronomic and seed quality traits, including seed yield, 100-seed weight, protein concentration, plant height, and days to maturity, and to identify oil quantitative trait loci (QTL) that are co-localized with the traits evaluated. A population of 203 F_4:6 recombinant inbred lines, derived from a cross between moderately high oil soybean genotypes OAC Wallace and OAC Glencoe, was developed and grown across multiple environments in Ontario, Canada, in 2009 and 2010. Among the 11 QTL associated with seed oil concentration in the population, which were detected using either single-factor ANOVA or multiple QTL mapping methods, the number of QTL that were co-localized with other important traits QTL were six for protein concentration, four for seed yield, two for 100-seed weight, one for days to maturity, and one for plant height. The oil-beneficial allele of the QTL tagged by marker Sat_020 was positively associated with seed protein concentration. The oil favorable alleles of markers Satt001 and GmDGAT2B were positively correlated with seed yield. In addition, significant two-way epistatic interactions, where one of the interacting markers was solely associated with seed oil concentration, were identified for the selected traits in this study. The number of significant epistatic interactions was seven for yield, four for days to maturity, two for 100-seed weight, one for protein concentration, and one for plant height. The identified molecular markers associated with oil-related QTL in this study, which also have positive effects on other important traits such as seed yield and protein concentration, could be used in the soybean marker breeding programs aimed at developing either higher seed yield and oil concentration or higher seed protein and oil concentration per hectare. Alternatively, selecting complementary parents with greater breeding values due to positive epistatic interactions could lead to the development of higher oil soybean cultivars.     

GC-MS分析香椿子石油醚提取物成分及其体外抗肿瘤活性

采用气相色谱-质谱法（GC-MS）及MTT法（MTT assay）对香椿〔Toona sinensis （A. Juss.） Roem.〕的果实香椿子的石油醚提取物组成成分和体外抗肿瘤活性进行研究。气相色谱-质谱法分析结果表明,从香椿子石油醚提取物的甲酯化与非甲酯化样品中共检测出108种成分,这些成分类型以萜类、烃类以及有机酸为主,从甲酯化样品中检测鉴定了67种,非甲酯化样品中检测鉴定了65种,24种为两个样品中的共有成分。在这些成分中,γ-谷甾醇、豆甾-4-烯-3-酮、亚油酸甲酯、棕榈酸甲酯等成分的相对含量均在8.0%以上。提取物对HepG2、THP-1、Hela、MCF-7均显示出一定的抑制作用,其中,对THP-1细胞在最大考察浓度的抑制率为（60.64±1.37）%。     

Lettuce seed germination: A phytochrome-mediated increase in the growth rate of lettuce seed radicles

Summary The rate of straight growth in mannitol of the axial portion of 2 different samples of photosensitive lettuce seeds was followed after red and far-red irradiation. If the growing half-seeds were classified into proportions of the sample that had undergone a growth increment greater than a certain, selected value, growth rate was found to be enhanced by red light and this enhancement paralleled the phytochrome-mediated increase in the rate of appearance of a geotropic curvature in seed axes, an effect which had been previously reported. The increase in the rate of growth of the radicle by red light was limited to early growth. Once extension has exceeded a certain value the rate of further growth is independent of the state of phytochrome.