Effects of Methyl Jasmonate on Shikonin and Dihydroechinofuran Production in Lithospermum Cell Cultures

Methyl jasmonate, when administered to Lithospermum erythrorhizoncell suspension cultures, was found to induce the productionof shikonin derivatives (the red naph-thoquinone pigments ofthe root) and dihydroechinofuran (an abnormal metabolite ofgeranylhydroquinone). Culture experiments showed that methyljasmonate caused a rapid increase in the activities of enzymesinvolved in the biosynthesis of shikonin such as p-hydroxybenzoategeran-yltransferase, which was followed by the rapid accumulationof dihydroechinofuran and the delayed production of shikonin.The induction patterns observed were similar to those elicitedby oligogalacturonides in Lithospermum cells, suggesting thatjasmonic acid or its derivative may act as a signaling moleculein the elicitation of shikonin biosynthesis. Interestingly,however, the copper ion, which is essential for inducing shikoninbiosynthesis by oligogalacturonides, was not required for shikonininduction by methyl jasmonate ¹Present address: Laboratory of Molecular & Cellular Biology,Department of Agricultural Chemistry, Kyoto University, Kitashirakawa,Kyoto, 606-01 Japan     

真菌诱导物在滇紫草细胞培养中对紫草色素形成的影响

在滇紫草细胞悬浮培养中,真菌诱导物可抑制细胞生长,促进紫草色素的合成。将培养６ｄ的曲霉菌丝体的粗提物以６００μｇ碳水化合物／５０ｍｌ培养液的浓度加入到处于指数生长初期的滇紫草细胞悬浮培养物中,诱导物促进紫草色素合成的作用最大,紫草色素含量为对照的两倍。经高压锅处理２０ｍｉｎ到２ｈ不影响诱导物的活性。真菌诱导物还影响了紫草色素各衍生物的相对含量。     

植物细胞培养生产黄酮类化合物研究进展

黄酮类化合物是中药的一种主要有效成分,本文综述了利用植物细胞培养方法合成黄酮类化合物的研究状况和各种环境条件对植物细胞生长和黄酮类化合物合成的影响。     

ABA Initiates Anthocyanin Production in Grape Cell Cultures

Abscisic acid (ABA) has a well-known positive impact on grape ripening, especially color development, but its role in the initiation of anthocyanin synthesis remains unclear. To elucidate this point, ABA treatment was applied to a simple Vitis vinifera model, consisting of Cabernet Sauvignon cell suspensions that do not spontaneously produce anthocyanins under laboratory conditions. Endogenous ABA levels, the expression of some genes in the upstream part of the anthocyanin pathway, and anthocyanin content were determined. Exogenous ABA treatment sharply increased cell ABA content and induced both structural and regulatory genes involved in anthocyanin production. These changes were promptly detected, as early as 6 h after ABA treatment, whereas anthocyanin production was observed only after 4 days in culture. These results demonstrate that ABA promotes anthocyanin synthesis in grape cell culture.     

Global Production Increased by Spatial Heterogeneity in a Population Dynamics Model

Spatial and temporal heterogeneity are often described as important factors having a strong impact on biodiversity. The effect of heterogeneity is in most cases analyzed by the response of biotic interactions such as competition of predation. It may also modify intrinsic population properties such as growth rate. Most of the studies are theoretic since it is often difficult to manipulate spatial heterogeneity in practice. Despite the large number of studies dealing with this topics, it is still difficult to understand how the heterogeneity affects populations dynamics. On the basis of a very simple model, this paper aims to explicitly provide a simple mechanism which can explain why spatial heterogeneity may be a favorable factor for production. We consider a two patch model and a logistic growth is assumed on each patch. A general condition on the migration rates and the local subpopulation growth rates is provided under which the total carrying capacity is higher than the sum of the local carrying capacities, which is not intuitive. As we illustrate, this result is robust under stochastic perturbations.     

The Production of Pyrethrins by Plant Cell and Tissue Cultures of Chrysanthemum cinerariaefolium and Tagetes Species

Pyrethrins, the most economically important natural insecticide, comprise a group of six closely related monoterpene esters. The industrial production is based on their extraction from Chrysanthemum cinerariaefolium (Pyrethrum) capitula. The world production of natural pyrethrins still falls short of global market demand stimulating the research in in vitro production as an alternative to conventional cultivation methods. The different biotechnological alternatives such as callus cultures, shoot and root cultures, plant cell suspension cultures, and bioconversion of precursors by means of enzymatic synthesis or genetically engineered microorganisms, as well as the progress achieved in methods for the identification and quantitation of insecticidal compounds have been reviewed. Although technology for plant cell culture exists, industrial applications have, to date, been limited due to both the low economical viability and technological feasibility at large scale. Bioconversion of readily available precursors looks more attractive, but more research is needed before this technology is used for the industrial production of pyrethrins.     

The Production of Pyrethrins by Plant Cell and Tissue Cultures of Chrysanthemum cinerariaefolium and Tagetes Species

Pyrethrins, the most economically important natural insecticide, comprise a group of six closely related monoterpene esters. The industrial production is based on their extraction from Chrysanthemum cinerariaefolium (Pyrethrum) capitula. The world production of natural pyrethrins still falls short of global market demand stimulating the research in in vitro production as an alternative to conventional cultivation methods. The different biotechnological alternatives such as callus cultures, shoot and root cultures, plant cell suspension cultures, and bioconversion of precursors by means of enzymatic synthesis or genetically engineered microorganisms, as well as the progress achieved in methods for the identification and quantitation of insecticidal compounds have been reviewed. Although technology for plant cell culture exists, industrial applications have, to date, been limited due to both the low economical viability and technological feasibility at large scale. Bioconversion of readily available precursors looks more attractive, but more research is needed before this technology is used for the industrial production of pyrethrins.     

Increased Production of IAA by Rhizoctonia solani is Induced by Culture Filtrate from Rice Suspension Cultures

To investigate the role of the plant hormones produced by fungi,we tried to construct a system to examine the interaction betweenRhizoctonia solani Kühn MAFF305219 and rice cells in suspensionculture (Oc). R. solani was previously found to produce IAA,with the main biosynthetic pathway via the indole-3-pyruvatepathway. The amount of IAA in the medium produced by R. solaniwas increased by cocultivation with rice cells (Oc) and by culturefiltrate (CF) of Oc. Further analysis revealed that the factor(s)that induced the enhanced accumulation of IAA was sensitiveto heat, to freezing and thawing and lyophilization, and themolecular weight was estimated to more than 10,000. These resultssuggest that the active agent(s) in the medium was (a) proteinor a proteinous substance. Among suspension cultures of variousplants, Oc and another line of rice cells (Ok) had the abilityto induce the accumulation of IAA in the fungal medium 4 h afterinoculation but other cultures of plant cells were ineffective.The promotive effect of rice CF on the accumulation of IAA wasalso observed with some strains of R. solani that belong toa different anastmosis group from MAFF305219. Thus, the accumulationof IAA was not related to the host specificity. (Received July 28, 1997; Accepted October 27, 1997)     

Mechanisms Behind the Inhibition of Lung Adenocarcinoma Cell by Shikonin

Shikonin, a natural naphthoquinone isolated from a traditional Chinese medicinal herb, can exert inhibitory effect on tumor cell growth. However, little has been known concerning the effect of shikonin on lung adenocarcinoma cell and underlying mechanisms. In the present study, we investigated the effect of shikonin on the proliferation, cell cycle arrest, and apoptosis in human lung adenocarcinoma cells. We found that shikonin significantly suppressed the proliferation of lung adenocarcinoma cells compared with control in dose- and time-dependent manner (P < 0.05). In the meantime, our results showed that shikonin markedly increased the proportion of A549 cells at stage G1 as well as induced apoptosis in A549 cells. Furthermore, suppressed CCND1 and elevated caspase3 and caspase7 expression levels at mRNA were found in this study, indicating that shikonin may inhibit the growth of lung adenocarcinoma cell by changing cell cycle and promoting cell apoptosis through the regulation of CCND1, caspase3, and caspase7. Although more studies are needed, this study suggests that shikonin has the potential to be used as an anti-cancer agent in the treatment of lung adenocarcinoma.     

Cell Growth and Shikonin Production of Arnebia euchroma in a Periodically Submerged Airlift Bioreactor

Arnebia euchroma was grown in a 2-l periodically submerged, airlift bioreactor (PSAB) in which the non-submerged (immobilization culture) and submerged (suspension culture) operations were controlled automatically. PSAB had advantages in improving cell growth, shikonin content, shikonin production and cell aggregation compared with suspension culture. Under the optimal submerged/non-submerged period of 10 min/15 h, the shikonin content (4.6%, w/w) and, cell dry mass (16.8 g/l) were 229 and 26% higher than those in suspension culture. Revisions requested 31 October 2005 and 6 December 2005; Revisions received 2 December 2005 and 13 January 2006     

紫草素及其衍生物的调控策略

紫草素及其衍生物在医药、食品、化妆品和印染等领域有着巨大的市场潜力,如何提高它们的产量已成为该领域研究的热点。综述了调控紫草素及其衍生物的合成和积累方法,主要包括高产细胞系的筛选、生产培养基的改良、外加物和外加条件的影响、基因工程调控、生物反应器的影响及分析和提取纯化技术等,并对有关紫草素及其衍生物今后的发展方向进行了展望。     

红豆杉细胞培养产物提取策略及其对紫杉醇的影响

研究了硫酸铈铵及原位提取对红豆杉细胞悬浮培养过程中细胞生长、紫杉醇合成及释放的影响。红豆杉细胞悬浮培养过程中培养第12d添加2mg/L硫酸铈铵能获得最大紫杉醇产量8．3mg/L,其中2．4mg/L释放到细胞外,分别为对照组的4倍及12倍。同时添加2mg/L硫酸铈铵、5％油酸(v/v)时胞外紫杉醇产量达到9mg/L,为对照组的45倍。将硫酸铈铵及原位提取与补料培养相结合,最高紫杉醇产量可达24．5mg/L,其中60％释放到胞外。     

Production of Axenic Gonyaulax Cultures by Treatment with Antibiotics

The effects of amphotericin B, chloramphenicol, dihydrostreptomycin sulfate, neomycin sulfate, polymyxin B sulfate, potassium penicillin G, and streptomycin sulfate (used singularly and in various combinations at different concentrations) on the growth and development of four marine dinoflagellates of the genus Gonyaulax and associated bacteria were studied. The combination of amphotericin B, dihydrostreptomycin, neomycin, and penicillin G was highly effective in eliminating bacteria and fungi without reducing dinoflagellate growth and provided a useful method for obtaining axenic cultures of two Gonyaulax species, G. catenella and G. excavata.     

条件培养液对红豆杉细胞Paclitaxel生产的促进作用

在两步法红豆杉（Taxus chinensis)细胞悬浮培养体系的生产阶段,加入从生长阶段悬浮培养物中制得的条件培养液（conditioned Medium,CM)既能促进细胞的生长,又能提高紫杉醇（paclitaxel)的产率,解决了生产培养时,细胞生长受抑制的问题,特别是,取自生长12天的细胞悬浮培养物的CM按体积分数为25％添加到新鲜生产培养基中时,可使细胞紫杉醇最高产量达28．5mg/L,细胞干重达32．3g/L,分别是对照的2．4倍和2．2倍,对CM中的蔗糖,果糖,NO3－和PO4－3等的含量的进行了分析。     

高产花色苷玫瑰茄细胞系的筛选

花色苷在植物中呈现粉红、红、紫红、紫等颜色,可以用作食品、药品及化妆品的着色剂,亦有药用价值。作为食品添加剂,颜色较合成色素自然,且安全无毒性。早在１９８７年,Ｍｉｚukami^[1]就建议用植物细胞培养物生产花色苷类代替合成色素。所有的植物培养细胞都是异源性的。各细胞之间产花色苷的能力相差很大^[2].因为产花色苷的细胞系带有颜色标记,所以容易识别并通过肉眼选择即可获得高产花色苷的细胞系。筛选的方法很多,如平板饲喂法^[3]、小细胞团法^[4]、细胞块法^[5]、肉眼观察直接挑选法及细胞分栋器法^[6]等。高产系花色苷的含量可增加几倍到几十倍,而且产量稳定。本文采用平板法及小细胞团法筛选高产花色苷的玫瑰茄(Hibiscus sabdariffa L.)细胞系。     

Ethylene Production by Plant Cell Cultures: Variations in Production during Growing Cycle and in Different Plant Species

Suspension cultures of Rosa sp., soybean (Glycine max L.), wheat (Triticum monococcum L.), sweet clover (Melilotus alba Desc.), Haplopappus gracilis Nutt., and rue (Ruta graveolens) produced ethylene. The amount varied with the species. The rate of formation in rose and Haplopappus cells paralleled growth but accelerated when the stationary phase was reached, after which the rate declined sharply. Light was not required for ethylene production. Exogenous ethylene could not replace 2,4-dichlorophenoxyacetic acid or naphthalineacetic acid in the cell cultures, and there was no stimulation of growth in the normal medium. Ethylene at 20 mm reduced growth of Ruta and rose cells by 30 and 20%, respectively. The amounts of ethylene produced by the cultures do not affect growth.     

Zymomonas mobilis Mutants with an Increased Rate of Alcohol Production

Two new derivatives of Zymomonas mobilis CP4 were isolated from enrichment cultures after 18 months of serial transfers. These new strains were selected for the ability to grow and produce ethanol rapidly on transfer into fresh broth containing ethanol and allyl alcohol. Ethanol production by these strains was examined in batch fermentations under three sets of conditions. Both new derivatives were found to be superior to the parent strain CP4 with respect to the speed and completeness of glucose conversion to ethanol. The best of these, strain YO2, produced 9.5% ethanol (by weight; 11.9% by volume) after 17.4 h compared with 31.8 h for the parent strain CP4. The addition of 1 mM magnesium sulfate improved ethanol production in all three strains. Two factors contributed to the decrease in fermentation time required by the mutants: more rapid growth with minimal lag on subculturing and the retention of higher rates of ethanol production as fermentation proceeded. Alcohol dehydrogenase isozymes were altered in both new strains and no longer catalyzed the oxidation of allyl alcohol into the toxic product acrolein. This loss of allyl alcohol-oxidizing capacity is proposed as a primary factor contributing to increased allyl alcohol resistance, although it is likely that other mutations affecting glycolysis also contribute to the improvement in ethanol production.