遮光胁迫下外源NO对盆栽长春花土壤养分含量及幼苗生长特征的影响

为探究遮光胁迫下施加外源NO供体（硝普钠,SNP）对土壤养分变化和长春花（Catharanthus roseus）幼苗生长的影响,设置4种处理（全光照生长为对照、外施SNP、遮光、遮光+外施SNP联合处理）,分析盆栽土壤中C、N、P养分含量和土壤理化指标,测定幼苗的株高、节间距、茎直径、叶长、叶宽、叶面积、全株鲜（干）质量。结果表明：遮光和外施NO联合处理下,土壤含水量、土壤pH和土壤有机碳含量显著增加（P<0.05）,而土壤中全氮、碱解氮、全磷和速效磷含量增加但不显著,土壤C/N和C/P值升高,且C/N值达到显著水平（P<0.05）;相关性分析结果表明：土壤pH与有机碳含量呈显著正相关性（P<0.05）,土壤有机碳含量与株高、叶片鲜质量之间呈显著正相关（P<0.05）。说明施加外源NO能促进遮光胁迫下土壤pH升高,改变土壤中有机碳含量,增加C/N值,改善土壤肥力,促进叶片生长发育,增加长春花的生物量,为长春花的科学平衡施肥和生物碱含量的积累提供一定依据。     

Hydroxylation and glycosylation of Δ9-tetrahydrocannabinol by Catharanthus roseus cell suspension culture

Δ⁹-tetrahydrocannabinol is the active constituent in Cannabis sativa, with reported analgesic, anti-emetic, anti-oxidative, neuroprotective, and anti-inflammatory activities. Δ⁹-THC has been used to treat a number of disease states including pain, anxiety, asthma, glaucoma, and hypertension. Poor water solubility of Δ⁹-THC greatly reduces its clinical effectiveness. Consequently, there is a need to modify the compound to increase its polarity and pharmaceutical efficacy. The aim of this study was to test the capability of Catharanthus roseus suspension cultured cells to convert Δ⁹-THC into more polar derivatives. The transformed metabolites were analyzed and isolated by HPLC. Structures of some new derivatives were proposed on the basis of molecular ion peaks and fragmentation patterns obtained from LC-MS and UV spectra obtained by HPLC, respectively. Δ⁹-THC was rapidly absorbed by Catharanthus roseus cultured cells and upon biotransformation new glycosylated and hydroxylated derivatives were isolated by preparative HPLC. In addition, cannabinol was detected as degradation product, including its glycosylated derivative. Based on these results, it is concluded that Catharanthus cultured cells have great potential to transform Δ⁹-THC into more polar derivatives and can be used for the large scale production of new cannabinoids, which can be a source of new compounds with interesting pharmacological profiles.     

Geminicoccus roseus gen. nov., sp. nov., an aerobic phototrophic Alphaproteobacterium isolated from a marine aquaculture biofilter

A Gram-negative, strictly aerobic, diplococcoid bacterium (strain D2-3^T) was isolated from the biofilter of a recirculating marine aquaculture system. Phylogenetic analysis of the 16S rRNA gene sequence of D2-3^T indicated that the new organism occupied a novel lineage within the -1 subclass of Proteobacteria and was related to the genera Rhodothalassium, Azospirillum, Craurococcus, Acidiphilium, and Tistrella. The highest sequence similarity (90.8%) of the 16S rRNA gene sequence of D2-3^T was to that of Candidatus “Alysiosphaera europaea”. D2-3^T was mesophilic, heterotrophic, required sea salt, and had a pH optimum of 8.0. Growth in the presence of light resulted in the formation of pink colonies, a 25% increased cell yield, and a slightly increased growth rate. D2-3^T contained carotenoids and low amounts of bacteriochlorophyll a. Membranes of D2-3^T contained b-type cytochromes. The G+C content of the DNA was 60.3±0.1 mol%. Phylogenetic, morphological, physiological, and biochemical analyses demonstrated that D2-3^T represented a new aerobic phototrophic genus, for which the name Geminicoccus roseus gen. nov., sp. nov. is proposed for the type species (D2-3^T=DSM 18922^T=ATCC BAA-1445^T).     

<Emphasis Type="Italic">Catharanthus roseus</Emphasis> L. plants and explants infected with phytoplasmas: alkaloid production and structural observations

Summary. The results of several experiments concerning the presence and composition of alkaloids in different tissues (stems, leaves, roots) of Catharanthus roseus L. plants and explants, healthy and infected by clover phyllody phytoplasmas, are reported. The alkaloids extracted and determined by the reverse phase high-pressure liquid chromatography were vindoline, ajmalicine, serpentine, vinblastine, and vincristine. The total alkaloid concentration was higher in infected plants than in the controls, in particular the increase of vinblastine in infected roots was very significant. The ultrastructural observations of infected roots showed alterations of the cell walls and of the nuclei. These results demonstrate that phytoplasmas, detected in all infected tissues by light fluorescence and transmission electron microscopy, play an important role on secondary metabolism of the diseased plants, modifying both the total content of alkaloids and their ratio.Correspondence and reprints: Dipartimento di Biologia Evolutiva e Funzionale, Universitá degli Studi di Parma, Parco Area delle Scienze 11A, 43100 Parma, Italy.     

Relation between dissolved oxygen concentration and ajmalicine production rate in high-density cultures of Catharanthus roseus

The relation between dissolved oxygen (DO) and the ajmalicine production rate of Catharanthus roseus was investigated in 15-L tank reactors at constant stirrer speed and gas flow rate. Below a DO concentration of 29% of air saturation the ajmalicine production rate was less than 0.06 mumol/g/d. Above a DO of 43% the ajmalicine production rate was constant at 0.21 mumol/g/d. Between a DO of 29% and 43% there was a strong relation between the ajmalicine production rate and the DO concentration. After a period of at least 12 days at DO /=57%. A kinetic equation is proposed for the relation between DO and the specific ajmalicine production rate. (c) 1995 John Wiley & Sons, Inc.     

Two-stage batch process for the production of ajmalicine by Catharanthus roseus: The link between growth and production stage

The link between the growth stage and the production stage in a two-stage batch process was investigated using (filtered) inocula from different periods of the stationary phase of the growth cycle. In the production stage, ajmalicine production by Catharanthus roseus in a 3-L stirred tank reactor was induced with a high glucose concentration (80 g/L). Ajmalicine production in cultures started with cells from the late stationary phase was five times higher than in cultures started with cells from the early stationary phase. After transfer to the production stage, cells from the early stationary phase showed a transient increase in respiration and enzyme induction, followed by culture browning. In contrast, cells in the late stationary phase showed a typical induction pattern: constant respiration, and permanent enzyme induction. A striking similarity between the geraniol-10-hydroxylase (G10H) activity and the ajmalicine accumulation profile could be observed in all cultures, suggesting that G 10H regulated ajmalicine production in this investigation. The intracellular nitrate concentration was significantly higher in the inoculum showing a high ajmalicine production than in the inoculum with a low production. Consequently, nitrate may act as a marker for the start of the production stage: as soon as the nitrate is depleted in the growth medium secondary metabolism can be induced. (c) 1995 John Wiley & Sons, Inc.     

土壤农杆菌转化的长春花冠瘿细胞培养

以土壤农杆菌C₅₈诱导的长春花冠瘿组织与从长春花茎、叶外植体诱导的愈伤组织进行比较,发现冠瘿组织在生长、总吲哚生物碱含量及药用成份阿吗碱含量等方面都优于愈伤组织。测定了光照、温度、蔗糖浓度及外加L－色氨酸前体等,对长春花冠瘿细胞的生长、总生物碱及阿吗碱含量的影响,为长春花冠瘿细胞培养生产吲哚生物碱的实际应用研究提供理论依据。     

Evidence for a Malate Transport into Vacuoles Isolated from Catharanthus roseus Cells

Malate uptake was investigated with vacuoles isolated from Catharanthus roseus cells. The uptake process showed saturation kinetics, was inhibited by organic anions, and was very strongly dependent on the pH of the medium. These data support the classical concept of an anion carrier or channel mechanism and suggest that the Hmal^? form was the transported species. Moreover, malate transport was stimulated by the proton gradient across the tonoplast. The H⁺ translocating enzymes ATPase and PPiase are able to favour malate uptake and, in combination, exert a synergistic effect on this transfer.