RESUMEN

Introduction: Taxus species contain the anticancer alkaloid paclitaxel, as well as other taxanes similar in structure and potentially in effect to paclitaxel. Tissue-specific distribution patterns and seasonal variations of taxanes in some Taxus species have been reported; however, it is still under-presented for the taxanes in Taxus cuspidata. Methods: The radial distributions of eight taxanes in the transverse surface of freeze-fixed T. cuspidata stems from the late summer and the spring seasons were investigated by cryo-time-of-flight secondary ion mass spectrometry and scanning electron microscopy (cryo-TOF-SIMS/SEM) visualization and liquid chromatography-mass spectrometry (LC-MS) quantitative analysis. By optical microscopic observation, seasonal differences in the amounts and distribution patterns of target taxanes were further characterized in specific tissues. Results and Discussion: The overall amount of taxanes was higher in the late summer than in the spring. Also, taxanes' radial distribution was generally found at higher concentration in the phloem, the cambium and lower level in the periderm, the latest-forming xylem, with different taxanes showing several patterns with distinction between seasons, which were considered related to seasonal plant physiological behaviors. In addition, the distribution of baccatin III (BAC) was investigated at the cellular level, which was regarded in specific cells suggesting its transport in the radial and axial directions in the T. cuspidata stem. Characterizing the microscopic distribution of taxanes in the T. cuspidata stem is expected to play a role in the further study of their biosynthesis and in planta behaviors.

RESUMEN

Objective: To study the compounds isolated from Penicillium HDS-Z-1E, an endophytic fungal strain isolated from Taxus cuspidata and their activation effect of catalase (CAT). Methods: The chemical constituents were isolated from Penicillium HDS-Z-1E, by using silica gel, Sephadex LH-20 and HPLC. The structural elucidations of five metabolites were elucidated on the basis of spectroscopic including 1H-NMR, 13C-NMR, HMBC and HSQC. Their activation sites of catalase have been investigated by molecular docking. Results: Five metabolites, compounds (1-5) were isolated from Penicillium HDS-Z-1E and identified as 4-hydroxy-4-methyltetrahydro-2H-pyran-2-one (1), 4-hydroxymethyl-5, 6-dihydro-pyran-2-one (2), 5, 6-dihydro-2-oxo-2H-pyran-4-carboxylic (3), N-acetyl-hydrazinobenzoic acid (4), and methyl 2-(2, 5-dihydroxyphenyl) acetate (5). Conclusion: Compound 3 is a new compound. Compounds 3 and 4 may have potential activators of catalase, providing a theoretical basis for the development of CAT activators.

RESUMEN

In this research, the cell growth, physiological, and biochemical reactions, as well as the paclitaxel production, of Taxus cuspidata suspension cells after treatment with polyethylene glycol (PEG), cyclodextrin (CD), or salicylic acid (SA) (alone or in combination) were investigated. To reveal the paclitaxel synthesis mechanism of T. cuspidata suspension cells under elicitor treatment, the transcriptomics of the Control group and P + C + S group (PEG + CD + SA) were compared. The results show that there were no significant differences in cell biomass after 5 days of elicitor treatments. However, the content of hydrogen peroxide (H2O2) and malondialdehyde (MDA), and the activities of phenylalanine ammonia-lyase (PAL) and polyphenol oxidase (PPO) after elicitor combination treatments were decreased compared with the single-elicitor treatment. Meanwhile, the antioxidant enzyme activity (superoxide dismutase (SOD), catalase (CAT), and peroxidase (PO)) and the contents of soluble sugar and soluble protein were increased after combination elicitor treatments. Additionally, the paclitaxel yield after treatment with the combination of all three elicitors (P + C + S) was 6.02 times higher than that of the Control group, thus indicating that the combination elicitor treatments had a significant effect on paclitaxel production in T. cuspidata cell suspension culture. Transcriptomics analysis revealed 13,623 differentially expressed genes (DEGs) between the Control and P + C + S treatment groups. Both GO and KEGG analyses showed that the DEGs mainly affected metabolic processes. DEGs associated with antioxidant enzymes, paclitaxel biosynthesis enzymes, and transcription factors were identified. It can be hypothesized that the oxidative stress of suspension cells occurred with elicitor stimulation, thereby leading to a defense response and an up-regulation of the gene expression associated with antioxidant enzymes, paclitaxel synthesis enzymes, and paclitaxel synthesis transcription factors; this ultimately increased the production of paclitaxel.

RESUMEN

To clarify the key factors constraining the maintenance of wild Taxus cuspidata populations and to develop conservation strategies and technical links for current populations, we investigated the renewal status and distribution patterns of wild T. cuspidata populations in the main distribution areas of China. We analyzed the effects of stand factors and human disturbance on population renewal and maintenance. The results showed that the overall regeneration of wild T. cuspidata populations was poor. The basal diameter and height class structure of renewed individuals showed an unhealthy state. 19% of the area was well regenerated. There were three types of regeneration, including poor regeneration with few adult trees, poor regeneration with many adult trees, and good regeneration with few adult trees. The communities in which T. cuspidata was found could be classified into Abies nephrolepis + Tilia amurensis forest, spinney forest, and Picea jezoensis var. microsperma + A. nephrolepis forest. The renewal number of A. nephrolepis + T. amurensis forest was significantly higher than that of spinney forest. Increased stand density and moderate human disturbance contributed to the regeneration of T. cuspidata. The regenerating T. cuspidata seedlings increased significantly when stand density increased from low to medium. The number of regenerating populations in moderately disturbed habitats was significantly higher than those in lightly disturbed habitats. Human disturbance and habitat were currently critical constraints to maintaining and regenerating wild T. cuspidata populations. The conservation of T. cuspidata should consider current status of population regeneration in each habitat patch to develop corresponding in situ conservation and regression conservation measures and focus on the influence of critical factors such as disturbances and habitat conditions.

Asunto(s)

Picea , Taxus , Tracheophyta , Adulto , Humanos , Bosques , Árboles , China

RESUMEN

Endophytic fungi occupy various ecological niches, which reinforces their diversity. As few studies have investigated the endophytic fungi of alpine conifers, we focused on four species of alpine conifers in this study-Abies nephrolepis, Pinus pumila, Taxus cuspidata var. nana, and Thuja koraiensis-and examined them for endophytic fungi. A total of 108 endophytic fungi were isolated. There were four taxa in A. nephrolepis, 12 in P. pumila, 18 in T. cuspidata var. nana, and 17 in T. koraiensis; these were divided into five classes: Agaricomycetes (3.2%), Dothideomycetes (29.0%), Leotiomycetes (15.0%), Sordariomycetes (41.9%), and Orbiliomycetes (1.6%). The most prevalent fungi were Sydowia polyspora (22.7%) and Xylariaceae sp. (22.7%) in P. pumila, Phomopsis juglandina (16.1%) in T. cuspidata var. nana, and Thuja-endophytes sp. 1 (70.0%) in T. koraiensis. However, there was no dominant species growing in A. nephrolepis. Some host plants were analyzed using next-generation sequencing. We obtained 4618 reads for A. nephrolepis and 2268 reads for T. koraiensis. At the genus level, the top three endophytic fungi were Ophiostomataceae_uc (64.6%), Nectriaceae_uc (15.5%), and unclassified organism (18.0%) in A. nephrolepis and Nectriaceae_uc (41.9%), Ophiostomataceae_uc (41.8%), and Magnaporthaceae_uc (9.2%) in T. koraiensis. Our results show that there are different communities of endophytic fungi among different host plants, even if the host plants are in the same region. Such ecological niches are important in terms of the ecological restoration of alpine conifers.

RESUMEN

Taxane compounds have attracted wide attention due to the basic chemical structure of taxol as an alternative anticancer drug. The full-scan tandem mass spectrometry (MS/MS) fragmentation behaviors of seven taxane compounds were studied. For taxanes of Sc-T and Sc-T-Xyl types, diagnostic product ions are originated from a cleavage in the ester bond of the C13 position and the C-O bond of the C7 position, and the subsequent fragmentation pattern is similar to those of M-type taxanes with the loss of different numbers of acetic acid moieties (AcOH), benzoic acid moieties (BzOH), and H2O molecules. A rapid (7 min) and one-step screening method of two-dimensional microscale carbon fiber and active carbon fiber columns combined with tandem mass spectrometry (2DµCFs-MS/MS) was developed for the screening of taxane compounds from Taxus cuspidata samples. Before MS/MS analysis, the 2DµCFs system can group the sample extract without any pretreatment into three chromatographic-type fractions of strong, medium, and weak polarity to avoid matrix interference, such as lipids and pigments. The 2DµCFs-MS/MS can also conduct qualitative and quantitative analysis of taxane compounds, which is evaluated by limits of detection ranging from 3 to 50 ng mL-1, limits of quantitation ranging from 10 to 150 ng mL-1, satisfactory recoveries from 75.2 to 112.2%, and reproducibilities with relative standard deviations from 1.4 to 11.7%.

Asunto(s)

Carbono , Espectrometría de Masas en Tándem , Fibra de Carbono , Fraccionamiento Químico , Cromatografía Líquida de Alta Presión/métodos , Taxoides

RESUMEN

UV-B radiation is a typical environmental stressor that can promote phytochemical accumulation in plants. Taxus species are highly appreciated due to the existence of bioactive taxoids (especially paclitaxel) and flavonoids. However, the effect of UV-B radiation on taxoid and flavonoid biosynthesis in Taxus cuspidata Sieb. et Zucc. is largely unknown. In the present work, the accumulation of taxoids and flavonoids in T. cuspidata plantlets was significantly induced by 12 and 24 h of UV-B radiation (3 W/m2), and a large number of significantly differentially expressed genes were obtained via transcriptomic analysis. The significant up-regulation of antioxidant enzyme- and flavonoid biosynthesis-related genes (phenylalanine ammonia lyase 1, chalcone synthase 2, flavonol synthase 1, and flavonoid 3', 5'-hydroxylase 2), suggested that UV-B might cause the oxidative stress thus promoting flavonoid accumulation in T. cuspidata. Moreover, the expression of some genes related to jasmonate metabolism and taxoid biosynthesis (taxadiene synthase, baccatin III-3-amino 3-phenylpropanoyltransferase 1, taxadiene-5α-hydroxylase, and ethylene response factors 15) was significantly activated, which indicated that UV-B might initiate jasmonate signaling pathway that contributed to taxoid enhancement in T. cuspidata. Additionally, the identification of some up-regulated genes involved in lignin biosynthesis pathway indicated that the lignification process in T. cuspidata might be stimulated for defense against UV-B radiation. Overall, our findings provided a better understanding of some potential key genes associated with flavonoid and taxoid biosynthesis in T. cuspidata exposed to UV-B radiation.

Asunto(s)

Vías Biosintéticas , Flavonoides/biosíntesis , Perfilación de la Expresión Génica/métodos , Tallos de la Planta/crecimiento & desarrollo , Taxoides/metabolismo , Taxus/genética , Cromatografía Líquida de Alta Presión , Regulación de la Expresión Génica de las Plantas/efectos de la radiación , Estrés Oxidativo , Proteínas de Plantas/genética , Tallos de la Planta/metabolismo , Tallos de la Planta/efectos de la radiación , RNA-Seq , Espectrometría de Masas en Tándem , Taxus/crecimiento & desarrollo , Taxus/metabolismo , Taxus/efectos de la radiación , Rayos Ultravioleta/efectos adversos

RESUMEN

In this study, the roots and rhizosphere soil of Abies koreana and Taxus cuspidata were collected from sites at two different altitudes on Mt. Halla. Ectomycorrhizal fungi (EMF) were identified by Illumina MiSeq sequencing. The proportion of EMF from the roots was 89% in A. koreana and 69% in T. cuspidata. Among EMF in rhizosphere soils, the genus Russula was the most abundant in roots of A. koreana (p < 0.05). The altitude did not affect the biodiversity of EMF communities but influenced fungal community composition. However, the host plants had the most significant effect on EMF communities. The result of the EMF community analysis showed that even if the EMF were isolated from the same altitudes, the EMF communities differed according to the host plant. The community similarity index of EMF in the roots of A. koreana was higher than that of T. cuspidata (p < 0.05). The results show that both altitude and host plants influenced the structure of EMF communities. Conifers inhabiting harsh sub-alpine environments rely strongly on symbiotic relationships with EMF. A. koreana is an endangered species with a higher host specificity of EMF and climate change vulnerability than T. cuspidata. This study provides insights into the EMF communities, which are symbionts of A. koreana, and our critical findings may be used to restore A. koreana.

RESUMEN

Taxus species have attracted much attention for their potency in cancer treatment. However, investigating the bioactivities of Taxus species is a complex task, due to their diversity, slow growth, and endangered state. The most important Taxus species in China are Taxus chinensis (T. chinensis), Taxus cuspidata (T. cuspidata), and Taxus × media (T. media), which mainly grow in the northeastern region. This article probes deep into the differences among the leaves of T. chinensis, T. cuspidata, and T. media, with the aid of gas chromatography-mass spectrometry (GC-MS). Through GC-MS, 162 compounds were detected in the samples and found to contain 35 bioactive metabolites. On this basis, 20 metabolites with significant bioactivities (antibiotic, antioxidant, anticancer, and antiaging effects) were identified via unsupervised learning of principal component analysis and supervised learning of partial least squares-discriminant analysis. The results show that T. media has the most prominent antibiotic, antioxidant, and anticancer effects, while T. cuspidata has the most diverse and abundant metabolites that slow down aging.

RESUMEN

BACKGROUND: Gymnosperms represent five of the six lineages of seed plants. However, most sequenced plant mitochondrial genomes (mitogenomes) have been generated for angiosperms, whereas mitogenomic sequences have been generated for only six gymnosperms. In particular, complete mitogenomes are available for all major seed plant lineages except Conifer II (non-Pinaceae conifers or Cupressophyta), an important lineage including six families, which impedes a comprehensive understanding of the mitogenomic diversity and evolution in gymnosperms. RESULTS: Here, we report the complete mitogenome of Taxus cuspidata in Conifer II. In comparison with previously released gymnosperm mitogenomes, we found that the mitogenomes of Taxus and Welwitschia have lost many genes individually, whereas all genes were identified in the mitogenomes of Cycas, Ginkgo and Pinaceae. Multiple tRNA genes and introns also have been lost in some lineages of gymnosperms, similar to the pattern observed in angiosperms. In general, gene clusters could be less conserved in gymnosperms than in angiosperms. Moreover, fewer RNA editing sites were identified in the Taxus and Welwitschia mitogenomes than in other mitogenomes, which could be correlated with fewer introns and frequent gene losses in these two species. CONCLUSIONS: We have sequenced the Taxus cuspidata mitogenome, and compared it with mitogenomes from the other four gymnosperm lineages. The results revealed the diversity in size, structure, gene and intron contents, foreign sequences, and mutation rates of gymnosperm mitogenomes, which are different from angiosperm mitogenomes.

Asunto(s)

Genoma Mitocondrial , Taxus/genética , Núcleo Celular , Cycadopsida/genética , Evolución Molecular , Intrones , Magnoliopsida/genética , Filogenia , Edición de ARN

RESUMEN

Taxus cuspidata S. et Z. is an excellent natural source of bioactive polysaccharides and has various biological activities. The objective of this study was to evaluate the effect of antidiabetic and antitumor activities of polysaccharides from Taxus cuspidata branches and leaves (TCBL) and to determine the optimum extraction technology of TCBL using a low-temperature and high-efficiency enzyme and ultrasound-assisted coupled extraction (EUCE) method. Optimal technology parameters were determined as follows: an extraction temperature of 51 °C, an extraction time of 33 min, a ratio of material to liquid of 1:19 (g:mL), and an enzyme concentration of 0.10 mg·mL-1. Under the optimized conditions, the polysaccharide yield from TCBL obtained by EUCE was 4.78% ± 0.18%. The four purified polysaccharides (Pe1, Pe2, Pe3, Pe4) from TCBL are mainly composed of arabinose, galactose, glucose, a small amount of xylose, and mannose. This composition was assessed by HPIC analysis. The antidiabetic activity and antitumor activity of polysaccharides from TCBL were assayed in vitro. Among the four purified polysaccharides from TCBL, purified Pe4 had the highest inhibitory capacity against α-glucosidase, and its IC50 value was 123.0 µg·mL-1. Pe1 had the highest antitumor capacity against MCF7 cells and HepG2 cells, with IC50 values of 169.0 and 132.0 µg·mL-1. Pe4 had the highest antitumor effect on human cervical cancer cells (Hela), and its IC50 value was 89.9 µg·mL-1. Pe4 polysaccharide demonstrated a good α-glucosidase inhibitory activity and antitumor capacity against Hela cells. Therefore, Pe4 polysaccharide from TCBL is a beneficial source of potential inhibitors of type II diabetes and human cervical cancer activity.

Asunto(s)

Fraccionamiento Químico , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Polisacáridos/aislamiento & purificación , Polisacáridos/farmacología , Taxus/química , Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/aislamiento & purificación , Antineoplásicos Fitogénicos/farmacología , Fraccionamiento Químico/métodos , Relación Dosis-Respuesta a Droga , Humanos , Monosacáridos/química , Extractos Vegetales/química , Hojas de la Planta/química , Polisacáridos/química , Análisis Espectral , Ondas Ultrasónicas

RESUMEN

BACKGROUND: Taxus cuspidata is well known worldwide for its ability to produce Taxol, one of the top-selling natural anticancer drugs. However, current Taxol production cannot match the increasing needs of the market, and novel strategies should be considered to increase the supply of Taxol. Since the biosynthetic mechanism of Taxol remains largely unknown, elucidating this pathway in detail will be very helpful in exploring alternative methods for Taxol production. RESULTS: Here, we sequenced Taxus cuspidata transcriptomes with next-generation sequencing (NGS) and third-generation sequencing (TGS) platforms. After correction with Illumina reads and removal of redundant reads, more than 180,000 nonredundant transcripts were generated from the raw Iso-Seq data. Using Cogent software and an alignment-based method, we identified a total of 139 cytochrome P450s (CYP450s), 31 BAHD acyltransferases (ACTs) and 1940 transcription factors (TFs). Based on phylogenetic and coexpression analysis, we identified 9 CYP450s and 7 BAHD ACTs as potential lead candidates for Taxol biosynthesis and 6 TFs that are possibly involved in the regulation of this process. Using coexpression analysis of genes known to be involved in Taxol biosynthesis, we elucidated the stem biosynthetic pathway. In addition, we analyzed the expression patterns of 12 characterized genes in the Taxol pathway and speculated that the isoprene precursors for Taxol biosynthesis were mainly synthesized via the MEP pathway. In addition, we found and confirmed that the alternative splicing patterns of some genes varied in different tissues, which may be an important tissue-specific method of posttranscriptional regulation. CONCLUSIONS: A strategy was developed to generate corrected full-length or nearly full-length transcripts without assembly to ensure sequence accuracy, thus greatly improving the reliability of coexpression and phylogenetic analysis and greatly facilitating gene cloning and characterization. This strategy was successfully utilized to elucidate the Taxol biosynthetic pathway, which will greatly contribute to the goals of improving the Taxol content in Taxus spp. using molecular breeding or plant management strategies and synthesizing Taxol in microorganisms using synthetic biological technology.

Asunto(s)

Regulación de la Expresión Génica de las Plantas , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Paclitaxel/biosíntesis , Análisis de Secuencia de Proteína/métodos , Taxus/genética , Transcriptoma , Secuencia de Aminoácidos , Filogenia , Reproducibilidad de los Resultados , Alineación de Secuencia , Taxus/metabolismo

RESUMEN

Tree bark represents an important source of medicinal compounds that may be useful for cancer therapy. In the current study, high-performance liquid chromatography with diode-array detection (HPLC-DAD) was used to determine the profile of the phenolic compounds of Catalpa speciosa, Taxus cuspidata, and Magnolia acuminata bark extracts. The antioxidant and anticancer bioactivities against different cancer cell lines were investigated. M. acuminata exerted significantly higher antioxidant activities in the diphenyl picrylhydrazine and ß-carotene-linoleic acid assays than the other species. In C. speciosa, novel profiles of phenolic acids (ferulic acid was the predominant compound) and catechin were detected. In T. cuspidata, six phenolic acids were detected; the predominant compounds were hydroxycaffeic acid and protocatechuic acid. In M. acuminata, two phenolic acids and three catechins were detected; catechin was the predominant compound. The three species exerted clear anticancer activity against MCF-7, HeLa, Jurkat, T24, and HT-29 cells, with the strongest activity found in the extracts from M. acuminata. No antiproliferative activity against normal cells was found. Flow cytometry revealed greater accumulation of necrotic and early/late apoptotic cells in various treated cancer cells than in untreated control cells, and protocatechuic acid induced a similar accumulation of necrotic cells to that of the bark extracts. Caspase-3 and -7 activity was increased in cancer cells treated with different bark extracts; the highest activity was found in the M. acuminata treatment. Our results suggested that the treatment of cancer cells with bark extracts of M. acuminata, C. speciosa, and T. cuspidata, and protocatechuic acid induced apoptosis, suggesting an association between anticancer activities and individual phenolic compounds.

Asunto(s)

Antineoplásicos/química , Antioxidantes/química , Neoplasias/tratamiento farmacológico , Fenoles/química , Antineoplásicos/aislamiento & purificación , Antineoplásicos/farmacología , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Bignoniaceae/química , Cromatografía Líquida de Alta Presión , Células HeLa , Humanos , Células MCF-7 , Magnolia/química , Fenoles/aislamiento & purificación , Fenoles/farmacología , Corteza de la Planta/química , Extractos Vegetales/química , Extractos Vegetales/farmacología , Taxus/química

RESUMEN

PREMISE OF THE STUDY: Taxus cuspidata (Taxaceae), which is well known for the effective anticancer metabolite paclitaxel (e.g., taxol), is an evergreen needle-leaved tree widely distributed in eastern Eurasia including Japan. We developed 15 microsatellite markers from this species and confirmed their utility for the dwarf variety nana, which is common in alpine regions along the Sea of Japan. METHODS AND RESULTS: Thirteen polymorphic loci were characterized for genetic variation in three populations of T. cuspidata. The number of alleles per locus ranged from 11 to 31, with an average of 18.5; the expected heterozygosity ranged from 0.78 to 0.95, with an average of 0.89. All loci were successfully amplified in T. cuspidata var. nana and showed high polymorphism. CONCLUSIONS: These markers will be useful for investigating speciation and range formation of T. cuspidata in Japan, and the results will provide crucial information for the conservation of Taxus species.

RESUMEN

A polyhydric alcohol (PAL) was isolated from Taxus cuspidata and its immunostimulatory activities were assessed. The primary monosaccharide composition of the PAL was determined to be glucose, where HPAEC analysis showed no significant amount of any other sugars. However, glycerol and xylitol were identified as the main sugar alcohols. Fourier-transform infrared (FT-IR) analysis indicated that the purified PAL is a complex glycitol, which structurally contains significant amount of hydroxyl groups. MALDI-TOF mass spectroscopy also demonstrated that PAL is a complex glycitol built in hexose polymerization. Enzyme linked immunosorbent assay showed that the PAL stimulates the release of the proinflammatory cytokines TNF-α and IL-6 in a dose-dependent manner. Furthermore, treatment of RAW 264.7 cells with PAL for 24h remarkably increased the phosphorylation levels of ERK, p38 and JNK in a dose-dependent manner, whereas the total protein levels of ERK (t-ERK), p38 (t-p38) and JNK (t-JNK) remained unchanged. These results clearly demonstrate that PAL stimulates the immune response in RAW 264.7 cells through the activation of MAPKs (ERK, p38 and JNK) signaling pathway. To the best of our knowledge, this is the first study to demonstrate the primary structure and immune-stimulating activities of PAL from the fruit of T. cuspidata.

Asunto(s)

Alcoholes/química , Alcoholes/farmacología , Factores Inmunológicos/química , Factores Inmunológicos/farmacología , Extractos Vegetales/química , Extractos Vegetales/farmacología , Taxus/química , Alcoholes/aislamiento & purificación , Animales , Línea Celular , Cromatografía Líquida de Alta Presión , Citocinas/biosíntesis , Factores Inmunológicos/aislamiento & purificación , Mediadores de Inflamación/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Macrófagos/metabolismo , Ratones , Peso Molecular , Monosacáridos/química , Óxido Nítrico/biosíntesis , Óxido Nítrico Sintasa de Tipo II/metabolismo , Extractos Vegetales/aislamiento & purificación , Polisacáridos/química , Polisacáridos/aislamiento & purificación , Solubilidad , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Espectroscopía Infrarroja por Transformada de Fourier , Alcoholes del Azúcar/química , Alcoholes del Azúcar/aislamiento & purificación , Alcoholes del Azúcar/farmacología

RESUMEN

Taxus cell suspension culture is a sustainable technology for the industrial production of paclitaxel (Taxol®), a highly modified diterpene anti-cancer agent. The methyl jasmonate (MJ)-mediated paclitaxel biosynthetic pathway is not fully characterized, making metabolic engineering efforts difficult. Here, promoters of seven genes (TASY, T5αH, DBAT, DBBT, PAM, BAPT, and DBTNBT), encoding enzymes of the paclitaxel biosynthetic pathway were isolated and used to drive MJ-inducible expression of a GUS reporter construct in transiently transformed Taxus cells, showing that elicitation of paclitaxel production by MJ is regulated at least in part at the level of transcription. The paclitaxel biosynthetic pathway promoters contained a large number of E-box sites (CANNTG), similar to the binding sites for the key MJ-inducible transcription factor AtMYC2 from Arabidopsis thaliana. Three MJ-inducible MYC transcription factors similar to AtMYC2 (TcJAMYC1, TcJAMYC2, and TcJAMYC4) were identified in Taxus. Transcriptional regulation of paclitaxel biosynthetic pathway promoters by transient over expression of TcJAMYC transcription factors indicated a negative rather than positive regulatory role of TcJAMYCs on paclitaxel biosynthetic gene expression.

RESUMEN

Plants of the genus Taxus are common ornamental shrubs that contain cardiotoxic alkaloids. Gross lesions consistent with heart failure are frequently reported in fatal cases; however, microscopic lesions in the heart have not been well characterized. The current report describes 2 related outbreaks in which 7 of 30, 250-kg calves died after confirmed exposure to clippings of Japanese yew (Taxus cuspidata). Three calves died 24 hr after initial exposure, with no significant gross or histologic lesions. Leaves of the yew plant were identified within the rumen contents, and Taxus alkaloids were confirmed by gas chromatography-mass spectrometry. Following the initial diagnosis, the yew clippings were burned. Two days later, the remaining calves were reintroduced to the enclosure. Within 24 hr, 3 additional calves began to show clinical signs of depression (3/3) or labored breathing (1/3), and by the fourth day, these 3 calves and an additional calf were found dead. Partially burnt yew leaves were found during close inspection of the enclosure. Two of 3 calves submitted for necropsy were severely autolyzed; the third had pulmonary edema and mild fibrinous pleural effusion. Histologic lesions in the latter included multifocal cardiac myocyte hypereosinophilia, sarcolemma fragmentation, pyknosis, karyolysis, myocyte loss, and a mild interstitial lymphoplasmacytic infiltrate with edema. Moderate fibrinosuppurative interstitial pneumonia was the only other significant finding. Cardiac changes were attributed to damage from the initial exposure to Taxus 6 days prior to death.

Asunto(s)

Enfermedades de los Bovinos/metabolismo , Cardiopatías/veterinaria , Intoxicación por Plantas/veterinaria , Taxus/metabolismo , Animales , Bovinos , Enfermedades de los Bovinos/patología , Resultado Fatal , Cromatografía de Gases y Espectrometría de Masas/veterinaria , Cardiopatías/metabolismo , Cardiopatías/patología , Histocitoquímica/veterinaria , Intoxicación por Plantas/metabolismo , Intoxicación por Plantas/patología , Taxoides/análisis , Taxus/toxicidad

RESUMEN

The effects of four elicitors, including 100 µmol/l MeJA (methyl jasmonate), 40 µl/l hydrogen peroxide (30%, w/w), 80 mg/l SA (salicylic acid) and 0.4 g/l F3 (fungal elicitor), on suspension cultures of Taxus cuspidata were studied. After addition of the above four elicitors, the enzyme activity of 10-DBAT (10-deacetylbaccatin III-10-O-acetyltransferase) was induced and reached its maximum of 5.47, 0.97, 3.30 and 6.82 U, respectively. After elicitation, the concentrations of cytochrome P450 monooxygenase were also induced to its maximum values of 0.069, 0.336, 0.321 and 0.193 nmol/ml, respectively. In addition, under the elicitation, the change in 10-DBAT activity was similar to that of cytochrome P450 monooxygenase concentration. The products of these two enzymes changed after the variety of the enzymes, and the taxol content increased through the cultivation.

RESUMEN

Objective: To increase the contents of Taxol and 10-deacetylbaccatin III in the callus of Taxus cuspidata through the application and combination of various plant growth regulators. Methods: Young stems of T. cuspidata were taken as expiants and then their effects of NAA, 2,4-D, and 6-BA on the accumulation of Taxol and 10-deacetylbaccatin III were examined with the method of orthogonal design. Results: NAA exhibited the largest effect on the yields of both total Taxol and 10-deacetylbaccatin III, and followed by 6-BA, 2,4-D. Conclusion: The optimal medium for production of total Taxol and total 10-deacetylbaccatin III is: B5+NAA 2.0 mg/L+2,4-D 0.5 mg/L+6-BA 0.2 mg/L.

RESUMEN

OBJECTIVE: To compare the content of taxol in stalk,leaf and callus of Taxus cuspidata.METHODS:Dichloromethane and water extract were extracted form T.cuspidata using methanol-chloroform(1 ∶ 1).HPLC method was used to determine the content of taxol.RESULTS: The content of taxol in stalk,leaf and callus of T.cuspidata were 0.018 30%,0.001 28%,0.000 66%,respectively.The content of taxol in the callus of T.cuspidata is higher than other two parts of it.CONCLUSION:This study is significance for the development and utilization of T.caspidate and its protection.