Taxus Wallichiana Zucc Research Articles

A new bicyclo [3.3.1]nonadienemethanol-type lignan from the leaves of Taxus wallichiana Zucc.

Following an α-glucosidase inhibitory activity-guided phytochemical study, a rare new bicyclo[3.3.1]nonadienemethanol-type lignan named 9,9′-oxidolanceolatanin B (1) and four known lignans, lanceolatanin B (2), pinoresinol (3), syringaresinol (4), and medioresinol (5), were isolated from a CHCl3-soluble fraction of Taxus wallichiana Zucc. leaves. The absolute configuration of 9,9′-oxidolanceolatanin B was elucidated based on NMR spectroscopic interpretation and ECD data. All isolated compounds 1–5 showed stronger α-glucosidase inhibitory potency than the positive control, acarbose. This data could support the conceit that Taxus wallichiana leaves could be a promising source for plant-based anti-diabetic drug discovery.

Using Historical Habitat Shifts Driven by Climate Change and Present Genetic Diversity Patterns to Predict Evolvable Potentials of Taxus wallichiana Zucc. in Future

Using Historical Habitat Shifts Driven by Climate Change and Present Genetic Diversity Patterns to Predict Evolvable Potentials of Taxus wallichiana Zucc. in Future

Investigation of Phytochemicals, Antimicrobial and Antioxidant Properties of Taxus wallichiana Found in Different Altitudes of Nepal

Indigenous people of hilly region have been using Taxus as a medicinal source for various diseases including cancer. Taxol, one of its main constituents, has been recognized and backed by scientific research; it has been used as a cancer cell suppressing biomolecule in chemo therapies. This study was carried out to collect data that would provide information on the influence of altitude on the production of metabolites and to investigate the phytochemicals, antimicrobial, and antioxidant properties present in Taxus wallichiana Zucc. found in Nepal. Needles were used for phytochemical extraction by Soxhlet method where methanol was used as a solvent. Some of the primary metabolites including carbohydrate and secondary metabolites such as alkaloids, flavonoids, tannins, phenols, saponins, coumarin & steroid were present. Quantitative estimation of the total flavonoid and phenol content of plant extracts were determined. TPC of the plant extract from Dolakha and Jajarkot were 68.745 ± 0.70 and 21.323 ± 1.15GAE/g in 100% concentration respectively. Similarly, TFC from Dolakha and Jajarkot were found to be 5.904 ± 0.024 and 6.275 ± 0.54 Quercetin equivalent per gram. Furthermore, their antioxidant (DPPH) and antimicrobial property was studied. The IC50 value for extract of T. wallichiana from Dolakha was 94.72 µg/mL and Jajarkot was 129 µg/mL, which was comparatively higher than the IC50 (14.54 µg/mL) of Quercetin. Additionally, the plant extract of concentrations of 700 and 950 mg/mL exhibited promising antibacterial properties in E. coli and S. aureus, except K. pneumonia. This study indicates that there is no significant role of altitude in metabolite production of T. wallichiana zucc. Nevertheless, it could be the potential drug due to considerable amounts of phenol and flavonoid contents including natural oxidant and antimicrobial properties.

Quantitative assessment of paclitaxel in Taxus wallichiana zucc. bark using HPTLC densitometry: Insights for germplasm selection and toxicological studies

Taxus wallichiana Zucc. [Synonym: Taxus baccata subsp. Wallichiana (Zucc.) Pilg.] or Himalayan Yew belonging to the evergreen gymnosperm family Taxaceae, grows in temperate regions and has been used in traditional medicines to treat cancer, bronchial disorders, colds, cough, etc. The plant is known to contain paclitaxel (PTX), a key bioactive component with anticancer properties. High-performance thin layer chromatography (HPTLC) was used in the present study to estimate paclitaxel from the bark of T. wallichiana obtained from 15 different geographical locations in the eastern and western Himalayas of India. HPTLC method was determined to be a reliable and efficient technique for the quantification of paclitaxel, offering rapid outcomes.The bark materials of T. wallichiana chemotype obtained from Sikkim (Samanden) contained the highest concentration of paclitaxel, followed by West Bengal (Sandakphu), Arunachal Pradesh (Dibang Valley), and respectively. The present study also examined the cytotoxicity and genotoxicity of the aqueous bark extract of the TW1 chemotype at various doses on the meristematic cells derived from the roots of Allium cepa L.. The bark extracts demonstrate considerable cytotoxic and mitotic properties. This study could be useful to the pharmaceutical industry for selecting elite (high paclitaxel-producing) chemotypes of T. wallichiana. The toxicological evaluation of the T. wallichiana chemotypes (using Allium cepa test) could also help to determine the genotoxic properties of the bark extract.

Phytochemicals, and antioxidants and antibacterial activities of Taxus wallichiana Zucc. root associated endophytic fungi

Phytochemicals, and antioxidants and antibacterial activities of Taxus wallichiana Zucc. root associated endophytic fungi

Habitat characteristics, population structure, distribution, and regeneration status of Taxus wallichiana in South Vietnam

Abstract. Hop NV, Xiong CC, Quy NV, Luong NT. 2023. Habitat characteristics, population structure, distribution, and regeneration status of Taxus wallichiana in South Vietnam. Nusantara Bioscience 15: 179-188. Taxus wallichiana Zucc. is an endangered plant species with high economic value, making it threatened to extinction. Biological information about this species is therefore essential for its conservation. This study is aimed to assess the habitat characteristics, population structure, distribution, and regeneration status of T. wallichiana in South Vietnam. Vegetation surveys were conducted from 2016 to 2021 in three regions in Lam Dong Province. The study revealed that it was distributed in regions with monsoon climate with a total extent of ??379.57 ha, with habitats located on the hillside, narrow shelves, dissected terrain, high slope, on feralite soil with light brown to yellow, red, the soil layer thickness over 0.3 m and in clusters or streaks. The study recorded 634 trees with a diameter at breast height (DBH)?25 cm and 137 trees with 6 cm ?DBH<25 cm. The trees were found primarily at 1401-1500 m above sea level in a mixed broadleaved-coniferous forest. The growth indicators of DBH and overall height (Hvn) developed strongly, but the stand volume was low owing to low tree density. The exponential function was the most suitable for simulating the relationship between Hvn and DBH. Seven thousand six hundred and seventy-three regenerating young individuals were recorded with low density. The density of the regenerating trees decreased as the tree height increased. The finding of this study suggests that T. wallichiana faces the danger of extinction, so solutions are required to conserve and develop this threatened species.

Effect of Methyl Jasmonate on the Growth and Biosynthesis of C13- and C14-Hydroxylated Taxoids in the Cell Culture of Yew (Taxus wallichiana Zucc.) of Different Ages.

The effects of methyl jasmonate (MeJ) on growth and taxoid formation in the cell culture of Taxus wallichiana were investigated to elucidate the specifics of phytohormone action in dedifferentiated plant cells in vitro. The characteristics of the same suspension cell culture were compared in 2017 (the «young» culture) and in 2022 (the «old» culture)-1.5 or 6 years after culture induction, respectively. MeJ (100 µM) is added to the cell suspension at the end of the exponential growth phase. Cell culture demonstrated good growth (dry weight accumulation 10-18 g/L, specific growth rate µ = 0.15-0.35 day-1) regardless of its «age», cultivation system, and MeJ addition. UPLC-ESI-MS analysis revealed the presence of C14-hydroxylated taxoids (yunnanxane, taxuyunnanine C, sinenxane C, and sinenxane B) in the cell biomass. The content of C14-OH taxoids increased from 0.2-1.6 mg/gDW in «young» culture to 0.6-10.1 mg/gDW in «old» culture. Yunnanxane was the main compound in «young» culture, while sinenxane C predominated in «old» culture. Without elicitation, small amounts of C13-OH taxoids (<0.05 mg/gDW) were found only in «young» cultures. MeJ addition to «young» culture had no effect on the content of C14-OH taxoids but caused a 10-fold increase in C13-OH taxoid production (up to 0.12-0.19 mg/gDW, comparable to the bark of yew trees). By contrast, MeJ added to «old» culture was not beneficial for the production of C13-OH taxoids but notably increased the content of C14-OH taxoids (1.5-2.0 times in flasks and 5-8 times in bioreactors). These findings suggest that hormonal signaling in dedifferentiated yew cells grown in vitro is different from that in plants and can be affected by the culture's age. This might be a result of the high level of culture heterogeneity and constant auto-selection for intensive proliferation, which leads to the predominant formation of C14-OH taxoids versus C13-OH taxoids and a modified cell response to exogenous MeJ treatment.

Production of Taxol by Endophytic Fungi Isolated from Roots of Himalayan Yew (Taxus wallichiana Zucc.)

Taxol® (generic name – Paclitaxel), the most promising chemotherapeuticagent was isolated from bark of different Taxus sp. As Taxus species arethreatened with extinction (endangered in Himalaya), thus it is imperativeto develop alternate and sustainable method for commercialization and scaleup production of paclitaxel. In this respect, physical and chemical parametersare effective and important key points for active compound production par-ticularly by using endophytic microbes. In the present study, five endophyticfungi isolated from the roots of Taxus wallichiana, are tested for paclitaxelproduction using biochemical and molecular methods. Subsequently, effectof physico-chemical parameters like temperature, pH, incubation time, andmedium constituents i.e., salt concentration, carbon and nitrogen sources onpaclitaxel production were also analyzed. Among isolates, two of the fungiviz. GBPI TWR F1 (Penicillium sp.) and GBPI TWR F5 (Aspergillus sp.)were found to be paclitaxel producing. The genomic DNA samples were sequenced to confirm the presence of two genes viz. 10-deacetylbaccatinIII-10-O-acetyl transferase (DBAT) and C-13 phenylpropanoid side chain-CoA acyltransferase (BAPT), implicated in paclitaxel biosynthesis. Boththe endophytes showed the amplicons of DBAT and BAPT genes. Resultsrevealed that after optimization of medium components and physical condi-tion, paclitaxel production was increased in both the endophytes, maximumpaclitaxel production i.e., 5.45 ± 0.98 mg/L was obtained by GBPI TWR F5(Aspergillus sp.) following 10 days of incubation at 15◦C in optimized S7liquid medium composition

Taxus wallichiana Zucc. (Himalayan Yew): A Medicinal Plant Exhibiting Antibacterial Properties.

Taxus wallichiana Zucc. or the Himalayan yew is a gymnosperm growing along the Himalayan region of India and adjacent countries. Traditionally, this plant was extensively used by indigenous people for folk medicines for treating various diseases such as fever, headache, diarrhoea, fractures, problems of the nervous system etc. It is also practiced in the Unani system of medicine. The plant is rich in various bio-organic compounds and natural products, such as hydrocarbons, glycosides, flavonoids, phenol, tannins, terpenoids etc. In this research work, an effort has been made to highlight the valuable properties of T. wallichiana. The present study was undertaken to evaluate the secondary metabolites (flavonoids, glycosides, phenols, saponins, tannins, terpenoids) and antibacterial potential of methanol extracts and the subsequent fractions of the leaves and fruit of Taxus wallichiana Zucc. In order to rationalise traditional use, methanol extracts from the leaves and fruit of Taxus wallichiana Zucc. were tested against five bacteria using the agar well diffusion method. Ciprofloxacin was used as a standard. All extracts and fractions displayed significant anti-microbial effects. Taxus wallichiana leaves and fruit methanolic extracts showed a maximum zone of inhibition with Bacillus subtilis, which is 18±0.0mm, and Staphylococcus aureus, 19±0.2mm. The methanolic extracts of the leaves of Taxus wallichiana tested positive for glycosides, flavonoids, phenol, tannins and terpenoids, whereas the T. wallichiana fruit tested positive for flavonoids, saponins and terpenoids. According to the research findings, it was identified that the methanol extract of Taxus wallichiana exhibited quite high anti-microbial activity as well as secondary metabolites, and with this quality, together with lots of its other values, this plant can very well become a source of medicine for the better management of a large number of diseases, including cancer, and value-added products.

Deciphering the Antimicrobial Potential of Taxus wallichiana Zucc: Identification and Characterization Using Bioassay-Guided Fractionation.

Taxus wallichiana Zucc. is a high valued medicinal plant and has been mainly studied for its anti-cancer properties. However, research on its other important biological activities, such as its antimicrobial potential, still needs attention. The focus of the present study is to investigate the antimicrobial activity of secondary metabolites of T. wallichiana needles against 3 different groups of microorganisms, i. e., bacteria, actinobacteria, and fungi. Bioactive compounds from T. wallichiana needles were separated through column chromatography, and, TLC-bioautography. Mobile phases were optimized using Snyder's selectivity triangle. Antimicrobial spots were fractionated and compounds were identified by gas chromatography-mass spectroscopy (GC/MS) and liquid chromatography-mass spectrometry (LC/MS). Functional groups were characterized using Fourier transform infrared spectrometry (FTIR) and nuclear magnetic resonance (NMR) was used to identify the molecular structures. GC/MS and LC/MS data analysis confirm the presence of fatty acids (arachidic acid, behenic acid, palmitic acid, and stearic acid), vitamins (nicotinamide), and alkaloids (cinchonine, timolol), aminobenzamides (procainamide), carbocyclic sugar (myoinositol), and alkane hydrocarbon (hexadecane), having antimicrobial activity in the needles of T. wallichiana. To the best of our knowledge, this is the first report on the isolation and characterization of antimicrobial compounds from the needles of Taxus wallichiana (Himalayan yew). The data obtained from the present study will be supportive to the new drug discoveries in modern medicine with various combinations of medicinal plant's active constituents that can be used for curing many diseases.

Cytotoxic terpenoids from the stem bark of Taxus wallichiana

Chemical study of the stem bark of Taxus wallichiana Zucc. afforded the isolation of two new cyclopenta[ b ]naphthalene terpenoids, wallichianones A ( 1 ) and B ( 2 ) and 13 taxane diterpenoids, baccatin III ( 3 ), 10-deacetylbaccatin III ( 4 ), baccatin IV ( 5 ), 1-dehydroxybaccatin IV ( 6 ), 1-deoxybaccatin VI ( 7 ), taxol ( 8 ), 10-deacetyltaxol ( 9 ), 7- epi -10-deacetyltaxol ( 10 ), taxol-7-xyloside ( 11 ), 7-xylosyl-10-deacetyltaxol C ( 12 ), cephalomannine ( 13 ), 10-deacetylcaphalomannine ( 14 ), and 7- epi -10-deacetylcephalomannine ( 15 ). Their structures were identified by comprehensive analyses of the spectroscopic methods, including NMR and mass spectra. The absolute configurations of 1 and 2 were clarified by time-dependent density functional theory (TD-DFT) electronic circular dichroism (ECD) spectroscopic analyses. Compounds 3 and 7–15 showed cytotoxicity against all five human cancer cell lines, including lung (SK-LU-1), liver (HepG2), breast (MCF7), skin (SK-Mel-2), and prostate (LNCaP), with IC 50 values ranging from 1.4 ± 0.2 to 88.1 ± 5.8 μM. Compounds 9–11 , 14 , and 15 were additionally cytotoxic against human embryonic kidney (HEK-293A) cell line (IC 50 = 14.5 ± 1.0–48.4 ± 1.0 μM), however, 13 was noncytotoxic toward this cell line. The positive control, ellipticine showed cytotoxicity against all the cell lines, with IC 50 values in a range of 1.5 ± 0.1–2.0 ± 0.3 μM. Preliminary analysis of the structural and cytotoxicity relationship of compounds 3–15 suggested that the phenylalanine substituent at C-13 may contribute an important role for the cytotoxicity of the taxane diterpenoids. • 15 compounds were isolated from Taxus wallichiana stem bark. • 2 new and 13 known compounds were identified by spectroscopic analyses. • Cytotoxicity on SK-LU-1, HepG2, MCF7, SK-Mel-2, LNCaP, HEK-293A cell lines.

Taxol and β-tubulins from endophytic fungi isolated from the Himalayan Yew, Taxus wallichiana Zucc.

Paclitaxel, better known as the anticancer drug Taxol®, has been isolated from several plant species and has been shown to be produced by fungi, actinomycetes, and even bacteria isolated from marine macroalgae. Given its cytostatic effect, studies conducted in the 1990's showed that paclitaxel was toxic to many pathogenic fungi and oomycetes. Further studies led to the idea that the differences in paclitaxel sensitivity exhibited by different fungi were due to differences in the β-tubulin protein sequence. With the recent isolation of endophytic fungi from the leaves and bark of the Himalayan Yew, Taxus wallichiana Zucc., and the availability of genomes from paclitaxel-producing fungi, we decided to further explore the idea that endophytic fungi isolated from Yews should be well-adapted to their environment by encoding β-tubulin proteins that are insensitive to paclitaxel. Our results found evidence of episodic positive/diversifying selection at 10 sites (default p-value threshold of 0.1) in the β-tubulin sequences, corresponding to codon positions 33, 55, 172, 218, 279, 335, 359, 362, 379, and 406. Four of these positions (i.e., 172, 279, 359, and 362) have been implicated in the binding of paclitaxel by β-tubulin or formed part of the binding pocket. As expected, all the fungal endophytes grew in different media regardless of the paclitaxel concentration tested. Furthermore, our results also showed that Taxomyces andreanae CBS 279.92, the first fungus shown to produce paclitaxel, is a Basidiomycete fungus as the two beta tubulins encoded by the fungus clustered together with other Basidiomycete fungi.

Antimicrobial compound production by pigment producing endophytic bacterium (Burkholderia sp. GBPI_TWL) isolated from Taxus wallichiana Zucc.

Antimicrobial compound production by pigment producing endophytic bacterium (Burkholderia sp. GBPI_TWL) isolated from Taxus wallichiana Zucc.

Annulohypoxylon sp. strain MUS1, an endophytic fungus isolated from Taxus wallichiana Zucc., produces taxol and other bioactive metabolites.

The online version contains supplementary material available at 10.1007/s13205-021-02693-z.

New Distributional Records of the Himalayan Yew in North Western Himalaya, India-A Consequence of Climate Change

Biological and ecological responses are apparent throughout the globe on plants and animals. In Himalaya, climate change greatly affected distribution and abundance of plants along latitude and elevation because temperature increased 2-5 times more than global average rate. Many treeline, ecotone and community studies are evidence to it, climatic conditions shape the ranges and distribution of species in Himalaya. The Himalayan Yew ( Taxus wallichiana Zucc.) is an endangered native high value medicinal plant, also shows the biological and ecological responses in the Himalayan region in context to global warming. The present study, investigated the distribution and abundance of Himalayan Yew in the Lahaul valley of the cold region.

Use of Root Microbiome in Conservation of Himalayan Yew (Taxus wallichiana Zucc.): An Ecofriendly Approach

The root microbiome, endophytes in particular, plays a crucial role in functioning of the host plant. These microorganisms, as a consortium, can be used as a part of the propagation protocol for producing healthy plants. Himalayan yew (Taxus wallichiana Zucc.) is an important medicinal plant species, mainly due to its known anticancer property. This species is listed in IUCN red list of threatened species and hence a priority candidate for conservation. Microscopic examination of T. wallichiana roots clearly indicates heavy colonization by mycorrhizae and other endophytes. The present study illustrates the application of the root-associated microbiome for raising healthy plantlets of T. wallichiana, using stem cuttings for field transfer, as an ecofriendly nursery practice.

Bioprospecting plant growth promoting endophytic bacteria isolated from Himalayan yew (Taxus wallichiana Zucc.)

The present study aims to investigate the endophytic bacteria, isolated from the roots of Taxus wallichiana Zucc. and designated as GBPI_TWL and GBPI_TWr, for their plant growth promoting traits. On the basis of phenotypic and molecular characters, the bacteria are identified as species of Burkholderia and Enterobacter, respectively. Both the bacteria could grow at wide range of temperature (5−40 °C, opt=25 °C) and pH (1.5–11.0, opt = 6–7), and tolerate salt concentration up to 12 %. While both the bacterial endophytes possessed siderophore, HCN, ammonia, and salicyclic acid producing abilities, GBPI_TWL showed IAA and ACC deaminase producing abilities, in addition. The bacteria were found to be potential phosphate solubilizers at wide temperature range (5−35 °C) by utilizing tricalcium, iron, and aluminium phosphate as substrate. Further, the bacterial isolates produced phytase and phosphatase enzymes in both acidic and alkaline conditions. Positive influence of the inoculation with the bioformulations of GBPI_TWL and GBPI_TWr was demonstrated on the test crops namely rice (Oryza sativa) and soybean (Glycine max) with respect to physico-chemical and plant growth parameters in net house experiments. The study will have implications in developing bioformulations, specifically for low temperature environments, in view of environmental sustainability.

Establishment of regenerative callus, cell suspension system, and molecular characterization of Taxus wallichiana Zucc. for the in vitro production of Taxol

Taxus wallichiana, an endangered Taxus species native to Nepal, is a source of Taxol (paclitaxel), an anticancer drug, which if produced via plant cell culture in vitro not only reduces the pressure on natural habitat but also allows continuous production throughout the seasons. Gamborg’s B5 medium was the culture medium to develop callus and suspension culture. Quantification of Taxol was carried out via high performance liquid chromatography (HPLC) using C18 column, water-acetonitrile mobile phase, and 1 ml/minute flowrate at 227 nm wavelength. Genetic variation among population was determined using the POPGEN32, version 1.31. Best callus response was in medium with 1 mg/l 2,4-D for both stem and needle explants. For cell suspension culture, callus from needle explants showed better response compared to stem. Fructose and sucrose proved to be the best carbon sources. Cell count increased as carbon concentration increased. 2,4-D at low concentration (5 mg/l) showed higher cell growth rate. Inoculum volume was directly proportional to cell count. Extra addition of macronutrients had inhibitory effect on cell count, but the effect was reversed for MgSO4. Using HPLC, for bark, the highest Taxol yield was 0.057 mg/g distilled water (DW), while for needle was 0.056 mg/g DW. For suspension cultured HPLC samples, Taxol yield ranged at 0.004%. Size range of amplified Deoxyribonucleic acids was between 200 and 2,000 bp. Thus, this investigation generated a concrete data for Taxus population of Nepal providing efficient protocols for establishment of callus and cell suspension culture with the aim of producing Taxol in in vitro conditions.

CHEMICAL CONSTITUENTS OF ETHYL ACETATE EXTRACT FROM Taxus wallichiana ZUCC. (TAXACEAE) LEAVES COLLECTED IN HA GIANG PROVINCE

Investigation on the chemical constituents of ethyl acetate extract from leaves of Taxus wallichiana Zucc. (Taxaceae), collected in Ha Giang province, has resulted in the isolation of three metabolites for the first time: sciadopitysin (1), β-sitosterol (2), and isoquercitrin (3). Their structures were determined by spectroscopic methods.

Ethnobotanical and pharmacological importance of Taxus wallichiana Zucc.

Taxus wallichiana Zucc. or the Himalayan Yew is a gymnosperm growing along the Himalayan region of Indian and adjoining countries. The plant is extensively used by local people for treatment of various diseases such as fever, headache, diarrhea, fractures, problems of nervous system etc. It also finds usage in Unani system of medicine. The plant is rich in various bioorganic compounds natural products such as hydrocarbons, terpene alcohols, terpenoids (including taxoids), organic acids etc. The plant has been explored for anti-inflammatory, analgesic, antipyretic, anticonvulsant, immunomodulatory, hepatoprotective and anticancer activity with satisfactory outcome. The pharmacological activity of the plant is largely due to the presence of large number of terpenoids. The bioactive constituents present in the plant interacts with a large number of biochemical pathways involved in inflammatory processes, cell division cycles and inhibits a number of enzymes to bring about its protective action against various diseases. In this review, an attempt have been made to highlight the beneficial properties of Taxus wallichiana in various levels of usage starting from its fundamental ethnobotanical use to pharmacological use involving both in-vitro and in-vivo studies. Insights into the molecular mechanisms of action of the active constituents in bringing about the beneficial activity have also been illustrated. The plant can very well become a source of medicine for better management of a large number of diseases including cancer.