80 results about "Taxus × media" patented technology

The invention relates to the taxus media culture technology, which belongs to the sticking culture technology. The taxus media culture technology comprises branch selection, nutrient solution soaking and soil sticking culture. The branch selection technology obtains the easiest surviving sticking branch selection standard of the taxus media, the selected and stuck branch soaking nutrient solution recipe and the environment conditions of the sticking soil substrate are very suitable for the survival of the taxus media. When the technology of the invention is used for propagation, the survival rate higher than 90 percent can be reached.

The invention pertains to agricultural planting technological field, in particular to a stereoscopic ecological cultivation method of taxus Media and cinnamomum camphora. The method comprises planting, field management, harvest and so on. Seedlings of the cinnamomum camphora with a height of 1.5-2m are chosen. Seedlings of the cinnamomum camphora and the taxus Media are planted at a south-north direction in a determinant. 3 rows of taxus are interplanted among each 2 rows of the cinnamomum camphora. The row spacing and planting distance of the taxus is 60cm-80cm, the row spacing between the taxus and the cinnamomum camphora is 1.0-1.2m and the planting distance of the cinnamomum camphora is 1.5-1.8m. The taxus Media is planted during the period from March to April, and the cinnamomum camphora can be planted at the same time, so as to form obvious difference in height with the seedlings of the taxus Media, thereby forming the stereoscopic planting mode. The cinnamomum camphora is transplanted or put into a plantlet market after 3 years of interplant, the whole trunk of the taxus or some of the leaves are harvested and used for extracting paclitaxel medicine. The invention can promote the survival rate and growth speed of the taxus Media so as to reduce the planting cost, thereby having better economic value.

The invention relates to a method for extracting essential oil of taxus media. The method comprises the following steps of: 1, preprocessing raw materials: removing impurity from tender leaves and / or branches of fresh taxus media and crushing the tender leaves and / or branches to obtain pieces or powder of the raw materials; 2, performing steam distillation: uniformly mixing the pieces or powder of the raw materials and common salt, and placing the mixture in a distiller for steam distillation; and 3, separating oil from water: performing high-speed centrifugal separation or standing separation on the collected oil-water mixture to obtain the essential oil of the taxus media. The method of the invention has the advantages of simple process, less equipment, low cost, no pollution, and special suitability for medium- and small-size enterprises.

The invention discloses a formula of a plant rooting solution. The formula comprises the following components: amino acids, nitrogen, phosphorus, potassium, calcium, naphthylacetic acid, indole butyric acid, ethyl alcohol, sugar and an abt rooting agent. Due to the addition of amino acids and trace elements such as nitrogen, phosphorus, potassium and the like, the cuttage survival rate is high. The cuttage survival rate can be significantly increased by soaking branches with difficulty in rooting in the rooting solution, for example, taxus media branches of the cuttage survival rate can reach above 85%, emergence and transplanting are fast, and the survival rate of transplanting is high and can reach above 90%.

The invention relates to an extraction method of taxol from branches and leaves of artificially cultivated yew. A plurality of methods, such as planting of stocky yew, cultivating of tissue and semi-synthesis and total synthesis of taxol, have been developed home and abroad so as to obtain taxol raw material; however, the tissue cultivating method has low yield and high broth price; and the total synthesis has extremely low yield, worse therapeutic effect than natural-source taxol, and a high cost; thereby being difficult for commercial application. The invention uses branches and leaves of artificially cultivated taxus media, Yunnan taxus, southern taxus or northeastern taxus as raw materials; the raw materials are marinated by carbinol and extracted by petroleum oil and propyl acetate in sequence; and normal-pressure normal-phase silica gel column chromatography or reverse-phase high-pressure liquid chromatography is used to obtain high-purity taxol with a purity being higher than 99.6%. The preparation method has simple process, low cost, recyclable solvent and eluant; therefore, the method is applicable to industrial production and protects plant resource.

The invention relates to a Taxus media cultivation method which comprises the following steps: (1) preparing before planting, (2) planting tree seedlings and (3) field management. The step (1) includes the following steps: selecting a cultivation field, and selecting cultivation seedlings. The step (2) includes the following steps: in March or April, planting the cultivation seedlings into the cultivation field with the row spacing and line spacing respectively of 55-65 cm, enabling soil moisture content in the cultivation field to be 30%-40%, and ensuring survival of cultivation seedlings to form small trees. The step (3) includes the following steps: weeding; conducting one-time topdressing to all small trees in April or May each year from the next year when planting is completed, concretely, topdressing bio-organic fertilizer 90-100 g, and topdressing compound fertilizer 50-60 g; spraying plant growth regulators to all small trees in April or May each year from the next year when planting is completed; and collecting new branches and leaves of the small threes in October or November each year from the next year when planting is completed, wherein the new branches and leaves are grown in the current year. By means of the Taxus media cultivation method, the cultivation seedlings can reliably survive, fast grow, and stably accumulate valid biomass.

The invention relates to a method for simultaneously extracting taxol and 10-DABIII (10-deacetyl baccatine III) from branches and leaves of Taxus media, which separately employs ultrasonic extraction, solvent lixiviation and Soxhlet extraction and comprises the following steps: concentrating an extracted methanol solution to be dry while depressurizing, extracting with n-hexane to remove pigment and other fat-soluble impurities, extracting with dichloromethane to obtain a final extraction liquid of taxol and 10-DABIII, and detecting the contents by HPLC (high-performance liquid chromatography). The method provided by the invention fully takes advantages of the different extraction methods to simultaneously extract taxol and 10-DABIII, improves the utilization rate of resources, and has the characteristics of simplicity, rapid extraction and high efficiency.

The invention relates to a cultivation and collection method for improving taxol content in taxus media branches and leaves, belonging to the technical field of medicinal plant cultivation. One-year-old or more than one-year-old taxus medias are cultivated in optimized soil with pH of 6.0 to 6.9 under sunny natural daylight; proper nitrogenous fertilizer, phosphate fertilizer and potash fertilizer are applied around summer begins and autumn begins each year, and newly growing branches and leaves in the current year are picked around winter begins, are dried in the shade or dried at the low temperature of 45 DEG C. According to the method provided by the invention, taxol content in taxus media branches and leaves can be improved by about 10 to 17%, and the quantity of branches of each taxus media is improved, thus effectively relieving shortage of medicine resource of taxus media, lowering the cost of medicine taxus media, and producing great economic benefit and social benefit.

The invention relates to a Taxus media rapid cutting propagation cultivation method, belonging to the field of asexual quick propagation technologies of plants. The rapid cutting propagation cultivation of Taxus media is realized through steps of branch pruning, proportioning of alcohol and 1-naphthylacetic acid and then mixing with water, soaking in soaking solution for 1-3 seconds after putting cuttings into the soaking solution, cutting, management, fertilization and transplantation. The Taxus media rapid cutting propagation cultivation method has the advantages that the time of soaking in nutrient solution is short, not only can the labor, the time and the cost be saved, but also the original variety nature and the inner quality of the Taxus media can be maintained to the utmost extent, the method is more economic propagation method, the survival rate of Taxus media branches can reach more than 98 percent and the method can be widely popularized.

The invention relates to a cultivation technology for Taxus media. The cultivation technology comprises branch selection, soaking of branches in nutrient solutions and cuttage cultivation in soil, to be specific, cutting Taxus branches into a plurality of sections, performing cuttage treatment on each section of Taxus branch to enable the Taxus branches to take root and sprout by using the regeneration capacity, wherein the length of each cutting is 10-15cm, and the bottom diameter of each cutting is 3-5cm; bundling up every 30 cuttings cut according to the specifications into one small bundle, and enabling lower cuts to be even and upper cuts to be uneven; soaking bundled branches in the nutrient solutions for 48 hours. According to the cultivation technology for the Taxus media, easy-to-survive cuttage branch selection standards of the Taxus media are achieved, and the selected formula of soaking cuttage branches in the nutrient solutions and the environmental condition of transplant soil matrix are quite suitable for survival of the Taxus. The survival rate of the Taxus can reach above 90% through the cultivation technology for the Taxus media.

The invention relates to the field of agricultural plant propagation, in particular to a method for promoting cutting rooting of taxus media. The method includes the following steps that firstly, a seedbed and a seedbed matrix are prepared; secondly, cuttings are selected; thirdly, the cuttings are processed; fourthly, cuttage is carried out; fifthly, management after cuttage is carried out. The steps of the method are synergistic to promote full absorption of nutrients and moisture of the cuttings, new roots are germinated quickly, the cutting survival rate of taxus media is increased, the survival rate is as high as 95% or above, the rotting period is 20-28 days, the cutting period is greatly shortened, workloads of later management are reduced, and the method has an active function on quickly reproducing taxus media and improving economic value.

The invention relates to an extracting method of a high-purity taxol and a derivative thereof, in particular to a method for extracting the high-purity taxol and the derivative thereof from Taxus Media branches and leaves, which comprises the steps of drying and grinding the Taxus Media branches and leaves, extracting with carbinol for three times, respectively decompressing the three extracted liquids and combining after recovering the carbinol, degreasing respectively with isometric petroleum ether (60-90 DEG C) and normal hexane and drying into dry paste at the temperature of not more than 50 DEG C. The extraction method has the advantages of complete extraction of the taxol and the derivative thereof, high purity and fewer impurities, and creates more favorable conditions for the follow-up industrialized or non industrialized purification and separation of medicines of taxol and derivative thereof.

The invention discloses a method for promoting fruit bearing of Taxus media. The method mainly includes the three steps of pollen collection, pollen compounding and pollination. With the method provided in the invention, under the premise of ensuring plant nutrient supply, the fruit bearing rate of Taxus media subjected to artificial pollination can be significantly enhanced, and can reach 70%-90%.

A novel method of raising seedlings by cutting with electromagnetic pulse for Taxus media is the method of raising the seedlings by cutting through the utilization of the electromagnetic pulse for assisting. The method is realized by the following technological processes: preparing a seedbed-burying a cathode screen and paving matrix-arranging an anode screen-clipping and cutting cutting slips-processing by an electric field- hardening seedling and transplanting, wherein in the electric field action environment, the phenomena of maturing and browning callus are restrained, and the formation time of root primordial is shortened to grow roots in 25 days; and under the condition of free of the electric field action environment, due to the maturing of the callus, the root primordium can break through the epidermis in about 45 days; and additionally, as the photosynthetic efficiency is improved, the number of root caps is obviously improved compared with that of the roots, so as to facilitate the survival of strong seedlings and transplanting. Compared with the prior art, both the rooting rate of cutting and the survival rate of transplanting reach to more than 95%.

The invention relates to a polysaccharide polyphenol plant sample RNA (ribonucleic acid) extraction method. By combining a CTAB (cetyltrimethylammonium bromide) process and a commercial kit provided by an OMEGA kit, the invention develops a high-efficiency high-quality RNA extraction method. The inspection proves that the method is applicable to various tissues of Neolamarckia cadamba, Camellia sinensis L., Eriobotrya japonica Lindl., Rosa chinensis, Litchi chinensis Sonn., Ginkgo biloba L., Opuntia ficus-indica L., Pinus taeda L., Aloe barbadensis Mill., Taxus media and other plants, and provides a favorable option for RNA extraction and preparation in molecular biology experiments in future, and the effect is good.

The invention discloses a taxus media non-test tube rapid propagation method which comprises the following steps of: (1) branch selection: selecting new branches growing on taxus X media in the same year and shearing to obtain two bundles of coarse and thin branches for standby; (2) seedbed substrate processing: deeply ploughing, unpacking, setting up a shed, disinfecting, sprinkling turfy soil and paving fine river sand; (3) rooting solution preparation: preparing amino acid, nitrogen, phosphorus, potassium, calcium, naphthylacetic acid, indole butyric acid, alcohol, white sugar and abt rooting agents; (4) branch soaking: soaking the fine branches obtained in the step (1) into rooting solution obtained in the step (3); (5) inserting; and (6) carrying out heat and moisture preservation on a seedbed. The taxus X media non-test tube rapid propagation method has the benefits that through carrying out technical innovation in aspects of branch selection, classification, seedbed substrate processing, rooting solution preparation and classified soaking, the purposes of high survival rate and high propagation speed are realized, the taxus X media can be quickly inserted and propagated on a large scale, and the mass propagation and the anticancer application of the taxus X media are facilitated.

The invention relates to a manufacturing method of a Taxus media water culture bonsai. The manufacturing method comprises the following steps: (1) selecting pest-free soil culture Taxus media seedlings with upright and plump plant shapes, washing soil on roots and cutting rotten roots, planting the seedlings in a seed bed, and supplementing leaf fertilizer for cultivation through leaf water supplementation; (2) after white new roots of the Taxus media which are 2-5 cm in length are grown out, transplanting in a water propagation seed bed, controlling the water temperature to be 21-25 DEG C, pumping water with a water pump for circulation so as to keep oxygen in water to be sufficient; (3) after the new roots of the Taxus media adapt to water environment, and taking out the Taxus media to put in a water culture flowerpot; (4) after the Taxus media is transplanted in water culture flowerpot, putting the water culture flowerpot in a greenhouse for curing for 20-40 days, wherein leaf spraying water supplementation is primary during the period, and the Taxus media is promoted to adapt to the new environment by spraying the leaf fertilizer and watering a nutrient solution. The manufacturing method is simple to operate and convenient to maintain, and the survival rate of indoor maintenance is improved.

The invention discloses a group of combined strains for highly producing baccatin III and a method for producing baccatin III, belonging to the technical field of biology. The combined strains for highly producing baccatin III are combined strains of baccatin III production strains and paclitaxel production strains, wherein the baccatin III production strains are Taxus chinensis var mairei endophytic fungi no.79, and paclitaxel production strains are Taxus media endophytic fungi Z58 or Taxus india endophytic Gliocladium SP. By sequentially carrying out independent culture and mixed culture on the baccatin III production strains and the paclitaxel production strains, the yield of baccatin III in an fermented object is substantially increased; by virtue of metabolic pathway complementation and a feedback inhibition theory, the yield of baccatin III is greatly increased and is far higher than that of an independent culture manner. According to the method, the industrial fermentation culture and production of baccatin III by virtue of endophytic fungi are realized, relatively cheap raw materials are provided for the production of paclitaxel by virtue of a semi-synthesis method, and the production cost is lowered; the dependence on the Taxus chinensis resource is reduced, and the damage to the environment is alleviated.

The invention discloses a method for increasing the yield of paclitaxel in an endophytic fungus fermentation product. The method comprises the steps that an endophytic fungus seed solution for producing paclitaxel is inoculated into an endophytic fungus solid state fermentation medium containing taxus media and valeriana jatamansi for culture, the mixture is then put into an alternating magnetic field environment for further culture, and a endophytic fungus solid state fermentation product is obtained; then the endophytic fungus seed solution for producing paclitaxel is inoculated into a liquid fermentation medium containing the endophytic fungus solid state fermentation product for culture, and the final endophytic fungus fermentation product is obtained. According to the method, raw materials are natural, no pollution is caused to the environment, a cycle is short, the yield is high, the method is suitable for industrial production, and compared with a common endophytic fungus fermentation product, the yield of paclitaxel in the endophytic fungus fermentation product is obviously increased.

The invention relates to a nucleotide sequence, a molecular probe and a method for identifying a taxus media seedling. The nucleotide sequence for identifying the taxus media seedling is represented as SEQ ID NO.1; upstream MHSF of the nucleotide molecular probe for identifying the taxus media seedling is represented as SEQ ID NO.2, and downstream MHSR is represented as SEQ ID NO.3; and by means of the nucleotide molecular probe MHSF / MHSR, taxus media is identified with a conventional PCR (polymerase chain reaction) method. The sample consumption is small, and the overall operation can be completed only by a small quantity of samples; accuracy and sensitivity are high; MHSF / MHSR is a specific molecular probe for the taxus media, and negative reaction is caused if T. chinensis or T. cuspidata is detected; and the method is simple, the PCR technology is adopted for detection, time is short, and the detection can be finished by half day.

The invention discloses an SNP molecular marker for identifying an European red bean fir or a Taxus media red bean fir, wherein the SNP molecular marker is positioned at a 450th basic group of the sequence shown in SEQ ID NO. 1, and if the 450th basic group is C, the SNP molecular marker is identified as the European red bean fir or the Taxus media red bean fir. The invention establishes a systemfor rapidly identifying the European red bean fir or the Taxus media red bean fir based on the SNP molecular marker, and thus the purpose of rapidly and accurately detecting the European red bean firor the Taxus media red bean fir is achieved, so that the SNP molecular marker is of great significance for species inspection at ports and the like.

The invention discloses a cultivation technique for Taxus media trees, and belongs to the technical field of plant cultivation. The cultivation technique comprises seedling growing and transplanting, wherein in seedling growing, cuttage is performed on the middle ten days of March and October, and river sand is taken as cuttage seedling growing soil; during cultivating, an organic fertilizer is applied according to different seasons, P and K fertilizers and a commercial boron fertilizer are applied appropriately, and illumination, temperature and the like are controlled; Taxus media can fruit in advance after cuttage planting management for about 3 years, the fruiting period is from April to December, the fruiting rate is high, and the fruiting time is prolonged.

The invention provides a solution capable of prompting rooting of taxus media. According to a formula, raffinose, saccharose and thiamine have the effects of promoting rooting of cottage plants; two traditional Chinese medicines, namely, astragalus membranaceus and liquorice are used in match with sodium humate, catechinic acid and citric acid to improve the effect of raffinose, saccharose and thiamine on promoting the rooting of the cottage plants, so that the rooting rate can be further increased, and the rooting rate of taxus media reaches more than 98%; meanwhile, a specific preparation process is performed to prepare the rooting solution, and the new roots of taxus media using the rooting solution are woody roots which are strong and high in environment adaptability and nutrition absorbency; as the woody roots increase, the resistance of seedling increases, and the survival rate of transplanting increases; in addition, the quantity of new tips is increased; leaves are the important places for the plants to synthesize nutrient; as the quantity of the new tips increases, more nutritional components can be synthesized by the plants, and therefore, the plants can be strong and strong in disease resistance.

The invention discloses a cutting nursery test method for Taxus media. The method comprises the following steps: selecting branches of proper cutting seedlings for carrying out cuttage test by taking the branches as cuttings; partitioning a test group into a slow soaking group and a quick soaking group for soaking; researching the actions of growth regulating substances on the rooting of the cuttings and the growth of seedlings respectively to obtain a most proper growth regulating substance; researching different concentrations of the growth regulating substances on the rooting of the cuttings and the growth of seedlings respectively to obtain a most proper concentration; researching the treatment time of the growth regulating substances on the rooting of the cuttings and the growth of seedlings respectively to obtain most proper treatment time; comparing the influences of two soaking treatment methods on the rooting of the cuttings and the growth of seedlings, and selecting a most proper treatment method; researching the relations between the types of the cuttings and the rooting of the cuttings to obtain a most proper cutting type. By adopting a scientific culture technology for cutting nursery for the Taxus media, the breeding and development of the Taxus media are facilitated.

A method for using Taxus media branches and leaves as a raw material to prepare high-purity 10-deacetyl baccatin III comprises the technological steps of (1) coarse extraction, (2) sedimentation, (3) adsorption, (4) dissolution, (5) crystallization and (6) recrystallization. According to the method, a crude product is dissolved by adopting different single solvents through macro-porous resin adsorption to remove a large amount of undissolved impurities, and then the high-purity 10-deacetyl baccatin III is obtained through multiple-time crystallization. The method is simple in process and low in cost and is beneficial to environmental protection, and operation is easy to master.

The invention relates to an extraction method of taxol from branches and leaves of artificially cultivated yew. A plurality of methods, such as planting of stocky yew, cultivating of tissue and semi-synthesis and total synthesis of taxol, have been developed home and abroad so as to obtain taxol raw material; however, the tissue cultivating method has low yield and high broth price; and the totalsynthesis has extremely low yield, worse therapeutic effect than natural-source taxol, and a high cost; thereby being difficult for commercial application. The invention uses branches and leaves of artificially cultivated taxus media, Yunnan taxus, southern taxus or northeastern taxus as raw materials; the raw materials are marinated by carbinol and extracted by petroleum oil and propyl acetate insequence; and normal-pressure normal-phase silica gel column chromatography or reverse-phase high-pressure liquid chromatography is used to obtain high-purity taxol with a purity being higher than 99.6%. The preparation method has simple process, low cost, recyclable solvent and eluant; therefore, the method is applicable to industrial production and protects plant resource.

The invention relates to a method for effectively reducing taxus media cell biosynthesis C-14 oxygenated taxane, which belongs to the technical field of genetic engineering. Through an meloidogynosis agrobacterium mediated binary vector transformation system, the method leads an antisense genetic expression vector constructed according to yew taxane 14Beta hydroxylase gene (referred to as 14OH) fragments into a Taxus media cultured cell. The method successfully obtains transgenic cells through an optimized transformation method. PCR-Southern hybridization, RT-PCR gene expression analysis and chemical component detection results have shown that the taxus media cell transgenic test succeeds, and the 14OH gen mRNA expression level in a transgenic cell system is significantly reduced. The method significantly inhibits the flowing of the taxus media cells to C-14 oxygenated taxoids, and is expected to lead metabolism to flow to C-13oxygenated taxoids including an anti-cancer drug taxol and the precursor baccatin III thereof, so as to provide a novel method for improving the biosynthesis yield of taxol.

Reported herein are methods for obtaining stably transformed callus in Taxus media ‘Hicksii’, including particularly methods that involve using needles, stem, or bark peel as explant material for transformation. Also provided are descriptions of several promoter activities in directing reporter gene expression in Taxus media cells, in particular cells in suspension cultures, callus and needles. Transgenic plants (e.g., Taxus plants), plant cells, cell lines, and tissues (including seeds) are also provided, in particular those that express one or more enzymes in a paclitaxel biosynthesis pathway.

The invention provides a taxus media grafting cultivation method. The technical core is that Chinese torreya seedlings of 1-2 years are used as stocks; taxus media of 1-2 years are used as cutting seedlings for carrying out grafting; and a method that cultivating is carried out after grafting is used for grafting cultivation of the taxus media. According to the method provided by the invention, the Chinese torreya seedlings are innovatively adopted as the stocks for grafting the taxus media; the Chinese torreya seedlings are used for overcoming the deficiency of stem rot; a characteristic thatthe disease resistance and the adaptive capacity on a growth environment of Chinese torreya are stronger than the disease resistance and the adaptive capacity of taxus media is utilized, so that theadult rate of the taxus media is improved to 93-97%; and grafting is carried out before cultivation in a field, so that the problem that the labor efficiency is low because grafting is carried out bysquatting in the field after the cutting seedlings survive is avoided.

The invention discloses a method for improving a rooting rate of taxus media tissue culture seedlings. The method comprises the following steps: (1) selection of explants: selecting 1-2-year-old semi-lignified robust branches of taxus media, cutting 3-5cm-long stem segments with axillary buds, and removing base needle leaves to obtain explants; (2) treatment of explants: after cleaning and disinfecting the explants, soaking the explants in an ABT1 solution, an NAA solution or an IBA solution for 5-30 min; and (3) rooting culture: inoculating the explants soaked in step (2) into a sugar-free 1 / 2 MS agar culture medium containing 0.08 g / L of mancozeb and carrying out light and shade alternate culture at 25+ / -2 DEG C, wherein an illumination intensity is 2,000-2,800Lx and an illumination duration is 12 h / d. The method provided is simple and controllable to operate, has a remarkable effect of promoting growth of a taxus media root system, and has a wide application prospect.