50 results about "Leaf cell" patented technology

The invention discloses a moisturizing and antioxidant composition containing a Campanumoea maculata extract and PDNR, and belongs to the technical field of cosmetics. The composition is prepared from the following components: an extract of barks of the calyx ampelopsis, an extract of fruits of annona squamosa, an extract of leaf cells of desert rose, an extract of fruits of fructus corni and polydeoxyribonucleotide (PDRN), and the composition has good moisturizing and anti-oxidation effects through the synergistic effect of the five components of the extract of the barks of the calyx ampelopsis, the extract of the fruits of the fructus corni and the polydeoxyribonucleotide (PDRN).

The invention relates to the field of molecular cytogenetics and potato ploidy breeding, in particular to a potato ploidy identification method for marking and counting chromosome centromere based on a potato CENH3 antibody and a ploidy detection kit. According to the method, the chromosome centromere is marked by specifically recognizing the antibody of the potato centromere nucleosome core histone CENH3 and combining the immunofluorescence technology, so that the accurate counting of the chromosome centromere is realized at the single cell level, and the ploidy of potatoes is judged. The method comprises the following steps: preparing a leaf cell suspension, sequentially incubating a potato CENH3 antibody and a labeled secondary antibody after immobilization, and observing and counting by a microscope after redyeing. The method has the advantages of high species specificity, high throughput and low cost, is not limited by special material taking parts of root tips or shoot tips and cell division periods, can accurately identify the uploid and aneuploid of potatoes, and provides an efficient tool for potato germplasm resource evaluation and cross breeding.

A fluid sensor device for measuring properties of a fluid is disclosed. The fluid sensor device includes a leaf cell sensor having a piezoelectric structure acting on a subdomain of the fluid that flows through the piezoelectric structure to create an intrinsic Helmholtz cavity response, and an enclosure enclosing the leaf cell sensor and including (i) a flowthrough shroud having an inlet that allows the fluid to enter the enclosure and pass across the leaf cell sensor, and a Helmholtz cavity wall that couples the intrinsic Helmholtz cavity response with an external acoustic field of the leaf sensor to increase a measurement sensitivity, (ii) a cylindrical housing having an outlet that allows the fluid to exit the enclosure, and (iii) a pressure feedthrough connector that transmits an electrical signal induced by the intrinsic Helmholtz cavity response to represent the properties of the fluid.

The invention discloses a method for inducing multiple types of somatic cells to change fate and directly develop into somatic embryos and application, and belongs to the technical field of plant genetic engineering. According to the invention, functional genes SPCH (AT5G53210) and ARF5 (AT1G19850) capable of promoting conversion of various types of somatic cells into the totipotent stem cells are found. Overexpression of the totipotent regulatory factor LEC2 can induce formation of somatic embryos on arabidopsis sinapis leaves, under the background, the functional genes are subjected to ectopic expression, more arabidopsis sinapis leaf cells can be converted into totipotent stem cells through SPCH overexpression without calluses, and therefore the totipotent stem cells can be used for inducing formation of the somatic embryos on the arabidopsis sinapis leaves. According to the method, more arabidopsis sinapis leaf cells can be transformed into totipotent stem cells, stem tip cells, hypocotyl cells and root cells can be transformed into totipotent stem cells, more arabidopsis sinapis leaf cells can be transformed into totipotent stem cells through ARF5 overexpression, the stem tip cells and the root cells can be transformed into totipotent stem cells, and finally somatic embryos are formed through induction.

The present invention belongs to the field of treatment of ulcerative colitis and provides an application of extracellular vesicles derived from Herba Euphorbiae in preparing a drug for preventing and treating ulcerative colitis. The application includes extraction and preparation of extracellular vesicles derived from Herba Euphorbiae and research on the preventive and therapeutic effects of extracellular vesicles derived from Herba Euphorbiae on ulcerative colitis in mice. The specific steps are as follows: Step 1, extracting leaf cells from Herba Euphorbiae: selecting fresh and healthy Herba Euphorbiae leaves as experimental materials and chopping them into pieces, adding the chopped samples into an enzymatic hydrolysis solution, ensuring that the samples are fully in contact with the enzymatic hydrolysis solution and incubating them, gently turning or stirring during the incubation to promote the enzymatic hydrolysis process, thereby isolating the leaf cells; Step 2, culturing the leaf cells of Herba Euphorbiae: culturing the leaf cells of Herba Euphorbiae in a culture medium; The present invention confirms that extracellular vesicles derived from Herba Euphorbiae can be used to treat ulcerative colitis, provides an experimental basis for the development of new drugs for the clinical prevention and treatment of IBD, and has good application prospects.

The invention relates to the field of transformation methods and application thereof, in particular to an agrobacterium tumefaciens-mediated pea leaf injection transient transformation method and application thereof, which comprises the following steps: introducing an exogenous gene into pea leaf cells through an injection method by using an agrobacterium tumefaciens-mediated gene transformation system in pea leaves, and realizing transient expression of the exogenous gene; according to the pea leaf injection transient transformation system based on agrobacterium tumefaciens mediation provided by the invention, through an optimized technical scheme, the problems of low transformation efficiency, unstable exogenous gene expression, high operation complexity and the like in the prior art are solved, and remarkable beneficial effects are brought to plant gene function research and commercial application; through an efficient, stable, simple and convenient pea leaf instantaneous transformation technology, many problems in the prior art are solved, the efficiency of gene function research and commercial application is improved, and a powerful technical support is provided for further development of plant molecular biology and genetic engineering technologies.

The invention provides a digital breeding-oriented Chinese cabbage leaf cell automatic segmentation and phenotype measurement system and method, and relates to the technical field of plant phenotype analysis. According to the system, joint coding of cell contours and tissue semantics is achieved through a feature extraction sub-network, scale alignment, global feature convergence and feature re-calibration, and pixel-level cell instance masks and tissue categories are output at the same time through mask decoding and category decoding. In combination with multi-tissue phenotypic parameter classification, in-tissue phenotypic index summarization and structured output, cell-level and tissue-level automatic quantitative analysis of upper epidermis, lower epidermis, fence tissues, sponge tissues, vascular bundles and the like is realized, and the efficiency and precision of multi-tissue cell screening and phenotypic statistics are remarkably improved. The method solves the problems that in the prior art, high-precision automatic segmentation and systematic phenotypic measurement for multi-tissue cells of the cross section of the Chinese cabbage leaf are insufficient, and digital breeding decision making is difficult to directly serve.

The present application relates to a kind of anti-aging composition and its application of brightening and shrinking pores.The composition includes the following components: halocyprid extract, purple magnolia extract, desert rose leaf cell extract, rain lily labiatae flower extract and white lily extract, and the mass ratio of the halocyprid extract, purple magnolia extract, desert rose leaf cell extract, rain lily labiatae flower extract and white lily extract is (0.2-2):(0.001-0.1):(0.001-0.1):(0.001-0.1):(0.001-0.1).The composition of the present application can effectively improve the clearance rate of collagenase, maintain the elasticity and tightness of skin, have good anti-aging and pore-shrinking effect, and can also effectively remove hydroxyl radicals, have good whitening and brightening effect.

The invention discloses a valve preparation method. The method comprises the following steps: performing decellularization treatment on a bovine pericardium material to be treated; enhancing the porosity of the decellularized bovine pericardium material through a physical and / or chemical method so as to promote the regeneration of native valve leaflet cells and tissues, and obtaining a composite material; and cutting the leaflet of the composite material to obtain the valve. The bovine pericardial valve which is high in durability, free of anticoagulation, excellent in biocompatibility, excellent in calcification resistance and low in immunogenicity can be prepared, and therefore the purpose of treating heart valve diseases is achieved.

The invention relates to an anti-aging, brightening and pore-shrinking composition and application thereof. The composition comprises the following components: an artemia extract, a magnolia liliiflora extract, a desert rose leaf cell extract, a lilium lanceolarium extract and a lilium fortunei extract. The mass ratio of the artemia extract to the magnolia liliiflora extract to the rosa deserticola leaf cell extract to the lilachium lanceolatum extract to the lilium fortunei extract is (0.2 to 2) to (0.001 to 0.1) to (0.001 to 0.1) to (0.001 to 0.1) to (0.001 to 0.1) to (0.001 to 0.1). The composition provided by the invention can effectively improve the collagenase clearance rate, maintain the elasticity and compactness of the skin, achieve good anti-aging and pore-shrinking effects, also can effectively remove hydroxyl radicals, and has good whitening and brightening effects.

The invention discloses a kind of HY4-25 and its application in the extraction of total flavonoids from lotus leaves. Before the total flavonoids are extracted from lotus leaves by ethanol ultrasonic extraction, the crude enzyme solution prepared by microbial fermentation is added for enzymatic hydrolysis. The enzyme-producing microorganism Aspergillus niger HY4-25 used is purposefully screened for being able to hydrolyze the cell wall of lotus leaves. The fermentation liquid cultivated by enzyme production contains a variety of hydrolases, which can synergistically hydrolyze substances such as cellulose and pectin in lotus leaves, damage the cell wall, and help dissolve flavonoids. The extraction yield of total flavonoids is significantly improved. The method for enzymatic hydrolysis of lotus leaves provided by the present invention can increase the yield of total flavonoids extracted from lotus leaves by 36.3% compared with the direct use of ethanol ultrasonic extraction method.

The utility model discloses a suspension type planting ball maintenance device which comprises an upper ball cover, a lower base is arranged at the bottom of the upper ball cover, and a suspension line is arranged on the inner wall of the upper ball cover. Through the arrangement and cooperative use of a first fishing line, a clamping rod and a moss ball layer in the suspension line, moss plants are a special group in the plant boundary and are the most original higher plants, and leaf cells of the moss plants are large colorless water absorption cells, have a very strong water absorption effect and can absorb moisture which is 15-20 times of the mass of the moss plants; in the prior art, the moss is good at absorbing water from the surrounding environment, especially water in the air, the moss is designed to utilize the strong water absorption characteristic of the moss and cooperate with a transparent glass cover container to create a high-humidity environment and achieve wireless transmission of the water, so that the moss is used for absorbing the water and transmitting the water to a planting matrix, and the moisture is kept well. The plants cannot be in a water shortage state; the upper part is connected by a thin transparent first fishing line, and the moss ball layer can absorb water from the air.

This invention discloses Chinese kale BoaFLZ35 The application of genes in promoting vegetative growth of plants. This invention utilizes a gene-based approach to construct a *Gynostemma pentaphyllum* plant. BoaFLZ35 Heterologous overexpression of the gene in Arabidopsis thaliana was used to observe and analyze its effects on vegetative growth traits such as root elongation and rosette leaf morphology in transgenic plants. The results showed that heterologous overexpression... BoaFLZ35 This gene can significantly increase root cell length, thereby promoting the elongation of the Arabidopsis taproot; simultaneously, it can also increase leaf cell volume, resulting in significantly larger rosette leaves in transgenic plants. Therefore, it is evident that... BoaFLZ35 This is an important gene in the FLZ family of plants that positively regulates vegetative growth. This invention provides new insights into the function of FLZ family genes in the genus Brassica and also provides important candidate genes for high-yield molecular breeding of Brassica vegetables such as Chinese kale.

The invention discloses an exocytosis protein subcellular localization method for mannitol-induced plasm-wall separation, and relates to the technical field of cytobiology, the method comprises the following specific steps: constructing a fusion expression vector: connecting a plant target exocytosis protein gene with a fluorescent protein gene through a gene cloning technology to obtain a fusion expression vector; inserting into a plant expression vector to obtain a recombinant expression vector containing the target extracellular secretory protein-fluorescent protein fusion gene; performing bacterial colony PCR identification and sequencing verification on the recombinant expression vector; by adopting an agrobacterium tumefaciens-mediated plant leaf instantaneous transformation technology, the fusion gene is rapidly expressed in plant cells, subsequent experiments can be carried out after culture is carried out for 2-3 days, the experiment period is greatly shortened, meanwhile, leaf cells are induced to be subjected to plasmon-wall separation through a gradient mannitol solution, positive cell screening and identification are not needed, and the application prospect is wide. And positioning can be realized by directly observing fluorescence signal distribution by using a laser confocal microscope.

The invention belongs to the technical field of gene engineering, and discloses application of a dehydrin gene in plant drought resistance. The dehydrin gene is separated and extracted from perennial ryegrass and is specifically an LpDHN1 gene, and the nucleotide sequence of the dehydrin gene is shown as SEQ ID NO: 1. The dehydrin gene positively regulates the drought resistance of the plant, and the drought resistance of the plant can be improved by overexpressing the dehydrin gene. After the LpDHN1 gene is heterologously expressed in arabidopsis thaliana, the survival rate of a transgenic plant under drought stress is remarkably improved, and the ion permeability and malondialdehyde content of leaf cells of the transgenic plant under the drought stress condition are remarkably reduced. The invention provides a novel germplasm material for cultivation of drought-resistant plant varieties and improvement of drought-resistant characters of plants.

A fluid sensor device for measuring properties of a fluid is disclosed. The fluid sensor device includes a leaf cell sensor having a piezoelectric structure acting on a subdomain of the fluid that flows through the piezoelectric structure to create an intrinsic Helmholtz cavity response, and an enclosure enclosing the leaf cell sensor and including (i) a flowthrough shroud having an inlet that allows the fluid to enter the enclosure and pass across the leaf cell sensor, and a Helmholtz cavity wall that couples the intrinsic Helmholtz cavity response with an external acoustic field of the leaf sensor to increase a measurement sensitivity, (ii) a cylindrical housing having an outlet that allows the fluid to exit the enclosure, and (iii) a pressure feedthrough connector that transmits an electrical signal induced by the intrinsic Helmholtz cavity response to represent the properties of the fluid.

The application discloses a method for preventing low-temperature freezing injury of sugarcane seedlings and relates to the technical field of sugarcane planting. The method for preventing freezing injury is that Bacillus amyloliquefaciens is uniformly mixed with base fertilizer and is applied together in the process of sugarcane planting, and a freezing prevention agent is sprayed in the sugarcane seedling stage, wherein the freezing prevention agent comprises 1-5 parts of sodium chloride, 100-200 parts of wheat extract and 2-10 parts of sodium caseinate according to weight fractions. The method for preventing freezing injury can increase the proline content in the cells of the sugarcane, reduce the cell membrane permeability, increase the osmotic pressure in the cells, prevent protoplasm from dehydrating and coagulating, and also increase the CAT, SOD and POD enzyme activities, so that the oxidation damage of oxygen negative ions to the cells and tissues is prevented, and the purpose of relieving the freezing injury of leaf cells is achieved.

The invention discloses a method suitable for evaluating the top-free flooding resistance of a tropical water lily stock plant and a fetal seedling, and relates to the field of adversity stress, and the water lily stock plant or the fetal seedling is subjected to top-free flooding treatment with different time lengths. Determining the damage condition by taking the leaf length, the leaf width, the leaf ear length, the leaf area, the stem length, the stem diameter, the chlorophyll content and the leaf cell structure of the normally growing and waterlogged plant as evaluation indexes, and calculating the leaf rot index and the flood tolerance index of the to-be-detected water lily according to the flood damage leaf rot area to realize evaluation. According to the method, the influence of waterlogging on the external phenotype and internal structure of the water lily is considered, the difference of waterlogging treatment on plant ages is considered, the waterlogging resistance is determined through phenotype and cell structure changes, the waterlogging resistance level of the tropical fetal water lily is determined, and the method has the remarkable advantages of being short in evaluation period, visual, reliable and the like. The method is beneficial for promoting innovation of water lily flooding-resistant germplasm and breeding of a flooding-resistant fetal water lily variety, and a new method is provided for research of a water lily flooding-resistant mechanism.

The application provides a crop water requirement dynamic calculation method and an intelligent irrigation system, and relates to the technical field of agricultural irrigation.The crop water requirement dynamic calculation method adopts a quantum dot fluorescence probe array to accurately obtain soil water activity with a 0.1mm spatial resolution, and the accuracy is improved by 3-5 times compared with that of a traditional sensor; terahertz spectrum technology is used to analyze leaf cell sap osmotic pressure, so that nondestructive detection of crop physiological water requirement signals is realized, destructive sampling is avoided, a multi-modal fusion model is used in combination with transfer learning and a quantum correction factor, the water requirement calculation error is less than or equal to 3%, the dynamic response speed is improved to the hour level, the blockchain technology ensures data authenticity, the flow control accuracy of a magneto-rheological actuator reaches + / - 2% FS, and the water saving rate is more than 30%.

The invention relates to the technical field of plant genetic engineering, and particularly discloses a radix bupleuri instantaneous transformation system and application thereof.The radix bupleuri instantaneous transformation system comprises an agrobacterium dip dyeing solution containing a target gene, and the agrobacterium dip dyeing solution is obtained by resuspending an agrobacterium culture containing the target gene through a 2-morpholine ethanesulfonic acid buffer solution; the OD600 of the agrobacterium dip dyeing liquid is 0.8 to 1.2. According to the radix bupleuri transient transformation system constructed by the invention, radix bupleuri leaf cell tissues are efficiently impregnated with agrobacterium tumefaciens, so that efficient transient expression of exogenous genes in radix bupleuri is realized. The operation process can be directly injected, the expression level is high, and the result is reliable.

The present invention relates to a method for mass-producing a target protein in a plant and, more specifically, to a method for producing high levels of protein in plant leaf cells, in which an expression cassette construct for high expression of a target gene and an expression cassette construct for high expression of p38 as a gene silencing suppressor, are prepared, a binary vector for transient expression in plant cells, having the construct together, is then prepared, and the binary vector is then subjected to agrobacterium-mediated transformation by using a single agrobacterium culture.

The present application relates to the field of plant molecular biology and genetic engineering technology, and particularly relates to a transient expression method of orchid plants and application. The method provided by the present application successfully breaks through the infection barrier of the thick cuticle of orchid plants by optimizing the core parameters of vacuum-assisted Agrobacterium infection, screening the infection strains and optimizing the composition of the infection solution. The high activity of the leaf cells can be maintained for more than 9 days after infection, and the exogenous genes can realize stable and efficient expression. The method provided by the present application is simple to operate, does not require special instruments and equipment, is convenient to obtain materials, is not limited by the flowering season, can be carried out throughout the year, and the complete experimental cycle can be completed within 7 days. The present application effectively solves the long-standing technical problems in the field of genetic transformation of orchid plants, and provides an efficient and convenient gene function transient verification technology platform for the functional genomics research, gene function verification, promoter activity detection and precise molecular breeding of orchid and other orchid plants.

The invention relates to a biological agent for controlling heavy metal ions and a preparation method thereof, and belongs to a biological agent. The biological agent is prepared from 50-90 kg of glucose powder, 260-350 kg of apple paste, 8-15 kg of 2000-mesh silicate mineral powder, 8-15 kg of EM microbial agent and 800-1200 L of water. The biological agent for controlling the heavy metal ions is reasonable in compatibility, and after the biological agent is sprayed to crop leaves, silicate mineral powder can directly enter leaf cells and then enter stem conduit tissues and root conduit tissues through leaf veins (leaf conduit tissues); heavy metal ions absorbed by a crop root system are rapidly adsorbed by silicate mineral powder distributed in root, stem and leaf conduit tissues, the silicate mineral powder becomes positively charged ionic groups, and the positively charged ionic groups and a large number of negatively charged phosphate radical ions absorbed by the root system in the root, stem and leaf conduit tissues are subjected to chelation reaction; the large molecular group is fixed in root and stem leaf duct tissues and cannot be transferred to the grains, so that the grains do not contain heavy metal elements.

The application describes a large scale process for the in vitro production of an olive cell culture. The application further describes a composition in a form of a powder comprising olive fruit / leaf cells grown in vitro. The cell culture of olive cells manufactured in vitro in a large scale according to the process of the invention comprises high level of verbascoside with undetectable levels of tyrosol, hydroxytyrosol, and oleuropein.

The application discloses a method for increasing drought adaptability of plant osmotic regulation substance accumulation, and belongs to the field of plant genetic engineering and drought-resistant molecular breeding. First, a plant to be transformed is combined with 13C-glutamic acid and 15N-choline double isotope labels to quantize carbon and nitrogen flow redistribution of proline synthesis pathways and glycine betaine synthesis pathways in leaf cells and vascular bundle companion cells, and to identify rate-limiting enzyme nodes. Second, a multi-cistronic plant expression vector is constructed. A first expression box is driven by a leaf-specific promoter and a drought response cis-element composite promoter to drive P5CS. A second expression box is driven by a companion cell-specific promoter and a drought response cis-element composite promoter to drive BADH. Positive transformants are obtained through Agrobacterium mediation or gene gun transformation and molecular screening. The expression quantity of the positive transformants is increased by 5 to 50 times under stress and falls back to a baseline within 72 hours of rehydration. Root activity is significantly improved, and yield is preserved.

The invention relates to the technical field of plant tissue culture, in particular to a medium-grain coffee embryogenic callus induction method, which comprises the following steps of: firstly, treating substances such as 5-azacytidine in darkness in an epigenetic reprogramming period; then, in a dedifferentiation stage, inducing callus through hormones such as 2, 4-D and the like; then, in the embryogenic induction period, the ratio of hormone to antioxidant is adjusted, and embryogenic differentiation is promoted; and finally, in an embryogenic maintenance period, adding a plurality of antioxidant and active substances, and optimizing culture conditions. According to the method, through multi-stage and multi-component fine regulation and control, the high-quality embryonic callus with the induction rate as high as 98%-99% can be obtained, the epigenetic regulation and control network effectively breaks through the epigenetic obstacle of differentiated leaf cells, and the induction rate is increased to 98%-99% from 30%-50% in the prior art.

The treatment method of the ginseng stem and leaf cell exosome-like vesicles provided by the embodiment of the invention comprises the following steps: step 1, crushing ginseng stems and leaves to prepare homogenate; step 2, sequentially carrying out multi-stage purification on the homogenate to obtain a vesicle layer; 3, based on the vesicle layer, stabilizing treatment is carried out, freeze-dried powder of the ginseng stem and leaf cell exosome-like vesicles is obtained, the freeze-dried powder of the ginseng stem and leaf cell exosome-like vesicles is obtained through stabilizing treatment in the step 3, and the technical effect that a high-stability product is obtained after the ginseng stem and leaf cell exosome-like vesicles are purified is achieved. Technical support is provided for batch production of high-quality ginseng stem and leaf extracellular vesicle products, and the method can be applied to industrial production.

A fluid sensor device (52) for measuring properties of a fluid is disclosed. The fluid sensor device includes a leaf cell sensor having a piezoelectric structure acting on a subdomain of the fluid that flows through the piezoelectric structure to create an intrinsic Helmholtz cavity response, and an enclosure enclosing the leaf cell sensor and including (i) a flowthrough shroud (501) having an inlet that allows the fluid to enter the enclosure and pass across the leaf cell sensor, and a Helmholtz cavity wall (502) that couples the intrinsic Helmholtz cavity response with an external acoustic field of the leaf sensor to increase a measurement sensitivity, (ii) a cylindrical housing having an outlet that allows the fluid to exit the enclosure, and (iii) a pressure feedthrough connector (507) that transmits an electrical signal induced by the intrinsic Helmholtz cavity response to represent the properties of the fluid.

The invention relates to the field of tea processing, in particular to black tea with a high amadol rearrangement product and a preparation method of the black tea. According to the preparation method of the black tea enriched with the Amadol rearrangement product, provided by the invention, the black tea enriched with the Amadol rearrangement product is obtained by performing pulse type light supplementing withering, rolling, fermentation and drying on high-amino-acid fresh tea leaves. Wherein pulse type light supplement in the withering process activates the activity of protease and amylase on one hand, and can increase the leaf cell breakage rate on the other hand, so that enrichment of theanine and soluble sugar in the leaves is fully promoted, and sufficient precursor substances are provided for subsequent Maillard reaction; therefore, the content of the amadol rearrangement product enriched in the finally obtained black tea is increased.

The present invention relates to a processing method of special flavor Kung Fu black tea, comprising: (1) selecting fresh leaves of small and medium leaf tea varieties suitable for making black tea as raw materials; (2) picking fresh leaves of small and medium leaf tea varieties from late March to early May; (3) performing temperature, light and wind withering treatment; (4) shaking treatment; (5) performing secondary withering; (6) ultrasonic crushing to promote the crushing of leaf cell membranes and vacuole membranes, and promote the full biochemical reaction between enzymes in the leaves and substrates (7) performing variable frequency short-time cold rolling; (8) performing low-temperature medium-humidity dynamic fermentation; (9) performing a primary drying operation by roller-hot air coupled rolling drying; (10) conveying the primary dried leaves treated in step (9) to a cooling room by a conveyor belt, and removing broken leaves and yellow leaves by a winnowing machine; (11) performing a low-temperature long-time hot air sufficient drying operation; (12) performing a temperature-controlled and humidity-controlled far-infrared aroma treatment on the tea sample obtained by the treatment in step (11), and after completion, a sweet and mellow black tea is obtained.