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1、植物的组织培养（Tissue culture of plants）Advantages of plant tissue culture1, small space, free from regional and seasonal restrictions.2. Cultivate virus-free crops3 、 short training period4. Special biochemical products can be obtained by using callus from tissue culture5. It can be propagated in short ti
2、me to save endangered plants6. Differentiate into necessary organs, such as roots and shoots7, solve some plants produce seeds of little or no problem,8, there is no variation, which can maintain all the genetic characteristics of the original female parent9, less investment, high economic benefits1
3、0, breeding methods, more varietiesTechnical terms for plant tissue cultureChinese Name: English Name: plant tissue culture plant tissue culture definition: in culture medium containing nutrients and plant growth substances in vitro plant tissues (organs or cells) and induced to grow into a complete
4、 plant technology. Subject: Cell Biology (first level discipline), cell culture and cell engineering (two discipline). This content is examined and approved by National Committee of scientists and techniciansmedical aircraftPlant tissue culture is a new technique of asexual reproduction based on the
5、 theory that plant cells are totipotent. Plant tissue culture is generalized in vitro, to meet the needs of the organization were isolated from plants. Organs or cells, protoplasts, through the aseptic operation, other products training to get the regeneration plants or production with economic valu
6、e under the condition of artificial control technology. The narrow sense refers to the plant tissue culture refers to the part of the organization, such as the formation of layers. The parenchyma. Mesophyll tissue. Endosperm culture regeneration, also refers to the callus culture generated from diff
7、erent organs in the process of culture, callus differentiation after plant regeneration.CatalogTheoretical originMethod for culturing plant tissueAdvantages of plant tissue culturePlant tissue culture is generally divided into the followingCollection of culture materialsPlant tissue culture stepThe
8、characteristics of plant tissue culture 1. The culture conditions can be controlled artificially2, short growth cycle, high reproduction rate3, convenient management, conducive to factory production and automatic controlTheoretical originMethod for culturing plant tissueAdvantages of plant tissue cu
9、lturePlant tissue culture is generally divided into the followingCollection of culture materialsPlant tissue culture stepThe characteristics of plant tissue culture 1. The culture conditions can be controlled artificially2, short growth cycle, high reproduction rate3, convenient management, conduciv
10、e to factory production and automatic controlEdit this paragraph, theoretical originIn 1830s, German botanist Schleiden and German zoologist who founded the Schwann cell theory, according to this theory, if cells and organisms like conditions, each cell should be able to live independently. In 1902,
11、 the theory of cell pluripotency by Haber Lent, a German botanist, was the theoretical basis for plant tissue culture. In 1958, an exciting news from the United States to the world, American botanist Christian Watt et al., with the carrot phloem cells were cultured, finally got the complete plant, a
12、nd the plant can blossom, confirmed Haberlandt more than 50 years ago on cells that. A simple process of plant tissue culture are as follows: splicing plant organs or tissues after dedifferentiation (also called dedifferentiation) - after the formation of callus differentiation of plant tissues or o
13、rgans after cultured and developed into a complete. The general process of plant tissue culture: under sterile conditions, the plant organs or tissues (such as bud, stem and root tips or anther) part of cutting down on artificial culture medium suitable for culture, these organs or tissues will unde
14、rgo mitosis, the formation of a new organization. However, this tissue does not differentiate but is a mass of parenchyma cells called calluses. Under suitable conditions such as light, temperature and certain nutrients and hormones, the calli begin to differentiate, producing various organs and tis
15、sues of the plant, and then developing into a complete plant. Plant sterile culture technique of plant tissue culture, plant cell totipotency is based on the theory of the plant body from the body organs such as root, stem, leaf, stem apex, flower and fruit etc.) organization (such as the formation
16、of layers, epidermis, cortex and pith cells of endosperm cells, etc.) or (such as the megaspore, microspore, somatic cells) and protoplasts in sterile and suitable artificial culture medium and light and temperature under artificial conditions could induce callus, adventitious buds, and finally form
17、 a complete plant subjectEdit this section of plant tissue culture methods1, the rapid propagation of non tube tissue micro tube rapid propagation technology of micro organization is the explants (leaf bud with a general requirements) placed in the room outside the ordinary sand culture medium, usin
18、g natural plant bud multiplication achieve the purpose of rapid propagation. A common plant can grow roots in 715 days. This technology has low investment and little operation. 2 、 tissue culture test tube tissue culture is the explant (namely, tissue, organ or cell) placed in test tubes and other c
19、ontainers, in sterile conditions for tissue culture, to obtain test tube seedlings.Edit the advantages of plant tissue culture in this paragraph1, small space, free from regional and seasonal restrictions. 2, cultivate virus-free crop culture to produce callus, 3 short cycle 4, available in the tiss
20、ue culture of special biochemical products 5, a short time to multiply, to save endangered plants, can induce the differentiation of the 6 needs of organs, such as roots and shoots 7, solve the problem of some plants produce seeds with little or no. 8, there is no variation, can keep the original pa
21、rent all the genetic characteristics of 9, less investment, high economic benefit, 10 modes of reproduction, variety trialEdit this paragraph, plant tissue culture is generally divided into the following1. Embryo culture refers to the in vitro axenic culture of mature or immature embryos isolated fr
22、om ovules. 2, organ culture refers to the plant roots, stems, leaves, flowers, fruit and other organs for in vitro aseptic explants, such as root and apical segments of the stem, stem apex, stem and leaf segments, leaf primordium, leaf, petiole, leaf and cotyledon, flower petals, stamens (anther and
23、 filament) and in vitro aseptic culture of ovule and ovary and fruit etc. Refers to the separation of the part of the plant tissue, 3 tissue culture (such as meristem, cambium, xylem, phloem, epidermis, cortex and endosperm tissue, pith parenchyma, etc.), or have induced callus as explants in vitro
24、aseptic culture. This is the narrow sense of plant tissue culture. 4. Cell culture refers to the in vitro axenic culture of individual free cells, such as the somatic cells separated from the tissues or the pollen cells, and the egg cells. 5. Protoplast culture. The axenic culture of protoplasts tha
25、t take the cell wall as explants.Edit this paragraph and collect the materialWe should select materials according to the purpose of training and select the principles: easy to induce and less bacteria. Select the sterile material inside the plant tissue. In this respect, materials should be taken fr
26、om sturdy plants, not with wounds or pests. On the other hand, on sunny days, it is better to collect materials at noon or in the afternoon, and never pick up materials on rainy days, cloudy days or wet dew. The tissues are generally sterile because they are self - disinfecting by vigorous plants an
27、d the tissues that breathe in clear weather. The disinfection of culture materials, plant materials selected from outside or inside, all sorts of microorganisms are found in varying degrees. Once these sources are contaminated with a culture medium, they can cause contamination of the culture medium
28、. Therefore, the plant material must be treated with a strict surface sterilization process and then sterilized to the culture medium. Reagent ethanol. Indole acetic acid (IAA) or 2, 4 - D (auxin analogs). Mercuric chloride (HgCl2) or sodium hypochlorite?. 6-? Benzylamino adenine (6-BA)? MS medium?
29、0.1 mol/L NaOH and 0.1 mol/L HCl medium preparation (1) callus inducing medium (MS medium sucrose content was 10 g/L, 2,4, D content is 2 mg/L, agar 10 g/L). (2) test medium: add IAA and 6 - BA in MS medium. Indole acetic acid (IAA) was first dissolved with a small amount of 0.1 mol/L NaOH, and 6- b
30、enzyl adenine was first dissolved with a small amount of 0.1 mol/L HCl, then diluted with distilled water and then added to the medium. Equipment training room, autoclave, bath, scalpel, flask, beaker (100mL), a Petri dish, cotton, inoculation box or super clean bench, analytical balance, long tweez
31、ers, scissors, flask, pipette, kraft paper. The medium was sterilized and the medium was added to the agar. The heat was dissolved and transferred to pH 5.8. The mixture was packed in 100 mL triangle flask and served at about 20 mL per bottle. The cooling medium solidified with a layer of a layer of
32、 kraft paper and weighing up the bottle (tube), and cotton string, and then in the autoclave at 121 (1 kg/cm 2) under 20 min sterilization. Remove the flask and place it on the table for cooling. All the equipment needed for inoculation (such as forceps, scalpel, scissors, etc.) and sterile water sh
33、ould be sterilized at the same time.Edit the plant tissue culture steps in this paragraphThe first step is to remove the unused material from the plant and carefully clean the needed parts, such as using a suitable brush. Cut the material into proper size, that is, the sterilizing container can be p
34、lanted properly. Wash the water under the tap water for a few minutes to a few hours, the washing time depends on the cleanliness of the material. Easy floating or small material, it can be washed in gauze bag. Running water is especially useful when polluted. When washing, you can add washing powde
35、r to wash, then rinse with clean water. The washing powder can remove the dirt which is slightly attached to the surface of the plant and remove the lipid substance so as to facilitate the direct contact of the sterilizing liquid. The ideal cleaning substance, of course, is surfactant Twain. The sec
36、ond step is to sterilize the surface of the material. To be completed in a super clean or inoculation box, prepare sterilized beaker, glass rods, 70% alcohol, disinfectant, sterile water, watches, etc. Soak 70% alcohol for 1030 seconds. Because alcohol has the effect of soaking the surface of plant
37、material, plus 70% alcohol penetrating power, it is also easy to kill plant cells, so the infiltration time can not be too long. There are some special materials, if real, bud, bag piece, Youbao bract at booting, multi scales of dormant buds and so on, as well as the main access to the interior of t
38、he material and is only 70% alcohol treatment time longer. The treated material is sterile and then stripped of the outer layer prior to evaporation of the alcohol. Internal material is used. The third step is to deal with the disinfectant. There are many kinds of surface sterilization agents, and 1
39、 - 2 kinds can be chosen according to the conditions. The fourth step is rinsed with sterile water, rinsed to each 3min, as the type of disinfectant, washing 3-l0 times. Sterile water washing is pay attention to side effects from plant cell killing disinfectant: alcohol permeability, tender easy mat
40、erial in alcohol chlorosis, so the soaking time should be short, to prevent alcohol kills plant cells. Aging materials, especially seeds, can be soaked in alcohol for a longer time, for example, seeds can be soaked for 5 minutes. The penetration of mercuric chloride is weak, generally soak for 10 mi
41、nutes or so, the plant material is not lethal. Bleaching powder easily leads to chlorosis of plant materials, so be careful with young materials. Adding Twain 20 or 80 (a wetting agent) with a concentration of 0.08 - 0.12% (a wetting agent) in the disinfectant solution can reduce the tension on the
42、surface of the plant material and achieve better disinfection effect.Edit the features of plant tissue culture in this paragraph1, the training conditions can be artificially controlledThe tissue culture of plant material is completely grown in medium and small man climate conditions, to get rid of
43、the adverse effects of changes in the nature of the four seasons, day and night, weather conditions, and even, on the growth of plants is favorable for stable annual production training.2, short growth cycle, high reproduction ratePlant tissue culture is due to the artificial control of culture cond
44、itions, according to different plants, different parts of the different requirements and provide different culture conditions, so the growth is faster. In addition, the plant is relatively small, often 20-30 days for a cycle. So, although the need for certain equipment and energy consumption of plan
45、t tissue culture, but because the plant material can produce according to geometric reproduction, so the overall cost is low, and can provide the same standard or high quality seedling virus-free seedling.3, convenient management, conducive to factory production and automatic controlPlant tissue cul
46、ture is in a certain place and environment, provide some artificial light, temperature, humidity, nutrition, hormones and other conditions, very conducive to highly intensive and high density of factory production, but also conducive to automatic control production. It is the development direction o
47、f industrialized farming in the future. Compared with pot planting and field cultivation, the utility model omits a series of complicated labor such as weeding, weeding, watering, fertilizing, preventing and controlling diseases and insect pests, and can greatly save manpower, material resources and land needed for planting in the field.9, 12, 15, 22, 5, 25, 15, 21