CN106717260B - Gentiana scabra seedling growing method and Gentiana scabra planting method - Google Patents
Gentiana scabra seedling growing method and Gentiana scabra planting method Download PDFInfo
- Publication number
- CN106717260B CN106717260B CN201611072300.6A CN201611072300A CN106717260B CN 106717260 B CN106717260 B CN 106717260B CN 201611072300 A CN201611072300 A CN 201611072300A CN 106717260 B CN106717260 B CN 106717260B
- Authority
- CN
- China
- Prior art keywords
- seedlings
- gentiana
- seeds
- culture medium
- illumination
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
- A01G9/14—Greenhouses
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/25—Greenhouse technology, e.g. cooling systems therefor
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Soil Sciences (AREA)
- Botany (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention relates to a seedling raising method of gentiana and a planting method of gentiana, which belong to the field of plant production, wherein sodium hypochlorite solution with the mass concentration of 10-20% is used for peeling and washing, peeled seeds are inoculated in MS culture medium containing perlite, indoleacetic acid with the mass concentration of 0.1-0.5mg/L, 6-benzylaminopurine with the mass concentration of 0.5-1mg/L and EM bacteria with the mass concentration of 0.01-0.05mg/L, the culture temperature is 15-25 ℃, the primary culture is carried out by irradiating at 3000lx illumination intensity of 2000-plus-energy for 10-12h every day, the obtained aseptic seedlings are inoculated in a secondary culture medium, the obtained rootless seedlings are transferred to a rooting culture medium to obtain gentiana seedlings, the training seedlings are cultivated in a greenhouse after the gentiana seedlings are trained, and the irradiation is carried out at 3000lx illumination of 2000-plus-energy for 10-12h every day, the mature gentiana scabra bunge is obtained by matching with field management, and the emergence rate and the seedling survival rate of the gentiana scabra bunge are high.
Description
Technical Field
The invention relates to the field of plant production, and particularly relates to a gentian flower seedling raising method and a gentian flower planting method.
Background
Gentian flower, named separately: elephantopus scaber, gentian, and kwan-yin herb. Gentiana scabra Bunge flower is bitter and cold, the bitter taste of traditional Chinese medicine is second to that of nux vomica, is superior to that of coptis root, is second to live, has multiple purposes of stomach destruction, is not easy to enter the mouth, is supplemented with liquorice to adjust the taste, and is commonly used for treating intractable migraine, head eczema, hypertension, acute conjunctivitis, rhinitis and the like.
Because the gentian flower has excellent medicinal value, the demand of the gentian flower in the market is continuously increased, the demand of the market is met only by picking wild gentian flower, the picking amount cannot meet the demand, and the survival risk of the gentian flower can be caused by excessive picking. In addition, the gentian peanuts grow in regions with higher altitude, the growing conditions are strict, the artificial seedling raising and cultivation difficulty is high, and the yield of the gentian peanuts cannot meet the market demand.
Disclosure of Invention
The invention aims to provide a seedling growing method of gentiana scabra bunge, which is characterized in that after seeds of gentiana scabra bunge are peeled, the seeds are artificially helped to break the peels, so that the germination time of the seeds is shortened; perlite is added into the primary culture medium to ensure that the seeds can breathe well in the primary culture medium, and the pH of the primary culture medium is kept at proper acidity, which is more favorable for the emergence of the seeds; the EM bacteria can make the aseptic seedlings stronger and increase the survival rate; after subculture and rooting culture, the culture time of the gentian flower seedlings from seeds to seedlings is short, and the emergence rate and the survival rate can be effectively improved.
Another object of the present invention is to provide a method for planting gentiana scabra bunge, which is characterized in that after seedlings of gentiana scabra bunge are acclimated, the seedlings of gentiana scabra bunge are more easily adapted to a planting environment, the survival rate of the seedlings of gentiana scabra bunge is increased, the planting yield of gentiana scabra bunge is increased, market demands are met, the demand pressure of wild gentiana scabra bunge is reduced, and extinction of gentiana scabra bunge is avoided to a certain extent.
The technical problem to be solved by the invention is realized by adopting the following technical scheme.
A seedling growing method of gentian flower comprises the steps of soaking seeds of gentian flower in a sodium hypochlorite solution with the mass concentration of 10% -20% for peeling, and washing the seeds with sterile water after peeling to obtain peeled seeds; inoculating the peeled seeds into a primary culture medium, and carrying out primary culture to obtain aseptic seedlings; taking cotyledons and hypocotyls of the aseptic seedlings as explants, inoculating the explants to a subculture medium, and carrying out subculture to obtain rootless seedlings; transplanting the rootless seedlings into a rooting culture medium, and carrying out rooting culture to obtain gentian flower seedlings; the primary culture temperature is 15-25 ℃, the illumination intensity is 2000-3000lx, the illumination time is 10-12h, and the primary culture medium is an MS culture medium added with perlite, indoleacetic acid with the mass concentration of 0.1-0.5mg/L, 6-benzylamino adenine with the mass concentration of 0.5-1mg/L and EM bacteria with the mass concentration of 0.01-0.05 mg/L.
A planting method of gentian flower is characterized in that gentian flower seedlings are cultured and acclimatized according to the gentian flower seedling culture method to obtain acclimatized seedlings; planting the domesticated seedlings in a greenhouse, controlling the illumination time in the greenhouse to be 10-12h and the illumination intensity to be 2000-3000lx, and matching with field management to obtain mature gentiana flowers.
The seedling raising method and the planting method of the gentian flower provided by the embodiment of the invention have the beneficial effects that: the rate of budding of the gentian flower seeds is accelerated by peeling the gentian flower seeds, and the primary culture medium is added with perlite, so that the respiration of the seeds can be facilitated, the appropriate acidity of the pH value of the primary culture medium is ensured, and the budding of the seeds is further accelerated; the EM can make the aseptic seedlings stronger and increase the survival rate of the aseptic seedlings; under the action of a series of tissue culture of subculture and rooting culture, the emergence rate and the survival rate of the gentian flower seeds are improved, and the adaptability of gentian flower seedlings is enhanced; the gentian flower seedlings are domesticated and then planted, the survival rate of the gentian flower seedlings can be further improved, the yield of gentian flowers is increased, the market demand is met, the picking amount of wild gentian flowers is reduced, and the danger of extinction of the gentian flowers is avoided.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The method for growing seedlings of gentiana scabra bunge and the method for planting gentiana scabra bunge according to the embodiment of the present invention will be specifically described below.
The embodiment of the invention provides a seedling raising method of gentiana scabra bunge, which is characterized in that seeds of gentiana scabra bunge are peeled by using a sodium hypochlorite solution, and the seeds are washed by using sterile water after peeling to obtain peeled seeds; and (3) carrying out primary culture, secondary culture and rooting culture on the peeled seeds to obtain the gentian flower seedlings.
The embodiment of the invention also provides a planting method of gentiana straminea, which is characterized in that domesticated seedlings are obtained after the gentiana straminea seedlings are domesticated; planting the domesticated seedlings in a greenhouse, controlling the illumination time in the greenhouse to be 10-12h and the illumination intensity to be 2000-3000lx, and matching with field management to obtain mature gentiana flowers.
Specifically, the seedling raising method of the gentian flower and the planting method of the gentian flower are carried out according to the following steps:
s1 peeling step:
placing seeds of gentian flower into a container capable of being sealed, such as: and (3) a PVC plastic bottle with a bottle cap, pouring a sodium hypochlorite solution with the mass concentration of 10% -20% into the container filled with the seeds, sealing the container, and soaking the seeds in the sodium hypochlorite solution with the mass concentration of 10% -20% for 5-10 min. The sodium hypochlorite solution can be used for removing the peel of the seed coat, sterilizing the seed, ensuring the successful tissue culture process and ensuring the higher rate of emergence of the seed.
Preferably, in-process soaking, can shake above-mentioned dress seed and sodium hypochlorite solution container, shake and sodium hypochlorite solution's dual function down, the not hard up degree of the crust of seed can increase to some extent.
Furthermore, before the sodium hypochlorite solution is poured into the container, a plurality of stones can be placed into the container, after the sodium hypochlorite solution is injected into the container and the container begins to vibrate, the stones inside can vibrate along with the vibration force, and in the collision of the stones and the seeds, part of seed coats can be loosened more quickly, so that the subsequent work of completely removing the seed coats is facilitated.
Specifically, after soaking, removing the sodium hypochlorite solution in the container, placing the soaked seeds in a mesh screen, washing the seeds with sterile water, pressing the seeds on the mesh screen for kneading while washing the seeds with sterile water, so that the outer skins of the seeds quickly fall off, and after the outer skins of the seeds fall off, continuously washing with sterile water for 20-30min to obtain the peeled seeds. The seeds soaked in the sodium hypochlorite solution are washed for a long time by using sterile water, so that the seeds can be effectively prevented from being damaged by the sodium hypochlorite, and a higher germination rate is ensured.
More preferably, the peeled seeds can also be laid on absorbent paper and sent into an ultraviolet sterilization box for ultraviolet induction, the wavelength of the ultraviolet induced ultraviolet is 250-254nm for example, and the time for ultraviolet induction is controlled to be 2-3 h. Through the ultraviolet induction, the peeled seeds can be further sterilized and disinfected, and the emergence rate of the seeds can be further improved.
S2 primary culture:
and (4) inoculating the peeled seeds on a primary culture medium, and carrying out primary culture to obtain aseptic seedlings.
The primary culture medium is prepared by adding perlite, indoleacetic acid (IAA) with mass concentration of 0.1-0.5mg/L, and 6-benzyl alcohol with mass concentration of 0.5-1mg/L into MS culture mediumAmino adenine (GA)3) 0.01-0.05mg/L EM bacteria, wherein the addition amount of the perlite is 10-20g added in 1L MS culture medium, and the particle size of the perlite is 6-8 mm; the MS culture medium is a commercial MS culture medium containing sucrose and agar. The indoleacetic acid can stimulate the division of cambium cells, stimulate the cell elongation of branches, inhibit the cell growth of roots, regulate the morphogenesis of callus, promote the quick germination of peeled seeds when being added into an MS culture medium, and improve the germination rate of the peeled seeds, but the control on the amount of the indoleacetic acid added is important, and the germination of the seeds can be inhibited if the amount of the indoleacetic acid added is too large; the 6-benzylamino adenine can improve differentiation and stress resistance and promote germination of seeds, and the 6-benzylamino adenine added in the MS culture medium is more than the indoleacetic acid so as to further promote the germination of peeled seeds; the EM bacteria comprise beneficial bacteria such as spores, saccharomycetes, lactic acid bacteria and the like, and can enhance the metabolism of plants, promote aseptic seedlings to be stronger and improve the survival rate; the perlite added in the culture medium can provide enough oxygen for the seeds inoculated in the culture medium, so that the seeds can be inoculated in the culture medium to breathe enough, if the seeds cannot breathe normally, the germination rate can be reduced, the pH acidity of the primary culture medium can be maintained, and the germination of the seeds is further promoted. When the indoleacetic acid, the 6-benzylamino adenine and the EM bacteria are mixed for use, the influence of each substance on the seed germination rate is improved, the survival rate of aseptic seedlings is increased, and the germination rate of peeled seeds is further improved.
The culture temperature during primary culture is 15-25 ℃, the illumination intensity is 2000-3000lx, the illumination time is 10-12h, and the environmental humidity during culture is 55-75%. Because the condition requirement of the growth of gentian flower to humidity and illumination is higher, so in order to improve the germination rate of seed when carrying out the primary culture, regulate and control the condition of cultivateing into the climatic condition that accords with its growth, temperature when the primary culture promptly is warm, and the illumination is less strong, and the illumination time is longer, and ambient humidity is higher, promotes the germination rate of seed.
S3 subculture:
inoculating cotyledon and hypocotyl of the aseptic seedling obtained after primary culture as an explant into a subculture medium, and carrying out subculture to obtain a rootless seedling.
Cutting cotyledon of aseptic seedling in primary culture to remove leaf tip, leaving leaf stalk, and cutting the leaf stalk into small blocks of 0.3cm × 0.3 cm; cutting hypocotyls of the aseptic seedlings into small sections of 0.3 cm; inoculating the cotyledon blocks and the hypocotyl sections on a subculture medium for subculture. The cotyledon and hypocotyl of the aseptic seedling developed from the seed are cut and then cultured again, so that the yield of the gentian flower seedling can be effectively increased, a plurality of high-quality seedlings can be cultured from one seed, and the yield of the seedling is greatly increased.
The subculture medium may be, for example, MS medium supplemented with 6-benzylamino adenine (6-BA) at a mass concentration of 0.1-0.5mg/L and Gibberellin (GA) at a mass concentration of 1-1.5mg/L3) And 0.5-1mg/L of indoleacetic acid (IAA), wherein the MS culture medium is a commercially available MS culture medium containing sucrose and agar. The 6-benzylamino adenine can promote cell division, increase cell growth and weight gain, promote the elongation growth of stems and leaves, inhibit aging, improve the damage resistance of plants and the like, and in the culture of the explant, the 6-BA is added to effectively improve the differentiation and stress resistance of the explant, so that the emergence rate of the explant can be improved; the IAA can stimulate the division of cambium cells, stimulate the cell elongation of branches, inhibit the cell growth of roots, regulate the morphogenesis of calluses, promote the rapid formation of rootless seedlings and improve the rate of emergence when added into an MS culture medium; GA3Accelerate cell division, promote the elongation growth of stems and leaves and accelerate the growth of rootless seedlings. Under the action of the three phytohormones, the phytohormones can act synergistically on the emergence of the explants, so that the emergence rate of the explants is further improved.
The culture temperature of the subculture is preferably 15-25 ℃, the illumination intensity is 2000-3000lx, the illumination time is 10-12h, and the culture environment humidity is 55-75%, and the temperature, the illumination condition and the environment humidity are close to the warm, weak-light and relatively humid environment condition of the original place of the gentian flower, so that the emergence rate of the explant can be ensured, the resistance of the rootless seedling cultured from the explant is improved, and the rootless seedling is stronger in growth vigor and higher in survival rate.
S4 rooting culture:
and (4) transplanting the rootless seedlings obtained by the subculture into a rooting culture medium, and carrying out rooting culture until the gentiana scabra seedlings are obtained.
The subculture is carried out by adding naphthylacetic acid (NAA) with a mass concentration of 1-1.5mg/L and Kinetin (KT) with a mass concentration of 0.5-1mg/L into MS culture medium, which is also commercially available MS culture medium containing sucrose and agar. NAA is a plant growth regulator for promoting the growth of plant root systems, has the functions of promoting cell division and expansion, and can enable rootless seedlings to grow adventitious roots quickly. KT can not only promote cell division, but also induce the differentiation and development of buds and increase the stomatal aperture, thereby promoting the carbon dioxide absorption of plants, increasing the photosynthesis of the plants, and facilitating the growth and rooting of the rootless seedlings with more nutrients.
The temperature during rooting culture is 15-25 ℃, and shading treatment is carried out before the rootless seedlings grow roots, so that the rootless seedlings can root, and the environmental humidity during rooting culture is 55-75%; meanwhile, the lower temperature and humidity conditions are close to the environment with higher altitude of the original place of the gentian flower, so that rooting of rootless seedlings is facilitated, stress resistance of the seedlings can be improved, and the survival rate of the seedlings is increased.
S5 planting gentian flower:
domesticating the gentiana scabra bunge seedlings cultivated according to the method to obtain domesticated seedlings, planting the domesticated seedlings in a greenhouse, and performing a series of field management to obtain mature gentiana scabra bunge.
The domestication refers to planting the gentiana seedlings in an illumination incubator, setting the illumination intensity at 2000-. Resistance training through carrying out high humidity environment and low humidity environment alternate cultivation to the seedling makes the gentian flower can adapt to the relatively higher growing environment of humidity in low altitude area and the relatively lower growing environment of humidity in high altitude area simultaneously, further promotes the survivability of gentian flower seedling after getting into the big-arch shelter and planting. The illumination conditions are close to the illumination conditions of the original place of the gentian flowers, the vigorous growth vigor of the gentian flower seedlings is guaranteed, the survival rate of transplanted seedlings is improved, and the gentian flower seedlings can better adapt to resistance training with different humidity. After the gentian flowers are transplanted into a greenhouse, the stock rate of the gentian flowers subjected to resistance training under different humidities is higher, and therefore the yield of the gentian flowers is increased.
When the gentian flower is planted in the greenhouse, the illumination intensity is preferably set to be 2000-3000lx, the illumination time is 10-12h, and farmyard manure, weed removal and other work can be matched, so that the gentian flower can grow more vigorously, and the yield of the gentian flower can be improved.
The method for growing seedlings of gentian flower and the method for growing gentian flower according to the present invention are described in further detail below with reference to examples.
Example 1
Filling gentiana scabra bunge seeds into a PVC plastic bottle, putting a small amount of stones into the PVC plastic bottle, then injecting a sodium hypochlorite solution with the mass concentration of 10%, tightly covering a bottle cap, soaking and shaking for 10min, removing the sodium hypochlorite solution, pouring the seeds in the bottle into a mesh screen, washing with sterile water, pressing the seeds on the mesh screen while washing, kneading until the seed coats are completely removed, and continuously washing with the sterile water for 20 min.
The peeled seeds which had absorbed dry water were placed in an ultraviolet sterilizing chamber and treated at a wavelength of 250nm for 3 hours.
The peeled seeds were inoculated into a primary medium containing 0.1mg/L indoleacetic acid, 0.5 mg/L6-benzylamino adenine, 0.05mg/L EM bacteria and 10g perlite having a size of 6mm, and cultured under conditions of a temperature of 25 ℃, an illumination intensity of 2000lx, an illumination time of 10h and a humidity of 55% until obtaining sterile seedlings.
Cutting cotyledon of aseptic seedling into 0.3cm × 0.3cm pieces, cutting hypocotyl into 0.3cm pieces, inoculating into subculture medium containing 0.1 mg/L6-benzylamino adenine, 1mg/L gibberellin and 0.5mg/L indoleacetic acid, respectively, and culturing at 15 deg.C, illumination intensity of 2000lx, illumination time of 12h and culture environment humidity of 55% until no-root seedling is obtained.
Transplanting the rootless seedlings into a rooting culture medium containing 1mg/L of naphthylacetic acid and 0.5mg/L of kinetin, and culturing under the conditions that the temperature is 15 ℃, the environmental humidity is 55 percent and the light is shielded until the gentiana rigescens seedlings are obtained.
And (3) cultivating the gentian flower seedlings in an illumination incubator, wherein the illumination intensity is 2000lx, the illumination time is 12h, the humidity of the illumination incubator is 1h as an interval, and the illumination incubator is alternately arranged according to 70% and 50% until domesticated seedlings are obtained.
And planting the domesticated seedlings in a greenhouse, ensuring that the illumination intensity in the greenhouse reaches 2000lx and the illumination time reaches 10h, and planting mature gentian flowers by matching with the work of applying farmyard manure, weeding and the like.
Example 2
Filling gentiana scabra bunge seeds into a PVC plastic bottle, adding a small amount of stones into the PVC plastic bottle, then injecting a sodium hypochlorite solution with the mass concentration of 20%, tightly covering a bottle cap, soaking and shaking for 5min, removing the sodium hypochlorite solution, pouring the seeds in the bottle into a mesh screen, washing with sterile water, pressing the seeds on the mesh screen while washing, kneading until the seed coats are completely removed, and continuously washing with the sterile water for 30 min.
The peeled seeds which had absorbed dry water were placed in an ultraviolet sterilizing chamber and treated at a wavelength of 254nm for 2 h.
The peeled seeds are inoculated in a primary culture medium containing 0.5mg/L indoleacetic acid, 1 mg/L6-benzylamino adenine, 0.01mg/L EM bacteria and 20g perlite with the size of 8mm, and cultured under the conditions of the temperature of 15 ℃, the illumination intensity of 3000lx, the illumination time of 12h and the humidity of 75 percent until sterile seedlings are obtained.
Cutting cotyledon of aseptic seedling into 0.3cm × 0.3cm pieces, cutting hypocotyl into 0.3cm pieces, inoculating into subculture medium containing 0.5 mg/L6-benzylamino adenine, 1.5mg/L gibberellin and 1mg/L indoleacetic acid, respectively, and culturing at 25 deg.C, illumination intensity of 3000lx, illumination time of 10 hr and culture environment humidity of 75% until no-root seedling is obtained.
Transplanting the rootless seedlings into a rooting culture medium containing 1.5mg/L of naphthylacetic acid and 1mg/L of kinetin, and culturing under the conditions that the temperature is 25 ℃, the environmental humidity is 75% and the light is shielded until the gentiana rigescens seedlings are obtained.
And (3) cultivating the gentian flower seedlings in an illumination incubator, wherein the illumination intensity is 3000lx, the illumination time is 10 hours, the humidity of the illumination incubator is 1 hour at intervals, and the illumination incubator is alternately arranged according to 70% and 50% until domesticated seedlings are obtained.
And planting the domesticated seedlings in a greenhouse, ensuring that the illumination intensity in the greenhouse reaches 3000lx and the illumination time reaches 12h, and planting mature gentian flowers by matching with the work of applying farmyard manure, weeding and the like.
Example 3
Filling gentiana scabra bunge seeds into a PVC plastic bottle, adding a small amount of stones into the PVC plastic bottle, then injecting a sodium hypochlorite solution with the mass concentration of 15%, tightly covering a bottle cap, soaking and shaking for 6min, removing the sodium hypochlorite solution, pouring the seeds in the bottle into a mesh screen, washing with sterile water, pressing the seeds on the mesh screen while washing, kneading until the seed coats are completely removed, and continuously washing with the sterile water for 25 min.
The peeled seeds which had absorbed dry water were placed in an ultraviolet sterilizing chamber and treated at a wavelength of 252nm for 2.5 h.
The peeled seeds are inoculated in a primary culture medium containing 0.3mg/L indoleacetic acid, 0.8 mg/L6-benzylamino adenine, 0.03mg/L EM bacteria and 15g perlite with the size of 7mm, and cultured under the conditions of the temperature of 20 ℃, the illumination intensity of 2500lx, the illumination time of 11h and the humidity of 65 percent until sterile seedlings are obtained.
Cutting cotyledon of aseptic seedling into 0.3cm × 0.3cm pieces, cutting hypocotyl into 0.3cm pieces, inoculating into subculture medium containing 0.3 mg/L6-benzylamino adenine, 1.2mg/L gibberellin and 0.8mg/L indoleacetic acid, respectively, and culturing at 20 deg.C, illumination intensity of 2500lx, illumination time of 11h and culture environment humidity of 65% until no-root seedling is obtained.
Transplanting the rootless seedlings into a rooting culture medium containing 1.2mg/L of naphthylacetic acid and 0.7mg/L of kinetin, and culturing under the conditions that the temperature is 20 ℃, the environmental humidity is 65 percent and the light is shielded until the gentian flower seedlings are obtained.
And (3) cultivating the gentian flower seedlings in an illumination incubator, wherein the illumination intensity is 2500lx, the illumination time is 11h, and the humidity of the illumination incubator is set at intervals of 1h alternately according to 70% and 50% until domesticated seedlings are obtained.
And planting the domesticated seedlings in a greenhouse, ensuring that the illumination intensity in the greenhouse reaches 2500lx and the illumination time reaches 11h, and planting mature gentian flowers by matching with the work of applying farmyard manure, weeding and the like.
Example 4
Filling gentiana scabra bunge seeds into a PVC plastic bottle, putting a small amount of stones into the PVC plastic bottle, then injecting a sodium hypochlorite solution with the mass concentration of 16%, tightly covering a bottle cap, soaking and shaking for 9min, removing the sodium hypochlorite solution, pouring the seeds in the bottle into a mesh screen, washing with sterile water, pressing the seeds on the mesh screen while washing, kneading until the seed coats are completely removed, and continuously washing with the sterile water for 24 min.
The peeled seeds which had absorbed dry water were placed in an ultraviolet sterilizing chamber and treated at a wavelength of 251nm for 2.2 h.
The peeled seeds were inoculated into a primary medium containing 0.2mg/L indoleacetic acid, 0.8 mg/L6-benzylamino adenine, 0.02mg/L EM bacteria and 12g perlite having a size of 6mm, and cultured under conditions of a temperature of 18 ℃, an illumination intensity of 2700lx, an illumination time of 12h and a humidity of 68% until obtaining sterile seedlings.
The cotyledon of the aseptic seedling is cut into small pieces of 0.3cm multiplied by 0.3cm, the hypocotyl is cut into small pieces of 0.3cm, the small pieces are respectively inoculated into a subculture medium containing 0.2mg/L of 6-benzylamino adenine, 1.3mg/L of gibberellin and 0.8mg/L of indoleacetic acid, and the subculture medium is cultured under the conditions that the temperature is 22 ℃, the illumination intensity is 2700lx, the illumination time is 11.5h and the culture environment humidity is 68% until the rootless seedling is obtained.
Transplanting the rootless seedlings into a rooting culture medium containing 1.4mg/L of naphthylacetic acid and 0.7mg/L of kinetin, and culturing under the conditions that the temperature is 22 ℃, the environmental humidity is 68% and the light is shielded until the gentian flower seedlings are obtained.
And (3) cultivating the gentiana scabra seedlings in an illumination incubator, wherein the illumination intensity is 2700lx, the illumination time is 12 hours, and the humidity of the illumination incubator is set at intervals of 1 hour alternately according to 70% and 50% until domesticated seedlings are obtained.
And planting the domesticated seedlings in a greenhouse, ensuring that the illumination intensity in the greenhouse reaches 2700lx and the illumination time reaches 12h, and planting mature gentian flowers by matching with the work of applying farmyard manure, weeding and the like.
The germination rates in the primary cultures of example 1, example 2, example 3, example 4 and control 1 were compared, wherein control 1 was treated in a similar manner to the examples, except that no EM bacteria were added to the primary culture medium of control 1. The survival rate of the aseptic seedlings (number of aseptic seedlings/number of seeds) x 100%, and the above 5 treatments were calculated for 100 seeds. The comparative results are shown in Table 1.
TABLE 1 emergence rate of seeds
| Group number | Example 1 | Example 2 | Example 3 | Example 4 | Control group 1 |
| Rate of emergence | 90％ | 93％ | 91％ | 89％ | 68％ |
As can be seen from the results of the comparison, the emergence rates of the 4 groups of examples were significantly higher than those of the control group 1. The result shows that the survival rate of aseptic seedlings can be obviously improved by adding EM (effective microorganisms) bacteria into the primary culture medium during seedling culture.
The germination rates of primary cultures of example 1, example 2, example 3, example 4 and control 2 were compared, wherein control 2 was treated in a similar manner to the examples, with the only difference that perlite was not added to the primary medium. The rate of emergence (number of aseptic seedlings/number of seeds) x 100%, and the above 5 treatments were calculated for 100 seeds. The comparative results are shown in Table 2.
TABLE 2 emergence rate of seeds
| Group number | Example 1 | Example 2 | Example 3 | Example 4 | Control group 2 |
| Rate of emergence | 90％ | 93％ | 91％ | 89％ | 72％ |
According to the comparison result, the emergence rate of the perlite seeds added in the primary culture medium is higher than that of the control group 2 without perlite. The fact that perlite is added into the primary culture medium is beneficial to the respiration of seeds and improves the rate of emergence.
The survival rates of seedlings planted in examples 1, 2, 3, 4 and 3 were compared, wherein control 3 was treated in a similar manner to the examples, with the only difference that no training of different ambient humidity was performed before the seedlings were planted in the greenhouse. The survival rate (number of seedlings surviving transplantation/total number of transplants) × 100%, and the above 5 treatments were calculated for 100 seedlings. The comparative results are shown in Table 3.
TABLE 3 survival rate of seedlings
| Group number | Example 1 | Example 2 | Example 3 | Example 4 | Control group 3 |
| Survival rate | 88％ | 89％ | 91％ | 93％ | 64％ |
According to the comparison result, the survival rate of the seedlings in the greenhouse in the 4 groups of embodiments is higher than that of the control group 3, which shows that the survival rate of the transplanted seedlings can be improved by performing different humidity training before transplanting the seedlings to the greenhouse.
In summary, according to the seedling raising method of gentiana straminea and the planting method of gentiana straminea of the embodiment of the present invention, the rate of germination of gentiana straminea seeds is accelerated by peeling the gentiana straminea seeds, and the seed respiration can be facilitated by adding perlite to the primary culture medium, and the acidity of the pH of the primary culture medium is maintained, so that the germination of the seeds is further accelerated; the EM can make the aseptic seedlings stronger and increase the survival rate of the aseptic seedlings; under the action of a series of tissue culture of subculture and rooting culture, the emergence rate and the survival rate of the gentian flower seeds are improved, and the adaptability of gentian flower seedlings is enhanced; the gentian flower seedlings are domesticated and then planted, the survival rate of the gentian flower seedlings can be further improved, the yield of gentian flowers is increased, the market demand is met, the picking amount of wild gentian flowers is reduced, and the danger of extinction of the gentian flowers is avoided.
The embodiments described above are some, but not all embodiments of the invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Claims (8)
1. A seedling growing method of gentiana scabra bunge is characterized in that seeds of gentiana scabra bunge are soaked in a sodium hypochlorite solution with the mass concentration of 10% -20% for peeling, and then washed by sterile water after peeling to obtain peeled seeds; inoculating the peeled seeds into a primary culture medium, and carrying out primary culture to obtain aseptic seedlings; taking cotyledons and hypocotyls of the aseptic seedlings as explants, inoculating the explants to a subculture medium, and carrying out subculture to obtain rootless seedlings; transplanting the rootless seedlings into a rooting culture medium, and carrying out rooting culture to obtain gentian flower seedlings; the primary culture temperature is 15-25 ℃, the illumination intensity is 2000-3000lx, the illumination time is 10-12h, and the primary culture medium is an MS culture medium added with perlite, indoleacetic acid with the mass concentration of 0.1-0.5mg/L, 6-benzylamino adenine with the mass concentration of 0.5-1mg/L and EM bacteria with the mass concentration of 0.01-0.05 mg/L; the temperature of the subculture is 15-25 ℃, the illumination intensity is 2000-3000lx, and the illumination time is 10-12 h.
2. A method of raising seedlings of gentiana macrophylla according to claim 1, wherein a time period for washing with the sterile water is 20 to 30 min.
3. The method for growing seedlings of gentiana scabra bunge according to claim 1, further comprising the step of performing ultraviolet induction on the peeled seeds before the seeds are inoculated into the primary culture medium, wherein the ultraviolet induction is to treat the peeled seeds for 2-3h under ultraviolet rays with the wavelength of 250-254 nm.
4. The method for raising seedlings of gentiana flower according to claim 1, wherein the temperature of rooting culture is 15 to 25 ℃, and the rootless seedlings transplanted in the rooting culture medium are subjected to shading treatment.
5. The method for raising seedlings of gentiana scabra bunge according to claim 1, wherein the environmental humidity of the primary culture, the secondary culture and the rooting culture is 55% -75%.
6. A method for raising seedlings of gentiana macrophylla according to claim 1, wherein the peeling with the sodium hypochlorite solution is performed in the following manner: and soaking the seeds in the sodium hypochlorite solution for 5-10 min.
7. The method for growing seedlings of gentiana scabra bunge according to claim 1, wherein the subculture medium is an MS medium to which 6-benzylaminopurine, gibberellin, and indoleacetic acid are added at mass concentrations of 0.1 to 0.5mg/L, 1mg to 1.5mg/L, and 0.5 to 1 mg/L; the rooting culture medium is an MS culture medium added with naphthylacetic acid with the mass concentration of 1mg-1.5mg/L and kinetin with the mass concentration of 0.5-1 mg/L.
8. A method for planting gentiana straminea, which is characterized in that seedlings of gentiana straminea cultured by the method for growing seedlings of gentiana straminea according to any one of claims 1 to 7 are domesticated, the seedlings of gentiana straminea are planted in an illumination incubator, the illumination intensity of the illumination incubator is 2000-3000lx, the illumination time is 10-12h, the humidity in the illumination incubator is alternately set according to 70% and 50% at one-hour intervals, and domesticated seedlings are obtained; and planting the domesticated seedlings in a greenhouse, controlling the illumination time in the greenhouse to be 10-12h and the illumination intensity to be 2000-3000lx, and matching with field management to obtain the mature gentiana scabra bunge.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611072300.6A CN106717260B (en) | 2016-11-29 | 2016-11-29 | Gentiana scabra seedling growing method and Gentiana scabra planting method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611072300.6A CN106717260B (en) | 2016-11-29 | 2016-11-29 | Gentiana scabra seedling growing method and Gentiana scabra planting method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106717260A CN106717260A (en) | 2017-05-31 |
| CN106717260B true CN106717260B (en) | 2021-01-08 |
Family
ID=58905397
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611072300.6A Active CN106717260B (en) | 2016-11-29 | 2016-11-29 | Gentiana scabra seedling growing method and Gentiana scabra planting method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106717260B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107006332B (en) * | 2017-06-14 | 2020-11-24 | 会东县天有老君滩种养专业合作社 | Artificial cultivation method of wild eagle tea |
| CN111466281A (en) * | 2019-12-25 | 2020-07-31 | 上林县温英成生态香蕉种植农民专业合作社 | High-survival-rate antiviral banana seedling cultivation method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104067939A (en) * | 2014-06-26 | 2014-10-01 | 广西壮族自治区药用植物园 | Tissue culture rapid propagation method of radix gentianae |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100199378A1 (en) * | 2006-11-20 | 2010-08-05 | Shing Kwok | Shade tolerance in plants |
| CN101530026B (en) * | 2009-03-10 | 2012-02-15 | 云南省农业科学院药用植物研究所 | Method for improving rate of emergence of Yunnan gentian seed |
| CN101627689B (en) * | 2009-08-03 | 2011-01-12 | 云南省农业科学院药用植物研究所 | Control method of gentiana rigescens spot blight |
| CN102626023B (en) * | 2012-04-01 | 2014-02-12 | 广西壮族自治区药用植物园 | Gentiana loureiri introduction and cultivation method and substrate |
| CN107027372A (en) * | 2016-11-16 | 2017-08-11 | 六盘水师范学院 | A kind of method for promoting red gentian seed to sprout and its seedling |
| CN106688367A (en) * | 2016-11-26 | 2017-05-24 | 全椒县悠然自得果蔬种植专业合作社 | Pollution-free watermelon planting method |
-
2016
- 2016-11-29 CN CN201611072300.6A patent/CN106717260B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104067939A (en) * | 2014-06-26 | 2014-10-01 | 广西壮族自治区药用植物园 | Tissue culture rapid propagation method of radix gentianae |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106717260A (en) | 2017-05-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101637092B (en) | Method for culturing large eucalyptus seedlings | |
| CN102119660B (en) | Method for rooting culture,seedling adaptation and transplantation of alpine rose tissue-cultured seedling in greenhouse | |
| CN105393919B (en) | Tissue culture and rapid propagation method for kadsura coccinea | |
| CN109258460A (en) | Micro-stem tip culture combines the breeding method of heat treatment acquisition Zengcheng honey chrysanthemum detoxic seedling | |
| CN101233807A (en) | Oriental China fir tissue culture seedling ex vitro rooting technique | |
| CN106258956B (en) | A kind of dendrobium candidum pedigree breeding and field transplantation method | |
| CN106386491A (en) | In vitro regeneration method of Begonia wallichiana Lehm | |
| CN110692521A (en) | High-seedling-rate cultivation method for stem tissue culture of polygala tenuifolia | |
| CN103430845A (en) | Strawberry tissue culturing method | |
| CN106258970A (en) | A kind of poison-removing method of Passifolra edulis | |
| CN106359087A (en) | Tissue culture quick-breeding seedling raising method for radix asparagi | |
| CN105941154A (en) | Comprehensive breeding method for superior winter jujube seedlings | |
| CN107509536A (en) | A kind of outer micro-grafting method of test tube of balsam pear tissue-cultured seedling | |
| CN106717260B (en) | Gentiana scabra seedling growing method and Gentiana scabra planting method | |
| CN102870579A (en) | Cutting propagation method of mono maple in seedling bags | |
| CN107041285A (en) | A kind of method of the live seedling of Dendrobidium huoshanness ball stem | |
| CN107135945B (en) | Tissue culture medium of linden tree and rapid propagation method thereof | |
| CN105993629A (en) | Grafting method for increasing eggplant yield | |
| CN106069748A (en) | The commercial tissue culture of sprout mating system of American red-maple splendidness in October | |
| CN105519448A (en) | Culture method of radix astragali tissue culture seedlings | |
| CN107646479A (en) | A kind of propagation method of five happinesses persimmon seedling | |
| CN107660450A (en) | A kind of method for culturing seedlings of big shed eggplant | |
| CN104521660A (en) | Root cutting cultivation method of jujube tree | |
| CN105075860B (en) | A kind of fleabane flower tissue culture method | |
| CN110574651A (en) | rice planting method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |