Erigeron breviscapus asexual propagation method

Abstract

The invention discloses an asexual propagation method for erigeron breviscapus, and belongs to the technical field of plant asexual culture. According to the method, erigeron breviscapus varieties with excellent characters are subjected to plant division treatment, small erigeron breviscapus plants obtained through plant division are put into a diluted potassium permanganate solution with the mass concentration of 95% to be soaked and properly turned over, and water culture is conducted after surface moisture is aired. According to the method disclosed by the invention, the separated plantlets can be subjected to water culture, the seedling recovery period is short, the growth vigor is strong, the tillering is fast, the plantlets are not easy to bloom early, the effective content is high, the yield is also high, the keep-alive rate is from 10% to more than 95%, and excellent characters of a female parent are also maintained, so that progeny plants and the female plant have the same quality and characteristics, heritable variation is reduced, and the survival rate is increased. And moreover, the method is simple and convenient to operate, relatively low in cost, relatively good in economic benefit and suitable for large-scale production.

Description

Technical Field

[0001] The invention relates to an asexual propagation method of Erigeron breviscapus, and belongs to the technical field of asexual culture of plants. Background Art

[0002] Yunnan Province is the main province where wild Erigeron breviscapus resources are distributed, and is also the main province for planting, processing, production and development of Erigeron breviscapus. It gathers 100% of the raw material production of Erigeron breviscapus, more than 97% of the supply of medicinal raw materials, and 100% of the production of extracts and concretions in the country, forming a complete industrial chain of planting, extraction and preparation production. It has been listed as one of the "five major natural series" of medicines in Yunnan Province and is given priority development.

[0003] Due to long-term excavation, the wild resources of Erigeron breviscapus are decreasing day by day, and the traditional breeding method is mainly based on direct seeding and broadcasting. Since the seeds are relatively small, there are limitations in the planting process, such as large seed usage, low germination rate, serious seedling shortage, low rate of strong seedlings, serious weed damage, low yield, low commodity rate, high labor input, and low production efficiency. In addition, Erigeron breviscapus is a strict self-incompatible plant, which means that it cannot reproduce offspring through self-pollination, which further limits its ability to reproduce naturally and leads to serious degradation of its seed source. Although some studies have successfully cultivated new Erigeron breviscapus varieties through systematic breeding methods, this method still requires a long time and a large amount of resource investment.

[0004] In plant cultivation, in order to reduce plant variation and maintain the excellent traits of the mother plant, asexual reproduction such as plant cuttings, layering, division and tissue culture has become a good choice. At present, the commonly used asexual reproduction methods of Erigeron breviscapus mainly include tissue culture and non-test tube high-efficiency rapid propagation culture. Due to the problems of low differentiation ability, few roots, easy browning, yellowing and vitrification in plant tissue culture, and the cultivated Erigeron breviscapus seedlings need specific culture medium and hormones to achieve 100% differentiation rate, rooting rate and induction rate, the later seedling training cycle is long, and the requirements for temperature, humidity and light are high; although non-test tube high-efficiency rapid propagation culture can reduce the variation rate, it also has problems such as large one-time investment, high cost, complicated technical steps, and poor technical transmission. Farmers cannot use it directly in production, and there are few projects that truly achieve the goal of industrialization.

[0005] Although the traditional division technology has been mature in the application of other plants, because other plants do not have too high requirements for the division method, the requirements for division of the gentian are relatively high, and the existing division method is relatively random, resulting in excessive wounds on the plants after cutting, increased corruption rate and browning rate, and the disinfection technology is not up to standard. The traditional soil cultivation root rot is serious and the survival rate is extremely low, so it is difficult to apply it on a large scale in production. Compared with soil culture technology, the hydroponic system can provide a more controllable and efficient environment to promote the rapid growth and reproduction of plants and optimize the nutritional status of plants. However, the existing hydroponic seedlings of the gentian are all obtained by explant callus culture, seed floating seedling technology or anther culture, and then transplanted. There are problems such as long initial cultivation time, high cost, complicated operation and uneven quality. At the same time, the selection of nutrient solution formula is also one of the key factors. Different plants have different nutritional requirements, so it is necessary to adjust the nutrient solution formula according to the growth characteristics and needs of specific plants. In addition, in the current research and application of hydroponic technology, there is no rapid propagation cultivation technology for Breviscapus to propagate by division through root suckers. Therefore, in view of the above problems, the method for hydroponic division and survival of Breviscapus needs to solve a series of problems such as the formula of nutrient solution suitable for Breviscapus, unified cutting technology, grasping the disinfection time, reducing the corruption rate and browning rate, improving the survival rate of plants, maintaining the excellent traits of the mother plant, and reducing the frequency of mutation. Summary of the invention

[0006] In view of the problems existing in the current cultivation of Erigeron breviscapus, the present invention provides an asexual propagation method of Erigeron breviscapus, and the specific steps are as follows:

[0007] (1) Clean the Erigeron chinensis, remove old roots and leaves, and divide the plants to obtain small plants.

[0008] The division method is one of the following three methods (selected according to the actual situation of Erigeron breviscapus):

[0009] When the tiller edge of the Erigeron breviscapus is clear (clear tiller edge means that the rhizomes of the Erigeron breviscapus are separate, but the roots are connected together), cut along the tiller edge.

[0010] When the tiller edge of the Erigeron breviscapus is unclear (the unclear tiller edge means that the rhizomes of the Erigeron breviscapus are completely connected together) and the diameter of the rhizome is greater than 1.5 cm, first divide the Erigeron breviscapus into two halves along the middle of the Erigeron breviscapus, then divide the Erigeron breviscapus into two halves again along the middle of each half, and finally divide the Erigeron breviscapus into 4 small plants.

[0011] When the tillering edge of the Erigeron breviscapus is unclear and the diameter of the rhizome is within 1.5 cm, divide the Erigeron breviscapus into two halves along the middle of the flower to obtain two small plants.

[0012] (2) The obtained plantlets are placed in a potassium permanganate solution diluted 250 times with a mass concentration of 95% and immersed and turned over, and then taken out and air-dried to dry the surface moisture.

[0013] (3) hydroponically cultivating the plantlets treated in step (2) until the plantlets grow new roots.

[0014] Preferably, the Erigeron breviscapus in step (1) is a plant of good variety, strong, with many tillers and free from diseases and insect pests.

[0015] Preferably, the tools used for division in step (1) need to be immersed in a 95% potassium permanganate solution diluted 250 times in advance for disinfection, and disinfected once each time they are used.

[0016] Preferably, the petiole and leaf blade of the plantlets obtained in step (1) retain a total of 5 to 6 cm, and each plantlet retains a taproot or fibrous root and 3 to 5 young leaves.

[0017] Preferably, if the plantlets obtained in step (1) do not retain taproots or fibrous roots, the roots of the plantlets need to be soaked in 1000-fold diluted naphthaleneacetic acid for 5 minutes before hydroponics, with the soaking depth being 3 to 4 cm.

[0018] Preferably, the soaking and stirring time in step (2) is 20 to 30 seconds.

[0019] Preferably, after the substrate used for hydroponics in step (3) is cleaned, it is then immersed in a 500-fold diluted carbendazim suspension for disinfection for 24 hours, and appropriately turned over several times in between to remove potential pathogens.

[0020] Preferably, in step (3), the roots of the small plants should be in contact with the nutrient solution, and the core leaves should not be buried, and all leaves can be sprayed with deionized water.

[0021] Preferably, the formula and dosage of the culture solution used in the hydroponic culture in step (3) are as follows:

[0022]

[0023] Preferably, the conductivity of the nutrient solution is between 2000 and 2500 μs / cm, and the dissolved oxygen content is 8 mg·L -1 .

[0024] Preferably, a carbendazim suspension concentrate is added to the nutrient solution in an amount of 2 g / L.

[0025] Beneficial Effects of the Invention

[0026] (1) The disinfectant of the present invention is selected from a large number of disinfectants to not only thoroughly disinfect plants but also cause little harm.

[0027] (2) The cutting method, cutting size, disinfection method and disinfection time described in the present invention can effectively reduce the browning rate and corruption rate of the plants.

[0028] (3) The plantlets obtained by division of the present invention can grow directly in a hydroponic environment, have a short seedling acclimatization period, strong growth potential, fast tillering, are not prone to early flowering, have high yields, and a survival rate ranging from 10% to over 95%.

[0029] (4) The present invention found that disinfecting the isolated plantlets with a 95% potassium permanganate solution diluted 250 times can effectively reduce the infection rate and browning rate of the plantlets and increase the survival rate.

[0030] (5) The culture method of the present invention can effectively increase the content of baicalin and total caffeic acid esters in Breviscapus breviscapus.

[0031] (6) The method of the present invention improves the reproductive efficiency and uniformity of the Erigeron breviscapus, while skipping the seed germination process, reducing the germination rate problem caused by inconsistent seed maturity. By adjusting the nutrient solution ratio, a 100% induction rate and rooting rate can be achieved, thereby improving the survival rate of the seedlings and maintaining the excellent traits of the mother plant, thereby ensuring that the offspring plants have the same quality and characteristics as the mother plant, reducing genetic variation, and making it more genetically stable.

[0032] (7) The method of the present invention increases the propagation speed and can efficiently multiply the number of original plants in geometric progression. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 A method of dividing plants with clear tillering edges.

[0034] Figure 2 The cutting method of Erigeron chinensis when the tillering edges are unclear and the rhizome is more than 1.5 cm.

[0035] Figure 3 The cutting method of Erigeron chinensis with unclear tillering edges and rhizomes less than 1.5 cm.

[0036] Figure 4 The morphology of the plantlet after cutting.

[0037] Figure 5 The plantlet morphology does not retain the taproot or fibrous roots of the mother plant. DETAILED DESCRIPTION

[0038] The technical solution of the present invention is further described below in conjunction with the accompanying drawings and through specific implementation methods. However, the following embodiments are only simple examples of the present invention and do not represent or limit the scope of protection of the present invention. The scope of protection of the present invention shall be subject to the claims.

[0039] Experimental Preparation

[0040] (1) A potassium permanganate solution with a mass concentration of 95% is prepared and diluted 250 times to obtain a potassium permanganate solution with a mass concentration of 95% diluted 250 times.

[0041] (1) The scissors and scalpels used for pruning and cutting need to be sterilized in advance by soaking in a 95% potassium permanganate solution diluted 250 times.

[0042] (2) The hydroponic medium should be expanded clay with slightly larger porosity. Before planting, it should be washed with clean water and then soaked in a 500-fold diluted carbendazim suspension for disinfection for 24 hours. The soil should be turned over several times in between to remove potential pathogens.

[0043] The carbendazim and potassium permanganate used in the experiment can be purchased from conventional channels.

[0044] Example 1

[0045] A method for asexual reproduction of Erigeron breviscapus, the specific steps are as follows:

[0046] (1) Plant selection and treatment

[0047] Give priority to selecting Erigeron crenata plants of excellent varieties, robustness, many tillers and no diseases or insect pests. Clean the soil at the roots of the plants with clean water, use scissors to remove diseased and old leaves, leaving only strong and tender leaves; use scissors to remove diseased and aging roots, leaving only strong and new roots; rinse the trimmed Erigeron crenata plants again with deionized water. To keep the Erigeron crenata plants hydrated, soak the rinsed Erigeron crenata plants in a container filled with deionized water for later use.

[0048] (2) Plant propagation and division

[0049] Take out the Erigeron breviscapus plants soaked in deionized water. When the tiller edge of the Erigeron breviscapus plants is clear, use a scalpel to cut along the tiller edge of the Erigeron breviscapus plants ( Figure 1 ), do not damage the core of the plant; large plants with unclear tillering edges and rhizomes with a diameter of more than 1.5 cm should be divided into quarters ( Figure 2 ), small plants with unclear tillering edges and rhizome diameters less than 1.5 cm were divided by bisection ( Figure 3 ), the main root or fibrous root of the mother plant should be retained during the tillering process. In order to reduce the evaporation of leaves, the length of the strong leaves of the small plant should be cut in half after cutting, and the petiole and leaves should be kept about 5 to 6 cm in total, and 3 to 5 complete new leaves should be retained ( Figure 4); the cut plantlets are placed in a 250-fold diluted 95% potassium permanganate solution and immersed and properly turned for 20 seconds, then taken out and air-dried to remove potential pathogens; during the cutting process, pests and diseased branches are promptly removed to avoid the occurrence of diseases and pests in the later stage.

[0050] The plantlets that have been cut and do not retain the taproot or fibrous roots of the mother plant ( Figure 5 ), after disinfection, soak the roots of the plants in 1000-fold diluted naphthaleneacetic acid for 5 minutes at a depth of 3 to 4 cm to promote the formation of callus and roots.

[0051] (3) Improved preparation and use of hydroponic nutrient solution

[0052] The improved formula and dosage of hydroponic nutrient solution are as follows:

[0053] Table 1 Formula of nutrient solution for hydroponic culture of Erigeron breviscapus

[0054] Element Type Dosage(mg / L) <![CDATA[Ca(NO 3 ) 2 ]]> 945 <![CDATA[KNO 3 ]]> 809 <![CDATA[NH 4 H 2 PO 3 ]]> 153 <![CDATA[MgSO 4 ·7H 2 Oh]]> 493 <![CDATA[Na 2 Fe-EDTA]]> 20 <![CDATA[H 3 BO 3 ]]> 2.86 <![CDATA[MnSO 4 ·4H 2 O4]]> 2.13 <![CDATA[ZnSO 4 ·7H 2 Oh]]> 0.44 <![CDATA[CuSO 4 ·5H 2 Oh]]> 0.08 <![CDATA[(NH 4 )6Mo 7 About 24 ·4H 2 O]]> 0.02 <![CDATA[H 2 Not. 3 ]]> 156

[0055] To prevent the occurrence of root and stem rot of Erigeron breviscapus, a carbendazim suspension should be added to the nutrient solution in the hydroponic box at a rate of 2g / L.

[0056] The conductivity of the hydroponic nutrient solution should be 2400μs / cm.

[0057] The dissolved oxygen content of the hydroponic nutrient solution is 8 mg·L, which can promote the growth and rooting of plant callus.

[0058] (4) Plant cultivation and fixation

[0059] The small plant obtained in step (2) is placed vertically in a hydroponic flower pot, and treated moist ceramsite is added. In order to absorb water, the roots of the plant must be in contact with the nutrient solution, and the core leaves cannot be buried.

[0060] After planting, to prevent the leaves from wilting, you can use a spray bottle to wet all the leaves, using deionized water.

[0061] (5) Daily management

[0062] LED fill light tubes that imitate natural light are installed on the hydroponic rack, and the lighting time is set to alternate between light and dark for 12 hours a day to promote photosynthesis of the Erigeron plants.

[0063] Check the condition of the plant's leaves regularly, and use tweezers to remove yellow and dead leaves in time to prevent gray mold.

[0064] Always observe the water level of the nutrient solution in the hydroponic box to avoid rapid evaporation of water and wilting of the plants.

[0065] The conductivity of the nutrient solution in the hydroponic box should be checked at any time to prevent the ion concentration of the nutrient solution from being too high, which may cause salt stress to the plants, hinder root growth, and even cause salt damage and root rot.

[0066] The nutrient solution in the hydroponic box should be replaced regularly to provide the nutrients needed by the plants.

[0067] When the indoor temperature exceeds 24℃, open the windows for ventilation every day.

[0068] (6) Pest and disease control

[0069] Under the conditions of propagation, diseases and pests are rare, and red spider mites, sharp-eyed fungus gnats, cabbage worms, whiteflies, gray mold, root and stem rot, etc. are occasionally seen; a few pests are removed manually, and solutions of cypermethrin, imidacloprid, etc. are sprayed in time, and sticky insect boards are used in combination; for gray mold and root and stem rot, the diseased plants can be removed, and 800 times diluted carbendazim wettable powder or 2400 times diluted metalaxyl soluble powder can be applied for prevention and control; spray 1000 times diluted carbendazim suspension once every 10 to 20 days. If the daily living environment conditions are good and no disease occurs, spraying is not required.

[0070] (8) Post-management

[0071] Obvious callus tissue will begin to appear after 5 to 7 days, and roots will begin to form after 10 days. After the plantlets have taken root, they can be transplanted into the soil as needed.

[0072] (9) Result Statistics

[0073] The present invention carries out division culture on 300 plants of Erigeron breviscapus. According to statistics, the survival rate of Erigeron breviscapus exceeds 95% when the method of the present invention is used to asexually propagate the Erigeron breviscapus.

[0074] Comparative Example 1

[0075] For comparison, the difference between this embodiment and embodiment 1 is that the Erigeron breviscapus plant with unclear tiller edges is not divided according to the size of the rhizome, but is divided according to the number of tillers of the Erigeron breviscapus. Except that the division method is different from that of embodiment 1, other methods are the same as those of embodiment 1. The survival rate of the resulting Erigeron breviscapus is shown in Table 2.

[0076] Table 2 The survival rate of Erigeron breviscapus with different ramet numbers

[0077]

[0078] It can be seen from Table 2 that if the division method described in the present invention is not used for division, the survival rate of Erigeron breviscapus is significantly reduced.

[0079] Comparative Example 2

[0080] For comparison, the difference between this embodiment and embodiment 1 is that the length of the petiole plus the leaf blade of the small plantlets after the cutting of the Erigeron breviscapus is different, and the number of intact new leaves retained is different. The other methods are the same as those in embodiment 1. The survival rate of the resulting Erigeron breviscapus is shown in Table 3.

[0081] Table 3 Survival rate of plants with different leaf lengths and numbers

[0082]

[0083] From the data in Table 3, it can be seen that when the length of the petiole and leaf of the small plant after the Erigeron breviscapus is cut is too short, the photosynthesis of the plant is reduced, and if it is too long, the water is easy to evaporate. After many experiments, it was found that the survival rate is highest when the petiole and leaf of the small plant are kept about 5 to 6 cm and 3 to 6 complete new leaves are kept.

[0084] Comparative Example 3

[0085] For comparison, the difference between this embodiment and embodiment 1 is that the fungicides used for sterilizing and disinfecting the plantlets are different, the sterilization time is different, and the other operations are the same as those in embodiment 1. After statistics, the browning rate, rooting rate and survival rate of different fungicides are shown in Table 4.

[0086] Table 4 Effects of different fungicides on the survival of Erigeron breviscapus

[0087]

[0088] From the data in Table 4, it can be seen that different fungicides have different sterilization effects on Erigeron breviscapus, and sterilization time that is too long or too short is not conducive to plant growth. Too long will easily lead to an increase in the browning rate of the plant, while too short will result in incomplete sterilization. After many experiments, it was found that small plants were completely sterilized without damaging the seedlings under the condition of using 95% potassium permanganate diluted 250 times for 20 seconds, with the lowest browning rate, the highest rooting rate and the highest survival rate.

[0089] After half a year, the excellent individual plants of No. 1 Erigeron breviscapus bred by our laboratory have been expanded from the first generation to the sixth generation, and from 1 plant to 85 plants. The survival rate of each generation is above 95%, and consistent excellent traits have been maintained.

[0090] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for asexual propagation of Erigeron breviscapus, characterized in that: The specific steps include: (1) Clean the Erigeron breviscapus, remove old roots and leaves, and divide the plants to obtain plantlets; The division method is one of the following three methods: When the tiller edge of the Erigeron breviscapus is obvious, cut along the tiller edge; When the tillering edge of the Erigeron breviscapus is not obvious and the diameter of the rhizome is greater than 1.5 cm, first divide the Erigeron breviscapus into two halves along the middle of the flower, then divide the flower into two halves again along the middle of each half, and finally divide the flower into 4 small plants; When the tillering edge of the Erigeron breviscapus is not obvious and the diameter of the rhizome is within 1.5 cm, the Erigeron breviscapus is divided into two halves along the middle of the Erigeron breviscapus to obtain two plantlets; (2) placing the obtained plantlets in a 95% potassium permanganate solution diluted 250 times, immersing and turning the solution, and then taking the plantlets out and air-drying the surface moisture; (3) Planting the plantlets treated in step (2) in a hydroponic flowerpot for cultivation until the plantlets grow new roots.

2. The asexual propagation method of Erigeron breviscapus according to claim 1, characterized in that: The tools used for division in step (1) need to be disinfected by soaking in a 95% potassium permanganate solution diluted 250 times, and disinfected once each time they are used.

3. The asexual propagation method of Erigeron breviscapus according to claim 1, characterized in that: The petiole and leaf blade of the plantlets obtained in step (1) are retained for a total of 5 to 6 cm, and each plantlet retains a taproot or fibrous root and 3 to 5 young leaves.

4. The asexual propagation method of Erigeron breviscapus according to claim 1, characterized in that: If the plantlets obtained in step (1) do not retain the taproot or fibrous roots, the roots of the plantlets need to be soaked in 1000-fold diluted naphthaleneacetic acid for 5 minutes before hydroponics, with the soaking depth being 3 to 4 cm.

5. The asexual propagation method of Erigeron breviscapus according to claim 1, characterized in that: The soaking and stirring time in step (2) is 20 to 30 seconds.

6. The asexual propagation method of Erigeron breviscapus according to claim 1, characterized in that: The formula and dosage of the culture solution used in the hydroponic culture in step (3) are as follows:

7. The asexual propagation method of Erigeron breviscapus according to claim 6, characterized in that: The conductivity of the nutrient solution is 2000-2500 μs / cm, and the dissolved oxygen content is 8 mg·L -1 .

8. The method for asexual propagation of Erigeron breviscapus according to claim 6, characterized in that: A carbendazim suspension concentrate was added to the nutrient solution in an amount of 2 g / L.

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