CN114521491B - Method for breaking transplanting dormancy of cymbidium tissue culture seedlings and shortening production period - Google Patents
Method for breaking transplanting dormancy of cymbidium tissue culture seedlings and shortening production period Download PDFInfo
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Abstract
The invention discloses a method for breaking the transplanting dormancy of tissue culture seedlings of cymbidium and shortening the production period, belonging to the field of agricultural biological planting. The tissue culture seedlings of the cymbidium sinensis are soaked by a specific root-extracting activating agent, dried and transplanted; after the Chinese orchid tissue culture seedlings are transplanted, a specific dormancy breaking agent is sprayed, so that the transplanting survival rate of the Chinese orchid tissue culture seedlings is effectively and stably improved, meanwhile, the dormant state in the transplanting process of the Chinese orchid tissue culture seedlings can be broken, the Chinese orchid tissue culture seedlings rapidly germinate and grow new buds, finally, the growth period from transplanting to blooming of the Chinese orchid tissue culture seedlings to finished products is obviously shortened, the production cost of the Chinese orchid tissue culture seedlings is obviously reduced, the production advantages are obvious, and the production and application of the Chinese orchid tissue culture seedlings can be promoted.
Description
The technical field is as follows:
the invention belongs to the field of agricultural biological planting, and particularly relates to a method for breaking the dormancy of tissue culture seedlings of cymbidium and shortening the production period.
Background art:
the national orchid is also called Chinese orchid, and the foreigners called east orchid or eastern orchid, usually refers to a part of the plant species of orchid (Cymbidiu) in the orchid family, and mainly includes 5 major species of cymbidium goeringii, comet orchid, cymbidium ensifolium, cymbidium sinense, cymbidium kanran and cymbidium kanran. The flower is pure and luxuriant, the flower fragrance is elegant, and the leaf state is beautiful. The Chinese orchid is a traditional flower in China, has a cultivation history of nearly thousands of years, and is widely popular with people due to high ornamental value, cultural value and medical practical value.
The national orchid tissue culture research starts in the last 80 th century, the national orchid tissue culture technology is researched for nearly 40 years, a basic technical system is established early, but the current national orchid tissue culture technology still has no major breakthrough, firstly, the transplanting survival rate of the national orchid tissue culture seedlings is unstable, secondly, the national orchid tissue culture seedlings are in a dormant state for 6-12 months after being transplanted, the flowering of the national orchid tissue culture seedlings is late due to the result, about 4-5 years are needed from bottle discharge to flowering, and the flowering is fast by adopting the plant division method for cultivation, so that the production and maintenance cost of the national orchid tissue culture seedlings is greatly increased compared with that of the traditional plant division propagation seedlings, and the commercialized production and application of the national orchid tissue culture seedlings are directly hindered.
In addition, the number of the national orchid varieties is small, but the national orchid varieties are suitable for common consumers. Some cymbidium varieties are suitable for mass consumption, but the yield and the commercialization degree are too low to meet the market demand. The method is limited by the existing difficulty of the national orchid commercialized tissue culture rapid propagation technology, so that the production period of the national orchid tissue culture seedling is too long, the production cost and the risk are too high, and the industrialized development of the national orchid tissue culture seedling does not form a complete industrial chain. Therefore, the difficulties that the transplanting survival rate of the tissue culture seedlings of the cymbidium sinense is unstable and the dormancy period is more than half a year after the transplanting is performed at present are solved on the basis of fully mastering the growth and development rules of the tissue culture seedlings of the cymbidium sinense.
In the patent literature, dingguang et al (2019) apply for a method for improving transplanting survival rate of hybrid tissue culture seedlings of orchid, and the invention patent utilizes the characteristic that the root system of a seedling in a water culture technology is directly contacted with a nutrient solution to absorb nutrients in the nutrient solution more quickly to transplant the hybrid seedlings for seedling recovery, thereby showing the effects of reducing cost, saving labor and obtaining a large amount of high-quality hybrid tissue culture seedlings of orchid. Roxburgh et al (2020) apply for a 'seedling hardening method for hybrid orchid tissue culture seedlings', the transplanting survival rate of the hybrid orchid tissue culture seedlings is effectively improved by selecting the hybrid orchid tissue culture seedlings which grow strongly and have rooted roots, cleaning and disinfecting the roots, soaking the roots in a nutrient solution for 20-24 hours, transplanting the seedlings into a seedling culture container filled with a seedling culture medium, culturing for 4-6 months to obtain medium and large seedlings, transplanting the medium and large seedlings into the culture medium on a culture field for hardening, and managing the hybrid orchid seedlings after watering and fertilizing. Zhang jin Jiang et al (2019) applied for 'a high-temperature high-humidity rapid propagation culture method of cymbidium sinense', the seedlings are hardened by gradually strengthening illumination to the seedlings, the seedlings are cleaned and disinfected, the seedlings are cultured under the conditions of 82-90% of air humidity and 25-30 ℃ of daytime temperature, the total time of illumination intensity 20000-30000lux is controlled to be 4-6 hours and 6-8 hours within 24 hours each day, a seedling culture tray is rotated by 60-180 degrees every 1-3 days, a culture medium is kept in a wet state by watering management every 3-5 days, watering and fertilizing are combined once every 2-3 times, and pesticide liquid is sprayed to cymbidium sinense every 25-28 days, so that high cymbidium sinense seedling rate is obtained, and the seedlings are strong and strong in adaptability. However, the above patent documents and other documents do not relate to the solution of the two difficulties in the current production and application of cymbidium tissue culture seedlings by using a root-enhancing activator and a dormancy-breaking agent.
The invention content is as follows:
the method strictly controls the rooting and strong seedling culture time of the cymbidium sinense tissue culture seedling bottle seedlings, the bottle seedlings are cleaned, disinfected, stressed by adding salt and then dried to improve the viability of the root systems of the cymbidium sinense tissue culture seedlings, and then the cymbidium sinense tissue culture seedlings are transplanted into a layered planting matrix, so that the requirement of the cymbidium sinense tissue culture seedlings on permeability conditions can be guaranteed, the rich nutrient components in the matrix can be absorbed by the root systems of the cymbidium sinense tissue culture seedlings, the key is that a dormancy breaking agent is applied to promote old seedlings to germinate new buds at the fastest speed, scientific water and fertilizer management is performed according to the growth and development rules of the new buds, finally, the transplanting survival rate of the cymbidium sinense tissue culture seedlings is high, the time from bottle emergence to flowering is obviously shortened, and the difficulty that the transplanting survival rate of the cymbidium sinense tissue culture seedlings is unstable and the dormancy stage is more than half a year after transplantation is solved.
The first purpose of the invention is to provide a method for breaking the dormancy of the tissue culture seedlings of the cymbidium and shortening the production period, the tissue culture seedlings of the cymbidium are soaked by a root-extracting activating agent, aired and then transplanted, the root-extracting activating agent is a mixed solution of 0.1-0.2% (mass fraction) of potassium permanganate and 0.3-0.6% (mass fraction) of sodium chloride; spraying dormancy breaking agent after transplanting the tissue culture seedling of Chinese orchid, wherein the dormancy breaking agent comprises 1/2-3/4MS, 0.05-0.1 mg/L6-BA, 0.25-1.0mg/L IBA and 50-100mg/LGA 3 、0.68-1.36g/L KH 2 PO 4 The pH value is 5.8-6.0.
Preferably, the method comprises the following steps:
(1) Soaking the tissue culture seedling of cymbidium sinensis in the root-extracting activating agent for 10-30min, placing in a shade and ventilated place, drying until the color of the root system of the tissue culture seedling turns white, and then transplanting;
(2) And (3) spraying the dormancy breaking agent 30 days after the tissue culture seedlings of the cymbidium sinensis are transplanted, spraying the dormancy breaking agent 1 time every 7-15 days, continuously spraying the dormancy breaking agent 3-5 times, and then carrying out growth promoting cultivation management.
More preferably, in the step (1), the cymbidium tissue culture seedling is a cymbidium tissue culture seedling bottle seedling which is cultured by selecting rooting strong seedlings for 60-90 days.
More preferably, in the step (1), the transplanting is to plant the above tissue culture seedlings on a layered filling matrix planting cup, wherein the middle and lower layers of the layered filling matrix planting cup are the pond sludge peanut shell mixed matrix, and the upper layer is the bark and orchid stone mixed matrix.
More preferably, in the step (2), when the sprout of the tissue culture seedling grows to 5.0 cm high and 2 leaves are unfolded, applying 3-5g of slow release fertilizer (N-P-K = 15-8-14) to each pot, and irrigating the root with 1000-1500 times of water-soluble fertilizer (N-P-K = 20-20-20) for 1 time every 7-10 days; the pots are changed every year in the second and third years.
The second purpose of the invention is to provide a root-extracting activator which is a mixed solution containing 0.1-0.2% (mass fraction) of potassium permanganate and 0.3-0.6% (mass fraction) of sodium chloride.
It is a third object of the present invention to provide a dormancy-breaking agent comprising 1/2-3/4MS, 0.05-0.1 mg/L6-BA, 0.25-1.0mg/L IBA, 50-100mg/LGA 3 、0.68-1.36g/L KH 2 PO 4 The pH value is 5.8-6.0.
Compared with the prior art, the invention has the beneficial effects that:
(1) The invention provides a method for breaking the dormancy of tissue culture seedlings of cymbidium, which can ensure that the cymbidium tissue culture seedlings can quickly recover from growth after being transplanted, and a dormancy breaking agent is sprayed in time 30 days after the cymbidium tissue culture seedlings are transplanted, so that the aged and dormant cymbidium tissue culture seedlings germinate 1-2 new buds to enter a quick growth stage; transplanting for 3-4 years, and flowering for 1-2 years earlier than conventional cymbidium tissue culture seedling culture.
(2) The transplanting survival rate of the tissue culture seedlings of the cymbidium sinense is stabilized while the sprouts grow rapidly, the transplanting survival rate can reach more than 95%, the production advantage is obvious, and therefore the commercialized and efficient production of the tissue culture seedlings of the cymbidium sinense can be realized.
(3) The used root-raising activating agent and dormancy-breaking agent have unique components, are simple and practical, have stable and obvious use effect and high application value.
The specific implementation mode is as follows:
the following examples are further illustrative of the present invention and are not intended to be limiting thereof. The specific test conditions and methods not indicated in the following examples are generally conventional means well known to those skilled in the art.
Example 1
1. Materials: tissue culture seedling of cymbidium ensifolium.
2. Selection of production place: the place which is convenient to traffic, faces the sun from the back to the north, is ventilated and transparent, has good drainage, low underground water level, sufficient fresh water source and power supply should be selected, and the place which is far away from the disease area of the national orchid virus disease and the intermediate host crop should be selected.
3. Construction of a planting shed: the general requirements of the planting shed are that the planting shed can prevent insects and diseases, and can timely adjust the environmental conditions of temperature, humidity, light and the like according to the growth requirements of the tissue culture seedlings of Chinese orchid. The width of the planting greenhouse is generally 7-8 meters, the height is 3 meters, the length can be determined according to the field and the requirement, a sidewalk (about 0.8 meter wide) is arranged in the middle of the greenhouse, and a buffer room is arranged at an entrance and an exit. The shed frame is made of bent galvanized pipe, and the outside of the shed frame is equipped with insect-proof gauze, plastic film and black shading net. The insect-proof net is made of plastic yarn, the size of the net is 40-60 meshes, and the insect-proof net is used for preventing pests from entering the planting shed to bite the tissue culture seedlings of the Chinese orchid and infecting diseases. The plastic film mainly plays a role in heat preservation and rain prevention. The shading net has the shading function, and equipment for water spraying, heating and brightening can be arranged in the greenhouse.
4. Preparing a planting matrix: the matrix comprises pond sludge, peanut shell, bark, and blue stone. Pond sludge and peanut shells which are rich in organic matters are firstly arranged at the middle part and the lower part of the planting cup (the volume ratio is 1: 1), a bark orchidite (the volume ratio is 1: 1) mixed matrix is used at the upper part of the planting cup, and new roots growing out quickly penetrate into the pond sludge to absorb nutrients, so that the aim of quick growth is fulfilled.
5. And (3) disinfection of the substrate: the background of the transplanting land and the transplanting matrix of the tissue culture seedlings of the Chinese orchid need to be disinfected and sterilized before use so as to ensure the healthy growth of the seedlings. The method can be used for steam disinfection, can thoroughly kill germs and pests, and can also be used for medicine disinfection, wherein formalin is the most common disinfectant, the stock solution can be diluted by 50 times with water and then evenly sprinkled on a nursery land or a culture medium until the nursery land or the culture medium is moistened, and then a plastic film is covered on the nursery land or the culture medium for retting for 4 to 5 days to open the nursery land or the culture medium to achieve the disinfection effect.
6. Selection of tissue culture seedlings of Chinese orchid in bottles: selecting a Chinese orchid tissue culture seedling bottle seedling which grows roots and strong seedlings and is cultured for 60-90 days, wherein the Chinese orchid tissue culture seedling bottle seedling is relatively strong in growth state and free of pollution, the number of leaves is 3-4, the leaves are dark green, 3-5 roots are grown, the root system is 2-5cm long, and the root system is strong and active.
7. Hardening and transplanting tissue culture seedlings: the Chinese orchid tissue culture seedling bottle seedlings with the rooting and strong seedling culture period of 60-90 days are moved to a place with sufficient scattered light, the seedlings are enabled to fully receive light, the bottle covers can be opened when necessary, the seedlings are enabled to fully contact with external environmental conditions, photosynthesis of the seedlings is facilitated, chlorophyll and fibrosis are enriched, air holes can be automatically adjusted to be opened and closed, the seedlings are enabled to be more robust, and hardening is carried out for 7-15 days. During transplanting, the rooted seedlings are taken out from a culture bottle, the attached culture medium is cleaned, and then the rooted seedlings are soaked in 0.1-0.2% (mass fraction) of potassium permanganate and 0.3-0.6% (mass fraction) of sodium chloride root-extracting activator for 10-30min and then placed in a cool and ventilated place to be aired until the color of the root systems of the tissue culture seedlings turns white, so that the root systems of the tissue culture seedlings of the cymbidium orchid can be exercised and improved in activity through salt and drought stress, and transplanting, field planting and survival and rapid growth are facilitated. When transplanting, the tissue culture seedlings are transplanted on a planting cup filled with the layered filling matrix, and the matrix is compacted, wherein the upper plane of the matrix is 1-2.0cm lower than the edge of the pot. And (3) watering thoroughly on the transplanting day until clear water flows out from the bottom of the flowerpot, spraying a broad-spectrum bactericide, keeping appropriate humidity and temperature and ventilation and illumination conditions, and spraying 3000 times of liquid by using a water-soluble fertilizer (N-P-K = 20-20-20) for 1 time every 7-15 days. In this example, two groups of root-raising activators with different concentrations are set for experiments, and the results are shown in table 1, and the survival rate of 30 days after transplantation can reach more than 95%.
TABLE 1 root-extracting vigor agent in different concentrations and its efficiency
8. Breaking dormancy of the temporary tissue culture seedling: 30 days after the tissue culture seedlings of the Chinese orchid are transplanted, the dormancy breaking agent (1/2-3/4 MS (only macroelements are changed to 1/2-3/4 times of the original), 0.05-0.1 mg/L6-BA, 0.25-1.0mg/L IBA and 50-100 mg/LGA) is sprayed 3 +0.68-1.36g/L KH 2 PO 4 pH 5.8-6.0), this example sets two different concentrations of dormancy-breaking agent for experiments, as shown in table 2. Every 7-15 daysSpraying for 1 time, and continuously spraying for 3-5 times.
TABLE 2 different concentrations of dormancy-breaking agents
In the present example, the composition of the MS medium is shown in Table 3:
TABLE 3MS Medium composition
The 1/2-3/4MS culture medium referred to in the examples refers to a culture medium prepared by reducing the amounts of macroelements in the MS culture medium to 1/2 and 3/4, respectively, while the concentrations of other components are unchanged.
9. Growth promoting cultivation management: when the sprout of the tissue culture seedling grows to 5.0 cm high and 2 leaves are unfolded, 3-5g of slow release fertilizer (N-P-K = 15-8-14) is applied to each pot, 1000-1500 times of water-soluble fertilizer (N-P-K = 20-20-20) is used for irrigating the root, and the root is irrigated for 1 time every 7-10 days. The survival rate of the tissue culture seedlings can reach more than 95 percent, the pots are changed once a year in the second and third years, and the seedlings enter the flowering period after being transplanted for 3 years, and bloom more than 300 days earlier than the conventional cymbidium tissue culture seedlings cultured without treatment.
The preparation method of the root-extracting vigor agent comprises the following steps: potassium permanganate and sodium chloride are dissolved in water, so that the concentration of the potassium permanganate is 0.1-0.2% (mass fraction) and the concentration of the sodium chloride is 0.3-0.6% (mass fraction).
The dormancy breaking agent is prepared by adding 0.05-0.1 mg/L6-BA, 0.25-1.0mg/L IBA, and 50-100mg/LGA into 1/2-3/4MS culture medium (only macroelements are changed to 1/2-3/4 times of original ones) 3 、0.68-1.36g/LKH 2 PO 4 Adjusting pH to 5.8-6.0 with 1N NaOH, and sterilizing.
Example 2
The experimental material is the tissue culture seedling of the white ink, the culture method and the steps are shown in the embodiment 1, the survival rate of the tissue culture seedling of the white ink after transplanting can reach more than 95%, and the seedling enters the flowering period after transplanting in 3 rd year, and blooms more than 300 days earlier than the conventional tissue culture seedling of the white ink without treatment.
Example 3
The experimental material is a hybrid tissue culture seedling of cymbidium ensifolium x cymbidium ensifolium, the culture method and the steps are shown in the embodiment 1, the survival rate of the hybrid tissue culture seedling of cymbidium ensifolium x cymbidium ensifolium after transplantation can reach more than 95%, and the hybrid tissue culture seedling of cymbidium ensifolium x cymbidium ensifolium after transplantation enters the flowering period after 3 years, and blooms more than 300 days earlier than the conventional hybrid tissue culture seedling of cymbidium ensifolium x cymbidium ensifolium without treatment.
Claims (2)
1. A method for breaking the transplanting dormancy of tissue culture seedlings of cymbidium and shortening the production period is characterized by comprising the following steps:
(1) Soaking the cymbidium tissue culture seedling in a root-extracting activator for 10-30min, placing in a cool and ventilated place, air-drying until the color of the root system of the tissue culture seedling turns white, and then transplanting;
(2) Spraying the dormancy breaking agent 30 days after the tissue culture seedlings of the cymbidium sinensis are transplanted, spraying the dormancy breaking agent 1 time every 7-15 days, continuously spraying the dormancy breaking agent 3-5 times, and then carrying out growth promoting cultivation management;
the root-extracting activator is a mixed solution of 0.1-0.2% by mass of potassium permanganate and 0.3-0.6% by mass of sodium chloride;
the dormancy breaking agent consists of the following components: 1/2-3/4MS, 0.05-0.1 mg/L6-BA, 0.25-1.0mg/L IBA, 50-100mg/LGA 3 、0.68-1.36 g/L KH 2 PO 4 The pH value is 5.8-6.0;
the transplanting is to plant the tissue culture seedlings on a layered filling matrix planting cup, the layered filling matrix planting cup is provided with a middle layer and a lower layer which are pond sludge peanut shell mixed matrixes, and an upper layer which is a bark and orchid stone mixed matrix;
the growth promoting cultivation management comprises the steps of applying slow release fertilizer N-P-K = 15-8-14-5 g to each pot when the new bud of the tissue culture seedling grows to be 5.0 cm high and 2 leaves are unfolded, and irrigating the roots with liquid with water soluble fertilizer N-P-K = 20-20-20-1000-1500 times, wherein the roots are irrigated for 1 time every 7-10 days; the pots are changed every year in the second and third years.
2. The method according to claim 1, wherein in step (1), the cymbidium tissue culture seedling is cymbidium tissue culture seedling bottle seedling selected from rooting strong seedling culture for 60-90 days.
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