CN107821162B - Large-scale production method of gypsophila paniculata plug seedlings - Google Patents
Large-scale production method of gypsophila paniculata plug seedlings Download PDFInfo
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Abstract
The invention provides a large-scale production method of gypsophila paniculata plug seedlings. The method comprises the steps of selecting newly germinated branch buds on robust branches as explants in 12 months, and finally obtaining high-quality gypsophila paniculata plug seedlings through induction, proliferation, seedling strengthening, rooting culture, out-of-bottle cuttage and rooting maintenance, plug seedling field planting and maintenance. The method is characterized in that the LED light formula adopted in the induction and proliferation stages is red light: blue light: white light 5:2: 3; the LED light formula adopted in the stages of strong seedling and rooting culture is red light: blue light: white light: compared with the traditional fluorescent lamp, the far-red light is 5:1:2:2, the electric energy is saved by 43.7-53%, and the increment rate and the seedling strengthening rate are respectively improved by 24.1% and 40%. After the cuttage outside the bottle, the rooted plantlets can be stored for a long time at low cost on the premise of only providing water, the flexible and sufficient supply of seedlings is ensured, a high-quality and high-efficiency planting and maintaining technology for the gypsophila paniculata plug seedlings is established, and the large-scale production of the gypsophila paniculata plug seedlings is realized.
Description
Technical Field
The invention belongs to the technical field of flower seedling propagation. In particular to a method for producing gypsophila paniculata plug seedlings.
Background
Caulis et folium Gypsophilae Paniculatae (Gypsophila paniculata L.), and its original name, Calophyllum paniculatum, Phyllostachys nigra, Cyanea incana, and caulis et folium Gypsophilae. Caryophyllaceae family stone flower genus perennial herb. Wild resources are distributed in areas such as Altai mountainous area and tasshcoul trunk in Xinjiang, and roots and stems can be used for medicine, so that the method is mainly used for cultivating cut flowers.
The planting area of the gypsophila paniculata in Yunnan is about 6000 mu, and the planting area and the yield both account for more than 90 percent of the total amount of the whole country. The existing gypsophila seedlings are mainly produced by two ways of cutting propagation and tissue culture propagation, wherein the cutting propagation way comprises the following steps: strong seasonality, low propagation coefficient, low efficiency and the like; the prior art of tissue culture propagation approach comprises: the tissue culture room has the problems of high energy consumption for artificial light supplement and temperature reduction, low seedling strengthening rate and the like. In addition, the supply of the gypsophila paniculata plug seedlings and the corresponding production technology are lacked, the traditional nutrition bag seedling culture is labor-consuming and time-consuming, and the transportation is extremely inconvenient. In this way, by screening the light formula, the tissue culture production energy consumption is reduced, the strong seedling rate is improved, a set of gypsophila paniculata plug seedling production technology is developed, the gypsophila paniculata plug seedling large-scale production is realized, and the method has strong practical application value and popularization prospect.
Disclosure of Invention
In order to solve the technical problems of high energy consumption of artificial light sources and air-conditioning cooling, low strong seedling rate and lack of gypsophila paniculata plug seedling production in the gypsophila paniculata breeding link, the invention provides a large-scale production method of gypsophila paniculata plug seedlings through the research and development of the links of gypsophila paniculata light source screening and evaluation, plug specification, matrix proportion and the like.
The technical scheme of the scale production method of the gypsophila paniculata plug seedlings provided by the invention comprises the following steps:
(1) selection and disinfection treatment of extraterrestrial bodies
Selecting newly-germinated branch buds on a robust branch of a Gypsophila paniculata plant with the diameter of more than 0.4 cm in 12 months per year, removing leaves, cutting and reserving 0.5-1.0cm of top buds, taking the cut and left top buds as Gypsophila paniculata explants after outer leaves are removed, and sterilizing the Gypsophila paniculata explants;
(2) induced culture of aseptic seedlings
Inoculating sterilized Gypsophila paniculata into an induction culture medium under aseptic conditions, and culturing at 24-26 deg.C under illumination of an LED combined light source IAnd (5) cultivating for 15-20 days to obtain sterile seedlings, wherein the illumination conditions of the LED combined light source I are as follows: LED red light: LED blue light: the illumination intensity ratio of the LED white light is 5:2:3, and the illumination intensity of a 25 cm position under the LED combined light source I is 55-57 mu mol.m-2·s-1The illumination time is 12 hours/day;
(3) proliferation culture
Under the aseptic condition, cutting the sterile plantlets induced in the step (2), transferring the cut sterile plantlets into a multiplication culture medium, and culturing for 18-23 d under the illumination condition of an LED combined light source I with the culture temperature of 22 +/-2 ℃ and the same as that of the step (2) to obtain clustered plantlets;
(4) strong seedling culture
Under the aseptic condition, cutting the cluster seedlings cultured in the step (3) into stem sections according to the nodes, inoculating the stem sections into a proliferation culture medium, and culturing for 25-30 d under the conditions that the culture temperature is 22 +/-2 ℃ and the illumination of an LED combined light source II is performed to obtain strong seedlings; the illumination conditions of the LED combined light source II are as follows: LED red light: LED blue light: LED white light: the illumination intensity ratio of the far red light of the LED is 5:1:2:2, and the illumination intensity of a 25 cm position under the LED combined light source II is 53-55 mu mol · m-2·s-1The illumination time is 12 hours/day;
(5) rooting culture
Cutting off a stem tip with the length of 1cm at the upper part of the strong seedling cultured in the step (4) under the aseptic condition, inoculating the cut stem tip into a rooting culture medium, and culturing for 7-10 days under the same culture temperature and illumination condition of an LED combined light source II as that in the step (4) to obtain a rooting seedling;
(6) cuttage outside bottle and rooting maintenance
Cuttage outside the bottle, illumination and humidity management
Before 3-4 months of rainy season every year, taking out the rooted seedlings cultured in the step (5), soaking the rooted seedlings in a carbendazim solution for 1-2 minutes, dipping the base parts of the rooted seedlings in a rooting agent, cutting the rooted seedlings into perlite substrates in seedling trays, spraying rooting water after cutting, putting the seedling trays of the rooted seedlings which are well cut into a greenhouse, covering an arch center above the seedling trays with a transparent colorless plastic film, building two layers of sunshade nets above the transparent colorless plastic film, wherein the shading rate of each layer of sunshade net is 70%, and keeping the relative humidity of air in the greenhouse to be more than 95%; after 3 days, removing the transparent colorless plastic film, controlling the relative humidity of the air in the greenhouse to be more than 90%, after 3-4 days, removing one layer of sunshade net, keeping the relative humidity of the air in the greenhouse to be more than 80%, and after 1 week, removing the other layer of sunshade net;
② moisture management
Placing the rooted seedling tray after cuttage in a greenhouse for 2-3 times per day until the seedling tray is placed in the greenhouse for 1 day to 10 days; watering and permeating water once every morning from day 11 to before transplanting of rooted seedlings;
(7) plug seedling planting and maintenance
And (3) when more than 50% of the rooted seedlings have 4-6 root systems with the root systems being more than 3cm in the step (6), transplanting the rooted seedlings into holes of a 128-hole plug filled with the matrix, watering the rooted seedlings with root fixing water for 1-2 times a day, spraying a compound fertilizer every 7-10 days, and when the seedlings are 4-5 cm high and have 10-11 leaves, obtaining the produced gypsophila paniculata plug seedlings.
Further, the gypsophila paniculata in the step (1) is a gypsophila paniculata which has obvious variety specific characters and good growth vigor.
Further, the step (1) of sterilizing the Gypsophila paniculata is to clean the Gypsophila paniculata in a 5% w/w detergent solution, sterilize the Gypsophila paniculata in a 0.16% w/w mercuric chloride solution for 9 to 10 minutes, sterilize the Gypsophila paniculata in a 0.25% w/w sodium hypochlorite solution for 8 to 9 minutes, and rinse the Gypsophila paniculata with sterile water for 3 to 4 times.
Further, the induction culture medium in the sterile seedling induction culture in the step (2) is as follows: 1.0-1.5 mg/L of MS +6-BA + 0.1mg/L of NAA0.1mg/L + 6.5-7.0 g/L of agar + 30g/L of white sugar, and the pH value is 5.5-5.8.
Further, the proliferation culture medium in the proliferation culture in the step (3) is the same as the proliferation culture medium in the strong seedling culture in the step (4), and the proliferation culture medium is as follows: 0.4-0.6 mg/L of MS +6-BA, 0-0.1 mg/L of NAA, 30g/L of sucrose and 6.5g/L of agar, and the pH value is 5.5-5.8.
Further, the rooting medium in the rooting culture in the step (5) is: 1/2MS + 0.2-0.5 mg/L NAA + 0.1-0.3 mg/L IAA + 30g/L sucrose + 6.5g/L agar, and the pH value is 5.5-5.8.
Further, the concentration of the carbendazim solution in the step (6) is 0.1% w/w.
Further, the rooting agent in the step (6) is 0.1mg/L IBA solution, the thickness of the perlite matrix in the seedling tray in the step (6) is 2.5-3.0 cm, and the cutting density of the rooted seedlings is 1.0cm multiplied by 1.5 cm.
Further, in the step (7), the substrate in the 128-hole tray holes is turf: the volume ratio of perlite is 5: 1.
Further, the compound fertilizer in the step (7) is a compound fertilizer solution containing 100mg/L, P25 mg/L of N and 110mg/L of K.
Compared with the prior art, the invention has the following main innovations and beneficial effects:
1. on the premise of ensuring the rapid tissue culture and propagation of gypsophila paniculata, high seedling strengthening rate and high seedling rate, the electric energy consumption is greatly reduced, the production safety of a tissue culture room is ensured, and the production cost is saved.
The power and luminous flux values of the proliferation light formula (illumination condition of the LED combined light source I) and the seedling strengthening light formula (illumination condition of the LED combined light source II) compared with the fluorescent lamp can be seen (table 2), compared with the comparison, the proliferation light formula and the seedling strengthening light formula provided by the invention provide more light quanta for tissue culture seedlings, the power consumption is only 56.3% and 47% of that of the fluorescent lamp respectively, and the power saving is 43.7% and 53%. In the tissue culture room, because the fluorescent lamp has high heat dissipation, even in winter, an air conditioning system of the tissue culture room is still in a refrigerating state, and some electric energy consumption can be generated. After the proliferation light formula and the seedling strengthening light formula are applied, the tissue culture and rapid propagation of gypsophila paniculata is ensured, the seedling strengthening rate is high, the seedling survival rate is high, the cooling load of an air conditioning system of a tissue culture room is obviously reduced due to the low heat dissipation of the LED, the electricity cost is saved, the load of an electrical system is reduced, and the production safety of the tissue culture room is ensured.
2. The reproduction rate and the strong seedling rate are improved.
According to the invention, the proliferation light formula (LED combined light source I illumination condition) and the seedling strengthening light formula (LED combined light source II illumination condition) adopted in the propagation stage (step (3) proliferation culture) ] and the seedling strengthening stage (step (4) seedling strengthening culture) ] respectively obviously improve the propagation rate and the seedling strengthening rate of seedlings, the propagation rate and the seedling strengthening rate are respectively improved by 24.1% and 40% compared with those of a common fluorescent light source, and the production efficiency is improved (see table 3 and table 4).
3. The developed out-of-bottle cuttage and rooting maintenance technology can preserve rooted plantlets for a long time at low cost, and ensures flexible and sufficient supply of seedlings.
According to the invention, the out-of-bottle cuttage and the maintenance are carried out in the step (6), and the rooted plantlets can be stored for a long time with low cost on the premise of only providing water after cuttage, so that the flexible and sufficient supply of the plantlets is ensured.
4. The time nodes of all the steps are properly controlled, and the efficient production and supply of the gypsophila paniculata plug seedlings are ensured.
Step (6), cuttage and maintenance are carried out outside the bottle, the cuttage is strictly controlled to be completed within 3-4 months, at the moment, rainy season does not come yet, air humidity is low and temperature is proper in sunny days, and the survival rate of cuttage outside the bottle is favorably improved (see table 5); in addition, the completion of the out-of-bottle cuttage in 3-4 months is beneficial to the completion of the hole tray seedling planting and maintenance in 5-6 months, ensures that the hole tray seedlings are supplied in large quantity in 6-7 months in the peak period of the market needing seedlings, the time point is well connected, the maintenance cost is reduced, the seedlings rooted in the perlite can be kept for more than 6 months at low cost (only watering is carried out to ensure the survival of the seedlings), and the sufficient and flexible supply of the hole tray seedlings is ensured.
5. The cultivated seedlings are strong and high-quality, so that the rooting rate of the cuttage outside the bottle reaches more than 93%, the seedling rate of the produced high-quality gypsophila hole tray seedlings reaches more than 98%, the production period is short, and a large amount of high-quality seedlings supplied to the market can be cultivated from a small amount of explants only in 7-8 months.
6. The method for producing the gypsophila paniculata plug seedlings is low in cost, high in quality, intensive and efficient, and greatly facilitates subsequent logistics transportation.
Compared with the nutrition bag seedling culture, the production link of the gypsophila paniculata plug seedlings produced by the invention obviously saves labor force and is greatly convenient for subsequent logistics transportation.
7. Establishes a high-quality and high-efficiency gypsophila paniculata plug seedling planting and maintaining technology.
By screening the plug specification and the matrix proportion, the step (7) adopts the proper plug specification and the matrix proportion, thereby obviously improving the plug seedling production efficiency and providing an intensive and efficient plug seedling production method; the 128-hole tray does not waste the matrix, and the seedlings have strong growth vigor, dark green leaf color, good quality and high production efficiency in unit area (see table 6).
Drawings
FIG. 1: the growth vigor of the gypsophila under the illumination culture condition of the LED combined light source II for strong seedling culture is compared with that under the light source of a fluorescent lamp. The right picture in FIG. 1 shows the growth of the gypsophila under the fluorescent lamp, with yellow leaves and slender stalks; the left figure shows the growth vigor of gypsophila under the illumination culture condition of the LED combined light source II for strong seedling culture, the leaf color is dark green, and the stem is thick and strong.
FIG. 2: the gypsophila paniculata plug seedlings (128 holes) produced by the method grow, and are robust and tidy.
Detailed Description
The present invention will be further illustrated by the following examples, but the present invention is not limited thereto, and conventional methods are not specifically illustrated in the examples.
Example 1
The implementation place is as follows: the Jiuxin base of the institute of agriculture and sciences, Yunnan province, is located in Jiuxin village of the town of Jiuxin county, Jiangxian city, Yunnan province, the altitude: 1680 m.
(1) Selection and disinfection treatment of extraterrestrial bodies
Selecting a full-scale extraterrestrial colonizer: in 18 days 12 months in 2013, newly-germinated branch buds on thick and strong branches with the diameter of more than 0.4 cm are picked from a plant of the gypsophila paniculata variety 'Yunxing 75' with obvious variety specific characters (the variety is white in flower, upright in plant shape and needle-shaped in leaves) and good growth vigor, leaves are removed, 0.5-1.0cm of terminal buds are cut and reserved, and 1-2 pieces of outer-layer leaves are peeled to obtain the gypsophila paniculata explant; the special characters of the variety are obvious, and the good growth vigor means that the quality characters such as flower color, plant shape, leaf shape and the like accord with the characteristics of the variety, and the plant grows healthily and robustly.
② sterilization treatment: the Gypsophila paniculata is cleaned in a 5% w/w detergent solution, sterilized in a 0.16% w/w mercuric chloride solution for 10 minutes, sterilized in a 0.25% w/w sodium hypochlorite solution for 8 minutes, and rinsed with sterile water for 3-4 times.
(2) Induced culture of aseptic seedlings
12 and 18 days in 2013, inoculating the Gypsophila paniculata which is sterilized by ② in the step (1) into an induction culture medium under the aseptic condition, and culturing for 20 days under the conditions that the culture temperature is 24-26 ℃ and the illumination intensity of an LED combined light source I is 5:2:3, wherein the illumination intensity of the LED combined light source I at a position of 25 cm under the LED combined light source I is 56 mu mol/m to obtain the aseptic plantlet after 1 month and 7 days-2·s-1The illumination time is 12 hours/day; the unit of the illumination intensity of the red light, the blue light and the white light of the LED is mu mol.m-2·s-1。
The induction culture medium is as follows: MS +6-BA 1.0mg/L + NAA0.1mg/L + agar 6.8g/L + white sugar 30g/L, and pH is 5.5-5.8.
(3) Proliferation culture
1 month and 7 days, cutting the sterile plantlets induced in the step (2) under aseptic conditions, transferring the cut sterile plantlets into the following proliferation culture media, and culturing for 23 days, namely 1 month and 30 days, under the illumination conditions of the culture temperature of 22 +/-2 ℃ and the LED combined light source I same as the step (2) to obtain a large amount of clustered plantlets; the proliferation culture medium is as follows: MS +6-BA 0.5mg/L + NAA0.1mg/L + sucrose 30g/L + agar 6.5g/L, and pH is 5.5-5.8.
(4) Strong seedling culture
1, 30 days in 1 month, cutting the cluster seedlings cultured by proliferation in the step (3) into stem sections according to nodes under the aseptic condition, inoculating into the proliferation culture medium in the step (3), and culturing for 25 days under the conditions that the culture temperature is 22 +/-2 ℃ and the illumination of an LED combined light source II is performed to obtain strong seedlings; the illumination conditions of the LED combined light source II are as follows: LED red light: LED blue light: LED white light: the illumination intensity ratio of the far red light of the LED is 5:1:2:2, and the illumination intensity of a position 25 cm below the LED combined light source II is 54 mu mol · m-2·s-1The illumination time was 12 hours/day. The unit of each illumination intensity of the LED red light, the LED blue light, the LED white light and the LED far-red light is mu mol.m-2·s-1。
(5) Rooting culture
2, 24 days in 2 months, cutting off a stem tip with the length of 1cm at the upper part of the strong seedling cultured in the step (4) under the aseptic condition, inoculating the cut stem tip into the following rooting culture medium, and culturing for 7 days under the same culture temperature and the illumination condition of an LED combined light source II as the step (4) to obtain a rooting seedling;
the rooting culture medium comprises: 1/2MS + NAA 0.4mg/L + IAA 0.2mg/L + sucrose 30g/L + agar 6.5g/L, and pH is 5.5-5.8.
(6) Cuttage outside bottle and rooting maintenance
Cuttage outside the bottle, illumination and humidity management
3 months and 3 days, before a rainy season comes, taking out the rooted seedlings cultured in the step (5), soaking the rooted seedlings in a carbendazim solution with the concentration of 0.1% w/w for 2 minutes, dipping the base parts of the rooted seedlings in a rooting agent, and then cutting the rooted seedlings into a perlite matrix in a seedling tray, wherein the thickness of the perlite matrix in the seedling tray is 2.5-3.0 cm, and the cutting density of the rooted seedlings is 1.0cm multiplied by 1.5 cm. The preparation of the 0.1% w/w carbendazim solution is 250 times that of 25% carbendazim wettable powder. The rooting agent is IBA solution with the concentration of 0.1mg/L, and the preparation method of the IBA solution with the concentration of 0.1mg/L comprises the following steps: weighing 0.1 g of IBA (indolebutyric acid) and dissolving in 100ml of NaOH solution with the concentration of 0.5mol/L, and adding purified water to reach the constant volume of 1000ml to obtain mother liquor with the concentration of 0.1 mg/ml; storing the mother liquor in a refrigerator at 4 ℃, taking 1ml of the mother liquor when needed, and using purified water to fix the volume to 1000ml to obtain the IBA solution with the concentration of 0.1 mg/L.
After cuttage, spraying root fixing water, putting a seedling tray of the rooted seedlings which are subjected to cuttage into a greenhouse, covering a transparent colorless plastic film on an arch center above the seedling tray, and building two layers of sunshade nets above the transparent colorless plastic film, wherein the shading rate of each layer of sunshade net is 70%, and the relative air humidity in the greenhouse is kept to be more than 95%; and after 3 days, removing the transparent colorless plastic film, controlling the relative air humidity in the greenhouse to be more than 90%, after 4 days, removing one layer of sunshade net, keeping the relative air humidity in the greenhouse to be more than 80%, and after 1 week, removing the other layer of sunshade net.
② moisture management
Placing the rooted seedling tray after cuttage in a greenhouse for the first day to the 10 th day, and watering for 2-3 times per day to permeate water; watering and permeating water once every morning from the 11 th day to before transplanting of rooted seedlings.
4.8 root systems with the length of more than 3cm exist after 30 days of cuttage, namely 4 months and 12 days, and the rooting rate is 94.7%.
The rooted seedlings can be stored in a seedling raising shed for 12 months in the year on the premise of properly watering (not fertilizing) and ensuring that the seedlings are not dehydrated and withered.
(7) Plug seedling planting and maintenance
And (4) for 2 days after 4 months, wherein more than 50% of the rooted seedlings in the step (6) have 4-6 root systems with the root systems of more than 3cm, and all the rooted seedlings in the seedling tray are transplanted to a seedling tray filled with turf: in 128-hole tray holes (the tray length is 52cm, the width is 26cm, the hole depth is 4cm) of a mixed matrix with the volume ratio of perlite being 5:1, after root water is poured, water is permeated for 1-2 times every day, and a compound fertilizer is sprayed once every 7-10 days, wherein the compound fertilizer is a solution containing N100 mg/L, P25 mg/L and K110 mg/L (the fluctuation range of the concentration is within 5 percent), and the preparation method of the compound fertilizer comprises the following steps: 54.21 kg of Ca (NO) are taken3)2·4H2O, 2.66 kg NH4NO322.51 kg KNO3And 10.93 kg KH2PO4Dissolving the fertilizer water mother liquor in 1000 liters of water to obtain fertilizer water mother liquor, and diluting the fertilizer water mother liquor by 100 times when needed to obtain solution containing N100 mg/L, P25 mg/L and K110 mg/L; transplanting for 30 days, namely 5 months and 2 days, wherein the height of the seedlings is 4-5 cm, 10-11 leaves are produced, namely the gypsophila paniculata plug seedlings, the high-quality seedling standard which can be listed is achieved, and the seedling rate is 98.0%.
Taking out the cutting seedlings in the step (6) from the matrix respectively at 23 days in 10 months and 26 days in 12 months, transplanting the cutting seedlings into the 128-hole plug tray, following the management, wherein the seedlings grow to 4-5 cm in 23 days in 11 months and 25 days in 1 month, 10-11 leaves are provided, the seedling reaches the standard of high-quality seedlings which can be listed, and the seedling rate is 97.8% and 97.6% respectively;
the light quality information of the above LED light sources is shown in table 1.
Table 1: light quality information of various light sources
| Spectral power distribution | Peak wavelength/nm | Wavelength half width/nm |
| LED Red light | 660 | 25 |
| LED blue light | 460 | 25 |
| LED far-red light | 715 | 25 |
| LED white light | 380 to 750 (wavelength range) | – |
Table 2: the light formula of the invention for proliferation and seedling strengthening has the advantages of luminous intensity and energy consumption compared with those of a common fluorescent lamp (/ m)2)
M in Table 22Which means that the light source irradiates an area of 1 square meter.
Table 2 shows that: the power consumption of the proliferation light formula (LED combined light source I) is only 56.3 percent of that of a fluorescent lamp, and the power consumption of the seedling strengthening light formula (LED combined light source II) is only 47 percent of that of the fluorescent lamp, which saves 43.7 percent of electric energy and 53 percent of electric energy compared with the common fluorescent lamp light source respectively. The light intensity of the two light formulas at a position of 25 cm under a light source is respectively 56 mu mol/s/m2、54μmol/s/m2All are higher than common fluorescent lamp lightThe illumination intensity of the source at the position of 25 cm below the light source not only saves energy consumption, but also ensures the growth of the gypsophila plant.
Table 3: influence of LED combined light source I on proliferation multiple of tissue culture seedlings of gypsophila paniculata
Table 4: influence of LED combined light source II on seedling strengthening rate of gypsophila paniculata tissue culture seedlings
Tables 3 and 4 show that: the proliferation rate of the proliferation light formula (LED combined light source I) is increased by 24.1% compared with that of the proliferation culture of a common fluorescent lamp light source, and the strong seedling rate of the strong seedling culture of the strong seedling light formula (LED combined light source II) is increased by 40% compared with that of the strong seedling culture of the common fluorescent lamp light source.
Table 5: comparison of external cutting effect of bottles in different months
Table 6: influence of different plug specifications on production efficiency of gypsophila paniculata plug seedlings
Examples 2 and 3 are the same as example 1 except that the measures listed in table 7 are different from example 1, and the description thereof is omitted.
Table 7: differences between example 2 and example 3 and example 1
Claims (7)
1. A large-scale production method of gypsophila paniculata plug seedlings is characterized by comprising the following steps:
(1) selection and disinfection treatment of gypsophila paniculata explants
Selecting newly-germinated branch buds on a robust branch of a gypsophila paniculata plant with the diameter of more than 0.4 cm in 12 months per year, removing leaves, cutting 0.5-1.0cm of terminal buds, taking the cut leaf buds as a gypsophila paniculata explant after the outer leaves are removed, and sterilizing the gypsophila paniculata explant;
(2) induced culture of aseptic seedlings
Inoculating the sterilized Gypsophila paniculata explants into an induction culture medium under the aseptic condition, and culturing for 15-20 days under the illumination condition of an LED combined light source I at the culture temperature of 24-26 ℃ to obtain aseptic seedlings, wherein the illumination condition of the LED combined light source I is as follows: LED red light: LED blue light: the illumination intensity ratio of the LED white light is 5:2:3, and the illumination intensity of a 25 cm position under the LED combined light source I is 55-57 mu mol.m-2·s-1The illumination time is 12 hours/day;
(3) proliferation culture
Under the aseptic condition, cutting the sterile plantlets induced in the step (2), transferring the cut sterile plantlets into a multiplication culture medium, and culturing for 18-23 d under the illumination condition of an LED combined light source I with the culture temperature of 22 +/-2 ℃ and the same as that of the step (2) to obtain clustered plantlets;
(4) strong seedling culture
Under the aseptic condition, cutting the cluster seedlings cultured in the step (3) into stem sections according to the nodes, inoculating the stem sections into a proliferation culture medium, and culturing for 25-30 d under the conditions that the culture temperature is 22 +/-2 ℃ and the illumination of an LED combined light source II is performed to obtain strong seedlings; the illumination conditions of the LED combined light source II are as follows: LED red light: LED blue light: LED white light: the illumination intensity ratio of the far red light of the LED is 5:1:2:2, and the illumination intensity of a 25 cm position under the LED combined light source II is 53-55 mu mol · m-2·s-1Time of illumination12 hours/day;
(5) rooting culture
Cutting off a stem tip with the length of 1cm at the upper part of the strong seedling cultured in the step (4) under the aseptic condition, inoculating the cut stem tip into a rooting culture medium, and culturing for 7-10 days under the same culture temperature and illumination condition of an LED combined light source II as that in the step (4) to obtain a rooting seedling;
(6) cuttage outside bottle and rooting maintenance
Cuttage outside the bottle, illumination and humidity management
Before 3-4 months of rainy season every year, taking out the rooted seedlings cultured in the step (5), soaking the rooted seedlings in a carbendazim solution for 1-2 minutes, dipping the base parts of the rooted seedlings in a rooting agent, cutting the rooted seedlings into perlite substrates in seedling trays, spraying rooting water after cutting, putting the seedling trays of the rooted seedlings which are well cut into a greenhouse, covering an arch center above the seedling trays with a transparent colorless plastic film, building two layers of sunshade nets above the transparent colorless plastic film, wherein the shading rate of each layer of sunshade net is 70%, and keeping the relative air humidity in the greenhouse to be more than 95%; after 3 days, removing the transparent colorless plastic film, controlling the relative humidity of the air in the greenhouse to be more than 90%, after 3-4 days, removing one layer of sunshade net, keeping the relative humidity of the air in the greenhouse to be more than 80%, and after 1 week, removing the other layer of sunshade net;
② moisture management
Placing the rooted seedling tray after cuttage in a greenhouse for 2-3 times per day until the seedling tray is placed in the greenhouse for 1 day to 10 days; watering and permeating water once every morning from day 11 to before transplanting of rooted seedlings;
(7) plug seedling planting and maintenance
When more than 50% of the rooted seedlings in the step (6) have 4-6 root systems with the root systems being more than 3cm, transplanting the rooted seedlings into holes of a 128-hole plug filled with a matrix, watering the rooted seedlings with root fixing water for 1-2 times a day, spraying a compound fertilizer every 7-10 days, and when the seedlings are 4-5 cm high and have 10-11 leaves, obtaining the produced gypsophila paniculata plug seedlings;
the induction culture medium in the induction culture of the aseptic seedlings in the step (2) is as follows: 1.0-1.5 mg/L of MS +6-BA + 0.1mg/L of NAA + 6.5-7.0 g/L of agar + 30g/L of white sugar, and the pH value is 5.5-5.8;
the proliferation culture medium in the proliferation culture in the step (3) is the same as the proliferation culture medium in the strong seedling culture in the step (4), and the proliferation culture medium is as follows: 0.4-0.6 mg/L of MS +6-BA, 0-0.1 mg/L of NAA, 30g/L of cane sugar and 6.5g/L of agar, wherein the pH value is 5.5-5.8;
the rooting culture medium in the rooting culture of the step (5) is as follows: 1/2MS + 0.2-0.5 mg/L NAA + 0.1-0.3 mg/L IAA + 30g/L sucrose + 6.5g/L agar, and the pH value is 5.5-5.8;
the rooting agent in the step (6) is 0.1mg/L IBA solution.
2. The large-scale production method of gypsophila paniculata seedlings according to claim 1, characterized in that:
the gypsophila paniculata in the step (1) is a gypsophila paniculata which has obvious special characters of varieties and good growth vigor.
3. The large-scale production method of gypsophila paniculata seedlings according to claim 1, characterized in that:
the step (1) of sterilizing the Gypsophila paniculata explant comprises the steps of cleaning the Gypsophila paniculata explant in a 5% w/w detergent solution, sterilizing the Gypsophila paniculata explant in a 0.16% w/w mercuric chloride solution for 9-10 minutes, sterilizing the Gypsophila paniculata explant in a 0.25% w/w sodium hypochlorite solution for 8-9 minutes, and rinsing the Gypsophila paniculata explant with sterile water for 3-4 times.
4. The large-scale production method of gypsophila paniculata seedlings according to claim 1, characterized in that:
the concentration of the carbendazim solution in the step (6) is 0.1% w/w.
5. The large-scale production method of gypsophila paniculata seedlings according to claim 1, characterized in that:
and (6) the thickness of the perlite matrix in the seedling tray is 2.5-3.0 cm, and the cutting density of the rooted seedlings is 1.0cm multiplied by 1.5 cm.
6. The large-scale production method of gypsophila paniculata seedlings according to claim 1, characterized in that:
in the step (7), the substrate in the 128-hole plug hole is turf: the volume ratio of perlite is 5: 1.
7. The large-scale production method of gypsophila paniculata seedlings according to claim 1, characterized in that:
the compound fertilizer in the step (7) is a compound fertilizer solution containing 100mg/L, P25 mg/L of N and 110mg/L of K.
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