CN112042521A

CN112042521A - Transplanting method for test-tube plantlet of rhizome plant

Info

Publication number: CN112042521A
Application number: CN202010796662.XA
Authority: CN
Inventors: 张华通; 张炜; 黄剑雄; 林云斌
Original assignee: Yangshan County Sanlianyang Ecological Agriculture And Forestry Development Co ltd
Current assignee: Yangshan County Sanlianyang Ecological Agriculture And Forestry Development Co ltd
Priority date: 2020-08-10
Filing date: 2020-08-10
Publication date: 2020-12-08

Abstract

The invention discloses a method for transplanting test-tube plantlets of rhizome plants, which comprises the following steps: s1, hardening and culturing tissue culture seedlings of rhizome plants; s2, cleaning and disinfecting the test-tube plantlets of the rhizomes, and airing the plantlets; s3, planting in a grading manner according to the growth condition of the test-tube plantlets of the rhizomes; s4, cultivation management after transplantation is carried out; s5, the test-tube plantlets of the rhizomes are grown into seedlings and are outplanted. The method introduces the sterilizing rooting powder in the transplanting process, is used in the tissue culture and industrialized seedling transplantation of polygonatum cyrtonema tissue culture of the rhizome test-tube plantlets, obtains high survival rate and seedling rate, accelerates the rooting and bud germination of the rhizome, plays a role in efficient sterilization, effectively prevents the generation of rhizome decay and the bite of underground pests, ensures the rooting and seedling of the rhizome, improves the transplanting survival rate of the rhizome test-tube plantlets, and is also suitable for other plants with meristem propagation.

Description

Transplanting method for test-tube plantlet of rhizome plant

Technical Field

The invention belongs to the technical field of plant cultivation, and particularly relates to a method for transplanting test-tube plantlets of a rhizome plant.

Background

Rhizome plants (such as Polygonatum cyrtonema Hua, potato Solanum tuberosum L., Zingiber officinale of fiscinale, Dioscorea zingiberensis C.H., Wright, Dioscorea opposita optifolia L.) bloom universally without forming seeds, or have few seeds and are difficult to germinate. Therefore, the seeds are difficult to be adopted for the large-scale seedling production. At present, except potatoes, the mass propagation of seedlings cannot be carried out by adopting plant tissue culture technology for most varieties. The rootstock meristem propagation is a relatively-numerous propagation method adopted at present, but the propagation method needs to consume a large amount of original rootstock materials, seedlings cultivated by the method are easy to degenerate, and the subsequent growth speed and the product quality of the seedlings are seriously influenced due to virus infection. In addition, the rootstocks adopted for the rootstock meristem propagation are very easy to cause rottenness and biting by underground pests due to the wounds left during cutting, thereby seriously affecting the seedling rate. Thus, the large-scale production of high-quality seedlings becomes a common problem of such plants.

Polygonatum cyrtonema Hua is sweet and neutral, and has the effects of invigorating qi, nourishing yin, invigorating spleen, moistening lung and tonifying kidney, and can be used for treating diseases caused by spleen deficiency, lung deficiency and kidney deficiency. It is recorded in the miscellaneous records of famous physicians. According to the regulation of pharmacopoeia of the people's republic of China, the medicinal polygonatum rhizome is dried rhizome of polygonatum kingianum, polygonatum rhizome or polygonatum cyrtonema of Liliaceae. Rhizoma polygonati is also a Chinese medicinal material approved by the national Wei Jian Wei and K. The sealwort has high economic value and medicinal value, can be widely applied to raw materials for producing new medicines, the field of chemical industry and the like, and has larger and larger market demand. However, wild polygonatum is slow in growth and breeding and low in survival rate, so that wild resources are scarce, and in recent years, wild polygonatum is excessively artificially harvested, the growth environment is damaged, seeds are disorderly introduced, and the like, so that the wild high-quality resources are increasingly reduced. In recent years, due to the fact that polygonatum is subjected to long-term yield pursuit, manual blind excavation and excessive mining and manual planting are not standard, so that wild polygonatum faces few seed sources, and the quality of polygonatum is reduced year by year. The shortage of excellent germplasm resources becomes the biggest bottleneck for the development of the polygonatum industry, the protection and the utilization of wild polygonatum resources become problems to be solved urgently, the breeding and the standardized cultivation are particularly urgent and important, and the method is an important means for solving the problem of the development of the polygonatum industry.

The method for industrially culturing seedlings by adopting tissue culture rootstocks is a unique and effective method for solving the problem of the bottleneck. However, no one has developed success at present, mainly because the polygonatum cyrtonema plants have special morphological structures, the fleshy root stems grow in soil, explants are difficult to sterilize and become sterile propagation materials, the root stems contain rich polysaccharides and other metabolic secondary substances and are easy to brown, and in addition, polygonatum cyrtonema has dormant biological characteristics, the number of buds of polygonatum cyrtonema needs to be continuously increased like most plant tissue culture in a bud multiplication mode, and the difficulty is very high. Therefore, the scientific, reasonable and effective method for tissue culture of the rhizome and large-scale seedling culture of the meristem is to be further researched.

Disclosure of Invention

The invention aims to provide a test-tube plantlet transplanting method for a rhizome plant.

The technical scheme adopted by the invention is as follows:

in a first aspect of the present invention, there is provided a method for transplanting a test-tube plantlet of a rhizome plant, comprising the steps of:

s1, hardening and culturing tissue culture seedlings of the rhizome plants to form rhizome test-tube seedlings;

s2, cleaning and disinfecting the test-tube plantlets of the rhizomes, and airing the plantlets;

s3, planting in a grading manner according to the growth condition of the test-tube plantlets of the rhizomes;

s4, performing cultivation management after transplantation to form test-tube plantlets with rootstocks;

s5, the test-tube plantlets of the rhizomes are grown into seedlings and are outplanted.

According to the method of the first aspect of the present invention, preferably, the conditions of the seedling exercising culture in the step S1 are: the temperature is 18.5-28.5 ℃, and the illumination intensity is 6000-8500 Lux; the sunshine duration is 8-11 h.

Hardening seedling shape to promote the culture process: preparing materials for hardening seedling and promoting root culture after the transfer on the superclean bench, immediately sealing the joint of a bottle cap and a bottle by using a preservative film on the same day after the transfer, and putting the bottle cap and the bottle into a seedling hardening plastic greenhouse for hardening seedling culture every other day.

The culture time for promoting the seedling hardening form is generally 40-60 days, and the specific time is determined according to actual conditions.

According to the method of the first aspect of the present invention, further, the seedling emergence criteria of the seedling exercising culture in step S1 are: (1) the color of the leaves is green, the number of the leaves reaches more than 3 or the leaves are withered and yellow without leaves; (2) the diameter of the rhizome is more than 2.5 cm; (3) the shape of the rhizome is obviously light green or green; (4) the growth is normal.

According to the method of the first aspect of the present invention, further, the specific method for cleaning and disinfecting the rhizomatic test-tube plantlets in step S2 is as follows: cleaning pollution-free seedlings with clean water, soaking the seedlings with a potassium permanganate solution, and then cleaning the seedlings with clean water again; cleaning polluted seedlings with clean water, soaking the polluted seedlings in a potassium permanganate solution, cleaning the polluted seedlings with clean water again, soaking the polluted seedlings in a sterilizing solution, and taking out the polluted seedlings.

Furthermore, after cleaning pollution-free seedlings with clean water, soaking the seedlings in a potassium permanganate solution for 7min, and then cleaning the seedlings with clean water again; cleaning polluted seedlings with clean water, soaking the polluted seedlings in a potassium permanganate solution for 10min, then cleaning the polluted seedlings with clean water again, soaking the polluted seedlings in a sterilization solution, and taking out the polluted seedlings.

More specifically, the test tube plantlets of the rootstock to be washed are separated into two types of non-contaminated plantlets and contaminated plantlets, and are washed separately to prevent cross contamination.

Taking out the test-tube plantlet from the rhizome: the trained tissue culture seedlings are gently picked out from the culture flask by using forceps (care is taken to avoid damaging the seedlings as much as possible), and then the seedlings are placed into a prepared clean plastic basin.

Cleaning the test-tube plantlets of the rhizomes: the test-tube plantlet of the rhizome needs to be cleaned by clean water for three times, firstly, the culture medium attached to the tissue culture seedling in the plastic basin is removed completely, a culture medium without residual base is needed, and the seedling is damaged as little as possible. And then cleaned twice with clean water.

Sterilizing pollution-free seedlings: and soaking the clean test tube seedling of the rhizome in 0.02 percent potassium permanganate solution for 7min, taking out, cleaning with clear water once, and cleaning residual potassium permanganate solution attached to the seedling.

Disinfecting the polluted seedlings: the cleaned test tube seedling of the rhizome is soaked in 0.02 percent potassium permanganate solution for 10min, then taken out, cleaned once by tap water, soaked in 1000 times of 70 percent carbendazim (or thiophanate methyl and the like) solution and then taken out.

And (4) placing the sterilized test-tube seedling of the rhizome in a ventilated and shady place for air drying until the moisture attached to the surface of the seedling just disappears.

According to the method of the first aspect of the present invention, further, the step S3 includes the following steps: the method is divided into a first stage, a second stage, a third stage and a stepless stage, and only the first stage, the second stage and the third stage are reserved;

first-stage: (1) the leaves are green, and the number of the leaves reaches more than 3; (2) the diameter of the rhizome is more than 3.5 cm; (3) the rhizome has obvious shape and is light green or green; (4) the growth is normal;

and (2) second stage: (1) the leaves are withered and yellow without leaves; (2) the diameter of the rhizome is 2.5-3.5 cm; (3) the rhizome has obvious shape and is light green or green; (4) the growth is normal;

third-stage: (1) the leaves are withered and yellow and fall off or have a certain number of leaves; (2) the diameter of the rhizome is 1.5-2.5 cm; (3) the rhizome has obvious shape and is light green or green; (4) the growth is normal.

According to the method of the first aspect of the present invention, in particular, the planting in step S3 is specifically performed by: wetting the test-tube plantlet of the rhizome, dipping the sterilizing and insect-preventing rooting powder on the surface, and planting the test-tube plantlet in the matrix.

More specifically, clean river sand (phi 0.2-3.0 mm) with proper thickness and good air and water permeability is required as a matrix for transplanting the test-tube plantlets of the rhizomes. And (3) disinfection: the matrix disinfectant is potassium permanganate, 50% sodium sulfadiazine and other soil disinfectant. The specific operation method is that 0.2 percent potassium permanganate solution is used for sprinkling to ensure that the matrix is uniformly wetted to the degree, and 500 times of 50 percent sodium diquat solution is used for sprinkling every other day.

The plants are planted in different grades and regions and are planted in river sand matrix, and the row spacing of the plants is 8 multiplied by 8 cm.

The substrate should cover the top of the rootstock, just the rootstock is buried in the substrate, and the leaf base cannot be covered, otherwise the seedling is rotten due to improper water spraying, and the seedling cannot be planted too shallowly, otherwise the growth of new roots of the seedling is not facilitated. After planting, the seedlings are lightly pinched by hands to ensure that the rhizome is fully contacted with the matrix, and the mixed solution of 1/4MS mineral elements, NAA0.2mg/L and 70 percent carbendazim (or thiophanate methyl and the like) 500 solution is sprayed.

According to the method of the first aspect of the invention, preferably 400-600 ppm IBA, 3-5 ppm 2.4-D, 20-40 ppm 6BA, 900-1100 ppm KH2PO4, 900-1100 ppm KNO3, 50-150 ppm FeSO 4.7H 20, 40-60 ppm H3BO3, 3-5 wt% nano silver ion solution, 4500-5500 ppm O, O-dimethyl- (2,2, 2-trichloro-1-hydroxyethyl) phosphonate and adhesive.

More preferably, it comprises per 1000g of unit weightThe following raw materials: 500ppm IBA, 4ppm 2.4-D, 30ppm 6BA, 1000ppm KH₂PO₄、1000ppm KNO₃、100ppm FeSO₄·7H₂0、50ppm H₃BO₃40 wt% nano silver ion solution, 5000ppm O, O-dimethyl- (2,2, 2-trichloro-1-hydroxyethyl) phosphonate and adhesive.

More preferably, the concentration of silver ions in the nano silver solution is 100 ppm; the adhesive is talcum powder; the mesh number of the talcum powder is 1000, and the mesh of the adhesive is to enable the rooting powder to be better adhered to the surface of the plant.

Further provides a preparation method of the rooting powder, and the preparation process of the rooting powder mainly comprises the following steps:

s11: weighing corresponding raw materials according to the proportion of any one of claims 1 to 4, dissolving and mixing, and fixing the volume to obtain a mixture A;

s12: drying the mixture A to obtain a mixture B;

s13: and crushing the mixture B into powder, and sieving the powder by using a soil sieve to obtain the rooting powder.

Preferably, the drying temperature in the step S2 is 65-75 ℃, the drying time is 2-4 h, and the drying is continuously repeated for 12-16 times.

According to the method of the first aspect of the present invention, further, the cultivation management of step S4 includes humidity management, light management, moisture management, temperature management, ventilation management, fertilizer management and pest control.

Specifically, according to the method of the first aspect of the present invention, the humidity is controlled to be 60-75% for the first 15-25 days after transplantation;

the illumination management is shading after transplantation, and the shading degree is 50-60% 35-45 days after transplantation;

the moisture management is that the substrate is always kept in a wet state;

the temperature management is to control the growth temperature to be 15-32 ℃, preferably 20-28 ℃;

the ventilation management is to maintain ventilation;

the fertilizer management comprises spraying a sterilization root-promoting nutrient solution 8-12 days before transplantation; further, the sterilization and root promotion nutrient solution is a mixed solution containing MS mineral elements, NAA, carbendazim or thiophanate methyl or zineb; furthermore, the sterilization root-promoting nutrient solution is a mixed solution containing 1/4MS mineral elements, 0.2mg/L NAA, carbendazim or thiophanate methyl or zineb.

The MS mineral elements are macroelements, microelements and iron salts in the MS culture medium mother liquor.

The disease and insect management is to spray a bactericide 18-22 days before transplantation; further, the bactericide is selected from at least one of carbendazim (N- (2-benzimidazole) -methyl carbamate), zineb (ethylene 1, 2-bis-thiocarbamate), thiophanate methyl (1, 2-bis (3-methoxycarbonyl-2-thioureido) benzene) and sodium diurethane (sodium p-dimethylamino-benzenediazosulfonate).

More specifically, 1, humidity management: after the test-tube plantlets of the rhizomes are planted, the management of a cultivation and domestication environment needs to be enhanced, generally, the humidity of about 20 days before the cultivation and domestication environment needs to be kept about 60-75%, ventilation and ventilation are kept, the humidity can be gradually reduced by ventilation according to the domestication condition, and the cultivation and domestication environment can be carried out according to the conventional management after the plantlets grow new roots and new buds.

2. Illumination management: the newly planted rootstock test-tube seedlings must be shaded (covered by a shading net of 70-85 percent), the illumination is too strong, the seedlings are easy to burn, high temperature and high humidity in a shed are easy to cause diseases due to the fact that ventilation is not performed, and the seedlings die seriously. After 40 days, the illumination is enhanced, and the shading degree is kept to be about 50-60%, so that the sealwort is more suitable for natural conditions when being planted in a field after being outplanted, and the field growth of sealwort is more facilitated.

3. Water content management: the moisture of the matrix must be kept in a wet state all the time, the moisture cannot be excessive, otherwise, seedlings are easy to mildew and rot, when the surface of stem is dry, the stem is wetted by spraying and often ventilated for cooling, attention is paid to observation, when the stem is dry, water is sprayed in time, when the stem is sprayed, a spraying mode is required, and the fact that a water faucet is directly opposite to flushing is avoided. The amount of the sprayed water is less firstly and more secondly, so that the substrate is kept in a wet state (the wet hand is pinched by hands but no water is dripped) at the position 1-3 cm away from the surface in the first 7d, and the growth of roots is promoted. After new roots grow out in about 25-30 days, the terminal buds start to be drawn high, seedlings start to survive and grow, and the water is sprayed to ensure that all substrates are wet (the wet hands are pinched by hands but no water is dripped).

4. Temperature management: the growth temperature of the sealwort is generally 15-32 ℃, the optimal growth temperature is 20-28 ℃, the growth is stopped when the temperature is lower than 15 ℃, the leaves are dried up below 10 ℃ and enter dormancy, and the sealwort grows slowly and is easy to burn when the temperature is higher than 32 ℃. Therefore, after the test tube plantlets of the rhizomes are planted, the temperature of the early-stage cultivation and domestication environment is preferably controlled to be 20-28 ℃, at the temperature, after about 20 days, the plantlets begin to grow new roots, adventitious buds grow or grow high, if the temperature requirement cannot be met, the temperature is controlled to be within the range of 15-32 ℃, otherwise, the test tube plantlets of the rhizomes are failed to be transplanted. In the later period, the management can be performed according to the routine management after about 30 days.

5. Ventilation management: through ventilation, the humidity and the temperature of the transplanting environment and the freshness of air can be adjusted, thereby being beneficial to the growth of seedlings and the control of diseases. The whole process of cultivation, domestication and planting needs to be always ventilated.

6. And (3) fertilizer management: 10 days before the test-tube plantlets of the rhizomes are planted, a mixed solution of a sterilization root-growth-promoting nutrient solution (formula: 1/4MS mineral elements + NAA0.2mg/L + 70% carbendazim (or thiophanate methyl or zineb) 500 solution) can be sprayed for 3 times, 1 time is carried out every 3 days, each time spraying is carried out until the surface layer of the matrix is wet, a new root grows out and grows to be high, water and fertilizer can be sprayed on the matrix, the water and fertilizer formula comprises potassium dihydrogen phosphate, potassium nitrate and calcium sulfate which are prepared into a water solution with the total concentration of 2500ppm according to the proportion of 1: 1: 0.5 and is applied every 7 days.

7. And (3) pest control: the bactericide is sprayed on the test-tube plantlet 20 days before transplantation, 1 time every 7 days, carbendazim, zineb, thiophanate-methyl, sodium dimesylate and the like can be used alternately, the bactericide is sprayed once every 10 days later, and after 60 days, the bactericide can be sprayed in time to prevent and treat diseases according to the occurrence condition of diseases. And (3) if diseases occur, timely removing the disease plants, spraying the pesticide once every 2-3 days, and continuously spraying for 2-3 times. The bactericide must be used interchangeably, and in particular, it may be used in combination of a plurality of kinds (the basic drug cannot be mixed with the acidic drug) and validamycin may be added.

According to the method of the first aspect of the present invention, further, the standard of the tube plantlet seedling emergence of the rhizome in step S5 is: (1) the leaves are full, the color of the leaves is dark green, the leaves are spread, and the number of the leaves reaches more than 5; (2) the root system is developed, the plant is healthy, and no diseases or insect pests exist; (3) the growth is normal and no distortion phenomenon exists; (4) the diameter of the rhizome is more than 4.0 cm.

Further, the test-tube plantlets with the rootstocks are cultivated, domesticated and planted in a plastic greenhouse for more than 1 year after the test-tube plantlets with the rootstocks are grown out of the nursery.

The invention has the beneficial effects that:

1. the transplanting method provided by the invention can be adopted in the plants propagated by the rootstocks (such as potatoes, ginger, small yellow ginger, Chinese yam and the like), solves the problem of low seedling rate of seedlings propagated by the rootstocks, simplifies the seedling management procedure and saves the labor cost. The seedling growth of the rhizome is efficiently ensured, the seedling growth of the rhizome is improved, the large-scale production of the Yangshan polygonatum cyrtonema seedlings becomes possible, and the bottleneck of the industrialized development of the Yangshan polygonatum cyrtonema is solved. The polygonatum sibiricum cultivated by tissue culture by adopting the method has the seedling rate of 90-95%, the rooting rate of 95-100% and the test-tube seedling transplanting survival rate of 91-98%; the method is adopted to cultivate high-quality Yangshan polygonatum cyrtonema rhizomes tissue culture seedlings 21590 by tissue culture and industrialized rhizome seedling culture production and rhizome meristem propagation. Aiming at the problems that wild polygonatum is excessively picked and an artificial cultivation and planting method is lacked at present, a tissue culture rhizome factory seedling raising method is provided.

2. The method introduces the sterilizing rooting powder in the transplanting process, is used in the transplanting of the polygonatum cyrtonema tissue culture industrialized seedling rhizome test-tube plantlets, obtains high survival rate and seedling rate, accelerates the rooting and bud germination growth of the rhizome, plays a role in efficient sterilization, effectively prevents the generation of rhizome decay and the biting of underground pests, ensures the rooting and seedling of the rhizome, improves the transplanting survival rate of the rhizome test-tube plantlets, and is also suitable for other plants with meristem propagation.

3. The transplantation matrix of the invention adopts river sand, is economical and convenient, has high transplantation survival rate and simple and convenient water control; the method can be popularized and applied to tissue culture industrial seedling culture of the rhizome plants, large-scale production and operation are realized, the economic benefit is obvious, the method leads the tissue culture test tube seedling transplantation technology in China, and the application prospect is wide.

Drawings

FIG. 1 shows the prepared and bottled rooting powder.

FIG. 2 tissue culture of Polygonatum cyrtonema rhizome transplanted out of a bottle.

FIG. 3 shows the rooting and germination conditions of Polygonatum cyrtonema rhizome after the test-tube plantlet is transplanted for 60 days.

FIG. 4 shows the growth of Polygonatum cyrtonema rhizome in every other 3 months after the test-tube plantlet is transplanted.

FIG. 5 shows the growth of tissue culture rhizome after transplanting and planting in 5 months every other year.

FIG. 6 shows the induction effect of potato explants.

FIG. 7 shows the formation of adventitious buds in potatoes.

FIG. 8 shows the subcultured strong bud of potato.

FIG. 9 shows rooting of potato shoots.

Figure 10 shows the induction effect of ginger explants.

FIG. 11 shows the formation of adventitious buds of ginger.

FIG. 12 shows the secondary proliferation of strong bud of ginger.

FIG. 13 shows rooting of ginger shoots.

Detailed Description

In order to enhance the understanding of the present invention, the present invention will be described in further detail with reference to the following examples, which are provided for the purpose of illustration only and are not intended to limit the scope of the present invention.

The nano silver solution is nano silver (Ag +) D stock solution which is purchased from materials science and technology ltd of Guangzhou city, and the content concentration of the silver ion component is 100 ppm.

Example 1

A test-tube plantlet transplanting method for rhizome plants comprises the following steps:

s1, hardening and culturing tissue culture seedlings of rhizome plants;

s4, cultivation management after transplantation is carried out;

Specifically, the method comprises the following steps: preparation steps before transplantation:

rhizome test-tube plantlet transplanting nursery selection

The garden for transplanting the rhizome of Polygonatum cyrtonema tissue culture industrial production seedling requires flat terrain, transparent periphery, sufficient illumination, good drainage, clean and pollution-free open areas, and the south side is required to be selected if the hillside is selected.

Rhizome test-tube plantlet transplanting facility preparation

1. The standard plastic greenhouse is preferably installed on condition, and the plastic greenhouse must be provided with various cultivation environment condition regulation and control facilities including a light regulation system, a humidity condition system, a ventilation regulation system, a spraying system and the like.

2. If the conditions are not satisfied, a temporary simple plastic shed can be adopted, but the temporary simple plastic shed needs to be rain-proof, and is easy to perform operations such as ventilation, illumination adjustment, temperature adjustment, humidity adjustment, water spraying, fertilization, pest control and the like.

Preparation of matrix for transplanting test-tube plantlets with rhizomes

1. Preparing a matrix: the matrix used as the transplanting matrix of the test-tube plantlet of the rhizome requires clean river sand (phi 0.2-3.0 mm) with proper thickness and good ventilation and drainage.

2. Matrix disinfection: the matrix disinfectant is potassium permanganate, 50% sodium sulfadiazine and other soil disinfectant. The specific operation method is that 0.2 percent potassium permanganate solution is used for sprinkling to ensure that the matrix is uniformly wetted to the degree, and 500 times of 50 percent sodium diquat solution is used for sprinkling every other day.

3. Laying a transplanting bed and filling a matrix: the ground of the transplanting shed is higher than the surrounding ground, drainage ditches are arranged around the transplanting shed, the height of the bed edge of the transplanting bed is about 15cm, the transplanting shed can be built by red bricks for construction, the transplanting shed is 1.2m wide, the length of the transplanting shed is specifically determined according to the length, and 40cm footpaths are paved by red bricks for construction at intervals between the bed and the bed. The transplanting bed is filled with transplanting substrates, and the filling height of the transplanting substrates is about 2cm lower than the bed edge.

S1, hardening off and culturing tissue culture seedlings of the rhizome plants/hardening off and shape promoting culture of the rhizome test tube seedlings

1. And (5) carrying out hardening-seedling shape forcing culture on the rhizoma polygonati rhizome test-tube seedlings in a plastic greenhouse. The seedling exercising method is characterized in that the illumination intensity is reasonably adjusted according to the principle of 'from weak to strong', the seedling is prevented from being burnt due to too strong light or the seedling grows badly due to too weak light, and the temperature, the humidity and the like of the exercising are timely adjusted according to the weather change.

2. Seedling hardening shape promoting culture conditions (seedling hardening plastic greenhouse): the temperature is 18.5-28.5 ℃, the illumination intensity is 6000-8500 Lux, and the illumination time is 8-11 h.

3. Hardening seedling shape to promote the culture process: preparing materials for hardening seedling and promoting root culture after the transfer on the superclean bench, immediately sealing the joint of a bottle cap and a bottle by using a preservative film on the same day after the transfer, and putting the bottle cap and the bottle into a seedling hardening plastic greenhouse for hardening seedling culture every other day.

4. The culture time for promoting the seedling hardening form is generally 40-60 days, and the specific time is determined according to actual conditions.

5. The seedling emergence quality standard of the test-tube plantlet of the rhizome is as follows: (1) the color of the leaves is green, the number of the leaves reaches more than 3 or the leaves are withered and yellow without leaves; (2) the diameter of the rhizome is more than 2.5 cm; (3) the shape of the rhizome is obviously light green or green; (4) the growth is normal.

S2, cleaning and disinfecting the test-tube plantlets of the rhizomes, and airing the plantlets

Taking out and cleaning test-tube plantlet of rhizome

1. Separating the test tube seedling of the rhizome to be cleaned into pollution-free seedling and pollution seedling, and cleaning separately to prevent cross contamination.

2. Taking out the test-tube plantlet from the rhizome: the trained tissue culture seedlings are gently picked out from the culture flask by using forceps (care is taken to avoid damaging the seedlings as much as possible), and then the seedlings are placed into a prepared clean plastic basin.

3. Cleaning the test-tube plantlets of the rhizomes: the test-tube plantlet of the rhizome needs to be cleaned by clean water for three times, firstly, the culture medium attached to the tissue culture seedling in the plastic basin is removed completely, a culture medium without residual base is needed, and the seedling is damaged as little as possible. And then cleaned twice with clean water.

Sterilizing and airing test-tube plantlet of rhizome

1. Sterilizing pollution-free seedlings: and soaking the clean test tube seedling of the rhizome in 0.02 percent potassium permanganate solution for 7min, taking out, cleaning with clear water once, and cleaning residual potassium permanganate solution attached to the seedling.

2. Disinfecting the polluted seedlings: the cleaned test tube seedling of the rhizome is soaked in 0.02 percent potassium permanganate solution for 10min, then taken out, cleaned once by tap water, soaked in 1000 times of 70 percent carbendazim (or thiophanate methyl and the like) solution and then taken out.

3. And (4) placing the sterilized test-tube seedling of the rhizome in a ventilated and shady place for air drying until the moisture attached to the surface of the seedling just disappears.

S3, planting in a grading manner according to the growth condition of the test-tube plantlet of the rhizome

1. Tissue culture seedling grading: the tissue culture seedlings after cleaning, sterilizing and airing are classified into one, two, three and stepless grades.

Stepless: except the first, second and third stages, the rest are all classified into stepless.

2. Carrying out rooting powder treatment operation and planting on the test-tube plantlets of the rhizomes: the plants are planted in different grades and regions and are planted in river sand matrix, and the plant-row spacing is 8 multiplied by 8 cm. During planting, after the test-tube plantlets of the rhizomes are wet, the sterilizing and insect-preventing rooting powder is dipped and adhered on the surface of the test-tube plantlets, a small thin layer of rooting powder is ensured to be uniformly adhered on the surface of each rhizome, and then the test-tube plantlets are planted on a transplanting bed. The substrate should cover the top of the rootstock, just the rootstock is buried in the substrate, and the leaf base cannot be covered, otherwise the seedling is rotten due to improper water spraying, and the seedling cannot be planted too shallowly, otherwise the growth of new roots of the seedling is not facilitated. After planting, the seedlings are lightly pinched by hands to ensure that the rhizome is fully contacted with the matrix, and the sterilization and root growth promoting nutrient solution (the formula of the sterilization and root growth promoting nutrient solution is 1/4MS mineral element, NAA0.2mg/L and 70 percent of carbendazim (or thiophanate methyl) 500 solution) is sprayed.

The component formula of the sterilizing insect-preventing rooting powder is shown in the following table 1.

TABLE 1 formulation and description of the special bactericidal insect-proof rooting powder

The preparation process of the tissue culture seedling-raising sterilizing insect-preventing rooting powder mainly comprises the following steps:

s1: weighing the corresponding raw materials:

1. weighing talcum powder: wearing a mask and disposable rubber gloves, weighing 990g of talcum powder by an electronic scale, and placing the talcum powder into a 40cm x 60cm stainless steel operating disk.

2.90% O, O-dimethyl- (2,2, 2-trichloro-1-hydroxyethyl) phosphonate was weighed: weighing 5g of 90% O, O-dimethyl- (2,2, 2-trichloro-1-hydroxyethyl) phosphonate, putting the weighed materials into a 1L plastic measuring cup, adding 100mL of purified water to dissolve the materials, uniformly pouring the materials onto the weighed talcum powder, and continuously stirring the materials by using a glass rod.

3. Nano silver (Ag)⁺) D, measuring stock solution: measuring 40mL of nano silver (Ag) by using measuring cylinder⁺) D, putting the stock solution into a 1L plastic measuring cup, diluting the stock solution to 200mL by using purified water, uniformly pouring the diluted stock solution onto the weighed talcum powder, and continuously stirring the mixture by using a glass rod.

4. Weighing other medicines, dissolving and fixing volume: the following were weighed separately with an electronic scale: 0.5g of indolebutyric acid is firstly dissolved by a proper amount of 1N NaOH solution; 0.004g of 2.4-dichlorophenoxyacetic acid is dissolved by using a proper amount of 1N NaOH solution; 0.03g of 6-benzylamino adenine is firstly dissolved by a proper amount of 1N HCl solution; 1g of monopotassium phosphate, 1g of potassium nitrate, 0.1g of ferrous sulfate and 0.05g of boric acid, dissolving the components in 50mL of purified water one by one in a 200mL beaker, pouring the dissolved components into a 200mL volumetric flask for constant volume, slowly and uniformly pouring the weighed talcum powder onto the weighed talcum powder, and continuously stirring the mixture by using a glass rod until the talcum powder is uniformly wetted and leveled.

S2: drying: the prepared talcum powder is put into an electric heating air blast drying box, a door is closed, the temperature is regulated and controlled to be 70 ℃, and drying is carried out at regular time for 3 hours. And (4) a drying process, namely checking the dry and wet degree of the talcum powder at the end of every 3 hours until drying. Generally, the moisture in the talcum powder can be dried only by continuously and repeatedly drying for 8 times.

S3: milling: taking the dried talcum powder out of the drying box, crushing the dried talcum powder into powder, sieving the powder by using a soil sieve of 400 meshes, putting the powder into a 20L plastic basin, continuously crushing the coarse talcum powder particles which cannot pass through the sieve, and rolling while sieving until all the talcum powder is sieved.

S4: packaging and storing: the sieved talcum powder is the tissue culture seedling cultivation sterilization insect prevention rooting powder, and the tissue culture seedling cultivation sterilization insect prevention rooting powder is put into a 500mL white plastic reagent bottle which is stuck with a label for recording the preparation date, the product name and the use method, sealed and stored in a shady, cool and ventilated and dry place.

S4, cultivation management after transplantation/cultivation and domestication management of test-tube seedlings of rhizomes

1. Humidity management: after the test-tube plantlets of the rhizomes are planted, the management of a cultivation and domestication environment needs to be enhanced, generally, the humidity of about 20 days before the cultivation and domestication environment needs to be kept about 60-75%, ventilation and ventilation are kept, the humidity can be gradually reduced by ventilation according to the domestication condition, and the cultivation and domestication environment can be carried out according to the conventional management after the plantlets grow new roots and new buds.

2. Illumination management: the newly planted rootstock test-tube seedlings must be shaded (covered by a shading net of 70-80 percent), the illumination is too strong, the seedlings are easy to burn, high temperature and high humidity in a shed are easy to cause diseases due to the fact that ventilation is not carried out, and the seedlings die seriously. After 40 days, the illumination is enhanced, and the shading degree is kept to be about 50-60%, so that the sealwort is more suitable for natural conditions when being planted in a field after being outplanted, and the field growth of sealwort is more facilitated.

6. And (3) fertilizer management: spraying 3 times with a sterilizing and root-growth-promoting nutrient solution (a mixed solution of 1/4MS mineral element, NAA0.2mg/L and 40% carbendazim (or thiophanate methyl and the like) 500 solution) 10 days before planting the test-tube plantlets of the rhizomes, wherein the spraying is carried out 1 time every 3 days until the surface layer of the matrix is wet. When new roots grow and grow high, water and fertilizer can be sprayed on the substrate, and the formula of the water and fertilizer is as follows: potassium dihydrogen phosphate, potassium nitrate and calcium sulfate are mixed according to the weight ratio of 1: 1: the ratio of 0.5 was adjusted to 2500ppm total concentration in water solution, every 7 days.

7. And (3) pest control: the bactericide is sprayed on the test-tube plantlet 20 days before transplantation, 1 time every 7 days, the bactericide such as carbendazim, haloxyfop, thiophanate-methyl, and sodium disulfate can be used alternately, the bactericide is sprayed once every 10 days later, and after 60 days, the bactericide can be sprayed in time to prevent and treat diseases according to the occurrence condition of diseases. And (3) if diseases occur, timely removing the disease plants, spraying the pesticide once every 2-3 days, and continuously spraying for 2-3 times. The bactericide must be used interchangeably, and in particular, it may be used in combination of a plurality of kinds (the basic drug cannot be mixed with the acidic drug) and validamycin may be added.

8. And (3) seedling emergence of the test-tube plantlet of the rhizome: 1. the quality standard of seedling emergence of test-tube plantlets of rhizomes is as follows: (1) the leaves are full, the color of the leaves is dark green, the leaves are spread, and the number of the leaves reaches more than 5; (2) the root system is developed, the plant is healthy, and no diseases or insect pests exist; (3) the growth is normal and no distortion phenomenon exists; (4) the diameter of the rhizome is more than 4.0 cm; (3) transplanting, cultivating, domesticating and planting in plastic greenhouse for more than 1 year.

Example 2

A method for transplanting a test-tube plantlet of a rhizome plant, the procedure of which is the same as that of example 1.

The formula of the bactericidal insect-proof rooting powder adopted in the embodiment comprises the following raw materials: 400ppm IBA, 3ppm 2.4-D, 20ppm 6BA, 900ppm KH₂PO₄、900ppm KNO₃、50ppm FeSO₄·7H₂0、40ppm H₃BO₃30mL of 100ppm Ag⁺Solution, 4500ppm O, O-dimethyl- (2,2, 2-trichloro-1-hydroxyethyl) phosphonate and an attachment agent.

Example 3

The formula of the bactericidal insect-proof rooting powder adopted in the embodiment comprises the following raw materials: 600ppm IBA, 5ppm 2.4-D, 40ppm 6BA, 1100ppm KH₂PO₄、1100ppm KNO₃、150ppm FeSO₄·7H₂0、60ppm H₃BO₃50mL of 100ppm Ag⁺Solution 5500ppm of O, O-dimethyl- (2,2, 2-tris)Chloro-1-hydroxyethyl) phosphonate and an adhesive.

Example 4 Polygonatum cyrtonema test tube plantlet transplanting Effect experiment

This example provides a comparative experiment of the transplantation effect of the sealwort flower tissue culture industrialized seedling test tube plantlet in the above examples 1 to 3.

1. And (3) experimental design: 4000 test-tube plantlets with rhizomes are selected, each test-tube plantlet comprises 1000 test-tube plantlets and a seedbed, the test-tube plantlets are divided into 1-3 groups and a control group, wherein the test-tube plantlet is transplanted in 3 months by using the transplanting method in the embodiment 1-3 of the invention for 1-3 groups, the control group is not used, the planting time after transplanting is 60 days, and the seedling rate and the growth condition of the test-tube plantlet are recorded.

2. Selecting test facilities: the temporary simple plastic shed is selected under the field environment condition, adopts rain-proof water, and is easy to perform operations such as ventilation, illumination adjustment, temperature adjustment, humidity adjustment, water spraying, fertilization, pest control and the like.

3. Preparing a transplanting bed: the ground of the transplanting shed is higher than the surrounding ground, drainage ditches are arranged around the transplanting shed, the height of the bed edge of the transplanting bed is 15cm, the transplanting shed is built into a width of 1.2m by red bricks for construction, the length of the transplanting shed is specifically determined according to the length, and 40cm footpaths are paved between beds by red bricks for construction. And filling dried and clean river sand on the transplanting bed to serve as a transplanting matrix, wherein the filling height of the transplanting matrix is about 2cm lower than the edge of the bed. And the substrate was disinfected with 0.2% potassium permanganate solution and 500 times 50% sodium diquat solution.

4. Preparing rootstock test-tube plantlets: taking out the bottled test tube seedling with rhizome after seedling hardening in a seedling hardening plastic greenhouse, cleaning, sterilizing, airing and grading.

5. Applying test planting of rooting powder of the test-tube plantlets of the rhizomes: first-level rhizome test-tube seedlings are uniformly selected and planted in different areas according to test design, and the row spacing of the seedlings is 8 multiplied by 8 cm. When planting, after wetting the rhizome test-tube plantlets, dipping and adhering the special sterilization and insect prevention rooting powder for transplanting the polygonatum multiflorum rhizome test-tube plantlets, ensuring that a small thin layer of rooting powder is uniformly adhered to the surface of each rhizome, and planting the control group on a transplanting bed without dipping and adhering.

6. Cultivating and domesticating the test-tube plantlets of the rhizomes: and after planting, strictly performing management such as humidity, illumination, moisture, temperature, ventilation, fertilizer content, pest control and the like according to the biological characteristics and ecological habits of polygonatum cyrtonema.

7. Statistical analysis of test data: after the test-tube plantlets are transplanted for 60 days, the seedling rate and the growth condition of the test-tube plantlets are subjected to statistical analysis, and the results are shown in table 2.

TABLE 2 Polygonatum sibiricum rhizome test-tube plantlet transplant seedling rate and growth status statistics

As can be seen from the data in Table 2, by using the transplanting method of examples 1 to 3, the rate of death due to rotting and damage due to insect bite of the transplanted rootstock test-tube plantlet is reduced, the rooting and seedling rate is remarkably improved, the rooting and sprouting of the shoots are accelerated, and the plant growth condition is good, as shown in the attached figures 2 to 4.

FIG. 2 shows the rhizome of Polygonatum cyrtonema after tissue culture and transplantation. The figure shows that the shape is formed, but the root is not formed, certain leaf buds grow out, and the rootstock grows under the artificially controlled environmental condition, so that the seedling is weak, and the seedling has no good capability of autonomously absorbing water and nutrients in the matrix.

FIG. 3 shows the rooting and germination conditions of Polygonatum cyrtonema rhizome after the test-tube plantlet is transplanted for 60 days. It can be seen that the root system is formed, the leaf bud is strong, large and tall, and has good capability of independently absorbing water and nutrients in the matrix.

FIG. 4 shows the growth of Polygonatum cyrtonema rhizome in 5 months every other year after the test-tube plantlet is transplanted. It can be seen that the growth is good at this time. The seedlings are tidy, the seedling rate is high, and the seedlings grow fast and robustly.

Example 5 Potato Tuber meristem growth experiment

This example provides comparative tests of the meristematic seedling raising effects of the potato tubers of the above examples 1 to 3, and verifies the effects of the meristematic seedling raising effects on the potato tubers based on the obtained test data, which mainly comprises the following steps:

1. and (3) experimental design: 2000 high-quality potato tubers are selected, each tuber is cut into 3-5 cm in size and contains 2-3 bud eyes, each group comprises 500 tubers and a seedbed, and each seedbed is divided into a test group 1-3 and a control group, wherein 1-3 groups adopt the special sterilization insect-prevention rooting powder formula for polygonatum cyrtonema tissue culture industrialized rhizome propagation and seedling culture in the embodiment 1-3 of the invention, a tuber meristem propagation and seedling culture test is carried out in 10 months, the control group does not adopt the formula, and the seedling rate and the growth condition of the tuber meristem propagation are recorded after 50 days of planting.

4. Preparing potato tubers: high-quality potato tubers without virus, disease and insect pests, which are used for producing seedlings, are purchased from the market and cut into small tubers with the size of 3-5 cm and 2-3 bud eyes.

5. Potato tuber meristem propagation seedling test: planting according to experimental design in different areas, wherein the row spacing of the plants is 10 multiplied by 10 cm. When the small potato tubers are wet, the special sterilization and insect prevention rooting powder for transplanting the polygonatum multiflorum rhizome test-tube plantlet is dipped and adhered, after a small thin layer of rooting powder is uniformly adhered to the surface of each rhizome, the control group is not dipped and adhered, and the rhizoma polygonati rhizome test-tube plantlet is planted on a transplanting bed.

6. Potato tuber meristem propagation seedling cultivation management: managing humidity, illumination, moisture, temperature, ventilation, fertilizer, pest control and the like strictly according to the biological characteristics and ecological habits of the potatoes after planting

7. Statistical analysis of test data: after the potato small tubers are planted for 50 days, the seedling rate and the growth condition of the potato small tubers are subjected to statistical analysis, and the results are shown in table 3.

TABLE 3 statistics of the seedling rate and growth condition of the potato meristematic propagation seedling

As can be seen from the data in Table 3, the use of the transplantation methods of examples 1 to 3 reduced the rate of death due to decay and damage due to insect bite of the planted potato tubers, significantly improved the rooting and seedling rate, accelerated the rooting and sprouting of the shoots, and good the growth of the plants.

FIG. 6 shows the induction effect of potato explants; FIG. 7 is the formation of adventitious buds of a potato; FIG. 8 shows the subcultured, proliferated and strengthened buds of potato; FIG. 9 shows rooting of potato shoots.

Example 6 meristem propagation and seedling raising experiment of ginger rhizome

This example provides comparative tests of the seedling effects of the foregoing examples 1 to 3 on the rhizome meristematic propagation of ginger, and based on the obtained test data, verifies the effect on the rhizome meristematic propagation of ginger, which mainly includes the following steps:

1. and (3) experimental design: selecting 2000 high-quality ginger roots, cutting each root into 3-5 cm in size, containing 1-2 bud eyes, dividing each root into 500 blocks and a seedbed into a test group 1-3 and a control group, wherein 1-3 groups adopt the special sterilization insect-prevention rooting powder formula for polygonatum cyrtonema tissue culture industrialized rhizome propagation and seedling culture in the embodiment 1-3 of the invention, carrying out a rhizome meristem propagation and seedling culture test in 5 months, and recording the seedling rate and growth condition of rhizome meristem propagation after planting for 50 days, wherein the control group does not adopt the formula.

4. Preparing ginger planting rootstocks: healthy and high-quality ginger roots without diseases and insect pests for seedling production are purchased from the market and cut into small roots with the size of 3-5 cm and 1-2 bud eyes.

5. Meristem propagation seedling test of ginger rhizome: planting according to experimental design in different areas, wherein the row spacing of the plants is 10 multiplied by 10 cm. When planting, after the small rhizome of the ginger is wet, dipping and sticking the special sterilization and insect prevention rooting powder for transplanting the polygonatum multiflorum rhizome test-tube plantlet, ensuring that a small thin layer of rooting powder is uniformly stuck on the surface of each rhizome, and planting the control group on a transplanting bed without dipping and sticking.

6. And (3) meristem propagation, seedling culture and management of ginger rootstocks: after planting, strictly according to the biological characteristics and ecological habits of the ginger, managing humidity, illumination, moisture, temperature, ventilation, fertilizer, pest control and the like

7. Statistical analysis of test data: after the ginger rhizome is planted for 50 days, the seedling rate and the growth condition of the ginger rhizome are subjected to statistical analysis, and the results are shown in table 4.

TABLE 4 statistics of seedling rate and growth condition of ginger rhizome meristem propagation seedling

As can be seen from the data in Table 4, by using the tissue culture seedling-raising, sterilizing and insect-preventing rooting powder in examples 1 to 3, the rate of death of small pieces of rhizome of ginger due to decay and damage due to insect bite is reduced, the rooting and seedling rate is remarkably improved, the rooting and bud germination are accelerated, and the plant growth condition is good.

FIG. 10 shows the induction effect of ginger explants; FIG. 11 shows the formation of adventitious buds of ginger; FIG. 12 shows the subcultured and proliferated strong bud of Zingiber officinale; FIG. 13 shows rooting of ginger shoots.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention.

Claims

1. A test-tube plantlet transplanting method for rhizome plants comprises the following steps:

2. The method according to claim 1, wherein the conditions of the seedling culture in the step S1 are as follows: the temperature is 18.5-28.5 ℃, and the illumination intensity is 6000-8500 Lux; the sunshine duration is 8-11 h.

3. The method according to claim 1, wherein the emergence criteria for forming rootstock tubes in step S1 are:

(1) the leaves are green, the number of the leaves reaches more than 3 or the leaves are withered and yellow and fall off; (2) the diameter of the rhizome is more than 2.5 cm; (3) the shape of the rhizome is obviously light green or green; (4) the growth is normal.

4. The method according to claim 1, wherein the specific method for cleaning and sterilizing the rootstock test-tube plantlets in the step S2 is as follows: taking a pollution-free test tube seedling of the rhizome, cleaning the test tube seedling with clear water, soaking the test tube seedling with a potassium permanganate solution, and then cleaning the test tube seedling with clear water again; cleaning polluted seedlings with clean water, soaking the polluted seedlings in a potassium permanganate solution, cleaning the polluted seedlings with clean water again, soaking the polluted seedlings in a sterilizing solution, and taking out the polluted seedlings.

5. The method according to claim 1, wherein the grading planting in step S3 is: dividing the test-tube plantlets of the rhizomes into a first stage, a second stage and a third stage, and planting the test-tube plantlets respectively; wherein,

6. The method as claimed in claim 1, wherein the specific operation of the classified planting in step S3 is: wetting the graded rootstock test-tube plantlets, dipping and adhering sterilizing insect-proof rooting powder on the surface, and planting the test-tube plantlets in river sand matrix.

7. The method as claimed in claim 6, wherein the bactericidal and insect-resistant rooting powder comprises the following raw materials: 400-600 ppm IBA, 3-5 ppm 2.4-D, 20-40 ppm 6BA, 900-1100 ppm KH₂PO₄、900～1100ppmKNO₃、50～150ppm FeSO₄·7H₂0、40～60ppm H₃BO₃3-5 wt% of nano silver ion solution, 4500-5500 ppm of O, O-dimethyl- (2,2, 2-trichloro-1-hydroxyethyl) phosphonate and an adhesive.

8. The method according to claim 1, wherein the cultivation management of step S4 includes humidity management, light management, moisture management, temperature management, ventilation management, fertilizer management, and pest control.

9. The method of claim 8, wherein the moisture management is 60-75% moisture maintenance for the first 15-25 days after transplantation;

the illumination management is shading after transplantation, the shading degree is 70-85%, and the shading degree is 50-60% after 35-45 days of transplantation;

the moisture management is that the substrate is always kept in a wet state;

the ventilation management is to maintain ventilation;

the fertilizer management comprises spraying a sterilization root-promoting nutrient solution 8-12 days before transplantation; further, the sterilization root-promoting nutrient solution is a mixed solution containing MS mineral elements, NAA and a bactericide; furthermore, the sterilization root-promoting nutrient solution is a mixed solution containing 1/4MS mineral elements, 0.2mg/L NAA, carbendazim and thiophanate methyl;

the pest control is to spray a bactericide 18-22 days before transplantation; further, the bactericide is selected from at least one of N- (2-benzimidazole) -methyl carbamate, ethylene 1, 2-dithio-zinc carbamate, 1, 2-di (3-methoxy carbon-2-thioureido) benzene and p-dimethylamino benzene diazo sodium sulfonate.

10. The method according to any one of claims 1 to 9, wherein the rhizome plant is polygonatum cyrtonema, potato, ginger.