CN110651715B - Industrial seedling growing method for actinidia arguta - Google Patents
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A—HUMAN NECESSITIES
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- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
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Abstract
The invention discloses an industrialized seedling method of actinidia arguta, which comprises the following steps: taking a sterile branch, and shearing the sterile branch into two parts, wherein one part is a terminal bud, and the other part is a stem section with a bud; inoculating the stem with the bud into a multiplication culture medium, culturing for 35-40 days to obtain stem with branches, shearing off the branches, and returning the branches to the previous step for repeated operation; inoculating the obtained terminal bud into a rooting culture medium, and culturing for 20-25 days to obtain a tissue culture seedling; then domesticating and hardening the seedlings for 5-7 days; taking out the tissue culture seedlings, cleaning the root culture medium, then transplanting the tissue culture seedlings into a plug tray filled with the matrix, and continuously culturing for 25-30 days and then planting. The seedling raising method is short in seedling raising time, is suitable for various actinidia arguta fruits, and has the advantage of wide applicability.
Description
Technical Field
The invention relates to the technical field of tissue culture seedling culture, in particular to an industrialized seedling culture method for actinidia arguta.
Background
Actinidia arguta (Actinidia arguta) belongs to Actinidia perennial vine plants of Actinidia of Actinidiaceae (Actinidiaceae), commonly known as fructus Choerospondiatis and Actinidia chinensis, and is mainly distributed in Liaoning, jilin, heilongjiang and Hebei. Is a new fruit with high nutritive value and great economic benefit. The ripe fruit is soft and juicy, has sweet and sour taste and delicious and fragrant taste. The vitamin C content of fresh fruit is 100-490 mg/100g, which is 20-80 times higher than apple, 5-12 times higher than orange and 30-140 times higher than pear. It also contains vitamin B, P, mineral nutrients such as calcium, magnesium, and phosphorus, and various amino acids required by human body. Has effects in nourishing, strengthening body constitution, promoting salivation, moistening lung, treating hypertension, angina pectoris, and hyperlipidemia, and inhibiting synthesis of carcinogen (nitrosomorpholine).
The cultivation method of actinidia arguta mainly comprises the steps of establishing a garden by using tissue culture seedlings and open field cuttage seedlings, using a greenhouse frame in a frame mode, and the like.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides an industrial seedling raising method for actinidia arguta, which can effectively solve the problem of long seedling raising time in the conventional seedling raising method, and is suitable for various actinidia arguta and has the advantage of wide applicability.
In order to achieve the purpose, the technical scheme adopted by the invention for solving the technical problems is as follows:
an industrialized seedling method of actinidia arguta comprises the following steps:
(1) Inoculating stem segment with single bud in enrichment medium, culturing at 22-28 deg.C under 1500-2500lux and 14-18 h/day for 35-40 days to obtain stem segment with branch, cutting branch, and cutting branch into two parts, one part is terminal bud with 2-3 leaves, and the other part is stem segment with single bud;
(2) Repeating the process in the step (1) on the stem sections with the single buds obtained in the step (1);
(3) Inoculating the terminal bud obtained in the step (1) into a rooting culture medium, and then culturing for 20-25 days at 22-28 ℃,1500-2500lux and 14-18 h/day of illumination time to obtain a tissue culture seedling;
(4) Domesticating and hardening the tissue culture seedlings obtained in the step (3) for 5-7 days;
(5) And (5) taking out the tissue culture seedlings in the step (4), cleaning a root culture medium, transplanting the tissue culture seedlings into a plug tray filled with a matrix, and continuously culturing for 25-30 days and then planting.
Further, the length of the terminal bud in the step (1) is 2.5-3.5cm, and the length of the stem segment is 0.8-1.4cm.
Further, the length of the terminal bud in the step (1) is 3cm, and the length of the stem section is 1cm.
Further, the proliferation culture medium in the step (1) consists of an MS culture medium, 6-BA, NAA, agar and sucrose or white sugar, wherein the mass concentration of the 6-BA is 0.8-1.4mg/L, the mass concentration of the NAA is 0.1-0.2mg/L, the mass concentration of the sucrose or white sugar is 20-40g/L, the mass concentration of the agar is 6-8g/L, and the pH value of the proliferation culture medium is 5.8-6.2.
Further, the multiplication culture medium in the step (1) consists of an MS culture medium, 6-BA, NAA, agar and sucrose or white sugar, wherein the mass concentration of the 6-BA is 1.0mg/L, the mass concentration of the NAA is 0.2mg/L, the mass concentration of the sucrose or white sugar is 30g/L, the mass concentration of the agar is 7.5g/L, and the pH value of the multiplication culture medium is 5.8-6.2.
Further, the rooting medium in the step (3) consists of a 1/2MS medium, IBA, agar and sucrose or white sugar, wherein the mass concentration of the IBA is 0.2-0.8mg/L, the mass concentration of the sucrose or white sugar is 10-40g/L, the mass concentration of the agar is 6-9g/L, and the pH value of the rooting medium is 5.8-6.2.
Further, the rooting medium in the step (3) consists of a 1/2MS medium, IBA, agar and sucrose or white sugar, wherein the mass concentration of the IBA is 0.7mg/L, the mass concentration of the sucrose or the white sugar is 30g/L, the mass concentration of the agar is 7.5g/L, and the pH value of the rooting medium is 5.8-6.2.
Further, the matrix in the step (5) is prepared by uniformly mixing coconut chaff, perlite and plant ash according to the volume ratio of 7.
Further, after the tissue culture seedlings are transplanted in the step (5), the plug tray is thoroughly poured by tap water, then the plug tray is placed on a tray filled with 1000-time diluted nutrient solution, and a moisturizing cover is covered on the tray.
The beneficial effects produced by adopting the scheme are as follows:
the tissue culture method is suitable for rapid propagation and seedling raising of a plurality of actinidia arguta varieties, has the advantage of wide applicability, and has the advantages of shorter seedling raising time, high survival rate and 100 percent of survival rate after transplanting compared with the conventional seedling raising method.
The method adopts the leafless stem with buds to carry out enrichment culture in the enrichment culture medium, the buds quickly grow into branches of 7-9cm within 35-40 days, the germination rate of the buds reaches 100 percent, the branches are cut and divided into two parts, the concentration of auxin and the like contained in the terminal bud part is higher, the terminal bud part can be directly placed in a rooting culture medium to be cultured into seedlings for transplantation, and the seedling culture period is greatly shortened; the rest stem section part is continuously used as a multiplication matrix of the bud to be cultured in a multiplication culture medium, and branches are grown by the multiplication culture, so that the circulation operation is realized, the utilization rate of the nursery stock is improved, and the industrialized mass production is realized.
Detailed Description
Example 1
An industrialized seedling method of actinidia arguta comprises the following steps:
(1) Taking stem segments of Actinidia arguta with single bud, inoculating the stem segments into a proliferation culture medium, culturing at 22 ℃ under the illumination condition of 1500lux and 14h/day for 35 days to obtain stem segments with 8cm branches, cutting off the branches, and cutting the branches into two parts, wherein one part is terminal buds with 2-3 leaves of 2.5cm in length, and the other part is stem segments with single bud of 0.8cm in length; wherein the enrichment culture medium consists of an MS culture medium, 6-BA, NAA, agar and sucrose or white sugar, the mass concentration of the 6-BA in the enrichment culture medium is 0.8mg/L, the mass concentration of the NAA in the enrichment culture medium is 0.1mg/L, the mass concentration of the sucrose or white sugar in the enrichment culture medium is 20g/L, the mass concentration of the agar in the enrichment culture medium is 6g/L, and the pH value of the enrichment culture medium is 5.8;
(2) Repeating the proliferation culture process in the step (1) on the single-bud stem section obtained in the step (1);
(3) Inoculating the terminal bud obtained in the step (1) into a rooting culture medium, and then culturing for 20 days under the conditions of 22 ℃,1500lux and 14 h/day of illumination time to obtain a new tissue culture seedling; wherein, the rooting culture medium consists of a 1/2MS culture medium, IBA, agar and sucrose or white sugar, the mass concentration of the IBA in the rooting culture medium is 0.2mg/L, the mass concentration of the sucrose or white sugar in the rooting culture medium is 10g/L, the mass concentration of the agar in the rooting culture medium is 6g/L, and the pH value of the rooting culture medium is 5.8;
(4) Adding distilled water with the height of 1cm into the tissue culture seedlings obtained in the step (3), then opening the bottle mouth, covering three quarters of the bottle mouth with a cover, and acclimatizing and hardening the seedlings for 5 days;
(5) Taking out the tissue culture seedlings in the step (4), cleaning a root culture medium, transplanting the tissue culture seedlings into a plug tray filled with coconut chaff, perlite and plant ash, thoroughly watering the plug tray by using tap water, placing the plug tray on a tray filled with 1000-time diluted nutrient solution, covering a cover on the tray for moisturizing, and continuously culturing for 25 days and then planting; wherein, the coconut husk, the perlite and the plant ash are uniformly mixed according to the volume ratio of 7.
Example 2
An industrialized seedling method of actinidia arguta comprises the following steps:
(1) Taking a stem section with a single bud of Actinidia arguta of Ribes Nintenwatt, inoculating the stem section into a proliferation culture medium, then culturing for 40 days under the illumination conditions of 28 ℃,2500lux and 18h/day to obtain a stem section with 9cm branches, cutting off the branches, and cutting the branches into two parts, wherein one part is a terminal bud with 2-3 leaves of length 3.5cm, and the other part is a stem section with a single bud of length 1.4 cm; wherein the proliferation culture medium consists of an MS culture medium, 6-BA, NAA, agar and sucrose or white sugar, the mass concentration of the 6-BA in the proliferation culture medium is 1.4mg/L, the mass concentration of the NAA in the proliferation culture medium is 0.2mg/L, the mass concentration of the sucrose or white sugar in the proliferation culture medium is 40g/L, the mass concentration of the agar in the proliferation culture medium is 8g/L, and the pH value of the proliferation culture medium is 6.2;
(2) Repeating the proliferation culture process in the step (1) on the single-bud stem section obtained in the step (1);
(3) Inoculating the terminal bud obtained in the step (1) into a rooting culture medium, and then culturing for 25 days under the conditions of 28 ℃,2500lux and 18 h/day of illumination time to obtain a new tissue culture seedling; wherein, the rooting culture medium consists of a 1/2MS culture medium, IBA, agar and sucrose or white sugar, the mass concentration of the IBA in the rooting culture medium is 0.8mg/L, the mass concentration of the sucrose or the white sugar in the rooting culture medium is 40g/L, the mass concentration of the agar in the rooting culture medium is 9g/L, and the pH value of the rooting culture medium is 6.2;
(4) Adding distilled water with the height of 1cm into the tissue culture seedlings obtained in the step (3), then opening the bottle mouth, covering three quarters of the bottle mouth with a cover, and acclimatizing and hardening the seedlings for 5 days;
(5) Taking out the tissue culture seedlings in the step (4), cleaning a root culture medium, transplanting the tissue culture seedlings into a plug tray filled with coconut chaff, perlite and plant ash, thoroughly watering the plug tray by using tap water, placing the plug tray on a tray filled with 1000 times diluted nutrient solution, covering a cover on the tray for moisturizing, continuously culturing for 30 days, and then planting; wherein, the coconut husk, the perlite and the plant ash are uniformly mixed according to the volume ratio of 7.
Example 3
An industrialized seedling method of actinidia arguta comprises the following steps:
(1) Taking a stem section of purple Sadowa Actinidia arguta with a single bud, inoculating the stem section into a proliferation culture medium, then culturing for 35 days under the illumination conditions of 25 ℃,2000lux and 169h/day to obtain a stem section with 8cm branches, cutting off the branches, and cutting the branches into two parts, wherein one part is a terminal bud with 2-3 leaves in a length of 3cm, and the other part is a stem section with a single bud in a length of 1 cm; wherein the enrichment culture medium consists of an MS culture medium, 6-BA, NAA, agar and sucrose or white sugar, the mass concentration of the 6-BA in the enrichment culture medium is 1.0mg/L, the mass concentration of the NAA in the enrichment culture medium is 0.2mg/L, the mass concentration of the sucrose or white sugar in the enrichment culture medium is 30g/L, the mass concentration of the agar in the enrichment culture medium is 7.5g/L, and the pH value of the enrichment culture medium is 6.0;
(2) Repeating the value-added culture process in the step (1) on the single-bud stem section obtained in the step (1);
(3) Inoculating the terminal bud obtained in the step (1) into a rooting culture medium, and then culturing for 23 days under the conditions of 25 ℃,2000lux and illumination time of 16 h/day to obtain a new tissue culture seedling; wherein, the rooting culture medium consists of a 1/2MS culture medium, IBA, agar and sucrose or white sugar, the mass concentration of the IBA in the rooting culture medium is 0.7mg/L, the mass concentration of the sucrose or white sugar in the rooting culture medium is 30g/L, the mass concentration of the agar in the rooting culture medium is 7.5g/L, and the pH value of the rooting culture medium is 5.8;
(4) Adding distilled water with the height of 1cm into the tissue culture seedlings obtained in the step (3), then opening the bottle mouth, covering three quarters of the bottle mouth with a cover, and acclimatizing and hardening the seedlings for 5 days;
(5) Taking out the tissue culture seedlings in the step (4), cleaning a root culture medium, transplanting the tissue culture seedlings into a plug tray filled with coconut chaff, perlite and plant ash, thoroughly watering the plug tray by using tap water, placing the plug tray on a tray filled with 1000-time diluted nutrient solution, covering a cover on the tray for moisturizing, and continuously culturing for 25 days and then planting; wherein, the coconut husk, the perlite and the plant ash are uniformly mixed according to the volume ratio of 7.
Example 4
An industrialized seedling method of actinidia arguta comprises the following steps:
(1) Taking a stem section of the red flame Actinidia arguta with single bud, inoculating the stem section into a multiplication culture medium, then culturing for 35 days under the illumination conditions of 25 ℃,2000lux and 169h/day to obtain a stem section with 8cm branches, cutting off the branches, and cutting the branches into two parts, wherein one part is a terminal bud with 2-3 leaves in a length of 2.8cm, and the other part is a stem section with a single bud in a length of 1.2 cm; wherein the enrichment culture medium consists of an MS culture medium, 6-BA, NAA, agar and sucrose or white sugar, the mass concentration of the 6-BA in the enrichment culture medium is 0.9mg/L, the mass concentration of the NAA in the enrichment culture medium is 0.2mg/L, the mass concentration of the sucrose or white sugar in the enrichment culture medium is 35.0g/L, the mass concentration of the agar in the enrichment culture medium is 8.0g/L, and the pH value of the enrichment culture medium is 6.0;
(2) Repeating the value-added culture process in the step (1) on the single-bud stem section obtained in the step (1);
(3) Inoculating the terminal bud obtained in the step (1) into a rooting culture medium, and then culturing for 23 days under the conditions of 25 ℃,2000lux and illumination time of 16 h/day to obtain a new tissue culture seedling; wherein, the rooting culture medium consists of a 1/2MS culture medium, IBA, agar and sucrose or white sugar, the mass concentration of the IBA in the rooting culture medium is 0.8mg/L, the mass concentration of the sucrose or the white sugar in the rooting culture medium is 25g/L, the mass concentration of the agar in the rooting culture medium is 8g/L, and the pH value of the rooting culture medium is 6.0;
(4) Adding distilled water with the height of 1cm into the tissue culture seedlings obtained in the step (3), then opening the bottle mouth, covering three quarters of the bottle mouth with a cover, and acclimatizing and hardening the seedlings for 5 days;
(5) Taking out the tissue culture seedlings in the step (4), cleaning a root culture medium, transplanting the tissue culture seedlings into a plug tray filled with coconut chaff, perlite and plant ash, thoroughly watering the plug tray by using tap water, placing the plug tray on a tray filled with 1000-time diluted nutrient solution, covering a cover on the tray for moisturizing, and continuously culturing for 25 days and then planting; wherein, the coconut husk, the perlite and the plant ash are uniformly mixed according to the volume ratio of 7.
Example 5
An industrialized seedling method of actinidia arguta comprises the following steps:
(1) Taking a stem section of 'Weiji' actinidia arguta with a single bud, inoculating the stem section into a proliferation culture medium, then culturing for 35 days under the illumination conditions of 25 ℃,2000lux and 169h/day to obtain a stem section with 8cm branches, cutting the branches, and cutting the branches into two parts, wherein one part is a terminal bud with 2-3 leaves in the length of 2.8cm, and the other part is a stem section with a single bud in the length of 1.2 cm; wherein the proliferation culture medium consists of an MS culture medium, 6-BA, NAA, agar and sucrose or white sugar, the mass concentration of the 6-BA in the proliferation culture medium is 1.2mg/L, the mass concentration of the NAA in the proliferation culture medium is 0.2mg/L, the mass concentration of the sucrose or white sugar in the proliferation culture medium is 25.0g/L, the mass concentration of the agar in the proliferation culture medium is 8.0g/L, and the pH value of the proliferation culture medium is 6.0;
(2) Repeating the value-added culture process in the step (1) on the single-bud stem section obtained in the step (1);
(3) Inoculating the terminal bud obtained in the step (1) into a rooting culture medium, and then culturing for 23 days under the conditions of 25 ℃,2000lux and 16 h/day of illumination time to obtain a new tissue culture seedling; wherein, the rooting culture medium consists of a 1/2MS culture medium, IBA, agar and sucrose or white sugar, the mass concentration of the IBA in the rooting culture medium is 0.6mg/L, the mass concentration of the sucrose or white sugar in the rooting culture medium is 35g/L, the mass concentration of the agar in the rooting culture medium is 8g/L, and the pH value of the rooting culture medium is 6.0;
(4) Adding distilled water with the height of 1cm into the tissue culture seedlings obtained in the step (3), then opening the bottle mouth, covering three quarters of the bottle mouth with a cover, and acclimatizing and hardening the seedlings for 5 days;
(5) Taking out the tissue culture seedlings in the step (4), cleaning a root culture medium, transplanting the tissue culture seedlings into a plug tray filled with coconut chaff, perlite and plant ash, thoroughly watering the plug tray by using tap water, placing the plug tray on a tray filled with 1000-time diluted nutrient solution, covering a cover on the tray for moisturizing, and continuously culturing for 25 days and then planting; wherein, the coconut husk, the perlite and the plant ash are uniformly mixed according to the volume ratio of 7.
Test examples
1. The rooting rate and the survival rate of transplanting after rooting culture of the seedlings in the examples 1 to 5 are respectively counted, and the specific results are shown in table 1.
Table 1: statistical table for rooting rate and transplanting survival rate of seedlings
According to the table, the seedling rooting rate is over 99 percent and reaches 100 percent at most when the seedling is cultured according to the method, the survival rate of the transplanted seedling is up to 100 percent, the seedling height of the seedling is over 28cm and reaches 35cm at most after 30 days of transplantation, and the seedling height is higher than that of the conventional seedling culture method after 90 days of propagation to transplantation.
Claims (1)
1. An industrialized seedling method of actinidia arguta is characterized by comprising the following steps:
(1) Taking a stem section of actinidia arguta with a single bud, inoculating the stem section into a proliferation culture medium, then culturing for 35 days under the illumination conditions of 25 ℃,2000lux, 169h/day to obtain a stem section with 8cm branches, cutting the branches, and cutting the branches into two parts, wherein one part is a terminal bud with 2-3 leaves in a length of 3cm, and the other part is a stem section with a single bud in a length of 1 cm; wherein the multiplication culture medium consists of an MS culture medium, 6-BA, NAA, agar and sucrose or consists of the MS culture medium, 6-BA, NAA, agar and white sugar, the mass concentration of the 6-BA in the multiplication culture medium is 1.0mg/L, the mass concentration of the NAA in the multiplication culture medium is 0.2mg/L, the mass concentration of the sucrose or the white sugar in the multiplication culture medium is 30g/L, the mass concentration of the agar in the multiplication culture medium is 7.5g/L, and the pH value of the multiplication culture medium is 6.0;
(2) Repeating the proliferation culture process in the step (1) on the single-bud stem section obtained in the step (1);
(3) Inoculating the terminal bud obtained in the step (1) into a rooting culture medium, and then culturing for 23 days under the conditions of 25 ℃,2000lux and illumination time of 16 h/day to obtain a new tissue culture seedling; wherein, the rooting culture medium consists of a 1/2MS culture medium, IBA, agar and sucrose or consists of a 1/2MS culture medium, IBA, agar and white sugar, the mass concentration of the IBA in the rooting culture medium is 0.7mg/L, the mass concentration of the sucrose or the white sugar in the rooting culture medium is 30g/L, the mass concentration of the agar in the rooting culture medium is 7.5g/L, and the pH value of the rooting culture medium is 5.8;
(4) Adding distilled water with the height of 1cm into the tissue culture seedlings obtained in the step (3), then opening the bottle mouth, covering three quarters of the bottle mouth with a cover, and acclimatizing and hardening the seedlings for 5 days;
(5) Taking out the tissue culture seedlings in the step (4), cleaning a root culture medium, transplanting the tissue culture seedlings into a plug tray filled with coconut chaff, perlite and plant ash, thoroughly watering the plug tray by using tap water, placing the plug tray on a tray filled with 1000-time diluted nutrient solution, covering a cover on the tray for moisturizing, and continuously culturing for 25 days and then planting; wherein, the coconut husk, the perlite and the plant ash are uniformly mixed according to the volume ratio of 7; the Actinidia arguta is Actinidia chinensis Franch, actinidia arguta Kiwi, or Actinidia polygama Kiwi.
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