Method for accelerating growth of peony tissue culture seedlings

Abstract

The invention provides a method for accelerating growth of peony tissue culture seedlings, and belongs to the technical field of rapid cultivation of peonies. The method comprises the steps of S1, tissue culture seedling transplantation, S2, low-temperature treatment and S3, repeated execution of the S1 and S2. Wherein in the step S1, rooting bottle seedlings are transplanted into nutrition bowls; when the height of the tissue culture seedlings transplanted into the nutrition pot reaches 10-12 cm, leaves are changed from green to yellow green, and dormant buds are formed at stem tips, the step S2 is executed for low-temperature treatment; the step S2 comprises the step of placing the tissue culture seedlings with completely developed dormant buds at low temperature to break dormancy. According to the method, the peony tissue culture seedlings are used as materials for the first time, the peony tissue culture seedlings grow for multiple rounds in one year by manually adjusting the culture environment of the tissue culture seedlings, the growth speed of the peony tissue culture seedlings is greatly increased, the time from bottle discharging to flowering of the tissue culture seedlings is shortened, and the application process of the tissue culture seedlings is accelerated.

Description

Technical Field

[0001] The invention relates to the technical field of rapid peony cultivation, and in particular to a method for accelerating the growth of peony tissue culture seedlings. Background Art

[0002] The statements in this section merely provide background art related to the present disclosure and do not necessarily constitute prior art.

[0003] Peony is a traditional famous flower in my country, known as "national beauty" and "king of flowers". In recent years, as the oil value, fresh cut flower value and health care function of peony are gradually excavated, peony has become an important economic plant that integrates ornamental, medicinal and edible health care. Its status in the flower industry is gradually improving, and the market prospect is broad. Peony is usually propagated by division and grafting, but due to season and land restrictions, the propagation speed is slow and the efficiency is low, which cannot meet the growing market demand. Therefore, using tissue culture technology for seedling propagation is the most feasible peony propagation method at present. However, the difficulty of rooting peony tissue culture seedlings and the low survival rate of transplanting have always been the main factors hindering its application in production practice. The researchers of the present invention have finally overcome the technical bottleneck of rooting and transplanting of peony tissue culture seedlings through years of research and development in recent years, with a rooting rate of about 90% and a transplanting survival rate of more than 80%, and a complete tissue culture rapid propagation technology system has been established.

[0004] However, peony tissue culture seedlings grow slowly after transplantation, similar to the growth habits in nature. Even in the artificial climate chamber, they only have a vigorous growth period of about 3 months, after which dormant buds appear, enter a dormant state, and stagnate in growth. In this case, tissue culture seedlings are usually transplanted to the field in autumn, and after being cooled in winter, dormancy is broken. In the spring of the second year, dormant buds germinate and enter the next round of growth. Under natural environmental conditions, it takes 3-4 years from the time the tissue culture seedlings are taken out of the bottle to flowering, which seriously reduces the promotion and application speed of tissue culture seedlings.

[0005] At present, most of the research on peony tissue culture rapid propagation technology is still on the rooting and transplanting of tissue culture seedlings. A complete and efficient tissue culture rapid propagation technology system has not yet been established. It is rare to obtain tissue culture seedlings that survive after transplantation, and there are few reports on the research on transplanting and training of tissue culture seedlings. How to shorten the dormancy cycle and achieve multiple rounds of growth in one year through artificial environment regulation has become a key issue in accelerating the application of peony tissue culture seedlings. Summary of the invention

[0006] In order to solve the above problems, the present invention provides a method for accelerating the growth of peony tissue culture seedlings. For the first time, peony tissue culture seedlings are used as materials, and the culture environment of the tissue culture seedlings is artificially adjusted to achieve multiple rounds of growth of the peony tissue culture seedlings in one year, thereby greatly improving the growth rate, shortening the time from the tissue culture seedlings being taken out of the bottle to flowering, and accelerating the application process of the tissue culture seedlings.

[0007] The present invention provides a method for accelerating the growth of peony tissue culture seedlings, comprising: step S1: transplanting tissue culture seedlings, step S2: low temperature treatment, step S3: repeatedly performing step S1 and step S2;

[0008] Wherein, step S1 comprises: transplanting the rooted bottle seedlings into the nutrient pot;

[0009] When the height of the tissue culture seedlings transplanted into the nutrient pots reaches 10-12 cm, the leaves change from green to yellow-green, and dormant buds are formed at the stem tips, step S2 of low temperature treatment is performed;

[0010] Step S2 includes: placing the tissue culture seedlings with fully developed dormant buds in low temperature to break dormancy.

[0011] Furthermore, the transplanting matrix in the nutrient pot includes: a mixture of peat and perlite in a volume ratio of 2:1-4:1, and 0.2-0.4 g / plant of a composite microbial agent.

[0012] Furthermore, the composite microbial agent includes any one or more of the following: Bacillus subtilis, Bacillus amyloliquefaciens, and Bacillus megaterium.

[0013] Furthermore, the step S1 also includes: in the initial 10 to 20 days after transplanting, the relative humidity of the artificial climate chamber is greater than or equal to 90%, the temperature is controlled at 16-20° C., the lighting time is 12 hours, and the lighting intensity is 3000-4000 Lux.

[0014] Further, when step S3 is executed and step S1 is executed again, the tissue culture seedlings transplanted into the nutrient pots in the previous round are transplanted into new nutrient pots; and the light intensity is replaced with 5000-6000 Lux.

[0015] Furthermore, the step S1 also includes: applying water-soluble fertilizer once every 10-15 days.

[0016] Furthermore, the ratio of fertilizer nutrients in the water-soluble fertilizer is N:P:K=20:20:20.

[0017] Furthermore, the temperature range of the low temperature treatment in step S2 is 2-4°C.

[0018] Furthermore, the time range of the low temperature treatment in step S2 is 40-50 days.

[0019] Furthermore, the low temperature treatment time in step S2 is preferably 45 days.

[0020] Compared with the prior art, the method for accelerating the growth of peony tissue culture seedlings provided by the present invention has the following beneficial effects: for the first time, peony tissue culture seedlings are used as materials, and the peony tissue culture seedlings are grown in multiple rounds in one year by artificially adjusting the culture environment of the tissue culture seedlings, which greatly improves the growth rate, shortens the time from the tissue culture seedlings leaving the bottle to flowering, and shortens the time from 3-4 years in the prior art to 1-2 years. The specific time from the bottle to flowering depends on how many rounds of growth are artificially performed before transplanting to the field. The method is simple to operate, easy to promote, accelerates the application process of tissue culture seedlings, and has high practical value. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] The accompanying drawings constituting a part of the present disclosure are used to provide a further understanding of the present disclosure. The illustrative embodiments of the present disclosure and their descriptions are used to explain the present disclosure and do not constitute an improper limitation on the present disclosure.

[0022] Figure 1 A photo of the tissue culture seedlings provided for the invention 45 days after transplantation;

[0023] Figure 2 This is a photo of the tissue culture seedlings provided by the present invention when they were transplanted 80 days ago;

[0024] Figure 3 A photo of dormant buds formed by the first round of growth provided by the present invention;

[0025] Figure 4 The tissue culture seedlings in the second round of growth provided by the present invention;

[0026] Figure 5 This is a photo of dormant buds formed during the second round of growth provided by the present invention. DETAILED DESCRIPTION

[0027] It should be noted that the following detailed descriptions are exemplary and are intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used in the present invention have the same meanings as those commonly understood by those skilled in the art to which the present invention belongs.

[0028] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit exemplary embodiments according to the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that the terms "including" and "having" and any of their variations are intended to cover non-exclusive inclusions, for example, a process or method comprising a series of steps or units is not necessarily limited to those steps clearly listed, but may include other steps that are not clearly listed or inherent to these processes or methods.

[0029] In the absence of conflict, the embodiments and features of the embodiments of the present invention can be combined with each other. The acquisition of all data in this embodiment is based on the legal application of data in compliance with laws and regulations and user consent.

[0030] Embodiment 1

[0031] The present invention provides a method for accelerating the growth of peony tissue culture seedlings, comprising the following steps:

[0032] 1. Transplanting of tissue culture seedlings

[0033] After the rooted seedlings are gradually adapted to the external environment, they are transplanted into a nutrient pot with a diameter of 8-9 cm. The transplanting matrix is ​​a mixture of peat and perlite with a volume ratio of about 3:1. At the same time, 0.3g / strain of composite microbial agent (including Bacillus subtilis, Bacillus amyloliquefaciens, and Bacillus megaterium) is added to the matrix to prevent stem rot of tissue culture seedlings and promote the production of capillary roots. The composite microbial agent is purchased from Henan Yangshao Biochemical Engineering Co., Ltd., and the trade name is "Yangshao Composite Microbial Agent". The environmental control of the artificial climate chamber is: the relative humidity is maintained above 90% for the first 10 to 20 days of transplanting, and then the humidity gradually decreases. The temperature can be kept constant or variable between 16 and 20°C, the lighting time is 12h, and the light intensity is 3500Lux. Apply water-soluble fertilizer (N:P:K=20:20:20) once every 15 days or so according to the humidity of the matrix. 1.5 months after transplanting, the tissue culture seedlings can grow to 7.5-8.5cm in height and have 1-2 new leaves (such as Figure 1 shown).

[0034] 2. Dormant bud formation

[0035] About 2 months after transplanting, the seedlings are 10-12 cm tall, the leaves gradually age, changing from green to yellow-green, and dormant buds can be seen at the tip of the stem (such as Figure 2 , Figure 3 At 2.5-3 months, the dormant buds gradually swell and develop completely, while the leaves of the tissue culture seedlings gradually turn yellow and enter the senescence stage.

[0036] 3. Low temperature treatment

[0037] The tissue culture seedlings with fully developed dormant buds are placed in a low temperature of 2-4℃ for about 45 days to break dormancy. When the dormant buds show signs of germination, the tissue culture seedlings are taken out and placed in the artificial climate chamber for cultivation again.

[0038] 4. Second round of growth

[0039] The dormant buds that have been stored in low temperature can germinate after about one week of cultivation in an artificial climate chamber (temperature controlled between 16 and 20°C, light duration of 12 hours, and light intensity increased to 5000 Lux). The leaves are significantly larger and thicker than those in the first round of growth (such as Figure 4 At this time, change the nutrient pot from 8-9cm to 12-13cm to provide more space for the second round of growth of the tissue culture seedlings. Apply water-soluble fertilizer (N:P:K=20:20:20) once every 15 days. After 2 months, the tissue culture seedlings can grow to 15-17cm in height, with 3-5 leaves, and the root system can be significantly enlarged and thickened.

[0040] 5. Secondary formation of dormant buds

[0041] At 2.5-3 months, the leaves of the tissue culture seedlings begin to turn yellow, and dormant buds can be seen at the tip of the stem. They gradually swell and become significantly larger than the dormant buds formed in the first round of growth. The leaves also gradually turn yellow and age. At this time, the stem has become lignified, and a few dormant buds have formed at the base of the stem (such as Figure 5 shown).

[0042] 6. Low temperature treatment

[0043] Repeat step 3 with the tissue culture seedlings.

[0044] 7. Three rounds of growth

[0045] The cultivation process is the same as step 4, and the nutrient pot is changed from 12-13cm to 16-17cm to provide more space for three rounds of growth of tissue culture seedlings. When the tissue culture seedlings grow for about 3 months, repeat steps 3 and 4, so that the cyclic growth of tissue culture seedlings can be achieved, and they can be transplanted to the field at any round of growth according to actual needs.

[0046] Embodiment 2

[0047] Patent document with application number CN202010129492.X provides a tissue culture propagation method for peony, and the transplanting method provided includes:

[0048] 1. Preparation for transplanting

[0049] The robust rooted seedlings with a root length of about 4-7 cm after 30-40 days of rooting culture were taken out from the culture room and cultured under diffuse light in a greenhouse at 20-22°C and the bottle caps were gradually removed. Transplantation was carried out after 5-7 days, the rooted seedlings were taken out, the culture medium at the roots was cleaned, and then transplanted into a mixed matrix containing peat soil, perlite, and vermiculite (3:1:1).

[0050] 2. Environmental Control

[0051] After transplanting, place it in a greenhouse with a temperature of 20-22°C and a relative humidity of more than 80%. The light cycle is 16 hours of light cultivation and 8 hours of dark environment cultivation in 24 hours a day, and the light intensity is 2000-3000Lux.

[0052] 3. Seedling hardening management

[0053] After 30 days, move the seedlings to an environment with a relative humidity below 80% to harden the seedlings, keeping the soil and water semi-dry.

[0054] 4. Growth cycle

[0055] Three months after transplanting, each plant has about 1-2 leaves and the seedlings grow vigorously; three years after transplanting, the plant height is about 30-50 cm and can bloom and bear fruit.

[0056] In the second embodiment, dormancy is not relieved by artificial low temperature treatment, but the plants rely on natural low temperature storage in winter. At the same time, there is no cyclic growth design, and the plants rely on the natural growth cycle only after a single transplant.

[0057] Embodiment 3

[0058] The transplanting methods provided in the paper "Optimization of the Micropropagation Technology System of 'Zhengwu' Peony" include:

[0059] 1. Preparation for transplanting

[0060] The rooted seedlings were placed in a 4°C refrigerator in the dark for 30 days to break dormancy, and then transferred to a bottle at room temperature for 1 week for seedling hardening; they were transplanted into a 8cm×8cm×9cm (length×width×height) plastic pot with a substrate of perlite: vermiculite: peat soil (volume ratio 1:1:1) sterilized at high pressure (121°C, 40min).

[0061] 2. Environmental Control

[0062] The cells were placed in an artificial climate chamber with a temperature of 20°C ± 1°C, a humidity of 70%, and a light intensity of 32.4 μmol·m -2 ·s -1 , the photoperiod is 14h per day.

[0063] 3. Seedling hardening management

[0064] The survival rate was calculated 60 days after transplanting (85% of the first-grade seedlings took root in one step). In the second year, the tissue culture seedlings were transferred to the greenhouse for growth and normal water and fertilizer management was carried out.

[0065] 4. Growth cycle

[0066] based on Figure 1 As shown in Figure 1, the tissue culture seedlings grew normally in the greenhouse in the second year after transplantation, but did not bloom at this time. Based on the flowering photos provided in another paper published by the author of this paper, "Establishment of the 'Noon' Peony Micropropagation Technology System", it can be seen that the method provided in Example 3 also requires the tissue culture seedlings to be transplanted into the field for three years before entering the flowering period.

[0067] Example 3 only has a single low temperature treatment (4°C low temperature dark treatment for 30 days), without a cyclic design. At the same time, it does not mention multiple rounds of growth regulation, relying on the natural growth rhythm.

[0068] In the first embodiment of the present invention, a method for accelerating the growth of peony tissue culture seedlings is provided. The trinity technical framework of "transplanting-low temperature dormancy release-cyclic growth" is adopted to solve the technical problems of slow growth and long dormancy period of peony tissue culture seedlings after transplanting. Compared with the transplanting methods provided in the second and third embodiments: the flowering cycle is shortened from 3-4 years to 1-2 years; multiple rounds of growth can significantly accelerate the industrial application of tissue culture seedlings, do a good job in disease prevention and control, and the comprehensive benefit can be increased to more than 30%; the program parameters are clear, providing a standardized template for the industrialized rapid propagation of peony.

[0069] Although the present invention is disclosed as above, the protection scope of the present invention is not limited thereto. Those skilled in the art may make various changes and modifications without departing from the spirit and scope of the present invention, and these changes and modifications will fall within the protection scope of the present invention.

Claims

1. A method for accelerating the growth of peony tissue culture seedlings, characterized in that: The method comprises: step S1: transplanting tissue culture seedlings, step S2: low temperature treatment, step S3: repeatedly performing step S1 and step S2; Wherein, step S1 comprises: transplanting the rooted bottle seedlings into the nutrient pot; When the height of the tissue culture seedlings transplanted into the nutrient pots reaches 10-12 cm, the leaves change from green to yellow-green, and dormant buds are formed at the stem tips, step S2 of low temperature treatment is performed; Step S2 includes: placing the tissue culture seedlings with fully developed dormant buds in low temperature to break dormancy.

2. The method for accelerating the growth of peony tissue culture seedlings as claimed in claim 1, characterized in that, The transplanting matrix in the nutrient pot includes: a mixture of peat and perlite in a volume ratio of 2:1-4:1 and 0.2-0.4 g / plant of a composite microbial agent.

3. The method for accelerating the growth of peony tissue culture seedlings as claimed in claim 2, characterized in that, The composite microbial agent includes any one or more of the following: Bacillus subtilis, Bacillus amyloliquefaciens, and Bacillus megaterium.

4. The method for accelerating the growth of peony tissue culture seedlings as claimed in claim 1, characterized in that: The step S1 also includes: in the initial 10-20 days after transplanting, the relative humidity of the artificial climate chamber is greater than or equal to 90%, the temperature is controlled at 16-20° C., the lighting time is 12 hours, and the lighting intensity is 3000-4000 Lux.

5. The method for accelerating the growth of peony tissue culture seedlings as claimed in claim 4, characterized in that: When step S3 is executed and step S1 is executed again, the tissue culture seedlings transplanted in the nutrient pots in the previous round are transplanted into new nutrient pots; the light intensity is replaced with 5000-6000 Lux.

6. The method for accelerating the growth of peony tissue culture seedlings as claimed in claim 1, characterized in that: The step S1 also includes: applying water-soluble fertilizer once every 10-15 days.

7. The method for accelerating the growth of peony tissue culture seedlings as claimed in claim 6, characterized in that: The ratio of fertilizer nutrient elements in the water-soluble fertilizer is N:P:K=20:20:

20.

8. The method for accelerating the growth of peony tissue culture seedlings as claimed in claim 1, characterized in that: The temperature range of the low temperature treatment in step S2 is 2-4°C.

9. The method for accelerating the growth of peony tissue culture seedlings as claimed in claim 1, characterized in that: The time range of the low temperature treatment in step S2 is 40-50 days.

10. The method for accelerating the growth of peony tissue culture seedlings as claimed in claim 9, characterized in that: The time for the low temperature treatment in step S2 is preferably 45 days.

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