Method for tissue culture and rapid propagation of aseptic seedlings of armeniaca mume

Abstract

The invention relates to the technical field of prunus mume, in particular to a method for tissue culture and rapid propagation of aseptic seedlings of prunus mume, and solves the problems that in the prior art, a traditional propagation mode is limited by seasons, the propagation efficiency is low, and consistency of offspring is difficult to guarantee; and large-scale application cannot be realized due to the problems of low regeneration frequency, slow growth and the like in early tissue culture attempts. The invention discloses a method for tissue culture and rapid propagation of aseptic seedlings of armeniaca mume. The method comprises the following steps: disinfecting and removing part of leaves; a culture medium based on 1 / 2 MS is prepared, and a plant tissue culture antibacterial agent is added for high-pressure sterilization treatment. Agricultural production is promoted, economic benefits of the flower industry are improved, development of plant molecular biology and genetic breeding science is promoted, ecological restoration is assisted, the urban landscaping level is improved, market requirements are met, the satisfaction degree and happiness of consumers are improved, domestic and overseas cultural exchange and cooperation can be enhanced through popularization of the breeding technology, and the method is worthy of popularization and application. And the competitiveness and influence of the Chinese flower industry in the international market are improved.

Description

Technical Field

[0001] The invention relates to the technical field of plum blossoms, in particular to a method for rapid propagation of green-calyx plum aseptic seedlings through tissue culture. Background Art

[0002] Plum blossoms (Prunus mume) are traditional Chinese famous flowers with a long history of cultivation. Among them, green-calyx plum blossoms are highly favored for their unique flower color and ornamental value. However, in the traditional propagation methods of green-calyx plum blossoms, such as cuttings, sowing, layering, etc., there are problems such as long propagation cycle, great influence of environmental factors, and unstable genetic traits of offspring. In recent years, although tissue culture technology has been increasingly used in plant propagation, the efficient regeneration system for green-calyx plum blossoms is still imperfect, which restricts its large-scale production and protection of germplasm resources.

[0003] Specifically, traditional propagation methods are limited by seasons, have low propagation efficiency, and are difficult to ensure the consistency of offspring; and early attempts at tissue culture have failed to achieve large-scale application due to problems such as low regeneration frequency and slow growth. Therefore, developing an efficient and stable method for rapid tissue culture propagation of green-calyx plum sterile seedlings is of great significance for improving propagation efficiency, shortening the propagation cycle, and maintaining genetic stability. Summary of the invention

[0004] The purpose of the present invention is to provide a method for rapid tissue culture propagation of green-calyx plum aseptic seedlings, which solves the problem that the traditional propagation method in the prior art is restricted by seasons, has low propagation efficiency, and is difficult to ensure the consistency of offspring; and early tissue culture attempts have failed to achieve large-scale application due to problems such as low regeneration frequency and slow growth.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] A method for rapid propagation of green-calyx plum aseptic seedling tissue culture, comprising selecting green-calyx plum annual branch with bud stem segments in the growth period as explants;

[0007] Pretreatment of explants, including disinfection and removal of some leaves;

[0008] Prepare a medium based on 1 / 2MS, add plant tissue culture antimicrobial agents and sterilize by high pressure;

[0009] The explants were inoculated into the starting medium containing different concentrations of 6-BA and the optimal starting medium formula was screened out.

[0010] Proliferation culture and rooting culture were carried out on the basis of the starting culture, and the concentration of I BA in the rooting medium was adjusted to obtain sterile seedlings of Prunus mume with high rooting efficiency.

[0011] Preferably, the pretreatment of the explant includes: placing the explant in 75% alcohol for disinfection for 30 seconds, then rinsing with sterile water for 3-5 times; then immersing and disinfecting in 2% NaClO for 2 minutes, and then rinsing with sterile water for 3-5 times until it is clean.

[0012] Preferably, the start-up culture medium is based on 1 / 2MS or MS, with different concentrations of 6-BA added, and the culture conditions are temperature 24°C, light intensity 220 μmol·m-2·s-1, humidity 40%, and photoperiod of 16 hours light / 8 hours dark.

[0013] Preferably, the rooting medium is based on 1 / 2MS, and different concentrations of I BA and a fixed concentration of 6-BA are added. By comparing the rooting effects under different I BA concentrations, the optimal rooting medium formula is screened out.

[0014] Preferably, the optimal rooting medium formula is 1 / 2MS+0.3mg / LI BA+0.2mg / L 6-BA, and its rooting rate reaches more than 95.5%, the average number of roots is not less than 5, and the average root length is not less than 4 cm.

[0015] Preferably, the method further comprises dripping detergent into a wide-mouth bottle to soak the explants, and rinsing under running water for 1-2 hours to further remove surface impurities and microorganisms.

[0016] Preferably, it is suitable for the rapid propagation of sterile seedlings of green-calyx plum and other plants of the same genus through tissue culture, and has broad application prospects and practical value.

[0017] The present invention has at least the following beneficial effects:

[0018] Promote agricultural production: The efficient and stable green-calyx plum breeding technology provides sufficient seedling resources for agricultural production, helps to expand the planting area of ​​green-calyx plum, and improves the economic benefits of the flower industry. Promote scientific research progress: The present invention has built a stable technical platform, which provides strong support for cutting-edge research such as genetic transformation and gene editing of green-calyx plum, and promotes the development of plant molecular biology and genetic breeding. Assist ecological restoration: As an ornamental plant and ecological tree species, green-calyx plum has important value in ecological restoration and greening beautification. The breeding technology of the present invention helps to quickly restore the damaged ecological environment and improve the level of urban greening. Meet market demand: With people's pursuit of a better life and the increase in demand for flower consumption, there is a strong market demand for high-quality green-calyx plum seedlings. The breeding technology of the present invention can meet the market demand for high-quality seedlings and improve consumer satisfaction and happiness. Promote cultural exchanges: As one of the important elements in Chinese traditional culture, the promotion of green-calyx plum's breeding technology helps to strengthen cultural exchanges and cooperation at home and abroad, and enhance the competitiveness and influence of China's flower industry in the international market. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying any creative work.

[0020] Figure 1 The structural schematic diagram of the main steps of the method for rapid propagation of green-calyx plum sterile seedlings tissue culture of the present invention DETAILED DESCRIPTION

[0021] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0022] Embodiment 1, refer to Figure 1 , a method for rapid propagation of green-calyx plum aseptic seedling tissue culture, comprising selecting green-calyx plum annual branch with bud stem segments in the growth period as explants;

[0023] Pretreatment of explants, including disinfection and removal of some leaves;

[0024] Prepare a medium based on 1 / 2MS, add plant tissue culture antimicrobial agents and sterilize by high pressure;

[0025] The explants were inoculated into the starting medium containing different concentrations of 6-BA and the optimal starting medium formula was screened out.

[0026] Proliferation culture and rooting culture were carried out on the basis of the starting culture, and the concentration of I BA in the rooting medium was adjusted to obtain sterile seedlings of Prunus mume with high rooting efficiency.

[0027] The present invention provides a method for rapid propagation of green-calyx plum aseptic seedling tissue culture, the method comprising the following steps:

[0028] Explant selection and pretreatment: Select annual green-calyx plum branches with bud stem segments in the growth period as explants. Collect materials in the morning or evening when the weather is cool. Cut stem segments with 1-2 buds, remove most of the leaves to reduce transpiration, place the explants in a wide-mouth bottle with an appropriate amount of water to maintain vitality, and conduct strict surface disinfection.

[0029] Preparation of culture medium: Use 1 / 2MS as the basic culture medium, add appropriate amount of sucrose, agar and plant tissue culture antimicrobial agent, adjust the pH value to the appropriate range and sterilize by high pressure to ensure a sterile environment. Add different concentrations of plant growth regulators such as 6-BA and I BA to the culture medium to optimize the effects of start-up culture and rooting culture.

[0030] Initiation culture: The pretreated explants were inoculated into 1 / 2MS or MS medium containing different concentrations of 6-BA, and cultured under suitable light, temperature and humidity conditions. The germination of axillary buds was observed and recorded regularly to screen out the optimal initiation culture medium formula.

[0031] Proliferation and rooting culture: Based on the initial culture, select healthy rootless seedlings for proliferation and rooting culture. By adjusting the concentration of IBA in the rooting medium, the most suitable medium formula for rooting of green-calyx plum sterile seedlings is screened to further improve the propagation efficiency.

[0032] According to the above embodiments, it can be known that: efficient reproduction: the present invention significantly improves the reproduction efficiency of green-calyx plum sterile seedlings and shortens the reproduction cycle by optimizing explant selection, culture medium formula and culture conditions.

[0033] Genetic stability: The genetic traits of the sterile green-calyx plum seedlings obtained using tissue culture technology are stable, which is conducive to maintaining the genetic characteristics of excellent varieties.

[0034] Strong environmental adaptability: The sterile seedling cultivation process is less affected by the external environment, reducing the risk of breeding failure due to environmental fluctuations.

[0035] Germplasm resource protection: This method provides an effective way for the long-term preservation of plum germplasm resources and helps the continuation of rare varieties.

[0036] Technical platform construction: It provides a stable and reliable technical platform for the genetic transformation of green-calyx plum, breeding of excellent varieties, and other research, and promotes the in-depth development of plum blossom research.

[0037] Example 2, refer to Figure 1 The pretreatment of the explants includes: placing the explants in 75% alcohol for disinfection for 30 seconds, then rinsing with sterile water for 3-5 times; then immersing and disinfecting them in 2% NaClO for 2 minutes, and then rinsing with sterile water for 3-5 times until they are clean.

[0038] The start-up culture medium is based on 1 / 2MS or MS, with different concentrations of 6-BA added, and the culture conditions are a temperature of 24°C, a light intensity of 220 μmol·m-2·s-1, a humidity of 40%, and a photoperiod of 16 hours of light / 8 hours of darkness.

[0039] The rooting medium is based on 1 / 2MS, and different concentrations of I BA and a fixed concentration of 6-BA are added. By comparing the rooting effects under different I BA concentrations, the optimal rooting medium formula is screened out.

[0040] The optimal rooting culture medium formula is 1 / 2MS+0.3mg / L BA+0.2mg / L 6-BA, and its rooting rate reaches more than 95.5%, the average number of roots is not less than 5, and the average root length is not less than 4 centimeters.

[0041] The method also includes dropping detergent into a wide-mouth bottle to soak the explants, and rinsing them under running water for 1-2 hours to further remove surface impurities and microorganisms.

[0042] The method is suitable for rapid propagation of sterile seedlings of green-calyx plum and other plants of the same genus through tissue culture, and has broad application prospects and practical value.

[0043] High efficiency: By optimizing the culture medium formula, selecting suitable plant growth regulators (such as 6-BA and I BA) and accurately controlling the culture conditions (such as light, temperature and humidity), the present invention significantly improves the reproduction efficiency of green-calyx plum sterile seedlings, so that a large number of high-quality seedlings can be obtained in a short time.

[0044] Stability: Tissue culture technology ensures a sterile environment during the breeding process, avoiding contamination by external microorganisms, thereby ensuring the stability and repeatability of the breeding process. At the same time, the genetic background of sterile seedlings is consistent, which is conducive to maintaining the genetic stability of the variety.

[0045] Environmental adaptability: Sterile seedlings grow under artificially controlled environmental conditions, which reduces the impact of natural environmental fluctuations on their growth and makes the breeding process more controllable. In addition, this method can be carried out in seasons that are not suitable for natural breeding, extending the breeding time window.

[0046] Germplasm resource protection: The present invention provides an effective means of preserving rare and endangered green-calyx plum germplasm resources. Through tissue culture, these precious germplasms can be propagated in large quantities, contributing to genetic breeding and biodiversity protection.

[0047] Flexibility: The technology platform of the present invention can be flexibly adjusted according to different varieties and different purposes. For example, by changing the culture medium formula or the type and concentration of plant growth regulators, directional propagation and optimization of different varieties of green-calyx plum can be achieved.

[0048] Beneficial effects:

[0049] Promote agricultural production: The efficient and stable green-calyx plum breeding technology provides sufficient seedling resources for agricultural production, helps to expand the planting area of ​​green-calyx plum and improve the economic benefits of the flower industry.

[0050] Promoting scientific research progress: This invention has built a stable technical platform, providing strong support for cutting-edge research such as genetic transformation and gene editing of green-calyx plum, and promoting the development of plant molecular biology and genetic breeding.

[0051] Helping ecological restoration: As an ornamental plant and ecological tree species, green-calyx plum has important value in ecological restoration and greening and beautification. The breeding technology of the present invention helps to quickly restore the damaged ecological environment and improve the level of urban greening.

[0052] Meeting market demand: With people's pursuit of a better life and increasing demand for flower consumption, the market demand for high-quality green-calyx plum seedlings is strong. The breeding technology of the present invention can meet the market demand for high-quality seedlings and improve consumers' satisfaction and happiness.

[0053] Promote cultural exchanges: As one of the important elements of Chinese traditional culture, the promotion of green-calyx plum breeding technology will help strengthen cultural exchanges and cooperation at home and abroad, and enhance the competitiveness and influence of China's flower industry in the international market.

[0054] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The above embodiments and descriptions only describe the principles of the present invention. The present invention may be subject to various changes and improvements without departing from the spirit and scope of the present invention. These changes and improvements fall within the scope of the present invention. The scope of protection claimed by the present invention is defined by the attached claims and their equivalents.

Claims

1. A method for rapid propagation of green-calyx plum sterile seedling tissue culture, characterized in that: The following steps are involved: One-year-old green-calyx plum branches with buds and stem segments in the growth period were selected as explants; Pretreatment of explants, including disinfection and removal of some leaves; Prepare a medium based on 1 / 2MS, add plant tissue culture antimicrobial agents and sterilize by high pressure; The explants were inoculated into the starting medium containing different concentrations of 6-BA and the optimal starting medium formula was screened out. Proliferation culture and rooting culture were carried out on the basis of the starting culture, and the concentration of IBA in the rooting medium was adjusted to obtain sterile seedlings of green-calyx plum with high rooting efficiency.

2. The method for rapid propagation of green-calyx plum aseptic seedlings by tissue culture according to claim 1, characterized in that: The explant pretreatment includes: placing the explant in 75% alcohol for disinfection for 30 seconds, then rinsing with sterile water for 3-5 times; then immersing and disinfecting with 2% NaClO for 2 minutes, and then rinsing with sterile water for 3-5 times until the explant is clean.

3. The method for rapid propagation of aseptic seedlings of green-calyx plum according to claim 1, characterized in that: The start-up culture medium is based on 1 / 2MS or MS, with different concentrations of 6-BA added, and the culture conditions are temperature 24°C, light intensity 220 μmol·m-2·s-1, humidity 40%, and a photoperiod of 16 hours of light / 8 hours of darkness.

4. The method for rapid propagation of green-calyx plum aseptic seedling tissue culture according to claim 1, characterized in that: The rooting medium is based on 1 / 2MS, and different concentrations of IBA and a fixed concentration of 6-BA are added. By comparing the rooting effects under different IBA concentrations, the optimal rooting medium formula is screened out.

5. The method for rapid propagation of green-calyx plum aseptic seedling tissue culture according to claim 4, characterized in that: The optimal rooting culture medium formula is 1 / 2MS+0.3mg / L IBA+0.2mg / L 6-BA, and its rooting rate reaches more than 95.5%, the average number of roots is not less than 5, and the average root length is not less than 4 centimeters.

6. The method for rapid propagation of green-calyx plum aseptic seedlings through tissue culture according to claim 5, characterized in that: The method also includes dropping detergent into a wide-mouth bottle to soak the explants, and rinsing them under running water for 1-2 hours to further remove surface impurities and microorganisms.

7. The method for rapid propagation of green-calyx plum aseptic seedlings through tissue culture according to claim 6, characterized in that: The method is suitable for rapid propagation of sterile seedlings of green-calyx plum and other plants of the same genus through tissue culture, and has broad application prospects and practical value.

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