Novel plant tissue culture support as well as preparation method and application thereof

Abstract

The invention belongs to the field of plant tissue culture, and particularly relates to a novel plant tissue culture support as well as a preparation method and application thereof. The novel plant tissue culture support comprises the following raw materials in parts by weight: 1-5 parts of sawdust, 1-2 parts of perlite, 1-3 parts of peat soil, 1-2 parts of coco coir and 0-1 part of moss. The preparation method comprises the following steps: mixing and stirring all raw materials to obtain a premix; and sterilizing the premix, thereby obtaining the product. Compared with the prior art, the method has the advantages that agar powder and carrageenan which are expensive in price are not used (actually, the agar powder and the carrageenan only have a supporting effect and do not contain any other nutrient substances), and the material cost of the culture medium is saved by about one third; the low-cost mixture matrix of the sawdust, the peat soil, the perlite, the coco coir and the sphagna is used, so that the material cost is saved, the tissue culture seedlings can adapt to the culture matrix for greenhouse production in advance, and the survival rate of the tissue culture seedlings is greatly increased.

Description

Technical Field

[0001] The invention belongs to the field of plant tissue culture, and in particular relates to a novel plant tissue culture support and a preparation method and application thereof. Background Art

[0002] In the process of exploring plant tissue culture, we have experienced the exploration stage (before the 1930s), the foundation stage (mid-1930s to the late 1950s) and the rapid development stage to date.

[0003] In the process of plant tissue culture, supports are indispensable. However, agar powder and carrageenan in the existing support composition only serve as coagulants and supports, but do not provide any nutrition. They are expensive, accounting for nearly one-third of the total value of culture medium medicines.

[0004] Therefore, based on this, the technical solution of the present invention is proposed. Summary of the invention

[0005] In order to solve the problems existing in the prior art, the present invention provides a novel plant tissue culture support, which comprises the following raw materials in parts by weight: 1-5 parts of sawdust, 1-2 parts of perlite, 1-3 parts of peat soil, 1-2 parts of coconut bran, and 0-1 part of moss.

[0006] Preferably, the novel plant tissue culture support comprises the following raw materials in parts by weight: 2 parts of sawdust, 1 part of perlite, 2 parts of peat soil, 1 part of coconut bran, and 1 part of moss.

[0007] Preferably, the particle size of the wood chips is 1 to 3 mm;

[0008] And / or, the particle size of the perlite is 0.5 to 1.5 mm;

[0009] And / or, in the peat soil, the length of the peat fibers is 1 to 5 mm;

[0010] And / or, the particle size of the coconut bran is 0.5-1.5 mm.

[0011] Based on the same technical concept, another solution of the present invention is to provide a method for preparing a novel plant tissue culture support, the preparation method comprising the following steps:

[0012] (1) mixing and stirring all raw materials to obtain a premix;

[0013] (2) sterilizing the premix to obtain the novel plant tissue culture support.

[0014] Preferably, in step (1), the mixing and stirring speed is 30 to 50 r / min, and the stirring time is 10 to 20 min.

[0015] Preferably, in step (2), the sterilization method is high temperature and high pressure sterilization.

[0016] Based on the same technical concept, another solution of the present invention is to provide a novel plant tissue culture support for use in plant tissue culture.

[0017] Preferably, the plants include flower plants and Chinese medicinal plants, more specifically:

[0018] Flower plants: orchids (Chinese orchids, cymbidium orchids, hybrid orchids, phalaenopsis, cattleya, oncidium, dendrobium officinale, roxburghii, etc.), anthurium, rose, rose, anthurium, lily, amaryllis, chrysanthemum, lisianthus, baby's breath, begonia, azalea, marantaceae, araceae, etc.

[0019] Chinese medicinal plants: ginseng, monk fruit, seven-fingered peach, nine-fingered peach, white striata, atractylodes, polygonatum, trichosanthes, pinellia, polygala, fritillaria, patchouli, sagittaria, etc.

[0020] It should be emphasized that when in use, the new plant tissue culture support needs to be added with the corresponding formula nutrient solution (which changes accordingly according to the type of plant) and can only be used for tissue culture inoculation after high temperature and high pressure sterilization.

[0021] The beneficial effects of the present invention are:

[0022] Compared with the existing method, the present invention does not use expensive agar powder and carrageenan (in fact, these two components only have a supporting function and do not contain any other nutrients), saving nearly one-third of the cost of culture medium materials; and the low-cost sawdust, peat soil, perlite, coconut bran, and sphagnum moss mixture matrix used in the present invention not only saves material costs, but also allows tissue culture seedlings to adapt to the cultivation matrix of greenhouse production in advance, thereby greatly improving the survival rate of tissue culture seedlings. DETAILED DESCRIPTION

[0023] To make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described in detail below. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other implementation methods obtained by ordinary technicians in this field without creative work belong to the scope of protection of the present invention.

[0024] Example 1

[0025] This embodiment provides a method for preparing a novel plant tissue culture support, the preparation method comprising the following steps:

[0026] (1) 2 parts of sawdust (particle size of 1-2 mm), 2 parts of perlite (particle size of 0.5-1 mm), 3 parts of peat soil (peat fiber length of 1-3 mm), and 1 part of coconut bran (particle size of 0.5-1 mm) were mixed and stirred at 30 r / min for 20 min until uniform to obtain a premix;

[0027] (2) The premix is ​​conventionally sterilized under high temperature and high pressure conditions to obtain the novel plant tissue culture support.

[0028] It should be noted that 1 portion is 10g, the same below.

[0029] Example 2

[0030] This embodiment provides a method for preparing a novel plant tissue culture support, the preparation method comprising the following steps:

[0031] (1) 1 part of sawdust (particle size of 1 to 3 mm), 2 parts of perlite (particle size of 0.5 to 1.5 mm), 3 parts of peat soil (peat fiber length of 1 to 5 mm), 1 part of coconut bran (particle size of 0.5 to 1.5 mm) and 1 part of moss were mixed and stirred at 50 r / min for 10 min until uniform to obtain a premix;

[0032] (2) The premix is ​​conventionally sterilized under high temperature and high pressure conditions to obtain the novel plant tissue culture support.

[0033] Example 3

[0034] This embodiment provides a method for preparing a novel plant tissue culture support, the preparation method comprising the following steps:

[0035] (1) 2 parts of sawdust (particle size of 2-3 mm), 1 part of perlite (particle size of 1-1.5 mm), 2 parts of peat soil (peat fiber length of 3-5 mm), 1 part of coconut bran (particle size of 1-1.5 mm) and 1 part of moss were mixed and stirred at 40 r / min for 15 min until uniform to obtain a premix;

[0036] (2) The premix is ​​conventionally sterilized under high temperature and high pressure conditions to obtain the novel plant tissue culture support.

[0037] Example 4

[0038] This embodiment provides a method for preparing a novel plant tissue culture support, the preparation method comprising the following steps:

[0039] (1) 1 part of sawdust (particle size of 2-3 mm), 2 parts of perlite (particle size of 1-1.5 mm), 2 parts of peat soil (peat fiber length of 3-5 mm), 2 parts of coconut bran (particle size of 1-1.5 mm) and 1 part of moss were mixed and stirred at 35 r / min for 20 min until uniform to obtain a premix;

[0040] (2) The premix is ​​conventionally sterilized under high temperature and high pressure conditions to obtain the novel plant tissue culture support.

[0041] Example 5

[0042] This embodiment provides a method for preparing a novel plant tissue culture support, the preparation method comprising the following steps:

[0043] (1) 2 parts of sawdust (particle size of 1 to 3 mm), 2 parts of perlite (particle size of 0.5 to 1.5 mm), 1 part of peat soil (peat fiber length of 2 to 4 mm), 2 parts of coconut bran (particle size of 1 to 1.5 mm) and 1 part of moss were mixed and stirred at 45 r / min for 20 min until uniform to obtain a premix;

[0044] (2) The premix is ​​conventionally sterilized under high temperature and high pressure conditions to obtain the novel plant tissue culture support.

[0045] Verification Example

[0046] (I) Using the novel plant tissue culture supports obtained in Examples 1 to 5 for rapid planting of Cymbidium orchid seedlings comprises the following steps:

[0047] (1) Select healthy Cymbidium orchid seedlings of 12 to 24 weeks old, keep the pseudobulb and peripheral roots under the conditions of a clean bench, remove the longer fleshy roots, and appropriately shorten the longer leaves;

[0048] (2) Transplanting the subcultured seedlings of Cymbidium orchids onto a culture medium containing a novel plant tissue culture support for light culture, and adding MS + TDZ 0.5-5.0 mg / L + NAA 0.1-2.0 mg / L + activated carbon 2 g / L + sugar 20 g / L to the planting culture medium;

[0049] (3) Place the inoculated culture bottle (bag) in a light culture room at 25±2°C and culture for 150 to 180 days.

[0050] The survival rate data of transplanted seedlings are shown in Table 1.

[0051] Table 1

[0052]

[0053]

[0054] Note: The control group is a traditional support containing agar powder and carrageenan; the mushroom bag refers to an organic mixture of sawdust, cottonseed hulls, corn cobs, etc. that are discarded after picking during the production process of bag-grown edible mushrooms, the same below.

[0055] (ii) using the novel plant tissue culture support obtained in Examples 1 to 5 for Bletilla striata varieties, comprising the following steps:

[0056] (1) Select robust Bletilla striata varieties, sterilize the explants, obtain tissue culture seedlings, take 6-week-old tissue culture seedlings, retain 1.0-2.0 cm of the main root under ultra-clean bench conditions, and remove the remaining peripheral roots; retain the main root and robust pseudobulbs, and remove the longer leaves;

[0057] (2) The seedlings were inoculated on a planting medium containing a novel plant tissue culture support with a moisture content of 65-80%, a sugar concentration of 20 g / L, and added with MS+NAA 1.0 mg / L, and cultured in a light culture room at 25±2°C for 5 weeks. The survival rate of the planted seedlings was finally measured to be above 98%.

[0058] (III) Using the novel plant tissue culture support obtained in Example 1 and the traditional support of the control group for Dendrobium officinale, comprising the following steps:

[0059] (1) Select healthy subculture seedlings of aseptically germinated Dendrobium officinale, select healthy clump seedlings under clean bench conditions, and plant them in culture medium containing novel plant tissue culture supports and traditional supports, respectively. In addition, the water content of the culture medium is 65-80%, the sugar concentration is 20 g / L, and MS+NAA 1.0 mg / L is added. The culture medium is placed in a light culture room at 25±2°C for 5 weeks, and the survival rate of the planted seedlings is above 95%.

[0060] (2) Acclimation of seedlings: The Dendrobium officinale was placed in a greenhouse in a bottle for acclimation for 10 days without opening the lid and for 3 days with opening the lid.

[0061] (3) Washing seedlings: Take the Dendrobium officinale out of the tissue culture bottle (bag), wash off the mixed materials on the roots with clean water, soak the roots in 1000 times thiabendazim solution, dry them and wait for planting.

[0062] (4) Planting: A seedbed transplanting comparison was conducted. About 3 cm of gravel was laid on the seedbed, and 10 cm of cultivation medium was laid on top of the gravel. The two groups of tissue culture seedlings were respectively transplanted on the seedbed. Half a month later, the effects of the two groups of planting culture media on the transplanting survival rate were statistically analyzed. The results are shown in Table 2.

[0063] Table 2 Survival rate of transplanting Dendrobium officinale

[0064] Group / two parallel experiments Example 1 Control group Group 1 98.0％ 72.5％ Group 2 98.5％ 72.1％

[0065] (IV) Using the novel plant tissue culture support obtained in Example 1 and the traditional support of the control group for five robust roxburghii tissue culture seedling varieties, comprising the following steps:

[0066] (1) Take 12-15 week old tissue culture seedlings, retain 1.0-2.0 cm of main roots under clean bench conditions, remove leaves, cut the golden thread vine into 1-3 cm stem segments, and ensure that each stem segment has an axillary bud. The cut stem segments are inoculated into culture medium containing new plant tissue culture supports and traditional supports, respectively. In addition, the sucrose concentration in the culture medium is 20 g / L, and MS+6-BA 0.5 mg / L+NAA 1.0 mg / L+banana juice 100 g / L+activated carbon 2 g / L is added. The culture medium is placed in a light culture room at 25±2°C for 16-20 weeks.

[0067] (2) Acclimation: Unopened tissue culture bottles (bags) are placed in a greenhouse for acclimation for 20 days, and opened bottles (bags) are placed in a greenhouse for acclimation for 3 days.

[0068] Different varieties and different supports showed different transplanting survival rates as shown in Table 3.

[0069] Table 3

[0070]

[0071]

[0072] The above is only a specific implementation of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be covered within the protection scope of the present invention. Therefore, the protection scope of the present invention shall be based on the protection scope of the claims, and any single use or change in proportion shall be an infringement.

Claims

1. A novel plant tissue culture support, characterized in that: The invention comprises the following raw materials in parts by weight: 1-5 parts of sawdust, 1-2 parts of perlite, 1-3 parts of peat soil, 1-2 parts of coconut bran and 0-1 part of moss.

2. The novel plant tissue culture support according to claim 1, characterized in that: The invention comprises the following raw materials in parts by weight: 2 parts of sawdust, 1 part of perlite, 2 parts of peat soil, 1 part of coconut husk and 1 part of moss.

3. The novel plant tissue culture support according to claim 1 or 2, characterized in that: The particle size of the sawdust is 1 to 3 mm; And / or, the particle size of the perlite is 0.5 to 1.5 mm; And / or, in the peat soil, the length of the peat fibers is 1 to 5 mm; And / or, the particle size of the coconut bran is 0.5-1.5 mm.

4. The method for preparing the novel plant tissue culture support according to any one of claims 1 to 3, characterized in that: The preparation method comprises the following steps: (1) mixing and stirring all raw materials to obtain a premix; (2) sterilizing the premix to obtain the novel plant tissue culture support.

5. The method for preparing the novel plant tissue culture support according to claim 4, characterized in that: In step (1), the mixing and stirring speed is 30 to 50 r / min, and the stirring time is 10 to 20 min.

6. The method for preparing the novel plant tissue culture support according to claim 4, characterized in that: In step (2), the sterilization method is high temperature and high pressure sterilization.

7. Use of the novel plant tissue culture support according to any one of claims 1 to 3 in plant tissue culture.

8. The use according to claim 7, characterized in that: The plants include flower plants and Chinese medicinal plants.

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