CN101278646A - How to make sterile bonsai - Google Patents

Abstract

A method for manufacturing an asepsis bonsai relates to a manufacture method of an ornamental gardening bonsai; that is, an aseptic seedling cultivated by isolated culture and decorative material are placed in an aseptic transparent container; as the water and nutrient are provided by a special culture medium, the aseptic seedling in an environment isolated from the outside can gradually grow and even blossom. Only by being placed in an environment with the normal temperature and under common scattered light, the asepsis bonsai can grow and develop in the transparent artware bottle container. People can observe the whole process of rootage, germination and blossoming. The aseptic seedling does not have any diseases and insect pests, does not need daily maintenance, and is environment-friendly, sanitary and beautiful; the ornamental period reaches more than 6 months. The asepsis bonsai not only is a recreation item, but also can help middle and primary school students recognize and understand plants.

Description

How to make sterile bonsai

technical field

The invention relates to a method for making ornamental gardening bonsai, in particular to placing sterile seedlings cultured in vitro and decorative materials in a sterile transparent container so that the sterile seedlings can grow or bloom in an environment isolated from the outside world. How to make fungus bonsai.

Background technique

Aseptic bonsai is a handicraft that uses sterile plants and decorations to form various miniature landscapes in containers. It does not require any maintenance, has the advantages of high novelty, elegant appearance and so on. It is an elegant work of art with good market development prospects.

There are a lot of successful varieties of the existing in vitro culture test-tube plantlets, and there are more than a dozen varieties of flowering in the culture bottle. Some of these varieties have a short flowering time in the bottle, and some varieties have a very low flowering rate. Some varieties of test-tube flowers have no ornamental value. Therefore, there are very few varieties that really bloom in the bottle, have a long flowering period, a high flowering rate, and are highly ornamental. The flowering period of miniature roses in Korea is only about a month. For making bonsai in aseptic container, there is no report at present. It is only seen that resin solids such as crystal mud are added to open containers, and plants are planted therein to make them survive for a short time. Because it is an open culture, it will inevitably cause water to evaporate, and it will stink after a long time, causing the roots to rot, so it needs to be managed regularly, which is one of the main reasons why crystal mudpot flowers are difficult to promote in the market.

Contents of the invention

The purpose of the present invention is to make aseptic bonsai, that is, to make aseptic seedlings and various decorative materials into various aseptic bonsai, and put the aseptic bonsai in a sterile transparent container to isolate the aseptic seedlings from the outside world. In an environment where water and nutrients are provided through a special medium, it gradually grows and even blooms.

The preparation method of aseptic bonsai of the present invention comprises explant disinfection, culture medium preparation and bonsai colonization, specifically comprises the following steps:

1. Disinfection of explants: the same as the routine disinfection of plant tissue culture. After washing the surface of the explant with washing powder, soak it in 75% alcohol for 30 seconds, soak it in 0.1% mercury liter for 5-20 minutes, wash it with sterile water for 3-5 times, and inoculate it. The explants are such as seeds, leaf buds, leaves... .

2. Medium preparation: According to the medium formula, first weigh agar powder and white sugar, add 300 ml of distilled water to boil, then add other ingredients in the formula in full; if you need to increase the color of the medium, you can also add various A food coloring, the total amount of coloring should not exceed 80mg/L; after dissolving and mixing the above ingredients, add distilled water to 1000ml, then adjust the pH to 5.8 with 3% NaOH or HCl; then, divide the medium into Seal in a transparent container. Sterilize under 1.1kg/cm high pressure for 20 minutes, cool and solidify for later use. Various compositions and consumption are as follows in the described culture medium:

Serial number Ingredient name Dosage mg/L Serial number Ingredient name Dosage mg/L

1 KNO ₃ 500～950 15 Niacin 0.5

2 NH ₄ NO ₃ 300～500 16 Pyridoxine hydrochloride 0.5

3 Mg(SO) ₄ 7H ₂ O 185～370 17 Thiamine hydrochloride 0.1

4 KH ₂ PO ₃ 85～170 18 Inositol 100

5 CaCl ₂ .2H ₂ O 220～440 19 Glycine 2

6 FeSO ₄ 7H ₂ O 27.8 20 White sugar 20000

7 Na ₂ -EDTA 37.3 21 Agar powder 6000

8 MnSO ₄ .4H ₂ O 22.3 22 6-benzyl adenine 0.1～3

9 ZnSO ₄ .7H ₂ O 8.6 23 Naphthalene acetic acid 0.1～3

10 CoCl ₂ .6H ₂ O 0.025 24 15% Paclobutrazol 0.01～5

11 CuSO ₄ .5H ₂ O 0.025 25 Spermidine 0～5

12 NaMoO ₄ .2H ₂ O 0.25

13 KI 0.83

14 H ₃ BO ₃ 6.2

The above chemical reagents are all dry solids and chemically pure; mg/L means the number of milligrams of drugs contained in 1 liter of medium. Wherein, 6-benzyl adenine was dissolved with 1N HCI. Naphthaleneacetic acid was dissolved in 95% alcohol.

3. Aseptic cultivation: Inoculate sterile vaccines and sterilized decorative materials into transparent containers under aseptic conditions, and then seal them. The sealing material, such as plastic wrap, or sealing film, or material that can sterilize and filter air, ...; the decorative material: such as rockery, coral; towers, pavilions, small bridges, small figures, and various animals made of ceramics; Bridges, flags, and various figures made of plastic... Decorative materials made of rockery, coral and pottery are sterilized with high temperature and high pressure, and decorative materials made of plastic are sterilized with chemical disinfectants before use.

The technology of the present invention has the following beneficial effects:

1. The culture medium formula of the present invention is compared with MS culture medium, and its N content reduces, and P, K content improves. It is beneficial to the flower bud differentiation of plants and promotes flowering (as shown in Table 1). 6-benzyl adenine and naphthalene acetic acid promote plant cell division and differentiation. 15% paclobutrazol can control the size of plants and promote early flowering (as shown in Table 2). Spermidine promotes flower bud differentiation and flowering in plants.

Food coloring is what adds to the beauty of the medium.

Table 1 Effects of different media on flowering in a feathery cockscomb vase

culture medium Number of inoculated buds Number of flowering plants Flowering rate (%) MS 60 10 16.7 1/2MS 60 13 21.7 Culture medium of the present invention 60 54 90.0

Note: Compared with MS and 1/2MS medium, the flowering rate is significantly improved when using the medium of the present invention, and the flowering rate reaches 90%.

Table 2 Effects of different concentrations of paclobutrazol on flowering in a feathery cockscomb vase ^*

15% Paclobutrazol (mg/L) The number of inoculated buds (pieces) Number of flowering plants (plants) Flowering rate (%) Flowering time (days) Average plant height (cm) 0 60 49 81.3 48 2.7 0.5 60 53 88.3 35 1.5 1 60 54 90.0 30 1.3 2 60 48 80.0 34 0.9

^* The medium is the medium of the present invention, except that paclobutrazol is different; the flowering time is the time from sowing to the emergence of flower buds. The test results showed that: with the increase of the concentration of paclobutrazol, the height of the plant was obviously reduced, but the flowering time was obviously advanced. For the flowering rate, the optimal concentration of paclobutrazol is 0.5 mg/L and 1.0 mg/L, and the flowering rate is above 88.0%.

2. The sterile bonsai of the present invention only needs to be placed in a normal temperature environment and under general scattered light, without any pests and diseases, and usually does not need any maintenance, and is environmentally friendly, hygienic and beautiful. The ornamental period is more than 6 months.

3. The sterile bonsai of the present invention is produced in a sterile, controllable environment, and is not limited by natural conditions, and can be produced every year.

4. The sterile bonsai of the present invention absorbs rich nutrient medium in a sterile, transparent process bottle container to make it grow and develop, and even bloom. People can observe the rooting, germination and flowering of plants during appreciation. This can be used as an extracurricular activity project for primary and middle school students, allowing them to know and understand plants.

5. Compared with the current technology, the sterile bonsai of the present invention has the most prominent point of reducing bonsai art and landscape art into sterile and transparent craft bottle containers. People can make various kinds of bonsai according to their own ideas. miniature artwork. This also provides a wider application space for the biotechnology industry.

6. The technology has high technological content, good market prospects, and is convenient for industrial production.

Description of drawings:

Fig. 1 is the aseptic bonsai of foliage plants, and the plant in the figure is red cage grass;

Fig. 2 is a kind of mixed aseptic bonsai, and the plants in the figure are colocasia, acacia and cockscomb.

Specific embodiments: In order to fully disclose the preparation method of the sterile bonsai of the present invention, it will be described below in conjunction with the examples.

Example 1: a kind of foliage plant aseptic bonsai (described foliage plant such as red pitcher plant, cocoon, oncidium, dendrobium orchid, variegated taro...), its preparation method is as follows:

1. Disinfection of explants: After washing the explants of plants (including seeds, leaf buds, leaves, etc.) on the surface of washing powder, soak them in 75% alcohol for 30 seconds, soak them in 0.1% mercury liter for 5-20 minutes, and then soak them in sterile water. Inoculate after washing 3-5 times;

2. Preparation of culture medium: according to the formula, first weigh agar powder and white sugar, add them to 300 ml of distilled water and boil them, then add the other ingredients in the formula in full, and add 3 mg/L of red food coloring; the above ingredients After dissolving and mixing, add distilled water to 1000 ml, and adjust the pH to 5.8 with 3% NaOH or 1N HCl, then divide the medium into transparent containers and seal them. Sterilize under high pressure of 1.1kg/ ^cm2 for 20 minutes, cool and solidify for later use. During autoclaving, high temperature and high pressure resistant decorative materials can be autoclaved at the same time. Described culture medium formula is as follows:

Serial number Ingredient name Dosage mg/L Serial number Ingredient name Dosage mg/L

1 KNO ₃ 950 15 Niacin 0.5

2 NH ₄ NO ₃ 450 16 Pyridoxine hydrochloride 0.5

3 Mg(SO) ₄ 7H ₂ O 185 17 Thiamine hydrochloride 0.1

4 KH ₂ PO ₃ 85 18 Inositol 100

5 CaCl ₂ .2H ₂ O 220 19 Glycine 2

6 FeSO ₄ 7H ₂ O 27.8 20 White sugar 20000

7 Na ₂ -EDTA 37.3 21 Agar powder 6000

8 MnSO ₄ .4H ₂ O 22.3 22 6-benzyl adenine 0.1

9 ZnSO ₄ .7H ₂ O 8.6 23 Naphthaleneacetic acid 0.5

10 CoCl ₂ .6H ₂ O 0.025 24 15% paclobutrazol 0.01

11 CuSO ₄ .5H ₂ O 0.025 25 Spermidine 0

12 NaMoO ₄ .2H ₂ O 0.25

13 KI 0.83

14 H ₃ BO ₃ 6.2

3. Aseptic cultivation: According to the shape and size of the container, select aseptic seedlings and decorative materials suitable for the container. On the purification workbench, according to the concept orientation of the bonsai to be made, the sterilized decorative material is first placed on the medium corresponding to the concept before the medium is completely solidified after high pressure. After the culture medium is completely solidified, inoculate various sterile seedlings on the culture medium at the corresponding position in the container, and finally seal the transparent container with a bottle cap or fresh-keeping film.

As shown in Figure 1, a sterile bonsai of a foliage plant can appreciate the growth process of the plant at room temperature and natural scattered light, and the viewing period can be more than 6 months.

Example 2: a kind of preparation method of ornamental plant aseptic bonsai, comprises the following steps: (described ornamental plant, as cockscomb, carnation, morning glory...):

Step 1 is the same as step 3 and the first embodiment, and the method for preparing medium in step 2 is as follows:

According to the formula of the culture medium, first weigh agar powder and white sugar, add them to 300 ml of distilled water and boil, then add the other ingredients in the formula; add lemon yellow food coloring 60mg/L; dissolve and mix the above ingredients Add distilled water to 1000 ml, and adjust the pH to 5.8 with 3% NaOH or 1N HCl, then divide the medium into transparent containers and seal them. Sterilize under high pressure of 1.1kg/ ^cm2 for 20 minutes, cool and solidify for later use. During autoclaving, high temperature and high pressure resistant decorative materials can be autoclaved at the same time. Described culture medium formula is as follows:

Serial number Ingredient name Dosage mg/L Serial number Ingredient name Dosage mg/L

1 KNO ₃ 500 15 Niacin 0.5

2 NH ₄ NO ₃ 300 16 Pyridoxine hydrochloride 0.5

3 Mg(SO) ₄ 7H ₂ O 370 17 Thiamine hydrochloride 0.1

4 KH ₂ PO ₃ 170 18 Inositol 100

5 CaCl ₂ .2H ₂ O 220 19 Glycine 2

6 FeSO ₄ 7H ₂ O 27.8 20 White sugar 20000

7 Na ₂ -EDTA 37.3 21 Agar powder 6000

8 MnSO ₄ .4H ₂ O 22.3 22 6-benzyl adenine 0.1

9 ZnSO ₄ .7H ₂ O 8.6 23 Naphthaleneacetic acid 1

10 CoCl ₂ .6H ₂ O 0.025 24 15% paclobutrazol 3

11 CuSO ₄ .5H ₂ O 0.025 25 Spermidine 1

12 NaMoO ₄ .2H ₂ O 0.25

13 KI 0.83

14 H ₃ BO ₃ 6.2

Example 3: a kind of mixed aseptic bonsai (the described mixed aseptic bonsai, as shown in Figure 2, the bonsai mixed with the aseptic seedlings of syringa, acacia and cockscomb), the preparation method may further comprise the steps:

Step 1 is the same as step 3 and the first embodiment, and the method for preparing medium in step 2 is as follows:

According to the formula of the culture medium, first weigh agar powder and white sugar, add them to 300 ml of distilled water and boil them, and then add the other ingredients in the formula; add 3 mg/L of blue food coloring, (or do not add coloring ), after dissolving and mixing the above ingredients, add distilled water to 1000 ml, then adjust the pH to 5.8 with 3% NaOH or 1N HCl, then divide the culture medium into transparent containers and seal them. Sterilize under high pressure of 1.1kg/ ^cm2 for 20 minutes, cool and solidify for later use. During autoclaving, high temperature and high pressure resistant decorative materials can be autoclaved at the same time. Described culture medium formula is as follows:

Serial number Ingredient name Dosage mg/L Serial number Ingredient name Dosage mg/L

1 KNO ₃ 500 15 Niacin 0.5

2 NH ₄ NO ₃ 300 16 Pyridoxine hydrochloride 0.5

3 Mg(SO) ₄ 7H ₂ O 370 17 Thiamine hydrochloride 0.1

4 KH ₂ PO ₃ 170 18 Inositol 100

5 CaCl ₂ .2H ₂ O 220 19 Glycine 2

6 FeSO ₄ 7H ₂ O 27.8 20 White sugar 20000

7 Na ₂ -EDTA 37.3 21 Agar powder 6000

8 MnSO ₄ .4H ₂ O 22.3 22 6-Benzyl adenine 0.5

9 ZnSO ₄ .7H ₂ O 8.6 23 Naphthaleneacetic acid 2

10 CoCI ₂ .6H ₂ O 0.025 24 15% paclobutrazol 1

11 CuSO ₄ .5H ₂ O 0.025 25 Spermidine 2

12 NaMoO ₄ .2H ₂ O 0.25

13 KI 0.83

14 H ₃ BO ₃ 6.2

Claims (5)

1, a kind of preparation method of sterilized miniascape comprises explant sterilization, medium preparation and aseptic cultivation, it is characterized in that

A, described culture medium preparation method according to culture medium prescription, take by weighing agar powder, white sugar earlier, add boil in 300 milliliters the distilled water after, number adds other compositions in the prescription for another example; Increase the color of medium if desired, can also add various food colorings in addition, the use total amount of pigment is no more than 80mg/L; More than the dissolving of various compositions mix back adding distil water to 1000 milliliter, transfer to pH5.8 with 3% NaOH or HCl again; Then, the medium branch is installed in the transparent vessel seal.Sterilization is 20 minutes under the 1.1kg/cm high pressure, and is standby after the cooled and solidified;

Various compositions and consumption are as follows in B, the described medium:

Sequence number composition title consumption mg/L sequence number composition title consumption mg/L

1 KNO ₃500～950 15 nicotinic acid 0.5

2 NH ₄NO ₃300～500 16 pyridoxine hydrochlorides 0.5

3 Mg (SO) ₄7H ₂O 185～370 17 thiamine hydrochlorides 0.1

4 KH ₂PO ₃85～170 18 inositols 100

5 CaCl ₂.2H ₂O 220～440 19 glycine 2

6 FeSO ₄7H ₂O 27.8 20 white sugar 20000

7 Na ₂-EDTA 37.3 21 agar powders 6000

8 MnSO ₄.4H ₂The misty purine 0.1～3 of O 22.3 22 6-benzyl glands

9 ZnSO ₄.7H ₂O 8.6 23 methyls 0.1～3

10 CoCl ₂.6H ₂O 0.025 24 15% paclobutrazol 0.01～5

11 CuSO ₄.5H ₂O 0.025 25 spermidine 0～5

12 NaMoO ₄.2H ₂O 0.25

13 KI 0.83

14 H ₃BO ₃ 6.2

C, described aseptic cultivation: the aseptic seedling and the ornament materials of sterilizing are inoculated in the transparent container under aseptic condition, seal then.

2, the preparation method of a kind of sterilized miniascape according to claim 1 is characterized in that adding various food colorings in medium.

3, the preparation method of a kind of sterilized miniascape according to claim 1 and 2 is characterized in that various compositions and consumption are as follows in the described medium:

Sequence number composition title consumption mg/L sequence number composition title consumption mg/L

1 KNO ₃500 15 nicotinic acid 0.5

2 NH ₄NO ₃300 16 pyridoxine hydrochlorides 0.5

3 Mg (SO) ₄7H ₂O 370 17 thiamine hydrochlorides 0.1

4 KH ₂PO ₃170 18 inositols 100

5 CaCI ₂.2H ₂O 220 19 glycine 2

6 FeSO ₄7H ₂O 27.8 20 white sugar 20000

7 Na ₂-EDTA 37.3 21 agar powders 6000

8 MnSO ₄.4H ₂The misty purine 0.1 of O 22.3 22 6-benzyl glands

9 ZnSO ₄.7H ₂O 8.6 23 methyls 1

10 CoCI ₂.6H ₂O 0.025 24 15% paclobutrazol 3

11 CuSO ₄.5H ₂O 0.025 25 spermidine 1

12 NaMoO ₄.2H ₂O 0.25

13 KI 0.83

14 H ₃BO ₃ 6.2

4, the preparation method of a kind of sterilized miniascape according to claim 2 is characterized in that the use total amount of pigment is no more than 80mg/L.

5, the preparation method of a kind of sterilized miniascape according to claim 1, but it is characterized in that joint filling material is preservative film or seals film or the material of disinfection filtering air.

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