CN106718712B - Method for transplanting and domesticating dendrobium officinale tissue culture seedlings - Google Patents
Method for transplanting and domesticating dendrobium officinale tissue culture seedlings Download PDFInfo
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Abstract
The invention discloses a method for transplanting and domesticating dendrobium officinale tissue culture seedlings, which comprises the following steps: peanut shell: water is mixed according to the mass ratio of 1: 1.5: 2-1: 2: 2, mixing the materials in proportion, fully fermenting the mixture to be used as a culture medium, paving the culture medium into a seedbed, and kicking out shallow trenches or digging pits on the seedbed; taking the rooted tissue culture seedling out of the culture bottle, washing the agar culture medium with clear water, spreading to dry the water on the surface of the plant, and planting the tissue culture seedling in a fixed shallow trench or a pit; sun-shading by a 50% sun-shading net, controlling the air temperature at 10-30 ℃ and the ground temperature at 12-28 ℃, and watering once in the morning and evening each day; and backfilling the matrix after 3-6 days of planting and fixedly planting the tissue culture seedlings, wherein the survival rate reaches over 95 percent after a fixed value is 15 days. The method reduces the in-bottle hardening-off step in the transplanting and domesticating process of the dendrobium officinale, simplifies the procedure, accelerates the fibrosis of tissue culture seedlings, promotes the transformation of plant nutrition types, improves the adaptability to the field environment in a short time, and improves the survival rate.
Description
Technical Field
The invention belongs to the field of agricultural production, and particularly relates to a method for transplanting and domesticating dendrobium officinale tissue culture seedlings.
Background
Dendrobium officinale is a perennial herb of the genus Dendrobium of the family Orchidaceae, also named as herba Dendrobii and herba Equiseti Ramosissimi are traditional and rare Chinese herbal medicines in China, and are recorded in the Chinese pharmacopoeia, and have the effects of tonifying stomach, promoting the production of body fluid, nourishing yin, clearing heat and the like. Modern pharmacological studies show that the dendrobium officinale contains components such as polysaccharide, alkaloid, amino acid and the like, has the effects of resisting tumors and aging, enhancing the immunity of the body, expanding blood vessels and resisting platelet aggregation, and is widely applied to clinical and traditional Chinese medicine compounds. Wild dendrobium officinale has strict requirements on natural ecological conditions, extremely low natural reproduction rate, slow growth and rare resources, and due to long-term mining, the ecological environment is deteriorated, a plurality of rare varieties are endangered and extincted, which are listed as key protection rare or endangered medicinal plants by the state, which seriously affects the development and utilization of dendrobium officinale.
The mass propagation of the dendrobium officinale mainly adopts a tissue culture propagation mode, the adaptability of the tissue culture seedlings of the dendrobium officinale cultured by propagation to the outside is weak, and a process of adapting to the outside environment is needed. The method adopted in the prior production is that the mixture is refined in a bottle for about one week and then planted in a matrix for acclimatization, so that the mixture gradually adapts to the external environment. The process has higher requirements on temperature, light, water, matrix and the like, because the tissue culture seedlings are transplanted from the culture bottles, the nutrition type is still in a semi-autotrophic type, part of roots are still in a fleshy state, the oxygen conveying capacity of the conduit is weak, and the adaptability is low; the tissue culture seedling roots are buried in the matrix too early, the roots are not fully fiberized, microorganism breeding infection is easily caused, the micro anaerobic environment exists in the matrix due to improper water management, root rot and seedling rot are easily caused, and part of planters are disastrous because the key technology of seedling hardening is not mastered.
In conclusion, the existing dendrobium officinale tissue culture seedling transplanting method has the advantages that the adaptability of the expanded and cultured dendrobium officinale tissue culture seedling to the outside is weak, the requirements on temperature, light, water, a matrix and the like are high, the breeding of microorganisms is easy to cause, and the phenomena of root rot and seedling rot are easy to generate.
Disclosure of Invention
The invention aims to provide a method for transplanting and domesticating dendrobium officinale tissue culture seedlings, which fully utilizes the characteristics of dendrobium officinale aerial roots and aims to solve the problems that in the transplanting and domesticating process of the dendrobium officinale tissue culture seedlings, the adaptability of the dendrobium officinale tissue culture seedlings to the outside is weak, the requirements on temperature, light, water, a matrix and the like are high, the breeding of microorganisms is easy to cause, root rot and seedling are easy to generate, the adaptability is weak and the survival rate is low. The moisture-keeping and air-permeable microenvironment is created manually, the fibrosis of roots and plants is accelerated, the transformation of the plant nutrition types is promoted, bactericides are reduced or even not used, the adaptability of the tissue culture seedlings to the field environment is improved in a short time, and the survival rate is improved; in addition, the method reduces the steps of hardening seedlings in bottles, simplifies the transplanting and domesticating procedures, and can realize large-scale transplanting and domestication of the dendrobium officinale tissue culture seedlings.
The invention is realized in such a way that the method for transplanting and domesticating the dendrobium officinale tissue culture seedlings comprises the following steps:
step one, preparation of a culture medium and a seedbed: wood chips are processed by the following steps: peanut shell: water is mixed according to the mass ratio of 1: 1.5: 2-1: 2: 2 for 50 days, and taking the mixture as a culture medium substrate after the mixture is fully fermented; placing the fermented substrate on an overhead seedling bed frame, wherein the thickness of the substrate is 15-20 cm, paving, and watering and wetting thoroughly;
step two, preparing tissue culture seedlings: taking out the tissue culture rooted seedlings from the culture bottle, cleaning the agar culture medium with clear water, spreading the agar culture medium on a ventilated and dry place, and airing the moisture on the surface of the plants;
step three, planting: fixing the plant planting on the inner surface of the shallow trench or the pit of the matrix by adopting shallow trench planting or pit planting without covering the matrix, so that part of roots are contacted with the matrix and the other part is exposed in the air; the planting distance is 3-5 cm, and the row spacing is 7-9 cm;
step four, planting management: sun-shading by a 50% sun-shading net, controlling the air temperature to be 10-30 ℃ and the ground temperature to be 12-28 ℃, respectively watering once in the morning and at the evening each day, keeping the water content of the culture medium to be 35% -60% and keeping the air humidity to be 50% -90%;
step five, setting a value: after 3-6 days of planting, the root tip of the tissue culture seedling is increased by 0.2-0.5 cm, then the substrate around the shallow trench or the pit can be filled back into the trench or the pit, and the substrate is lightly pressed to fix the root on the substrate. The thickness of the covering substrate is suitable for covering the roots and standing the seedlings;
step six, constant value management: sun-shading by a 50% sun-shading net, controlling the air temperature to be 10-30 ℃ and the ground temperature to be 12-28 ℃, respectively watering once in the morning and at the evening each day, keeping the moisture of the matrix to be 35% -60% and keeping the air humidity to be 50% -90%; the survival rate of the tissue culture seedlings reaches more than 95 percent after the fixed value is 15 days.
Furthermore, the method for shallow trench planting comprises the following steps:
kicking a shallow trench with the depth of 1.5-3 cm and the width of 2.5-4.5 cm on the substrate of the seedbed, fixing the tissue culture seedling in the shallow trench under light pressure without covering the substrate, making part of the root contact the surface of the substrate, and exposing part of the root in the air.
Further, the pit planting method comprises the following steps:
digging 1.5-3 cm deep pits with diameter of about 2cm on the substrate of the seedbed, slightly pressing the tissue culture seedling into the pits for fixing the root part without covering the substrate, so that part of the root part is contacted with the surface of the substrate, and part of the root part is exposed in the air.
Further, the fixed value: an interval of 3-6 days is formed between the fixed value and the planting, after the time period, part of aerial roots of the dendrobium officinale can keep moisture in the concave part of the matrix, the part exposed in the air can fully absorb oxygen, after the root tip of the tissue culture seedling is increased by 0.2-0.5 cm, the matrix around the shallow trench or the pit can be filled back into the trench or the pit, and the matrix is lightly pressed to fix the roots on the matrix; the thickness of the covering substrate is suitable for just covering the roots and standing the seedlings.
The method reduces the in-bottle hardening-off step in the transplanting and domesticating process of the dendrobium officinale, simplifies the procedure, accelerates the fibrosis of tissue culture seedlings, promotes the transformation of plant nutrition types, improves the adaptability to the field environment in a short time, and improves the survival rate. The reason why the survival rate of the dendrobium officinale tissue culture seedlings can be obviously, simply and conveniently improved is as follows: the characteristics of make full use of dendrobii officmalis caulis aerial root for through the ventilative microenvironment of manual work building moisture retention for root and plant fibrosis promote the plant nutrition type and change, reduce and do not use germicide even, promote the adaptability of tissue culture seedling to the field environment in the short time, the scale of being convenient for is planted.
Drawings
Fig. 1 is a flowchart of a method for transplanting and domesticating a dendrobium officinale tissue culture seedling provided by an embodiment of the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The following detailed description of the principles of the invention is provided in connection with the accompanying drawings.
Referring to fig. 1, the method for transplanting and domesticating the dendrobium officinale tissue culture seedlings provided by the embodiment of the invention comprises the following steps:
s101, selecting robust tissue culture seedlings with roots;
s102, preparation of a culture medium and a seedbed: wood chips are processed by the following steps: peanut shell: water is mixed according to the mass ratio of 1: 1.5: 2-1: 2: 2 for 50 days, and taking the mixture as a culture medium after the mixture is fully fermented. The cultivation substrate is laid on the air to be made into a seedbed with the thickness of 15 cm-20 cm, and the seedbed is thoroughly watered for standby.
S103, preparing tissue culture seedlings: taking the rooted tissue culture seedling out of the culture bottle, cleaning the agar culture medium with clear water, spreading the agar culture medium on a ventilated and dry place, and airing the moisture on the surface of the plant;
s104, fixing the plant planting on the inner surface of a shallow ditch or a pit of the substrate by adopting shallow ditch planting or pit planting without covering the substrate;
s105, planting management: shading by a 50% shading net, controlling the air temperature to be 12-30 ℃, controlling the ground temperature to be 12-28 ℃, watering once a day in the morning and at the evening respectively, and keeping the culture medium moist;
s106, constant value: after 3-6 days of planting, after the root tip length of the tissue culture seedling is 0.2-0.5 cm, filling the matrix around the shallow trench or the pit back into the trench or the pit, lightly pressing the matrix, and fixedly planting the tissue culture seedling on the culture medium;
s107, constant value management: the sun is shaded by a 50% sunshade net, the air temperature is controlled to be 10-30 ℃, the ground temperature is 12-28 ℃, watering is carried out once in the morning and at the evening each day, the moisture of the matrix is kept to be 35% -60%, and the air humidity is kept to be 50% -90%. The survival rate of the tissue culture seedlings can reach more than 95 percent.
Furthermore, the method for shallow trench planting comprises the following steps:
kicking a shallow trench with the depth of 1.5-3 cm and the width of 2.5-4.5 cm on the substrate of the seedbed, fixing the tissue culture seedling in the shallow trench under light pressure without covering the substrate, making part of the root contact the surface of the substrate, and exposing part of the root in the air.
Further, the pit planting method comprises the following steps:
digging 1.5-3 cm deep pits with diameter of about 2cm on the substrate of the seedbed, slightly pressing the tissue culture seedling into the pits for fixing the root part without covering the substrate, so that part of the root part is contacted with the surface of the substrate, and part of the root part is exposed in the air.
The width and depth of the kicked shallow trench or the size and depth of the nest cavity are convenient for fixing the tissue culture seedlings according to the size of the tissue culture seedlings, and meanwhile, the sunken part of the substrate can keep certain humidity to prevent the sun; the light pressure plays a role in fixing, and simultaneously the root of the tissue culture seedling is in certain contact with the substrate; without covering the matrix, most of the roots of the tissue culture seedlings are exposed in the air, so that the occurrence of insect pests and the growth of harmful microorganisms can be reduced, the fibrosis of the roots and plants is accelerated, the transformation of the plants from semi-autotrophic nutrition type to autotrophic nutrition type is promoted, and the adaptability to the field environment is improved in a short time.
Further, the fixed value: an interval of 3-6 days is formed between the fixed value and the planting, after the time period, part of aerial roots of the dendrobium officinale can keep moisture in the concave part of the matrix, the part exposed in the air can fully absorb oxygen, after the root tip of the tissue culture seedling is increased by 0.2-0.5 cm, the matrix around the shallow trench or the pit can be filled back into the trench or the pit, and the matrix is lightly pressed to fix the roots on the matrix; the thickness of the covering substrate is suitable for just covering the roots and standing the seedlings.
The application of the principles of the present invention will now be described in further detail with reference to specific embodiments.
Example 1:
preparing a culture medium and a seedbed: wood chips are processed by the following steps: peanut shell: water is mixed according to the mass ratio of 1: 1.5: 2 for 50 days, and taking the mixture as a culture medium substrate after the mixture is fully fermented. And (3) placing the fermented substrate on an overhead seedling bed frame, wherein the thickness of the substrate is 18-20 cm, paving, and watering and drenching thoroughly.
Preparing tissue culture seedlings: taking out the tissue culture rooted seedlings from the culture bottle, cleaning the agar culture medium with clear water, and spreading the agar culture medium at a ventilated and dry place to dry the water on the surface of the plants.
Planting: the plant is planted and fixed on the inner surface of the shallow ditch or the pit of the substrate by adopting shallow ditch planting or pit planting, the substrate is not covered, a part of roots are contacted with the substrate, and a part of roots are exposed in the air.
Planting management: the sun is shaded by a 50% sunshade net, the air temperature is controlled to be 15-22 ℃, the ground temperature is 15-23 ℃, watering is carried out once in the morning and at the evening each day, the moisture of the matrix is kept to be 35% -60%, and the air humidity is kept to be 40% -55%.
Setting the value: after 5 days of planting, after the root tip of the tissue culture seedling is increased by 0.2-0.5 cm, filling the matrix around the shallow trench or the pit back into the trench or the pit, and lightly pressing the matrix to fix the root on the matrix. The thickness of the covering substrate is suitable for just covering the roots and standing the seedlings.
Constant value management: the sun is shaded by a 50% sunshade net, the air temperature is controlled to be 15-23 ℃, the ground temperature is 15-24 ℃, watering is carried out once in the morning and at the evening each day, the water content of the culture medium is kept to be 35% -60%, and the air humidity is kept to be 53% -87%. The survival rate of the tissue culture seedlings reaches 98 percent after the fixed value is 15 days.
Example 2:
preparing a culture medium and a seedbed: wood chips are processed by the following steps: peanut shell: water is mixed according to the mass ratio of 1: 2: 2 for 50 days, and taking the mixture as a culture medium substrate after the mixture is fully fermented. Placing the fermented substrate on an overhead seedling bed frame, wherein the thickness of the substrate is 15-18 cm, paving, and watering and wetting thoroughly.
Preparing tissue culture seedlings: taking out the tissue culture rooted seedlings from the culture bottle, cleaning the agar culture medium with clear water, and spreading the agar culture medium at a ventilated and dry place to dry the water on the surface of the plants.
Planting: the plant is planted and fixed on the inner surface of the shallow ditch or the pit of the substrate by adopting shallow ditch planting or pit planting, the substrate is not covered, a part of roots are contacted with the substrate, and a part of roots are exposed in the air.
Planting management: the sun is shaded by a 50% sunshade net, the air temperature is controlled to be 10-30 ℃, the ground temperature is 12-28 ℃, watering is carried out once in the morning and at the evening each day, the moisture of the matrix is kept to be 35% -60%, and the air humidity is kept to be 50% -90%.
Setting the value: after 3 days of planting, the root tip of the tissue culture seedling is increased by 0.2-0.3 cm, the matrix around the shallow trench or the pit can be filled back into the trench or the pit, and the matrix is lightly pressed to fix the root on the matrix. The thickness of the covering substrate is suitable for just covering the roots and standing the seedlings.
Constant value management: the sun is shaded by a 50% sunshade net, the air temperature is controlled to be 22-25 ℃, the ground temperature is 22-28 ℃, watering is carried out once in the morning and at the evening each day, the water content of the culture medium is kept to be 35% -60%, and the air humidity is kept to be 48% -90%. The survival rate of the tissue culture seedlings reaches 95 percent after the fixed value is 15 days.
Example 3:
preparing a culture medium and a seedbed: wood chips are processed by the following steps: peanut shell: water is mixed according to the mass ratio of 1: 2: 2 for 50 days, and taking the mixture as a culture medium substrate after the mixture is fully fermented. Placing the fermented substrate on an overhead seedling bed frame, wherein the thickness of the substrate is 15-18 cm, paving, and watering and wetting thoroughly.
Preparing tissue culture seedlings: taking out the tissue culture rooted seedlings from the culture bottle, cleaning the agar culture medium with clear water, and spreading the agar culture medium at a ventilated and dry place to dry the water on the surface of the plants.
Planting: the plant is planted and fixed on the inner surface of the shallow ditch or the pit of the substrate by adopting shallow ditch planting or pit planting, the substrate is not covered, a part of roots are contacted with the substrate, and a part of roots are exposed in the air.
Planting management: the sun is shaded by a 50% sunshade net, the air temperature is controlled to be 10-30 ℃, the ground temperature is 12-28 ℃, watering is carried out once in the morning and at the evening each day, the moisture of the matrix is kept to be 30% -55%, and the air humidity is kept to be 50% -90%.
Setting the value: after 6 days of planting, the root tip of the tissue culture seedling is increased by 0.2-0.5 cm, the substrate around the shallow trench or the pit can be filled back into the trench or the pit, and the substrate is lightly pressed to fix the root on the substrate. The thickness of the covering substrate is suitable for just covering the roots and standing the seedlings.
Constant value management: the sun is shaded by a 50% sunshade net, the air temperature is controlled to be 22-25 ℃, the ground temperature is 22-28 ℃, watering is carried out once in the morning and at the evening each day, the water content of the culture medium is kept to be 35% -60%, and the air humidity is kept to be 50% -92%. The survival rate of the tissue culture seedlings reaches 96 percent after the fixed value is 15 days.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (1)
1. A method for transplanting and domesticating dendrobium officinale tissue culture seedlings is characterized by comprising the following steps:
step one, preparation of a culture medium and a seedbed: wood chips are processed by the following steps: peanut shell: water is mixed according to the mass ratio of 1: 1.5: 2-1: 2: 2 for 50 days, and taking the mixture as a culture medium substrate after the mixture is fully fermented; placing the fermented substrate on an overhead seedling bed frame, wherein the thickness of the substrate is 15-20 cm, paving, and watering and wetting thoroughly;
step two, preparing tissue culture seedlings: taking out the tissue culture rooted seedlings from the culture bottle, cleaning the agar culture medium with clear water, spreading the agar culture medium on a ventilated and dry place, and airing the moisture on the surface of the plants;
step three, planting: fixing the plant planting on the inner surface of the shallow trench or the pit of the matrix by adopting shallow trench planting or pit planting without covering the matrix, so that part of roots are contacted with the matrix and the other part is exposed in the air; the planting distance is 3-5 cm, and the row spacing is 7-9 cm;
step four, planting management: sun-shading by a 50% sun-shading net, controlling the air temperature to be 10-30 ℃ and the ground temperature to be 12-28 ℃, respectively watering once in the morning and at the evening each day, keeping the water content of the culture medium to be 35-60% and the air humidity to be 50-90%, and avoiding leaf withering of the tissue culture seedlings;
fifthly, an interval of 3-6 days is reserved between planting and planting, in the period of time, part of aerial roots of the dendrobium officinale can keep moisture in the concave part of the matrix, the part exposed in the air can fully absorb oxygen, after the root tip of the tissue culture seedling is increased by 0.2-0.5 cm, the matrix around the shallow trench or the pit can be filled back into the trench or the pit, and the matrix is lightly pressed to fix the roots on the matrix; the thickness of the covering substrate is suitable for covering the roots and standing the seedlings;
step six, field planting management: sun-shading by a 50% sun-shading net, controlling the air temperature to be 10-30 ℃ and the ground temperature to be 12-28 ℃, respectively watering once in the morning and at the evening each day, keeping the matrix water content to be 35-60% and the air humidity to be 50-90%, and avoiding leaf withering of the tissue culture seedlings; the survival rate of the tissue culture seedlings after 15 days of field planting reaches more than 95 percent;
the method of shallow trench planting comprises the following steps:
kicking a shallow trench with the depth of 1.5-3 cm and the width of 2.5-4.5 cm on the seedbed substrate, fixing the tissue culture seedling in the shallow trench under light pressure without covering the substrate, enabling part of the root to contact the surface of the substrate, and exposing part of the root in the air;
the pit planting method comprises the following steps:
digging 1.5-3 cm deep pits with diameter of about 2cm on the substrate of the seedbed, slightly pressing the tissue culture seedling into the pits for fixing the root part without covering the substrate, so that part of the root part is contacted with the surface of the substrate, and part of the root part is exposed in the air.
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| CN101352140A (en) * | 2008-08-22 | 2009-01-28 | 李明焱 | Dendrobium officinale substrate and planting method |
| CN102960226A (en) * | 2012-12-07 | 2013-03-13 | 农定霖 | Dendrobium officinale Kimura et Migo tissue culture seedling acclimatization method |
| CN103039225A (en) * | 2012-11-26 | 2013-04-17 | 李奋勇 | Floating exercising seedling and cultivating method of dendrobium officinale tissue culture seedling |
| CN103314857A (en) * | 2013-07-08 | 2013-09-25 | 重庆市秀山红星中药材开发有限公司 | Domestication cultivation method of dendrobium candidum tissue culture seedling |
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| CN101352140A (en) * | 2008-08-22 | 2009-01-28 | 李明焱 | Dendrobium officinale substrate and planting method |
| CN103039225A (en) * | 2012-11-26 | 2013-04-17 | 李奋勇 | Floating exercising seedling and cultivating method of dendrobium officinale tissue culture seedling |
| CN102960226A (en) * | 2012-12-07 | 2013-03-13 | 农定霖 | Dendrobium officinale Kimura et Migo tissue culture seedling acclimatization method |
| CN103314857A (en) * | 2013-07-08 | 2013-09-25 | 重庆市秀山红星中药材开发有限公司 | Domestication cultivation method of dendrobium candidum tissue culture seedling |
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