CN111011213A - Anoectochilus roxburghii culture method for improving drying rate - Google Patents
Anoectochilus roxburghii culture method for improving drying rate Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
In order to solve the problems of poor plant growth and low light energy utilization rate in the existing anoectochilus roxburghii culture process in the prior art, the invention provides an anoectochilus roxburghii culture method for improving the dry reduction rate. The anoectochilus formosanus culture method comprises preparing culture medium; inoculating; culturing: culturing under the condition of red and blue mixed illumination; culturing for three months, wherein the illumination time is 7.5-8.5h/d in the first month, 9.5-10.5h/d in the second month, and 10.5-12.5h/d in the third month. The invention has the beneficial effects that: the invention adopts red and blue light to replace the traditional white light, can improve the volume of plants and has better growth vigor. In addition, the effective time of the plants for absorbing the illumination in the first month is 7.5-8.5h/d, the effective time of the plants for absorbing the illumination in the second month is 9.5-10.5h/d, and the effective time of the plants for absorbing the illumination in the third month is 10.5-12.5h/d, so that the illumination time is set to be corresponding duration, and the electric energy is saved.
Description
Technical Field
The invention belongs to the technical field of anoectochilus formosanus, and particularly relates to an anoectochilus formosanus culture method for improving the dry-folding rate.
Background
Anoectochilus formosanus is a plant of Anoectochilus in Orchidaceae, with round and shield-shaped leaves, thin rhizome, slender leaf stalk, and brown color, and can climb. Anoectochilus roxburghii can be used as medicine at each part, and has the effects of clearing heat and cooling blood, dispelling wind and promoting diuresis, detoxifying, relieving pain, relieving cough and the like.
At present, in the process of culturing anoectochilus formosanus, the anoectochilus formosanus is cultured under the white light condition, so that the plant has small volume, poor growth vigor and low drying rate, in addition, under the existing culture condition, the illumination time every day is fixed and too long, the light energy of the anoectochilus formosanus can not be absorbed in the exceeding time, so that the light energy is seriously wasted, and the light energy utilization rate is low.
Disclosure of Invention
In order to solve the problems of poor plant growth vigor and low drying rate in the existing anoectochilus roxburghii culture process, the invention provides an anoectochilus roxburghii culture method for improving the drying rate.
On the one hand, the invention provides a method for culturing anoectochilus formosanus with improved dry breaking rate, which comprises the following steps:
preparing a culture medium: the culture medium comprises 1900g/l potassium nitrate, 1650g/l ammonium nitrate, 170g/l monopotassium phosphate, 370 magnesium sulfate, 440g/l calcium chloride dihydrate, 0.83g/1 potassium iodide, 6.2g/l boric acid, 22.3g/l manganese sulfate, 8.6g/l zinc sulfate, 0.25g/l sodium molybdate, 0.025g/l copper sulfate, 0.025g/l cobalt chloride, 37.3g/l disodium edetate, 27.8g/l ferrous sulfate, 100g/l inositol, 0.5g/l nicotinic acid, 0.5g/l pyridoxine hydrochloride, 0.1g/l thiamine hydrochloride, 2g/l glycine and 30g/l sucrose; the pH of the medium is 5.7;
inoculation: inoculating the anoectochilus formosanus tissue culture seedling into a culture medium under the aseptic condition;
culturing: placing the inoculated tissue culture seedlings in a culture chamber, and culturing under the condition of red and blue mixed illumination; culturing for three months, wherein in the first month, the ratio of red light to blue light is 12: 1; in the second month, the ratio of the red light to the blue light is 5: 1; in the third month, the ratio of red light to blue light is 12: 1.
Furthermore, the illumination time is 7.5-8.5h/d in the first month, 9.5-10.5h/d in the second month and 10.5-12.5h/d in the third month. The effective time of the plants for absorbing the illumination in the first month is 7.5-8.5h/d, the effective time of the plants for absorbing the illumination in the second month is 9.5-10.5h/d, and the effective time of the plants for absorbing the illumination in the third month is 10.5-12.5h/d, so that the illumination time is set to be corresponding duration, and the electric energy is saved.
Further, in the culture process, a red and blue mixed lamp is adopted to provide red and blue mixed illumination for the tissue culture seedlings; the red and blue mixed lamp comprises a lamp holder and 78 × N light emitting diodes fixed on the lower surface of the lamp holder, wherein N is more than or equal to 1; the lower surface of the lamp holder is provided with a lampshade buckled on the outer side of the light-emitting diode, and the lampshade is composed of single covers which correspond to the light-emitting diodes one by one; the lampshade consists of a red fixed cover body, a red detachable cover body, a blue fixed cover body and a blue detachable cover body; the red fixed cover body is formed by 65 × N red single covers, the red detachable cover body comprises 7 × N red single covers, the blue fixed cover body is formed by 6 × N blue single covers, and the blue detachable cover body comprises 7 × N blue single covers; the red fixed cover body and the blue fixed cover body are fixed on the lower surface of the lamp holder; the red cover body and the blue cover body can be dismantled through the mode of tearing open and set up in turn at the lower surface of lamp stand, and the red cover body that can be dismantled sets up the lower surface at the lamp stand in first period and third cycle, at the second cycle, the blue cover body that can be dismantled sets up the lower surface at the lamp stand.
Further, the lamp holder is circular; 78 light-emitting diodes are arranged from inside to outside into a first circle to a fourth circle, the first circle is provided with 7 light-emitting diodes, the second circle is provided with 15 light-emitting diodes, the third circle is provided with 24 light-emitting diodes, and the fourth circle is provided with 32 light-emitting diodes; the blue detachable cover body and the red detachable cover body are respectively composed of a central disc and 7 single covers which are in one-to-one correspondence with the first circle of light-emitting diodes, the central disc and the 7 single covers are connected through connecting rods, and the central disc is detachably arranged on the lower surface of the center of the lamp holder; the blue fixed cover body is composed of 6 independent single covers, and the 6 blue single covers of the blue fixed cover body are evenly and distributively buckled on the third circle of light-emitting diodes; the red fixed cover body is formed by 65 independent single covers and is buckled on the rest light-emitting diodes.
Furthermore, a magnet is arranged on the central disc and is adsorbed in the center of the lower surface of the lamp holder through magnetic force.
Further, the proportion of the red light and the blue light is as follows: 2: 0.9-2: 1.1.
Further, the proportion of the red light and the blue light is as follows: 4.8: 1-5.2: 1.
Further, the proportion of the red light and the blue light is as follows: 11.5: 1-12.5: 1.
The invention has the beneficial effects that: the ratio of red light to blue light is 12: 1, the first month is the key period of rooting, and the ratio of red light to blue light is 12: 1, which is favorable for rooting; in the second month, the ratio of the red light to the blue light is 5: 1, the second month is a key period of plant height, and the ratio of the red light to the blue light is 5: 1, so that the plant height is accelerated, and the protein content is improved; in the third month, the ratio of the red light to the blue light is 12: 1, which is the key period for plant thickening and leaf area enlargement, and the ratio of the red light to the blue light is 12: 1, which is more beneficial to plant thickening and leaves are more luxuriant. Compared with the traditional natural light culture, the volume of the plant can be improved, the growth vigor is better, and the dry-turning rate is higher.
Drawings
FIG. 1 is a schematic structural view of the lower surface of a red-blue hybrid lamp when a red detachable cover is disposed on the lower surface of a lamp holder;
FIG. 2 is a schematic structural diagram of the lower surface of the red-blue mixed lamp when the blue detachable cover body is arranged on the lower surface of the lamp holder;
fig. 3 is a schematic structural diagram of a red detachable cover body.
Reference numerals: 1-lamp holder, 21-red single cover, 22-blue single cover, 3-central disk, 4-connecting rod and 5-magnet.
Detailed Description
The invention is further explained below with reference to the drawings and the specific embodiments.
The first embodiment is as follows:
the embodiment provides a method for culturing anoectochilus formosanus with improved dry breaking rate, which comprises the following steps:
preparing a culture medium: the culture medium comprises 1900g/l potassium nitrate, 1650g/l ammonium nitrate, 170g/l monopotassium phosphate, 370 magnesium sulfate, 440g/l calcium chloride dihydrate, 0.83g/l potassium iodide, 6.2g/l boric acid, 22.3g/l manganese sulfate, 8.6g/l zinc sulfate, 0.25g/l sodium molybdate, 0.025g/l copper sulfate, 0.025g/l cobalt chloride, 37.3g/l disodium edetate, 27.8g/l ferrous sulfate, 100g/l inositol, 0.5g/l nicotinic acid, 0.5g/l pyridoxine hydrochloride, 0.1g/l thiamine hydrochloride, 2g/l glycine and 30g/l sucrose; the pH of the medium is 5.7;
inoculation: inoculating the anoectochilus formosanus tissue culture seedling into a culture medium under aseptic conditions.
Culturing: placing the inoculated tissue culture seedlings in a culture chamber, and culturing under the condition of red and blue mixed illumination; the ratio of red light to blue light is 12: 1 in the first month after three months of culture, the first month is the key period of rooting, and the ratio of red light to blue light is 12: 1, which is favorable for rooting; in the second month, the ratio of the red light to the blue light is 5: 1, the second month is a key period of plant height, and the ratio of the red light to the blue light is 5: 1, so that the plant height is accelerated, and the protein content is improved; in the third month, the ratio of the red light to the blue light is 12: 1, which is the key period for plant thickening and leaf area enlargement, and the ratio of the red light to the blue light is 12: 1, which is more beneficial to plant thickening and leaves are more luxuriant.
The illumination time is 7.5-8.5h/d in the first month, 9.5-10.5h/d in the second month and 10.5-12.5h/d in the third month.
In the culture process, a red and blue mixed lamp is adopted to provide red and blue mixed illumination for the tissue culture seedlings; the red and blue mixed lamp comprises a lamp holder and 78 × N light emitting diodes (the × represents a multiplication sign) fixed on the lower surface of the lamp holder, wherein N is more than or equal to 1; the lower surface of the lamp holder is provided with a lampshade buckled on the outer side of the light-emitting diode, and the lampshade is composed of single covers which correspond to the light-emitting diodes one by one; the lampshade consists of a red fixed cover body, a red detachable cover body, a blue fixed cover body and a blue detachable cover body; the red fixed cover body is composed of 65 × N red single cover bodies 21, the red detachable cover body comprises 7 × N red single cover bodies 21, the blue fixed cover body is composed of 6 × N blue single cover bodies 22, and the blue detachable cover body comprises 7 × N blue single cover bodies 22; the red fixed cover body and the blue fixed cover body are fixed on the lower surface of the lamp holder, the red detachable cover body and the blue detachable cover body are alternately arranged on the lower surface of the lamp holder in an opening mode, and the red detachable cover body is arranged on the lower surface of the lamp holder in the first period and the third period, so that the ratio of red light to blue light is 12: 1; in the second period, the blue detachable cover body is arranged on the lower surface of the lamp holder, and the ratio of the red light to the blue light is 5: 1.
Referring to fig. 1 to 3, as a preferred scheme in the present embodiment: the lamp holder 1 is circular; 78 light-emitting diodes are arranged from inside to outside into a first circle to a fourth circle, the first circle is provided with 7 light-emitting diodes, the second circle is provided with 15 light-emitting diodes, the third circle is provided with 24 light-emitting diodes, and the fourth circle is provided with 32 light-emitting diodes;
the blue detachable cover body is composed of a central disc 3 and 7 blue single covers 22 which are in one-to-one correspondence with the first circle of light-emitting diodes, and the central disc 3 is connected with each blue single cover 22 through a connecting rod 4; the red detachable cover body consists of a central disc 3 and 7 red single covers 21 which are in one-to-one correspondence with the first circle of light-emitting diodes; the central disc 3 is connected with each red single cover 21 through a connecting rod 4; the central disc 3 is detachably arranged on the lower surface of the center of the lamp holder 1; the central disc 3 can be detachably connected in a buckling mode, a threaded mode and the like, as a feasible specific scheme, the central disc 3 is provided with the magnet 5, and the magnet is adsorbed in the center of the lower surface of the lamp holder 1 through magnetic force.
As shown in fig. 2, the blue fixed cover body is composed of 6 independent blue single covers 22, and the 6 blue single covers 22 of the blue fixed cover body are evenly and distributively fastened on the third ring of light emitting diodes; namely, there are 3 red cell covers 21 between two adjacent blue cell covers 22.
The red fixed cover body is composed of 65 independent red single covers 21 and is buckled on the rest light-emitting diodes.
Since the proportions of red and blue light are different at different periods in this embodiment, the change is made. The existing red and blue mixed lamp proportion is fixed; if the existing red and blue lamps are adopted for irradiation, the lamps need to be changed in a large scale at different periods; or in different periods, transferring the anoectochilus formosanus to a light room with a corresponding proportion, wherein the transfer scale is large; both cases are more workload. Adopt the mixed lamp of red blue in this embodiment, only need can dismantle the cover body setting with red on the lamp panel in first stage and third stage, can dismantle the cover body setting with blue on the lamp panel in the second stage, it is convenient to change, and it is fast to change.
Example two:
under the same conditions as the rest, the ratio of red light to blue light is different from that of the first embodiment: 2: 0.9-2: 1.1. Under the condition, the drying rate of the anoectochilus formosanus is highest.
Example three:
under the same conditions as the rest, the ratio of red light to blue light is different from that of the first embodiment: 4.8: 1-5.2: 1. Under the condition, the length of the anoectochilus formosanus is the highest.
Example four:
under the same conditions as the rest, the ratio of red light to blue light is different from that of the first embodiment: 11.5: 1-12.5: 1. Under this condition, the effect is next to that of the first embodiment.
The above examples are further illustrated by the following experiments:
experimental materials: anoectochilus roxburghii variety "Fujian 2013010A" tissue culture seedling is provided by Jinhua city agricultural institute, and is identified as an authentic Anoectochilus roxburghii by the institute of traditional Chinese medicine in Zhejiang province. 7 light source processes are provided, namely a fluorescent lamp (CK), a red and blue light mixed light source (R1B12, R2B1, R5B1, R7B1, R12B1, the illumination condition described in the first embodiment), wherein R1B12 represents that the ratio of red light to blue light is 1: 12, R represents red light, B represents blue light, and the rest is analogized in sequence.
The experimental conditions are as follows: the experiment was carried out in the farming academy of Jinhua city at 3/1/2018 using 250mL glass round bottles as culture containers and the culture medium used was the one described in example I. Under aseptic conditions, tissue culture seedlings were inoculated into the culture medium, 10 stem segments were inoculated per bottle of seedlings, uniform illumination intensity was employed, and each light source treatment was repeated 4 times. And placing the inoculated tissue culture seedlings in a culture chamber for culture at the culture temperature of 25 ℃ for 90 days in total, wherein the illumination time of the 1 st to 30 th days is 8h/d, the illumination time of the 31 st to 60 th days is 10h/d, and the illumination time of the 61 st to 90 th days is 12 h/d.
Measurement items and methods: inoculating 90 bottles of 7 layers of anoectochilus roxburghii tissue culture seedlings for each treatment, after 90 days, randomly taking 3 bottles of seedlings from each layer, and measuring the morphological indexes of 5 seedlings; the same treatment and mixed sampling method are adopted for measuring the physiological indexes. Wherein the plant height (from the base of the stem to the growing point) is measured with a ruler; the stem thickness (1 cm below the cotyledon) was measured with a vernier caliper; the number of leaves; the leaf area (2 nd leaf from top to bottom) is scanned and measured by using EPSON Expression 11000 XL; the maximum root length (from root base to growth point) was measured with a ruler, and 3 were averaged; measuring the total fresh mass and the total dry mass by using an electronic balance; the content of Anoectochilus roxburghii glycoside is determined by high performance liquid chromatography, the content of total flavone is determined by ultraviolet spectrophotometry, the content of soluble protein is determined by Coomassie brilliant blue G-250 staining method, and the content of soluble sugar is determined by anthrone colorimetric method.
The data measured are shown in the following table:
from the above experimental data it can be seen that:
under the illumination conditions described in example one, the plant height, diameter, root number, root length, leaf number, leaf area, and fresh weight average per plant were the highest or nearly the highest, indicating that Anoectochilus roxburghii grew the best and the volume per plant was the largest under the conditions described in example one.
Except for the illumination condition conditions of example one; under the condition of R5B1, the plant height is highest under the treatment of R5B1, the stem thickness, the root number, the root length, the leaf area and the fresh weight average of a single plant are second to those under the treatment of R12B1, and the overall highest index proportion is the largest; the maximum number of the leaves treated by R1B12 is 4, and R5B1 times; the drying rate difference of anoectochilus formosanus among the treatments is the largest, from large to small, R2B1 & gtCK & gtR 12B1 & gtR 1B12 & gtR 7B1 & gtR 5B1, and the drying rate difference of anoectochilus formosanus under the treatment of R2B1 is obviously higher than that of other treatments and is 8.98%.
The present invention is not limited to the above-described alternative embodiments, and various other forms of products can be obtained by anyone in light of the present invention. The above detailed description should not be taken as limiting the scope of the invention, which is defined in the claims, and which the description is intended to be interpreted accordingly.
Claims (8)
1. A method for culturing anoectochilus formosanus with improved dry recovery rate is characterized by comprising the following steps:
preparing a culture medium: the culture medium comprises 1900g/l potassium nitrate, 1650g/l ammonium nitrate, 170g/l monopotassium phosphate, 370 magnesium sulfate, 440g/1 calcium chloride dihydrate, 0.83g/l potassium iodide, 6.2g/1 boric acid, 22.3g/l manganese sulfate, 8.6g/l zinc sulfate, 0.25g/l sodium molybdate, 0.025g/l copper sulfate, 0.025g/l cobalt chloride, 37.3g/l disodium edetate, 27.8g/l ferrous sulfate, 100g/l inositol, 0.5g/l nicotinic acid, 0.5g/l pyridoxine hydrochloride, 0.1g/l thiamine hydrochloride, 2g/l glycine and 30g/l sucrose; the pH of the medium is 5.7;
inoculation: inoculating the anoectochilus formosanus tissue culture seedling into a culture medium under the aseptic condition;
culturing: placing the inoculated tissue culture seedlings in a culture chamber, and culturing under the condition of red and blue mixed illumination; culturing for three months, wherein in the first month, the ratio of red light to blue light is 12: 1; in the second month, the ratio of the red light to the blue light is 5: 1; in the third month, the ratio of red light to blue light is 12: 1.
2. The method for culturing Anoectochilus roxburghii with improved dry folding rate as claimed in claim 1, wherein the illumination time is 7.5-8.5h/d in the first month, 9.5-10.5h/d in the second month, and 10.5-12.5h/d in the third month.
3. The method for culturing Anoectochilus roxburghii with improved dry recovery rate as claimed in claim 1, wherein in the culturing process, a red-blue mixed lamp is used to provide red-blue mixed illumination for the tissue culture seedlings;
the red and blue mixed lamp comprises a lamp holder and 78 × N light emitting diodes fixed on the lower surface of the lamp holder, wherein N is more than or equal to 1; the lower surface of the lamp holder is provided with a lampshade buckled on the outer side of the light-emitting diode, and the lampshade is composed of single covers which correspond to the light-emitting diodes one by one; the lampshade consists of a red fixed cover body, a red detachable cover body, a blue fixed cover body and a blue detachable cover body; the red fixed cover body is formed by 65 × N red single covers, the red detachable cover body comprises 7 × N red single covers, the blue fixed cover body is formed by 6 × N blue single covers, and the blue detachable cover body comprises 7 × N blue single covers; the red fixed cover body and the blue fixed cover body are fixed on the lower surface of the lamp holder; the red cover body and the blue cover body can be dismantled through the mode of tearing open and set up in turn at the lower surface of lamp stand, and the red cover body that can be dismantled sets up the lower surface at the lamp stand in first period and third cycle, at the second cycle, the blue cover body that can be dismantled sets up the lower surface at the lamp stand.
4. The method for culturing Anoectochilus roxburghii with improved drying rate as claimed in claim 3, wherein the lamp holder is circular; 78 light-emitting diodes are arranged from inside to outside into a first circle to a fourth circle, the first circle is provided with 7 light-emitting diodes, the second circle is provided with 15 light-emitting diodes, the third circle is provided with 24 light-emitting diodes, and the fourth circle is provided with 32 light-emitting diodes;
the blue detachable cover body and the red detachable cover body are respectively composed of a central disc and 7 single covers which are in one-to-one correspondence with the first circle of light-emitting diodes, the central disc and the 7 single covers are connected through connecting rods, and the central disc is detachably arranged on the lower surface of the center of the lamp holder;
the blue fixed cover body is composed of 6 independent single covers, and the 6 blue single covers of the blue fixed cover body are evenly and distributively buckled on the third circle of light-emitting diodes;
the red fixed cover body is formed by 65 independent single covers and is buckled on the rest light-emitting diodes.
5. The method as claimed in claim 4, wherein the central plate is provided with a magnet and is magnetically attached to the center of the lower surface of the lamp holder.
6. The method for culturing Anoectochilus roxburghii with improved drying rate as claimed in claim 1, wherein the ratio of red light to blue light is: 2: 0.9-2: 1.1.
7. The method for culturing Anoectochilus roxburghii with improved drying rate as claimed in claim 1, wherein the ratio of red light to blue light is: 4.8: 1-5.2: 1.
8. The method for culturing Anoectochilus roxburghii with improved drying rate as claimed in claim 1, wherein the ratio of red light to blue light is: 11.5: 1-12.5: 1.
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