CN113728922B - Preparation method of houttuynia cordata planting seedlings - Google Patents
Preparation method of houttuynia cordata planting seedlings Download PDFInfo
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- CN113728922B CN113728922B CN202111086612.3A CN202111086612A CN113728922B CN 113728922 B CN113728922 B CN 113728922B CN 202111086612 A CN202111086612 A CN 202111086612A CN 113728922 B CN113728922 B CN 113728922B
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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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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
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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
- A01H4/002—Culture media for tissue culture
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
Abstract
The invention provides a preparation method of houttuynia cordata planting seedlings, which comprises the following steps: preparing penicillium citrinum spore suspension, extracting tissue of the houttuynia cordata, performing aseptic treatment on the tissue to obtain an explant of the houttuynia cordata, placing the explant in the penicillium citrinum spore suspension and performing shake culture to obtain a bacterium-carrying explant, transferring the bacterium-carrying explant into an aseptic culture medium, culturing to obtain a bacterium-carrying seedling, and culturing the bacterium-carrying seedling to obtain a houttuynia cordata planting seedling. The embodiment of the invention provides a preparation method of a houttuynia cordata planting seedling, which is characterized in that a houttuynia cordata explant and a penicillium citrinum fungal spore suspension are mixed and cultured under an aseptic condition, so that penicillium citrinum fungi and the houttuynia cordata explant can fully symbiotic, and the obtained planting seedling has a developed root system and vigorous growth of plants in a production and planting process.
Description
Technical Field
The invention relates to the technical field of houttuynia cordata seedling culture, and particularly relates to a preparation method of a houttuynia cordata seedling.
Background
The houttuynia cordata is one of the traditional Chinese medicines, can be used for treating lung carbuncle, purulent vomiting, phlegm heat, cough and asthma and other symptoms, and is also a favorite vegetable in southwest regions. With the continuous improvement of living standard, the residents have higher pursuit on the yield and the quality of the houttuynia cordata.
However, due to the nature of houttuynia cordata and the limitations of traditional breeding techniques, the yield of houttuynia cordata in a unit area of land is still not high, and the problem that the yield of houttuynia cordata cannot meet the needs of the public still exists, and how to improve the yield of houttuynia cordata needs to be solved urgently. Therefore, a preparation method of the houttuynia cordata planting seedlings is provided.
Disclosure of Invention
The invention aims to provide a preparation method of houttuynia cordata planting seedlings, which can at least partially overcome the defects in the prior art.
According to one aspect of the invention, the preparation method of the houttuynia cordata seedlings comprises the following steps:
preparing penicillium citrinum fungal spore suspension;
extracting tissue of the houttuynia cordata, and performing aseptic treatment on the tissue to obtain an explant of the houttuynia cordata;
placing the explant in the penicillium citrinum spore suspension, and performing shake culture at the temperature of 16-30 ℃ for not less than 0.5 hour to obtain a bacterium-carrying explant;
transferring the explant with bacteria into a sterile culture medium, and culturing to obtain houttuynia cordata seedlings with bacteria;
and culturing the houttuynia cordata seedling with bacteria to obtain the houttuynia cordata planting seedling.
Preferably, the spore concentration of the penicillium citrinum spore suspension is 102CFU/ml to 106CFU/ml.
Preferably, the spore concentration of the penicillium citrinum spore suspension is 103CFU/ml.
Preferably, the time of the shake culture is 0.5 to 2 hours.
Preferably, the culturing the houttuynia cordata seedling with bacteria to obtain the houttuynia cordata planting seedling comprises the following steps:
preparing sterile culture soil;
and (3) planting the houttuynia cordata seedling with bacteria into the sterile culture soil, and culturing for 12-16 days under the condition of avoiding direct strong light to obtain the houttuynia cordata seedling.
Preferably, the step of planting the cordate houttuynia seedlings with bacteria into the sterile culture soil comprises the step of selecting the cordate houttuynia seedlings with bacteria with the height of 3-7 cm;
and (3) planting the houttuynia cordata seedling with bacteria into the sterile culture soil, wherein the planting depth is 2-3cm. .
Preferably, the preparing of the sterile culture soil includes:
selecting yellow soil with the organic matter content of 500-2000 g/kg, the clay particle silicon-aluminum rate of 2.0-2.5 and the silicon-aluminum rate of 1.8-2.2, drying and crushing the yellow soil, and performing steam sterilization for not less than 60 minutes to obtain the sterile culture soil.
Preferably, the culturing for 12 to 16 days under the condition of avoiding direct irradiation of strong light to obtain the houttuynia cordata planting seedlings comprises the following steps:
culturing for 12-16 days under the condition of avoiding direct irradiation of strong light, and irrigating the houttuynia cordata seedling with bacteria at a spore concentration of not less than 10 every other day 2 CFU/ml of Penicillium citrinum spore suspension enables the Penicillium citrinum fungal spores inoculated to each houttuynia cordata seedling with bacteria to be not lower than 20000CFU, and the houttuynia cordata seedling is obtained.
Preferably, the extracting the explant of the houttuynia cordata plant comprises:
and extracting at least one of germinated seeds and plant stem tips of the houttuynia cordata to serve as the explant.
Preferably, the sterile medium is selected from 1/2MS medium or MS medium.
The embodiment of the invention provides a preparation method of a houttuynia cordata planting seedling, wherein a houttuynia cordata explant and a penicillium citrinum fungal spore suspension are mixed and cultured under an aseptic condition, so that penicillium citrinum fungi and the houttuynia cordata explant can fully symbiotic, and the obtained planting seedling has a developed root system and vigorous plant growth in a production and planting process.
Drawings
Other features, objects and advantages of the invention will become more apparent upon reading of the detailed description of non-limiting embodiments with reference to the following drawings:
FIG. 1 is a flow chart for preparing houttuynia cordata seedlings according to the present invention;
FIG. 2 is a flow chart of the cultivation of seedlings with bacteria into houttuynia cordata seedlings according to the present invention;
FIG. 3 is experimental data of experimental examples 1 to 4 according to the present invention and comparative example 1.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings and examples. It is to be understood that the specific embodiments described herein are merely illustrative of the relevant invention and not restrictive of the invention. For convenience of description, only portions related to the invention are shown in the drawings.
It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict.
The present application will be described in further detail with reference to the following drawings and examples. It is to be understood that the specific embodiments described herein are merely illustrative of the invention and are not to be construed as limiting the invention. It should be noted that, for convenience of description, only the portions related to the present invention are shown in the drawings.
The Penicillium citrinum in the present invention is a fungus of the order chitinales, family californiaceae, named Penicillium citrinum. The spores mainly refer to fungus propagation organs from penicillium citrinum hyphae, and the specific acquisition mode can be that the penicillium citrinum fungi are cultured by using a culture medium, the hyphae are scraped after the hyphae grow out, and the spores are obtained by the steps of dilution, filtration and the like, or the spores are obtained by the methods of air filtration and the like under the field condition, and the spores belong to the fungus propagation organs of the penicillium citrinum.
The planting seedling in the invention mainly refers to a seedling body which has certain plant height and activity and can be planted in suitable soil to obtain a mature plant of the houttuynia cordata, so that the houttuynia cordata seedling body can be conveniently packaged and transported while keeping some special shapes, and is beneficial to large-scale popularization.
The Houttuynia cordata in the invention is a plant which is named Houttuynia cordata Thunb in Houttuynia of Saururaceae of pepper order.
The invention provides a preparation method of houttuynia cordata seedlings, as shown in figure 1, comprising the following steps:
s101: preparing penicillium citrinum spore suspension;
s102: extracting tissue of the houttuynia cordata, and performing aseptic treatment on the tissue to obtain an explant of the houttuynia cordata;
s103: placing the explant in the penicillium citrinum spore suspension, and performing shake culture at 16-30 ℃ for not less than 0.5 h to obtain a bacterium-carrying explant;
s104: transferring the explant with bacteria into a sterile culture medium, and culturing to obtain the seedling with bacteria of houttuynia cordata;
s105: culturing the houttuynia cordata seedling with bacteria to obtain the houttuynia cordata seedling.
In the step S101, the penicillium citrinum spore suspension may be prepared by culturing penicillium citrinum in a culture medium known in the art, such as a potato dextrose agar culture medium or a glucose culture medium, scraping the mycelia after growing, mixing the mycelia with sterile water, filtering the warp cloth, and collecting the filtrate to obtain a penicillium citrinum spore suspension, or by dissolving and centrifuging the culture containing penicillium citrinum. The spore concentration of Penicillium citrinum spore suspension is not less than 10 2 CFU/ml, preferably 10 2 CFU/ml to 10 6 CFU/ml, more preferably 10 3 CFU/ml. The proper spore concentration of the penicillium citrinum spore suspension is beneficial to planting enough spore quantity on the explant in the subsequent explant bacterium carrying process, and the interference of the future growth of the explant caused by the overhigh spore concentration is avoided. Through experimental research, 10 is found 2 CFU/ml to 10 6 CFU/ml is a better range of spore concentration of the penicillium citrinum suspension.
In the step S102, the tissue from which the houttuynia cordata is extracted may preferably be the root tip, stem segment, stem tip, young leaf or young embryo from which the houttuynia cordata is extracted, and these parts have strong plant cell differentiation capability, which is beneficial for the formation of complete seedlings by the subsequent culture. The method for performing the aseptic treatment on the tissue of the houttuynia cordata in the treatment step S102 may be sterilization using alcohol, mercuric chloride or bleaching powder, and the specific sterilization operation is well known to those skilled in the art and will not be described herein again. The extraction method in step S102 may be cutting, picking, pinching, or the like, which is well known to those skilled in the art to extract the tissue of the houttuynia cordata, and is not described herein.
It should be noted that the processing S101 and the processing S102 may be interchanged without having a sequential execution order, and fig. 1 is only a flow processing step of a preferred implementation manner, and the sequential order of the processing S101 and the processing S102 is not limited. In the implementation mode shown in fig. 1, the time for the extracted tissue of the houttuynia cordata to wait for shaking culture can be reduced, and the activity of the tissue of the houttuynia cordata can be improved.
In the treatment S103, the shake culture time of the explant in the Penicillium citrinum spore suspension can be preferably 0.5-2 hours, so that the culture time is sufficient and the probability of damage of the explant caused by too long shake culture time is reduced.
In the step S104, the sterile medium used may be an MS medium designed by Murashige and Skoog in 1962 and containing major elements such as ammonium nitrate, potassium nitrate, calcium chloride, magnesium sulfate, potassium dihydrogen phosphate, and the like, and trace elements such as potassium iodide, boric acid, magnesium sulfate, and the like, which are well known to those skilled in the art, or may be a 1/2MS medium with half of the content of each element in the MS medium, so that the explant with bacteria can be cultured to have bacteria relatively quickly, and the operations such as controlling temperature and humidity, supplementing electrolyte, and the like in the seedling culture process are all operations of plant cloning or plant tissue culture which are well known to those skilled in the art, and will not be described herein again.
In the process S105, the purpose of culturing the seedlings with bacteria to the seedlings for planting is to culture the seedlings with lower plant height in the culture medium to have a certain plant height and activity, and to be planted in suitable soil to obtain the seedling body of the mature plant of houttuynia cordata, which can be conveniently packaged and transported while maintaining some special shapes.
The specific implementation manner of processing the seedlings with bacteria cultured in S105 to obtain the planted seedlings can be as follows as shown in fig. 2:
s1051: preparing sterile culture soil;
s1052: and (3) planting the seedlings with bacteria into sterile culture soil, and culturing for 12-16 days under the condition of avoiding direct strong light to obtain the houttuynia cordata planting seedlings.
One way of implementing the preparation of the culture soil of the bacteria in the process of S1051 may be: yellow soil with the organic matter content of 500-2000 g/kg, the clay grain silicon-aluminum rate of 2.0-2.5 and the silicon-iron-aluminum rate of 1.8-2.2 is selected, dried and crushed, and steam sterilized for no less than 60 minutes to obtain the sterile culture soil. Experiments prove that the yellow soil meeting the coefficient can better promote the growth of the seedlings with bacteria into the houttuynia cordata planting seedlings meeting the conditions through the fertility, the water storage performance and the air permeability
In the treatment S1051, preferably, seedlings with bacteria with plant heights of 3cm to 7 cm can be selected, so that the culture survival rate is higher; and (5) transplanting the seedlings with the bacteria into the sterile culture soil prepared in the step (S1050), wherein the planting depth is 2-3cm. Culturing for 12-16 days under the condition of avoiding direct irradiation of strong light, and irrigating the seedling with bacteria at a spore concentration of not less than 10 every other day for further increasing symbiotic relationship between Penicillium citrinum and herba Houttuyniae 2 The CFU/ml Penicillium citrinum spore suspension ensures that the Penicillium citrinum fungal spores inoculated on average to each bacterial-bearing seedling are not lower than 20000CFU, so that the obtained houttuynia cordata planting seedlings have better properties.
Experimental example 1
Culturing Penicillium citrinum with potato glucose agar medium with diameter of 7 cm, scraping off mycelium with sterile scraper after the mycelium grows out, mixing the mycelium with pure water, filtering to obtain Penicillium citrinum spore suspension, and adjusting the Penicillium citrinum spore suspension to concentration of 10 2 CFU/ml。
Extracting stem tip of herba Houttuyniae with a concentration of 0.5 cm, and sterilizing with 75% ethanol to obtain explant.
Placing the explant at 10 2 Culturing in CFU/ml Penicillium citrinum spore suspension at 24 deg.C for 1 hr under shaking to obtain explant with bacteria.
Transferring the explant with bacteria into a 1/2MS sterile culture medium, and culturing to obtain the seedling with bacteria.
Selecting bacteria-carrying seedlings with the plant height of 3-7 cm, transplanting yellow soil with the organic matter content of 1500 g/kg, the clay particle silicon-aluminum rate of 2.0 and the silicon-aluminum rate of 2.0, performing steam sterilization for 90 minutes, and culturing for 14 days under the condition of avoiding direct irradiation of strong light to obtain the houttuynia cordata planting seedlings.
Experimental example 2
Culturing Penicillium citrinum with potato glucose agar medium with diameter of 7 cm, scraping off mycelium with sterile scraper after the mycelium grows out, mixing the mycelium with pure water, filtering to obtain Penicillium citrinum spore suspension, and adjusting the Penicillium citrinum spore suspension to concentration of 10 3 CFU/ml。
Extracting stem tip of herba Houttuyniae with a concentration of 0.5 cm, and sterilizing with 75% ethanol to obtain explant.
Placing the explant at 10 2 Culturing under shaking in CFU/ml Penicillium citrinum spore suspension at 24 deg.C for 1 hr to obtain explant with bacteria.
Transferring the explant with bacteria into a 1/2MS sterile culture medium, and culturing to obtain the seedling with bacteria.
Selecting bacteria-carrying seedlings with the plant height of 3-7 cm, transplanting yellow soil with the organic matter content of 1500 g/kg, the clay grain silicon-aluminum rate of 2.0 and the silicon-iron-aluminum rate of 2.0, sterilizing for 90 minutes by steam, and culturing for 14 days under the condition of avoiding direct irradiation of strong light to obtain the houttuynia cordata planting seedlings.
Experimental example 3
Culturing Penicillium citrinum with potato glucose agar medium to obtain 7 cm diameter mycelium, scraping off the mycelium with sterile scraper, mixing the mycelium with pure water, filtering to obtain Penicillium citrinum spore suspension, and adjusting the Penicillium citrinum spore suspension to concentration of 10 6 CFU/ml。
Extracting stem tip of herba Houttuyniae with a concentration of 0.5 cm, and sterilizing with 75% ethanol to obtain explant.
Placing the explant at 10 2 Culturing in CFU/ml Penicillium citrinum spore suspension at 24 deg.C for 1 hr under shaking to obtain explant with bacteria.
Transferring the explant with bacteria into a 1/2MS sterile culture medium, and culturing to obtain the seedling with bacteria.
Selecting bacteria-carrying seedlings with the plant height of 3-7 cm, transplanting yellow soil with the organic matter content of 1500 g/kg, the clay grain silicon-aluminum rate of 2.0 and the silicon-iron-aluminum rate of 2.0, sterilizing for 90 minutes by steam, and culturing for 14 days under the condition of avoiding direct irradiation of strong light to obtain the houttuynia cordata planting seedlings.
Experimental example 4
Culturing Penicillium citrinum with potato glucose agar medium with diameter of 7 cm, scraping off mycelium with sterile scraper after the mycelium grows out, mixing the mycelium with pure water, filtering to obtain Penicillium citrinum spore suspension, and adjusting the Penicillium citrinum spore suspension to concentration of 10 2 CFU/ml。
Extracting stem tip of herba Houttuyniae with a concentration of 0.5 cm, and sterilizing with 75% ethanol to obtain explant.
Placing the explant at 10 2 Culturing under shaking in CFU/ml Penicillium citrinum spore suspension at 24 deg.C for 1 hr to obtain explant with bacteria.
Transferring the explant with bacteria into a 1/2MS sterile culture medium, and culturing to obtain the seedling with bacteria.
Selecting the seedling with bacteria with the plant height of 3-7 cm, transplanting yellow soil with the organic matter content of 1500 g/kg, the clay particle silicon-aluminum rate of 2.0 and the silicon-aluminum rate of 2.0 and steam sterilization for 90 minutes, culturing for 14 days under the condition of avoiding direct irradiation of strong light, and irrigating the seedling with bacteria with spore concentration not lower than 10 every other day 2 CFU/ml of Penicillium citrinum spore suspension enables the average Penicillium citrinum spore inoculated to each germ-carrying seedling to be not lower than 20000CFU, and the houttuynia cordata planting seedling is obtained.
Comparative example 1
Extracting stem tip of herba Houttuyniae with a concentration of 0.5 cm, and sterilizing with 75% ethanol to obtain explant.
The explants were transferred to 1/2MS sterile medium and cultured to obtain seedlings.
Selecting seedlings with the plant height of 3-7 cm, transplanting yellow soil with the organic matter content of 1500 g/kg, the clay grain silicon-aluminum rate of 2.0 and the silicon-aluminum rate of 2.0, sterilizing for 90 minutes by steam, and culturing for 14 days under the condition of avoiding direct irradiation of strong light to obtain the houttuynia cordata planting seedlings.
The test method comprises the following steps: selecting 20 houttuynia cordata seedlings prepared in experimental examples 1 to 4 with the average plant height of 10 cm and the houttuynia cordata seedlings prepared in comparative example 1, selecting 100 plants in total, transferring yellow soil with the organic matter content of 1500 g/kg, the clay particle silicon-aluminum rate of 2.0 and the silicon-aluminum rate of 2.0, the soil thickness of 50 cm, the cultivation depth of 5 cm, keeping the air temperature of 25 ℃ and the humidity of 80%, watering 30 ml of each plant averagely every other day, supplementing 10 mg of potassium dihydrogen phosphate to each plant, growing for 30 days under the condition that the daily average natural illumination time is 12 hours, measuring the plant height and the root length of the plant obtained by planting the houttuynia cordata seedlings in each experimental example and each comparative example, counting the average plant height and the average root length of each group of experimental examples and comparative examples, and calculating the result is shown in figure 3.
As can be seen from fig. 3, the average plant height of the houttuynia cordata seedlings obtained in experimental examples 1 to 4 ranges from 5.67 cm to 7.17 cm and the average root length ranges from 4.11 cm to 6.68 cm after being cultured for 30 days under the above-mentioned culture conditions, which are much larger than the average plant height of the houttuynia cordata seedlings prepared in comparative example 1 of 4.56 cm and the average root length of 3.11 cm, which fully proves that the mixture of the penicillium citrinum fungi has a promoting effect on the growth of the houttuynia cordata plants.
Meanwhile, after the houttuynia cordata planting seedlings prepared in the experimental example 4 are cultured for 30 days under the above-mentioned culture conditions, the average plant height is 7.17 cm, the average root length is 6.68 cm, and the average root length is slightly larger than that of the experimental examples 1-3, so that in the process of culturing the bacterial seedlings to the houttuynia cordata planting seedlings, the continuous supplement of the penicillium citrinum fungal spores has a certain promotion effect on the growth of the subsequent plants.
The above description is only a preferred embodiment of the application and is illustrative of the principles of the technology employed. It will be appreciated by those skilled in the art that the scope of the invention herein disclosed is not limited to the particular combination of features described above, but also encompasses other arrangements in which any combination of the above features or their equivalents is incorporated without departing from the spirit of the invention. For example, the above features may be replaced with (but not limited to) features having similar functions disclosed in the present application.
Claims (5)
1. The preparation method of the houttuynia cordata planting seedlings is characterized by comprising the following steps:
preparation of spore concentration of 10 2 CFU/ml to 10 6 CFU/ml Penicillium citrinum spore suspension;
extracting a 0.5 cm shoot tip of the houttuynia cordata, and performing aseptic treatment on the tissue to obtain an explant of the houttuynia cordata;
placing the explant in the penicillium citrinum spore suspension, and performing shake culture at the temperature of 16-30 ℃ for 0.5-2 hours to obtain a bacterium-carrying explant;
transferring the explant with bacteria into a sterile culture medium, and culturing to obtain seedlings with bacteria;
culturing the bacteria-carrying seedlings to obtain houttuynia cordata planting seedlings, and the method specifically comprises the following steps:
preparing sterile culture soil, namely selecting yellow soil with 500-2000 g/kg of organic matter content, 2.0-2.5 of clay particle silicon-aluminum rate and 1.8-2.2 of silicon-iron-aluminum rate, drying and crushing the yellow soil, and performing steam sterilization for no less than 60 minutes to obtain the sterile culture soil;
and (3) planting the seedling with bacteria into the sterile culture soil, and culturing for 12-16 days under the condition of avoiding direct strong light to obtain the houttuynia cordata planting seedling.
2. The method for preparing houttuynia cordata seedlings according to claim 1, wherein the spore concentration of the penicillium citrinum spore suspension is 10 3 CFU/ml。
3. The method for preparing houttuynia cordata seedlings according to claim 1, wherein the step of planting the seedlings with bacteria into the sterile culture soil comprises the following steps:
selecting the bacteria-carrying seedlings with the plant heights of 3cm to 7 cm;
and (3) planting the seedlings with the bacteria into the sterile culture soil, wherein the planting depth is 2-3cm.
4. The method for preparing houttuynia cordata thunb seedlings according to claim 1, wherein the culture for 12 to 16 days under the condition of avoiding direct irradiation of strong light comprises the following specific steps:
culturing for 12-16 days under the condition of avoiding direct irradiation of strong light, and irrigating the germ-carrying seedlings with spore concentration not lower than 10 every other day 2 CFU/ml of Penicillium citrinum spore suspension enables the average Penicillium citrinum spore inoculated to each bacterium-carrying seedling to be not less than 20000CFU, and the houttuynia cordata planting seedling is obtained.
5. The method for preparing a houttuynia cordata plantlet according to any one of claims 1 to 4, wherein the sterile medium is selected from a 1/2MS medium or an MS medium.
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| CN113728922A (en) | 2021-12-03 |
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