CN114424743A - Method for ex-vitro rooting of tissue culture seedlings of Chinese roses in courtyard - Google Patents
Method for ex-vitro rooting of tissue culture seedlings of Chinese roses in courtyard Download PDFInfo
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- CN114424743A CN114424743A CN202111491121.7A CN202111491121A CN114424743A CN 114424743 A CN114424743 A CN 114424743A CN 202111491121 A CN202111491121 A CN 202111491121A CN 114424743 A CN114424743 A CN 114424743A
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- 240000008254 Rosa chinensis Species 0.000 title claims abstract description 32
- 235000000664 Rosa chinensis Nutrition 0.000 title claims abstract description 32
- 238000000034 method Methods 0.000 title claims abstract description 28
- 239000011159 matrix material Substances 0.000 claims abstract description 25
- 235000000100 Hibiscus rosa sinensis Nutrition 0.000 claims abstract description 20
- 235000016785 Rosa della China Nutrition 0.000 claims abstract description 20
- 238000007789 sealing Methods 0.000 claims abstract description 19
- 239000000758 substrate Substances 0.000 claims abstract description 13
- 238000012258 culturing Methods 0.000 claims abstract description 11
- 239000003415 peat Substances 0.000 claims description 17
- 239000002689 soil Substances 0.000 claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 235000019362 perlite Nutrition 0.000 claims description 12
- 239000010451 perlite Substances 0.000 claims description 12
- 238000005520 cutting process Methods 0.000 claims description 11
- 238000005286 illumination Methods 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 8
- 239000008223 sterile water Substances 0.000 claims description 8
- 239000012528 membrane Substances 0.000 claims description 3
- 238000002791 soaking Methods 0.000 claims 1
- 230000001954 sterilising effect Effects 0.000 abstract description 7
- 229930195732 phytohormone Natural products 0.000 abstract description 2
- 238000000338 in vitro Methods 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 6
- 238000011282 treatment Methods 0.000 description 5
- 238000009736 wetting Methods 0.000 description 4
- 230000008901 benefit Effects 0.000 description 3
- 238000004080 punching Methods 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 241000220317 Rosa Species 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 229930192334 Auxin Natural products 0.000 description 1
- 235000004789 Rosa xanthina Nutrition 0.000 description 1
- 241000220222 Rosaceae Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000000540 analysis of variance Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000002363 auxin Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 238000005553 drilling Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000001932 seasonal effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
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Classifications
-
- 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
- A01G31/00—Soilless cultivation, e.g. hydroponics
-
- 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
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
- A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
-
- 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
- A01G24/28—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing peat, moss or sphagnum
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Soil Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Inorganic Chemistry (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention relates to a method for ex-vitro rooting of tissue culture seedlings of China roses in a courtyard, belonging to the technical field of China rose propagation and comprising the following steps of: (1) preparing and sterilizing a substrate; (2) preparing a substrate for filling; (3) rooting culture outside the bottle; (4) sealing; (5) and (5) culturing. The method utilizes the mode of combining the ex-vitro rooting matrix and the 126-hole tray to carry out the rooting culture of the courtyard Chinese rose, simplifies the tissue culture process, saves the tissue culture space, reduces the use of phytohormone, and has higher rooting rate.
Description
Technical Field
The invention belongs to the technical field of Chinese rose propagation, and particularly relates to a method for ex-vitro rooting of tissue culture seedlings of courtyard Chinese roses.
Background
China Rose (Rose chinensis Jacq) is a Rose plant of the Rosaceae family, is known as 'queen in flowers', has great ornamental value and great commercial value, is deeply loved by people and is widely cultivated all over the world. The garden China rose of Meiang company is widely used for landscaping, yard beautification and the like due to the large flower, elegant and graceful flower type and rich flower color. At present, the commercial production of China roses mainly adopts cuttage, but has few cutting slips and low propagation coefficient, and meanwhile, the quality of seedlings bred by cuttage is easy to reduce due to the accumulation of viruses. The tissue culture overcomes the defect of low propagation coefficient of the conventional propagation method, maintains the excellent properties of the variety, obtains a large number of nursery stocks with consistent growth and development in a short period, and has the advantages of short production period, no seasonal limitation, remarkable economic benefit and the like.
At present, the tissue culture seedling of the Chinese rose is generally rooted by adding auxin into a culture medium or dipping the seedling into a rooting agent and then inserting the seedling into a matrix for rooting, the process is complex, and the production cost is high.
Disclosure of Invention
In order to overcome the problems in the background technology, the invention provides a method for the ex-bottle rooting of tissue culture seedlings of the courtyard China rose, which is characterized in that the rooting culture of the courtyard China rose is carried out by combining an ex-bottle rooting substrate with a 126-hole tray, the tissue culture process is simplified, the tissue culture space is saved, the use of phytohormones is reduced, and the rooting rate is high.
In order to realize the purpose, the invention is realized by the following technical scheme:
the method for rooting the tissue culture seedlings of the Chinese roses in the courtyard outside the bottle comprises the following steps:
(1) matrix: the matrix is prepared from peat soil and perlite according to a certain proportion, the matrix is watered and moistened and uniformly stirred before use, the humidity is 65% -70%, and the matrix is packaged in a plastic bag made of high-temperature resistant materials and sterilized for 40 minutes at high temperature and high pressure;
(2) preparing a matrix by subpackaging: filling the sterilized substrate in the step (1) into a 126-hole tray in a clean bench;
(3) and (3) rooting culture outside the bottle: cutting off terminal buds of cluster bud seedlings which grow well in the bottle and are 5-6cm high, directly inserting the cut terminal buds into the split-packaged matrix in the step (2), transferring the 126-hole tray into a sterile culture box matched with the cut terminal buds, and adding 300mL of sterile water along the edge of the box;
(4) sealing: placing the culture box on a film sealing machine, and sealing the film;
(5) culturing: and (5) placing the culture box after the membrane sealing in an environment with the temperature of 25 +/-2 ℃ and illumination for 8h for culturing for 30 d.
Preferably, the cut seedlings in step (3) have a length of 1-1.5cm, the bases are cut flat, and the cut seedlings are inserted into a plug tray of 1cm × 1cm × 1 cm.
Preferably, the culture box is reusable, and is not required to be disposed of when first used, and is soaked in 0.2% KMn4 for more than 8h before use when used again.
Preferably, the matrix in the step (1) is pure peat soil or peat soil: perlite =1:1, pure perlite.
The invention has the beneficial effects that:
1. the method adopts the ex-bottle rooting technology for the garden Chinese rose, can combine the rooting and domestication of the tissue culture seedlings, can simplify the tissue culture procedure of the garden Chinese rose, shortens the production period of the tissue culture seedlings and improves the propagation coefficient.
2. When the method is used for inducing the tissue culture seedlings of the courtyard China rose to root outside the bottle, no hormone is used, the production cost can be reduced, and the method provides assistance for green production of the courtyard China rose.
3. The invention simplifies the tissue culture process, saves the tissue culture space, shortens the tissue culture period, realizes the rooting rate outside the bottle of more than 80 percent, and has important significance for improving the industrialization level of the courtyard Chinese rose.
Drawings
FIG. 1 is a schematic view of the external rooting of the courtyard China rose.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings to facilitate understanding of the skilled person.
Example 1
A method for rooting tissue culture seedlings of China roses in vitro in a courtyard comprises the following steps:
(1) pouring pure peat soil thoroughly with water, moistening thoroughly, keeping humidity at about 65%, placing into plastic bag made of high temperature resistant material, and sterilizing at 121 deg.C and 1.01KPa for 40 min.
(2) Filling the sterilized substrate in the step (1) into a 126-hole tray on a clean bench and compacting.
(3) Cutting off terminal buds of cluster bud seedlings with good growth in the bottle and 5-6cm high to obtain seedlings with the length of 1-1.5cm, directly inserting the seedlings into the sterilized and packaged matrix in the step (2), transferring the plug tray into a culture box, and adding 300mL of sterile water along the edge of the box.
(4) The culture box is placed on a film sealing machine, and the film is sealed by using 36 lines of laser punched films.
(5) And (5) placing the culture box processed in the step (4) in an environment with 25 +/-2 ℃ and 8h of illumination for culturing for 30 d.
Example 2
A method for rooting tissue culture seedlings of China roses in vitro in a courtyard comprises the following steps:
(1) pouring pure peat soil thoroughly with water, moistening thoroughly, keeping humidity at about 65%, placing into plastic bag made of high temperature resistant material, and sterilizing at 121 deg.C under 1.01KPa for 40 min.
(2) Filling the substrate sterilized in the step (1) into a 126-hole tray on a clean bench and compacting.
(3) Cutting off terminal buds of cluster buds with the height of 5-6cm and the length of 1-1.5cm, directly inserting the cut terminal buds into the sterilized and packaged matrix in the step (2), transferring the plug tray into a culture box, and adding 300mL of sterile water along the edge of the box.
(4) The culture box is placed on a film sealing machine, and 15 lines of laser punched films are used for sealing the films.
(5) And (5) placing the culture box processed in the step (4) in an environment with 25 +/-2 ℃ and 8h of illumination for culturing for 30 d.
Example 3
A method for rooting tissue culture seedlings of China roses in vitro in a courtyard comprises the following steps:
(1) mixing peat soil: perlite (volume ratio) =1:1, thoroughly watering the substrate with water, fully wetting, keeping the humidity at about 65%, placing into a plastic bag made of high temperature resistant material, and sterilizing at 121 deg.C and 1.01KPa for 40 min.
(2) Filling the sterilized substrate in the step (1) into a 126-hole tray on a clean bench and compacting.
(3) Cutting off terminal buds of cluster buds with the height of 5-6cm and the length of 1-1.5cm, directly inserting the cut terminal buds into the sterilized and packaged matrix in the step (2), transferring the plug tray into a culture box, and adding 300mL of sterile water along the edge of the box.
(4) The culture box is placed on a film sealing machine, and the film is sealed by using 36 lines of laser punched films.
(5) And (5) placing the culture box processed in the step (4) in an environment with 25 +/-2 ℃ and 8h of illumination for culturing for 30 d.
Example 4
A method for rooting tissue culture seedlings of China roses in vitro in a courtyard comprises the following steps:
(1) mixing peat soil: perlite (volume ratio) =1:1, thoroughly watering the substrate with water, fully wetting, keeping the humidity at about 65%, placing into a plastic bag made of high temperature resistant material, and sterilizing at 121 deg.C and 1.01KPa for 40 min.
(2) Filling the peat soil sterilized in the step (1) into a 126-hole plug tray on a clean bench, and compacting.
(3) Cutting off terminal buds of cluster bud seedlings with good growth in the bottle and the height of 5-6cm to obtain seedlings with the length of 1-1.5cm, and directly inserting the seedlings into the sterilized and packaged matrix in the step (2). The plate was transferred to a culture box and 300mL of sterile water was added along the edge of the box.
(4) The culture box is placed on a film sealing machine, and 15 lines of laser punched films are used for sealing the films.
(5) And (4) placing the culture box processed in the step (4) in an environment with 25 +/-2 ℃ and 8h of illumination for culturing for 30 d.
Example 5
A method for rooting tissue culture seedlings of China roses in vitro in a courtyard comprises the following steps:
(1) pouring pure perlite with water thoroughly, wetting thoroughly, keeping humidity at about 65%, placing into plastic bag made of high temperature resistant material, and sterilizing at 121 deg.C and 1.01KPa for 40 min.
(2) Filling the sterilized substrate in the step (1) into a 126-hole tray on a clean bench and compacting.
(3) Cutting off terminal buds of cluster buds with the height of 5-6cm and the length of 1-1.5cm, directly inserting the cut terminal buds into the sterilized and packaged matrix in the step (2), transferring the plug tray into a culture box, and adding 300mL of sterile water along the edge of the box.
(4) The culture box is placed on a film sealing machine, and 15 lines of laser punched films are used for sealing the films.
(5) And (4) placing the culture box processed in the step (4) in an environment with 25 +/-2 ℃ and 8h of illumination for culturing for 30 d.
Example 6
A method for rooting tissue culture seedlings of China roses in vitro in a courtyard comprises the following steps:
(1) pouring pure perlite with water thoroughly, wetting thoroughly, keeping humidity at about 65%, placing into plastic bag made of high temperature resistant material, and sterilizing at 121 deg.C and 1.01KPa for 40 min.
(2) Filling the sterilized substrate in the step (1) into a 126-hole tray on a clean bench and compacting.
(3) Cutting off terminal buds of cluster buds with the height of 5-6cm and the length of 1-1.5cm, directly inserting the cut terminal buds into the sterilized and packaged matrix in the step (2), transferring the plug tray into a culture box, and adding 300mL of sterile water along the edge of the box.
(4) The culture box is placed on a film sealing machine, and the film is sealed by using 36 lines of laser punched films.
(5) And (5) placing the culture box processed in the step (4) in an environment with 25 +/-2 ℃ and 8h of illumination for culturing for 30 d.
Analysis of experiments
To illustrate the effects of the present invention, the following comparative tests were performed, as shown in table 1:
TABLE 1 Effect of different substrates and gas-permeable membranes on the in vitro survival Rate
TABLE 2 analysis of variance of ex-vitro rooting results with different treatments
From the tables 1 and 2, it can be seen that the survival rate of the courtyard China rose with pure peat soil as the matrix is 80-81% with the highest survival rate, and the 0.05 level is significantly higher than that of other matrixes, and the root volume of the treatment with pure peat soil as the matrix is significantly higher than that of other matrixes at the levels of 0.01 and 0.05; the total root length between treatments is between 9.81 and 21.10cm, wherein the total root length of C5 (perlite + peat soil laser punching 15 rows of breathable films) is the largest and is significantly different from other treatments on the level of 0.01 and 0.05; the number of roots has no significant difference among the treatments, the number of the roots treated by C5 (perlite + peat soil laser punching 15 rows of breathable films) is the largest, and is 47, and the number of the roots treated by C3 (pure peat soil laser punching 15 rows of breathable films) is the smallest, and is 36.
The results of the examples show that the best method for the external rooting of the Chinese rose in the courtyard is to use pure peat soil as a matrix and 36-line laser drilling breathable film as a sealing film.
Finally, it is noted that the above-mentioned preferred embodiments illustrate rather than limit the invention, and that, although the invention has been described in detail with reference to the above-mentioned preferred embodiments, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the scope of the invention as defined by the appended claims.
Claims (4)
1. A method for ex-vitro rooting of tissue culture seedlings of Chinese roses in a courtyard is characterized by comprising the following steps of: the method comprises the following steps:
(1) matrix: the matrix is prepared from peat soil and perlite according to a certain proportion, the matrix is watered and moistened and uniformly stirred before use, the humidity is 65% -70%, and the matrix is packaged in a plastic bag made of high-temperature resistant materials and sterilized for 40 minutes at high temperature and high pressure;
(2) preparing a matrix by subpackaging: filling the sterilized substrate in the step (1) into a 126-hole tray in a clean bench;
(3) and (3) rooting culture outside the bottle: cutting off terminal buds of cluster bud seedlings which grow well in a bottle and are 5-6cm high, inserting the cut terminal buds into the split-packaged matrix in the step (2), transferring a 126-hole disc into a sterile culture box matched with the cut terminal buds, and adding 300mL of sterile water along the edge of the box;
(4) and (3) sealing: placing the culture box on a film sealing machine, and sealing the film;
(5) culturing: and (5) placing the culture box after the membrane sealing in an environment with the temperature of 25 +/-2 ℃ and illumination for 8h for culturing for 30 d.
2. The method for ex-vitro rooting of tissue culture seedlings of courtyard China roses according to claim 1, characterized in that: and (4) cutting the seedlings cut in the step (3) to be 1-1.5cm long, and flatly cutting the base parts of the seedlings, and inserting the seedlings into a plug tray of 1cm multiplied by 1 cm.
3. The tissue culture seedling outside the bottle of the courtyard China rose according to claim 1The rooting method is characterized by comprising the following steps: the culture box can be repeatedly used, and is not required to be treated when being used for the first time, and 0.2% KMn is required to be used before being used for the second time4Soaking for more than 8 h.
4. The method for ex-vitro rooting of tissue culture seedlings of courtyard China roses according to claim 1, characterized in that: the matrix in the step (1) is pure peat soil or peat soil: perlite =1:1, pure perlite.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111491121.7A CN114424743A (en) | 2021-12-08 | 2021-12-08 | Method for ex-vitro rooting of tissue culture seedlings of Chinese roses in courtyard |
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| CN202111491121.7A CN114424743A (en) | 2021-12-08 | 2021-12-08 | Method for ex-vitro rooting of tissue culture seedlings of Chinese roses in courtyard |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115581201A (en) * | 2022-08-26 | 2023-01-10 | 云南省农业科学院花卉研究所 | Diploid rose F induced by stem segment as explant 1 -61 plant regeneration method |
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| CN110786240A (en) * | 2018-11-22 | 2020-02-14 | 江苏省林业科学研究院 | Ex-vitro rooting method for callicarpa chinensis tissue culture seedlings |
| CN113728845A (en) * | 2021-09-17 | 2021-12-03 | 贵州省林业科学研究院 | Method for rooting of Ardisia mamillata Hance tissue culture seedlings outside bottles |
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2021
- 2021-12-08 CN CN202111491121.7A patent/CN114424743A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110786240A (en) * | 2018-11-22 | 2020-02-14 | 江苏省林业科学研究院 | Ex-vitro rooting method for callicarpa chinensis tissue culture seedlings |
| CN113728845A (en) * | 2021-09-17 | 2021-12-03 | 贵州省林业科学研究院 | Method for rooting of Ardisia mamillata Hance tissue culture seedlings outside bottles |
Non-Patent Citations (1)
| Title |
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| 赵春莉等: "‘红双喜’月季组培苗瓶外生根技术研究", 《东北农业科学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115581201A (en) * | 2022-08-26 | 2023-01-10 | 云南省农业科学院花卉研究所 | Diploid rose F induced by stem segment as explant 1 -61 plant regeneration method |
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Application publication date: 20220503 |