CN112042540A - Rapid propagation method of hydrangea macrophylla - Google Patents
Rapid propagation method of hydrangea macrophylla Download PDFInfo
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- CN112042540A CN112042540A CN202010967730.4A CN202010967730A CN112042540A CN 112042540 A CN112042540 A CN 112042540A CN 202010967730 A CN202010967730 A CN 202010967730A CN 112042540 A CN112042540 A CN 112042540A
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- stem
- hydrangea macrophylla
- axillary
- culture medium
- tips
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- 235000014486 Hydrangea macrophylla Nutrition 0.000 title claims abstract description 50
- 244000267823 Hydrangea macrophylla Species 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 24
- 239000001963 growth medium Substances 0.000 claims abstract description 17
- 230000035755 proliferation Effects 0.000 claims abstract description 17
- 241000196324 Embryophyta Species 0.000 claims abstract description 15
- 230000006698 induction Effects 0.000 claims abstract description 11
- 230000035784 germination Effects 0.000 claims abstract description 9
- 238000005520 cutting process Methods 0.000 claims abstract description 8
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 8
- 239000012883 rooting culture medium Substances 0.000 claims abstract description 7
- 230000008929 regeneration Effects 0.000 claims abstract description 4
- 238000011069 regeneration method Methods 0.000 claims abstract description 4
- 239000011159 matrix material Substances 0.000 claims abstract description 3
- 238000005286 illumination Methods 0.000 claims description 24
- 229920001817 Agar Polymers 0.000 claims description 9
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 9
- 229930006000 Sucrose Natural products 0.000 claims description 9
- 239000008272 agar Substances 0.000 claims description 9
- 229960004793 sucrose Drugs 0.000 claims description 9
- 241001092080 Hydrangea Species 0.000 claims description 8
- 230000001939 inductive effect Effects 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 4
- 230000001954 sterilising effect Effects 0.000 claims description 4
- 239000000758 substrate Substances 0.000 claims description 4
- 239000005720 sucrose Substances 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 3
- 239000010451 perlite Substances 0.000 claims description 3
- 235000019362 perlite Nutrition 0.000 claims description 3
- 239000002689 soil Substances 0.000 claims description 3
- 239000010455 vermiculite Substances 0.000 claims description 3
- 235000019354 vermiculite Nutrition 0.000 claims description 3
- 229910052902 vermiculite Inorganic materials 0.000 claims description 3
- 229960002523 mercuric chloride Drugs 0.000 claims description 2
- 230000002062 proliferating effect Effects 0.000 abstract 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 239000008223 sterile water Substances 0.000 description 3
- 239000003630 growth substance Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 241000448032 Hydrangea bretschneideri Species 0.000 description 1
- 101150034699 Nudt3 gene Proteins 0.000 description 1
- 241000220151 Saxifragaceae Species 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 230000036544 posture Effects 0.000 description 1
- 230000001902 propagating effect Effects 0.000 description 1
- 230000001932 seasonal effect Effects 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention relates to a rapid propagation method of hydrangea macrophylla, which comprises the following steps: (1) taking young and tender stem segments and stem tips of annual new branches of hydrangea macrophylla as explants, and carrying out disinfection treatment; (2) cutting tender stem segments of annual new branches of the hydrangea macrophylla treated in the step (1) into stem segments with at least one axillary bud and stem tips, inoculating the stem segments and the stem tips into an axillary bud germination induction culture medium, and allowing the axillary buds to germinate and the stem tips to grow in an extending mode; (3) when the axillary buds are elongated and the stem tips reach a certain length in the step (2), cutting the axillary buds and the stem tips, inoculating the axillary buds and the stem tips into a proliferation culture medium, and proliferating the axillary buds and the stem tips to obtain a large number of proliferation buds; (4) inoculating the proliferation bud in the step (3) to a rooting culture medium for induction culture to form a tissue culture seedling; (5) transplanting the tissue culture seedlings obtained in the step (4) into a seedling hardening matrix for culture to obtain a regeneration plant of the hydrangea macrophylla with strong roots and stems. The invention has the advantages that: has the advantages of high propagation speed and high propagation coefficient.
Description
Technical Field
The invention belongs to the technical field of forestry asexual propagation, and particularly relates to a rapid propagation method of hydrangea macrophylla.
Background
Hydrangea macrophylla (Hydrangea bretschneideri Dipp), also known as Hydrangea macrophylla, Saxifragaceae (Saxfragaceae) Hydrangea (Hydrangea) shrub, 1-3m high; growing on valley stream side or hillside forest; distributed in inner Mongolia, Hebei, Shanxi, Ningxia, Gansu, Qinghai, Henan provinces and other provinces; it is good for warm and humid environment, and has strong adaptability to cold and drought.
China has a lot of rare plants, which are important wealth in the national ornamental plant resource treasury, some wild plants have many kinds of resources, wide distribution, strong adaptability and strong resistance, beautiful postures and beautiful flowers have unique ornamental value, and the wild plants are used for indoor and outdoor arrangement and beautification to enrich the life of people.
The hydrangea ornamental plant resources are rich, the flower types and the flower colors are gorgeous and varied, foreign hydrangea original seeds are few, but the breeding work starts earlier, and the hydrangea varieties planted in China are almost all introduced from foreign countries, which is not matched with the rich hydrangea germplasm resources in China. Along with the modern construction of cities, the plant beautifying environment and the ornamental value are more and more important. However, in arid northwest areas, the traditional ornamental plants are limited in variety and cannot meet the aesthetic requirements of the public. The wild ornamental plants are utilized in propagation expansion, so that local resources, natural features and local characteristics can be displayed, different visual feelings can be brought to people, and the purpose of water-saving resource cultivation can be achieved.
Disclosure of Invention
The invention aims to provide a rapid propagation method of the hydrangea macrophylla based on the defects of the prior art, which takes annual tender stem segments and stem tips of the hydrangea macrophylla as explants, induces axillary buds and axillary buds to proliferate and induces root systems of the proliferated buds to form by using MS and 1/2MS culture media and biological growth regulators such as 6-BA, IBA, NAA and the like with different concentrations, and refines seedlings to realize the rapid propagation of the hydrangea macrophylla.
The purpose of the invention is realized by the following technical scheme:
a rapid propagation method of hydrangea macrophylla is characterized by comprising the following steps: the method comprises the following steps:
(1) taking annual young stem segments and stem tips of the hydrangea macrophylla as explants, and carrying out sterile disinfection treatment;
(2) cutting the annual tender stem segments and stem tips of the hydrangea macrophylla treated in the step (1) into hydrangea stem segments and stem tips with at least one axillary bud, inoculating the hydrangea macrophylla stem segments and stem tips into an axillary bud germination induction culture medium, and germinating the axillary buds and extending the stem tips to grow;
(3) when the axillary buds and the stem tips in the step (2) are extended to a certain length, cutting and inoculating the axillary buds and the stem tips into an axillary bud and stem tip proliferation culture medium to proliferate the axillary buds and the stem tips to obtain a large number of proliferation buds;
(4) inoculating the proliferation bud in the step (3) into a rooting culture medium for induction culture to form a tissue culture seedling;
(5) transplanting the tissue culture seedlings obtained in the step (4) into a seedling hardening matrix for culture to obtain a regeneration plant of the hydrangea macrophylla with strong roots and stems.
The disinfection treatment in the step (1) comprises the following steps: sterilizing annual young stem and stem tip of hydrangea macrophylla with alcohol for 30-60s, and sterilizing with 0.1% mercuric chloride for 8-10 min.
The culture medium for inducing the axillary bud germination in the step (2) comprises the following components: MS, 1.5-2.0 mg/L, IAA 0.3.3-0.5 mg/L of 6-BA, 30g/L of cane sugar and 6g/L of agar, and the pH value is controlled between 5.8 and 6.0.
And (3) carrying out germination culture on axillary buds and stem tips by using the axillary bud germination induction culture medium under the conditions of the temperature of 25 +/-1 ℃, the illumination intensity of 2500 Lx and the illumination time of 12 h/d.
The axillary bud and stem tip proliferation culture medium in the step (3) comprises the following components: MS, 0.1-0.5 mg/L, IBA 0.1.1-0.5 mg/L of 6-BA, 30g/L of cane sugar and 6g/L of agar, and the pH value is controlled between 5.8 and 6.0.
And performing proliferation culture of axillary buds by using the axillary bud and stem tip proliferation culture medium under the conditions of 25 +/-1 ℃, the illumination intensity of 2500 Lx and the illumination time of 12 h/d.
The rooting culture medium in the step (4) comprises the following components: 1/2MS, IBA 0.1-1.0 mg/L or 1/2MS, NAA1.0-2.0 mg/L, sucrose 30g/L, agar 6g/L, pH controlled between 5.8-6.0.
And (3) using the rooting culture medium to perform rooting induction culture of the proliferation buds under the conditions of the temperature of 23 +/-2 ℃, the illumination intensity of 1800 Lx and the illumination time of 12 h/d.
The hardening-seedling substrate in the step (5) comprises the following components: the ratio of the turfy soil to the vermiculite to the perlite is 1:1: 1.
The seedling exercising substrate is used for exercising and culturing the seedlings under the conditions that the temperature is 23 +/-2 ℃, the illumination intensity is 1800 Lx and the illumination time is 12 h/d.
The invention has the advantages that: has the advantages of high propagation speed and high propagation coefficient, is not limited by seasons and propagation materials, and can obtain a large number of plants in a short time; can breed the hydrangea macrophylla in a large number under the manual control environment, for the potted plant flowers in the north, fresh cut flower and seasonal greening market provide a large amount of seedlings, reduce the cost of flower traders, also provide theoretical and technical support for the further research of the hydrangea macrophylla.
Detailed Description
The features of the present invention and other related features are further described in detail below by way of examples to facilitate understanding by those skilled in the art:
example (b): in the embodiment, the method for quickly propagating the hydrangea macrophylla includes the following steps of taking annual tender stem segments and stem tips of the hydrangea macrophylla as explants, inducing axillary buds to proliferate and inducing root systems to form and hardening seedlings by using an MS culture medium and biological growth regulators such as 6-BA, IBA and NAA with different concentrations to realize quick propagation of the hydrangea macrophylla, and specifically including the following steps:
(1) taking annual young stem segments and stem tips of the hydrangea macrophylla as explants, and removing the dirt on the surfaces of the selected stem segments by using a soft brush with a detergent solution before disinfection. Cutting the stem into about 6cm, placing into a beaker, adding two drops of Tween 2.0, covering the opening of the beaker with gauze, and washing with running water for 30 min. After draining, the explant is washed by sterile water on a super-clean workbench for 3 times, each time for 1 min, then soaked in 75% alcohol for 30-60s, washed by the sterile water for 3 times, then soaked by 0.1% mercury bichloride for 8-10min, the bottle is continuously stirred in the soaking process, and then washed by the sterile water for 3-5 times, thereby realizing the sterile disinfection treatment of the explant.
(2) Cutting the stem sections treated in the step (1) into stem sections with 1-2 axillary buds, and inoculating the stem sections in the following culture media: MS, 1.0-2.0 mg/L, IAA 0.1.1-0.5 mg/L of 6-BA, 30g/L of sucrose and 6g/L of agar, wherein the pH value is controlled to be 5.8-6.0, the temperature is 25 +/-1 ℃, the illumination intensity is 2500 Lx, the illumination time is 12h/d, and axillary buds begin to germinate after being cultured for 5 d.
(3) When the axillary bud is elongated to 2-3cm in step (2), the cut is inoculated in the following medium: MS, 0.1-0.5 mg/L, IBA 0.1.1-0.5 mg/L of 6-BA, 30g/L of cane sugar and 6g/L of agar, wherein the pH value is controlled to be 5.8-6.0, the temperature is 25 +/-1 ℃, the illumination intensity is 2500 Lx, and the illumination time is 12h/d, axillary buds are proliferated in the culture process, and a large number of proliferated buds can be obtained after 30 d.
(4) Inoculating the proliferated shoots in step (3) in the following medium: 1/2MS, IBA.1-0.5 mg/L or NAA1.0-2.0 mg/L, sucrose 30g/L and agar 6g/L, and inducing and culturing for 30d under the conditions of pH 5.8-6.0, temperature 23 + -2 deg.C, illumination intensity 1800 Lx and illumination time 12h/d to form tissue culture seedling.
(5) Transplanting the tissue culture seedlings in the step (4) into the following matrixes: turfy soil: vermiculite perlite =1:1:1 culturing for 30d under the conditions of 23 +/-2 ℃ of temperature, 1800 Lx of illumination intensity and 12h/d of illumination time, and then growing into a regeneration plant of the hydrangea macrophylla with strong roots and stems.
The rapid propagation method of the hydrangea macrophylla in the embodiment has the disinfection survival rate of 60.0%, the axillary bud induction rate of 90.0%, the multiplication coefficient of 10.7, the rooting rate of 80.0% and the hardening and transplanting survival rate of 86.7%. The propagation speed in the culture process is high, the propagation coefficient is large and is not limited by seasons and propagation materials, a large number of plants can be obtained in a short time, and the method has important significance for mass propagation and production of the hydrangea macrophylla.
Although the above embodiments have been described in detail with respect to the purpose of illustrating the invention, it will be apparent to those skilled in the art that various changes and modifications can be made therein without departing from the scope of the invention as defined in the appended claims.
Claims (10)
1. A rapid propagation method of hydrangea macrophylla is characterized by comprising the following steps: the method comprises the following steps:
(1) taking annual young stem segments and stem tips of the hydrangea macrophylla as explants, and carrying out sterile disinfection treatment;
(2) cutting the annual tender stem segments and stem tips of the hydrangea macrophylla treated in the step (1) into hydrangea stem segments and stem tips with at least one axillary bud, inoculating the hydrangea macrophylla stem segments and stem tips into an axillary bud germination induction culture medium, and germinating the axillary buds and extending the stem tips to grow;
(3) when the axillary buds and the stem tips in the step (2) are extended to a certain length, cutting and inoculating the axillary buds and the stem tips into an axillary bud and stem tip proliferation culture medium to proliferate the axillary buds and the stem tips to obtain a large number of proliferation buds;
(4) inoculating the proliferation bud in the step (3) into a rooting culture medium for induction culture to form a tissue culture seedling;
(5) transplanting the tissue culture seedlings obtained in the step (4) into a seedling hardening matrix for culture to obtain a regeneration plant of the hydrangea macrophylla with strong roots and stems.
2. The method for rapid propagation of hydrangea macrophylla as claimed in claim 1, wherein: the disinfection treatment in the step (1) comprises the following steps: sterilizing annual young stem and stem tip of hydrangea macrophylla with alcohol for 30-60s, and sterilizing with 0.1% mercuric chloride for 8-10 min.
3. The method for rapid propagation of hydrangea macrophylla as claimed in claim 1, wherein: the culture medium for inducing the axillary bud germination in the step (2) comprises the following components: MS, 1.5-2.0 mg/L, IAA 0.3.3-0.5 mg/L of 6-BA, 30g/L of cane sugar and 6g/L of agar, and the pH value is controlled between 5.8 and 6.0.
4. The method for rapid propagation of hydrangea macrophylla as claimed in claim 3, wherein: and (3) carrying out germination culture on axillary buds and stem tips by using the axillary bud germination induction culture medium under the conditions of the temperature of 25 +/-1 ℃, the illumination intensity of 2500 Lx and the illumination time of 12 h/d.
5. The method for rapid propagation of hydrangea macrophylla as claimed in claim 1, wherein: the axillary bud and stem tip proliferation culture medium in the step (3) comprises the following components: MS, 0.1-0.5 mg/L, IBA 0.1.1-0.5 mg/L of 6-BA, 30g/L of cane sugar and 6g/L of agar, and the pH value is controlled between 5.8 and 6.0.
6. The method for rapid propagation of hydrangea macrophylla as claimed in claim 5, wherein: and performing proliferation culture of axillary buds by using the axillary bud and stem tip proliferation culture medium under the conditions of 25 +/-1 ℃, the illumination intensity of 2500 Lx and the illumination time of 12 h/d.
7. The method for rapid propagation of hydrangea macrophylla as claimed in claim 1, wherein: the rooting culture medium in the step (4) comprises the following components: 1/2MS, IBA 0.1-1.0 mg/L or 1/2MS, NAA1.0-2.0 mg/L, sucrose 30g/L, agar 6g/L, pH controlled between 5.8-6.0.
8. The method for rapid propagation of hydrangea macrophylla as claimed in claim 7, wherein: and (3) using the rooting culture medium to perform rooting induction culture of the proliferation buds under the conditions of the temperature of 23 +/-2 ℃, the illumination intensity of 1800 Lx and the illumination time of 12 h/d.
9. The method for rapid propagation of hydrangea macrophylla as claimed in claim 1, wherein: the hardening-seedling substrate in the step (5) comprises the following components: the ratio of the turfy soil to the vermiculite to the perlite is 1:1: 1.
10. The method for rapid propagation of hydrangea macrophylla as claimed in claim 9, wherein: the seedling exercising substrate is used for exercising and culturing the seedlings under the conditions that the temperature is 23 +/-2 ℃, the illumination intensity is 1800 Lx and the illumination time is 12 h/d.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202010967730.4A CN112042540A (en) | 2020-09-15 | 2020-09-15 | Rapid propagation method of hydrangea macrophylla |
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| CN202010967730.4A CN112042540A (en) | 2020-09-15 | 2020-09-15 | Rapid propagation method of hydrangea macrophylla |
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| CN112042540A true CN112042540A (en) | 2020-12-08 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110972947A (en) * | 2019-12-19 | 2020-04-10 | 美尚生态景观股份有限公司 | Culture medium and culture method for hydrangea-polar bear tissue culture |
| CN115868411A (en) * | 2023-03-03 | 2023-03-31 | 云南聚佰贤科技有限公司 | Tissue culture and rapid propagation method of hydrangea strigosa |
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|---|---|---|---|---|
| US20190059313A1 (en) * | 2017-08-28 | 2019-02-28 | Ronaldus Wilhelmus VLASVELD | Methods for producing a standard form Hydrangea plants and cuttings thereof |
| CN110972947A (en) * | 2019-12-19 | 2020-04-10 | 美尚生态景观股份有限公司 | Culture medium and culture method for hydrangea-polar bear tissue culture |
| CN111587784A (en) * | 2020-01-19 | 2020-08-28 | 内蒙古农业大学 | Rapid propagation method of hydrangea macrophylla |
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2020
- 2020-09-15 CN CN202010967730.4A patent/CN112042540A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20190059313A1 (en) * | 2017-08-28 | 2019-02-28 | Ronaldus Wilhelmus VLASVELD | Methods for producing a standard form Hydrangea plants and cuttings thereof |
| CN110972947A (en) * | 2019-12-19 | 2020-04-10 | 美尚生态景观股份有限公司 | Culture medium and culture method for hydrangea-polar bear tissue culture |
| CN111587784A (en) * | 2020-01-19 | 2020-08-28 | 内蒙古农业大学 | Rapid propagation method of hydrangea macrophylla |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110972947A (en) * | 2019-12-19 | 2020-04-10 | 美尚生态景观股份有限公司 | Culture medium and culture method for hydrangea-polar bear tissue culture |
| CN115868411A (en) * | 2023-03-03 | 2023-03-31 | 云南聚佰贤科技有限公司 | Tissue culture and rapid propagation method of hydrangea strigosa |
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Application publication date: 20201208 |