CN112772418A - Method for tissue culture and rapid propagation of dendrobium nobile flower buds - Google Patents
Method for tissue culture and rapid propagation of dendrobium nobile flower buds Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 34
- 240000004638 Dendrobium nobile Species 0.000 title claims abstract description 24
- 241000196324 Embryophyta Species 0.000 claims abstract description 34
- 241001523681 Dendrobium Species 0.000 claims abstract description 32
- 230000006698 induction Effects 0.000 claims abstract description 31
- 238000005520 cutting process Methods 0.000 claims abstract description 15
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 13
- 230000035784 germination Effects 0.000 claims abstract description 10
- 229930191978 Gibberellin Natural products 0.000 claims abstract description 7
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000003448 gibberellin Substances 0.000 claims abstract description 7
- 238000005070 sampling Methods 0.000 claims abstract description 6
- 230000000249 desinfective effect Effects 0.000 claims abstract description 5
- 239000007943 implant Substances 0.000 claims abstract description 3
- 239000001963 growth medium Substances 0.000 claims description 22
- 230000035755 proliferation Effects 0.000 claims description 17
- 239000002609 medium Substances 0.000 claims description 8
- 239000012883 rooting culture medium Substances 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 5
- 239000012882 rooting medium Substances 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- PRPINYUDVPFIRX-UHFFFAOYSA-N 1-naphthaleneacetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CC=CC2=C1 PRPINYUDVPFIRX-UHFFFAOYSA-N 0.000 description 9
- 229930006000 Sucrose Natural products 0.000 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 description 9
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 6
- NWBJYWHLCVSVIJ-UHFFFAOYSA-N N-benzyladenine Chemical compound N=1C=NC=2NC=NC=2C=1NCC1=CC=CC=C1 NWBJYWHLCVSVIJ-UHFFFAOYSA-N 0.000 description 6
- 239000000701 coagulant Substances 0.000 description 6
- 239000008223 sterile water Substances 0.000 description 6
- 230000001954 sterilising effect Effects 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 230000004083 survival effect Effects 0.000 description 5
- 230000001133 acceleration Effects 0.000 description 4
- 238000002054 transplantation Methods 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 239000004471 Glycine Substances 0.000 description 3
- 241000234295 Musa Species 0.000 description 3
- 235000018290 Musa x paradisiaca Nutrition 0.000 description 3
- 230000003044 adaptive effect Effects 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- JTEDVYBZBROSJT-UHFFFAOYSA-N indole-3-butyric acid Chemical compound C1=CC=C2C(CCCC(=O)O)=CNC2=C1 JTEDVYBZBROSJT-UHFFFAOYSA-N 0.000 description 3
- 229940064880 inositol 100 mg Drugs 0.000 description 3
- 229960002523 mercuric chloride Drugs 0.000 description 3
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 238000002791 soaking Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 230000034303 cell budding Effects 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000011031 large-scale manufacturing process Methods 0.000 description 2
- XMTQQYYKAHVGBJ-UHFFFAOYSA-N 3-(3,4-DICHLOROPHENYL)-1,1-DIMETHYLUREA Chemical compound CN(C)C(=O)NC1=CC=C(Cl)C(Cl)=C1 XMTQQYYKAHVGBJ-UHFFFAOYSA-N 0.000 description 1
- 240000007576 Dendrobium bigibbum Species 0.000 description 1
- 241001673954 Magnolia sieboldii Species 0.000 description 1
- 241000233855 Orchidaceae Species 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
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- 239000005293 duran Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 1
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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
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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- Botany (AREA)
- Environmental Sciences (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a method for tissue culture and rapid propagation of dendrobium nobile flower buds, which comprises the following steps: (1) selection and disinfection of explants: (2) induction culture of cluster buds; (3) carrying out enrichment culture on cluster buds; (4) rooting culture; (5) transplanting test-tube seedlings; wherein the specific process of selecting and disinfecting the external implant in the step (1) comprises the following steps: selecting healthy dendrobium autumn plant as explant object, topping and pinching branches of the healthy dendrobium autumn plant in the previous year, and smearing gibberellin solution on the hidden bud position of the branches of the healthy dendrobium autumn plant in the current year for accelerating germination; sampling when the hidden bud is promoted to sprout and grow to 1-2cm and the new bud has 3-5 sections; cutting stem segments with nodes and stem tips after disinfection as explants for standby; in the enrichment culture of the cluster buds in the step (3), the culture period of the enrichment culture and the culture period of the subculture are both 25-35 d. The method can improve propagation speed, and is convenient for high-efficiency and large-scale tissue culture production.
Description
Technical Field
The invention relates to the technical field of tissue culture and rapid propagation, in particular to a method for tissue culture and rapid propagation of dendrobium nobile flower buds.
Background
The dendrobium nobile lindl is evergreen dendrobium nobile, is a plant of dendrobium genus of orchidaceae family, and is also called butterfly dendrobium nobile, duran, dendrobium nobile, magnolia sieboldii and the like. The dendrobium nobile lindl is a flower popular in the China cut flower market in recent years, has various varieties, beautiful flower shape, bright color, long flowering phase and higher ornamental value, increases the market demand of cut flowers and potted flowers year by year in China, and has urgent demand on high-quality seedlings.
At present, domestic and foreign researches on the fast propagation technology of dendrobium autumn are mainly focused on using stem buds as explants to propagate separate seedlings or germinating seeds to obtain seedlings. However, stem buds are used as explants, browning is easily caused, the pollution incidence rate is high, the character separation degree of the progeny of the dendrobium nobile cultivated by seeds is high, the difference between individuals is large, and plants with consistent characters are difficult to realize.
In Chinese patent with application number CN200410077729.5 entitled "tissue culture and rapid propagation method of high quality seedling of Dendrobium sp", it is proposed that a new germ of Dendrobium sp is used as an explant, and a culture medium is used for carrying out cluster bud proliferation and obtaining the seedling; the method establishes a propagation technology taking the germ plasm new bud of the dendrobium nobile as an explant, but the propagation efficiency and the transplanting survival rate are still not ideal, and the defect of relatively complicated operation process exists.
The invention provides a propagation method for subsequent rooting culture without strong seedling culture after propagation in Chinese patent with application number of CN201710282269.7 and name of 'a tissue culture rapid propagation method of dendrobium autumn' seedlings, and the method overcomes the defects of relatively complicated seedling culture operation process, low efficiency, poor resistance of tissue culture seedlings and low transplanting survival rate of dendrobium autumn seedlings in the prior art. However, in the propagation method, the word propagation culture of the cluster buds still needs a longer culture period, so that the propagation speed is slow, and the mass tissue culture is not facilitated.
Disclosure of Invention
The invention aims to solve the technical problem of providing a method for tissue culture and rapid propagation of dendrobium nobile flower buds, improving the propagation speed and facilitating efficient and large-scale tissue culture production.
In order to solve the technical problems, the technical scheme of the invention is as follows:
a method for tissue culture and rapid propagation of dendrobium nobile flower buds comprises the following steps:
(1) selecting and disinfecting explants;
(2) induction culture of cluster buds: inoculating the explant sterilized in the step (1) into a cluster bud induction culture medium for cluster bud induction culture;
(3) and (3) propagation culture of the cluster buds: cutting cluster buds obtained by induction in the step (2) into cluster bud groups with 3-6 buds, and then inoculating the cluster bud groups into a proliferation culture medium for proliferation culture; then, replacing a new multiplication culture medium for carrying out subculture multiplication culture for 1-2 times, and then continuously culturing for 10-15 days to obtain robust cluster buds;
(4) rooting culture: cutting the robust cluster buds obtained in the step (3) into single buds, and inoculating the single buds on a rooting culture medium for culture;
(5) transplanting test-tube seedlings: hardening the bottle seedlings obtained in the step (4), and transplanting;
particularly, the specific process of selecting and disinfecting the external implant in the step (1) comprises the following steps: selecting healthy dendrobium autumn plant as explant object, topping and pinching branches of the healthy dendrobium autumn plant in the previous year, and smearing gibberellin solution on the hidden bud position of the branches of the healthy dendrobium autumn plant in the current year for accelerating germination; sampling when the hidden bud is promoted to sprout and grow to 1-2cm and the new bud has 3-5 sections; after disinfection, the stem segment with node and the stem tip are cut out as explant for further use.
In the enrichment culture of the cluster buds in the step (3), the culture period of the enrichment culture and the culture period of the subculture are both 25-35 d.
Preferably, in the step (1), the branches of the healthy dendrobium nobile mother plant are subjected to girdling treatment while topping and pinching are performed.
Preferably, the cluster bud induction medium, the multiplication medium and the rooting medium all contain 0.1-0.5 g/LPVP.
Preferably, the cluster bud induction medium, the multiplication medium and the rooting medium all contain 0.3-0.4 g/LPVP.
Preferably, the size of the clump in step (3) is 0.4-0.7 cm.
Preferably, the size of the single bud in the step (4) is 1.8-2.5 cm.
According to the technical scheme, the healthy dendrobium autumn-dendrobium mother plant with vigorous growth is selected as an explant object, and branches of the healthy dendrobium autumn-dendrobium mother plant are subjected to topping and pinching treatment in the previous year, or subjected to girdling treatment and gibberellin solution smearing on the branches of the healthy dendrobium autumn-dendrobium mother plant after topping and pinching treatment, so that the hidden bud germination of the branches of the healthy dendrobium autumn-dendrobium mother plant is promoted, and an ideal explant is obtained.
The invention has the beneficial effects that:
according to the technical scheme, when the dendrobium autumn explant is selected, the hidden bud on the dendrobium autumn branch is subjected to germination acceleration treatment, and a new bud after the germination acceleration treatment and final germination is used as a tissue culture object; in the prior art, when the dendrobium nobile explant is selected, the germinated new bud is directly selected as a tissue culture object; the novel bud after the hidden bud germination acceleration is selected as an explant tissue culture object, the culture period of the process is obviously shortened in the propagation culture process of the cluster buds, the increment coefficient is as high as 6.0-6.6, and the robust cluster buds can be obtained in a short period, so that the culture time is shortened for the whole propagation culture process, the propagation culture efficiency is improved, and the efficient large-scale production is facilitated.
Detailed Description
The following examples further illustrate embodiments of the present invention. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
Example 1
A method for tissue culture and rapid propagation of dendrobium nobile flower buds comprises the following steps:
(1) selection and disinfection of explants: selecting healthy dendrobium autumn plant as explant object, topping and pinching branches of the healthy dendrobium autumn plant in the previous year, and smearing gibberellin solution on the hidden bud position of the branches of the healthy dendrobium autumn plant in the current year for accelerating germination; sampling when the hidden bud is promoted to sprout and grow to 1-2cm and the new bud has 3-5 sections; cleaning with sterile water, sequentially soaking in alcohol and mercuric chloride solution for sterilization, and then washing with sterile water, and cutting stem segments with nodes and stem tips as explants for later use;
(2) induction culture of cluster buds: inoculating the explant subjected to the sterilization treatment in the step (1) into a cluster bud induction culture medium for cluster bud induction culture; after culturing for 15-23d, cluster buds are induced continuously, and the bud induction rate is 84%; wherein, the cluster bud induction culture medium comprises the following components: MS + 6-benzylaminopurine +3.0mg/L + naphthylacetic acid 0.2mg/L + white sugar 30g/L + agar powder 5.0 mg/L.
(3) And (3) propagation culture of the cluster buds: cutting the cluster buds obtained by induction in the step (2) into cluster bud groups with 3-6 buds, wherein the size of the cluster bud groups is 0.4-0.7cm, and then inoculating the cluster bud groups into a proliferation culture medium for proliferation culture for 35 days, wherein the proliferation coefficient reaches 6.0-6.6; then, replacing a new multiplication culture medium to carry out subculture multiplication culture for 1 time, wherein the culture period is 35d, and obtaining robust cluster buds; then continuing culturing for 10 days to obtain robust cluster buds; wherein, the components of the proliferation culture medium are as follows: MS + white sugar 30000mg/L + coagulant 6300mg/L + 6-benzylaminopurine 2.0mg/L + naphthylacetic acid 0.1mg/L + PVP0.5g/L.
(4) Rooting culture: cutting the robust cluster buds obtained in the step (3) into single buds, wherein the size of the single buds is 1.8-2.5cm, and inoculating the single buds on a rooting culture medium for culture; the complete plant with the plant height of 6.5-8.0cm, the root length of 2.8-3.7cm and the root number of 4-6 is obtained in the culture period of 67d, and the rooting rate is 100.0 percent; wherein, the components of the rooting culture medium are as follows: MS + glycine 2.0mg/L + inositol 100mg/L + white sugar 20000mg/L + coagulant 6300mg/L + indolebutyric acid 0.3mg/L + naphthylacetic acid 0.1mg/L + active carbon 500mg/L + and banana puree 50000 mg/L.
(5) Transplanting test-tube seedlings: and (5) hardening the bottle seedlings obtained in the step (4) in a greenhouse until the bottle seedlings are adaptive to a cultivation environment, and transplanting. The statistical survival rate of transplanting 2 months after transplanting is 98%.
Example 2
A method for tissue culture and rapid propagation of dendrobium nobile flower buds comprises the following steps:
(1) selection and disinfection of explants: selecting healthy dendrobium autumn mother plants with vigorous growth as explant objects, topping and pinching branches of the healthy dendrobium autumn mother plants in the previous year, simultaneously performing girdling treatment on the branches of the healthy dendrobium autumn mother plants, and smearing gibberellin solution on the non-budding positions of the branches of the healthy dendrobium autumn mother plants in the current year for pregermination; sampling when the hidden bud is promoted to sprout and grow to 1-2cm and the new bud has 3-5 sections; cleaning with sterile water, sequentially soaking in alcohol and mercuric chloride solution for sterilization, and then washing with sterile water, and cutting stem segments with nodes and stem tips as explants for later use;
(2) induction culture of cluster buds: inoculating the explant subjected to the sterilization treatment in the step (1) into a cluster bud induction culture medium for cluster bud induction culture; cluster buds are induced continuously after 22 days of culture, and the bud induction rate is 83%; wherein, the cluster bud induction culture medium comprises the following components: MS + 6-benzylaminopurine +3.0mg/L + naphthylacetic acid 0.2mg/L + white sugar 30g/L + agar powder 5.0mg/L + PVP0.3g/L.
(3) And (3) propagation culture of the cluster buds: cutting the cluster buds obtained by induction in the step (2) into cluster bud groups with 3-6 buds, wherein the size of the cluster bud groups is 0.4-0.7cm, and then inoculating the cluster bud groups into a proliferation culture medium for proliferation culture for 30 days, wherein the proliferation coefficient reaches 6.0-6.6; then, replacing a new multiplication culture medium to carry out subculture multiplication culture for 1 time, wherein the culture period is 30d, and obtaining robust cluster buds; then continuing to culture for 15d to obtain robust cluster buds; wherein, the components of the proliferation culture medium are as follows: MS + white sugar 30000mg/L + coagulant 6300mg/L + 6-benzylaminopurine 2.0mg/L + naphthylacetic acid 0.1mg/L + PVP0.3g/L.
(4) Rooting culture: cutting the robust cluster buds obtained in the step (3) into single buds, wherein the size of the single buds is 1.8-2.5cm, and inoculating the single buds on a rooting culture medium for culture; the complete plant with the plant height of 6.5-8.0cm, the root length of 2.8-3.8cm and the root number of 4-6 is obtained after the culture period is 65d, and the rooting rate is 100.0 percent; wherein, the components of the rooting culture medium are as follows: MS + glycine 2.0mg/L + inositol 100mg/L + white sugar 20000mg/L + coagulant 6300mg/L + indolebutyric acid 0.3mg/L + naphthylacetic acid 0.1mg/L + active carbon 500mg/L + and banana puree 50000 mg/L.
(5) Transplanting test-tube seedlings: and (5) hardening the bottle seedlings obtained in the step (4) in a greenhouse until the bottle seedlings are adaptive to a cultivation environment, and transplanting. The statistical survival rate of transplantation is 99% 2 months after transplantation.
Example 3
A method for tissue culture and rapid propagation of dendrobium nobile flower buds comprises the following steps:
(1) selection and disinfection of explants: selecting healthy dendrobium autumn mother plants with vigorous growth as explant objects, topping and pinching branches of the healthy dendrobium autumn mother plants in the previous year, simultaneously performing girdling treatment on the branches of the healthy dendrobium autumn mother plants, and smearing gibberellin solution on the non-budding positions of the branches of the healthy dendrobium autumn mother plants in the current year for pregermination; sampling when the hidden bud is promoted to sprout and grow to 1-2cm and the new bud has 3-5 sections; cleaning with sterile water, sequentially soaking in alcohol and mercuric chloride solution for sterilization, and then washing with sterile water, and cutting stem segments with nodes and stem tips as explants for later use;
(2) induction culture of cluster buds: inoculating the explant subjected to the sterilization treatment in the step (1) into a cluster bud induction culture medium for cluster bud induction culture; after 20 days of culture, cluster buds are induced continuously, and the bud induction rate is 81 percent; wherein, the cluster bud induction culture medium comprises the following components: MS + 6-benzylaminopurine +3.0mg/L + naphthylacetic acid 0.2mg/L + white sugar 30g/L + agar powder 5.0mg/L + PVP0.3g/L.
(3) And (3) propagation culture of the cluster buds: cutting the cluster buds obtained by induction in the step (2) into cluster bud groups with 3-6 buds, wherein the size of the cluster bud groups is 0.4-0.7cm, and then inoculating the cluster bud groups into a proliferation culture medium for proliferation culture for 25 days, wherein the proliferation coefficient reaches 6.0-6.6; then, replacing a new multiplication culture medium to carry out subculture multiplication culture for 1 time, wherein the culture period is 35d, and obtaining robust cluster buds; then continuing culturing for 18d to obtain robust cluster buds; wherein, the components of the proliferation culture medium are as follows: MS + white sugar 30000mg/L + coagulant 6300mg/L + 6-benzylaminopurine 2.0mg/L + naphthylacetic acid 0.1mg/L + PVP0.3g/L.
(4) Rooting culture: cutting the robust cluster buds obtained in the step (3) into single buds, wherein the size of the single buds is 1.8-2.5cm, and inoculating the single buds on a rooting culture medium for culture; the whole plant with the plant height of 7.0-8.0cm, the root length of 3.0-4.0cm and the root number of 4-6 is obtained after the culture period of 70d, and the rooting rate is 100.0 percent; wherein, the components of the rooting culture medium are as follows: MS + glycine 2.0mg/L + inositol 100mg/L + white sugar 20000mg/L + coagulant 6300mg/L + indolebutyric acid 0.3mg/L + naphthylacetic acid 0.1mg/L + active carbon 500mg/L + and banana puree 50000 mg/L.
(5) Transplanting test-tube seedlings: and (5) hardening the bottle seedlings obtained in the step (4) in a greenhouse until the bottle seedlings are adaptive to a cultivation environment, and transplanting. The statistical survival rate of transplantation is 99% 2 months after transplantation.
In conclusion, the novel bud after the hidden bud germination acceleration is selected as the explant tissue culture object, the culture period of the process is obviously shortened in the propagation culture process of the cluster buds, the propagation coefficient is as high as 6.0-6.6, and the robust cluster buds can be obtained in a short period, so that the culture time is shortened for the whole propagation culture process, the propagation culture efficiency is improved, and the efficient large-scale production is facilitated.
The embodiments of the present invention have been described in detail with reference to the examples, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, and the scope of protection is still within the scope of the invention.
Claims (6)
1. A method for tissue culture and rapid propagation of dendrobium nobile flower buds comprises the following steps:
(1) selecting and disinfecting explants;
(2) induction culture of cluster buds: inoculating the explant sterilized in the step (1) into a cluster bud induction culture medium for cluster bud induction culture;
(3) and (3) propagation culture of the cluster buds: cutting cluster buds obtained by induction in the step (2) into cluster bud groups with 3-6 buds, and then inoculating the cluster bud groups into a proliferation culture medium for proliferation culture; then, replacing a new multiplication culture medium for carrying out subculture multiplication culture for 1-2 times, and then continuously culturing for 10-15 days to obtain robust cluster buds;
(4) rooting culture: cutting the robust cluster buds obtained in the step (3) into single buds, and inoculating the single buds on a rooting culture medium for culture;
(5) transplanting test-tube seedlings: hardening the bottle seedlings obtained in the step (4), and transplanting;
the method is characterized in that: the specific process of selecting and disinfecting the external implant in the step (1) is as follows: selecting healthy dendrobium autumn plant as explant object, topping and pinching branches of the healthy dendrobium autumn plant in the previous year, and smearing gibberellin solution on the hidden bud position of the branches of the healthy dendrobium autumn plant in the current year for accelerating germination; sampling when the hidden bud is promoted to sprout and grow to 1-2cm and the new bud has 3-5 sections; cutting stem segments with nodes and stem tips after disinfection as explants for standby;
in the enrichment culture of the cluster buds in the step (3), the culture period of the enrichment culture and the culture period of the subculture are both 25-35 d.
2. The method for tissue culture and rapid propagation of flower buds of dendrobium nobile according to claim 1, wherein in the step (1), the branches of the healthy mother plant of dendrobium nobile are subjected to girdling treatment while topping and pinching are performed.
3. The method for tissue culture and rapid propagation of floral buds of dendrobium nobile as claimed in claim 1 or 2, wherein the multiple bud induction medium, the propagation medium and the rooting medium all contain 0.1-0.5 g/LPVP.
4. The method for tissue culture and rapid propagation of floral buds of dendrobium nobile as claimed in claim 3, wherein the multiple shoot induction medium, the enrichment medium and the rooting medium all contain 0.3-0.4 g/LPVP.
5. The method for tissue culture and rapid propagation of flower buds of dendrobium nobile as claimed in claim 1, wherein the size of the clump bud mass in step (3) is 0.4-0.7 cm.
6. The tissue culture and rapid propagation method for dendrobium nobile flower buds according to claim 1, wherein the size of the single bud in the step (4) is 1.8-2.5 cm.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117084173A (en) * | 2023-09-19 | 2023-11-21 | 中国热带农业科学院热带作物品种资源研究所 | Method for culturing dendrobium candidum tissue culture seedlings by using tissue culture bags |
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| US20070006338A1 (en) * | 2005-05-16 | 2007-01-04 | Fabbri Bradon J | Corn plants and seed enhanced for asparagine and protein |
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| CN117084173B (en) * | 2023-09-19 | 2024-08-20 | 中国热带农业科学院热带作物品种资源研究所 | Method for culturing dendrobium candidum tissue culture seedlings by using tissue culture bags |
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