CN106665355A - Thunia alba tissue culture seedling culture method - Google Patents
Thunia alba tissue culture seedling culture method Download PDFInfo
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- CN106665355A CN106665355A CN201611157500.1A CN201611157500A CN106665355A CN 106665355 A CN106665355 A CN 106665355A CN 201611157500 A CN201611157500 A CN 201611157500A CN 106665355 A CN106665355 A CN 106665355A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
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- Botany (AREA)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention discloses a thunia alba tissue culture seedling culture method, which comprises the following steps of (1) culture medium preparation; (2) explants preparation; (3) inoculation culture; (4) domestication. By using the method provided by the invention for thunia alba tissue culture seedling culture, the germination rate reaches 90 percent or higher; the breeding coefficient reaches 3 to 5; the characteristics of high breeding coefficient and high breeding speed are realized; in addition, the bred seedlings have the plant features identical to the parent body; the resistance is high. The problem of difficult breeding of the thunia alba is greatly solved. The scaled and factory production of the thunia alba are facilitated.
Description
Technical field
The present invention relates to plant tissue culture seedlings-raising technical field, and in particular to a kind of Herba Thuniae Albae tissue cultivating and seedling method.
Background technology
Herba Thuniae Albae, is the herb of orchid Radix Crotalariae szemoensis orchid Thunia alba (Lindl.) Rchb.f., also known as logical orchid, thrum,
Dianthus chinensis, rock bamboo.Perennial draft of growing nonparasitically upon another plant, high about 30~50 centimetres of fibrous root majorities, meat shape, canescence or light brown.Rock
It is Radix Crotalariae szemoensis sweet in the mouth, mild-natured, according to《Yunnan Chinese herbal medicine》、《Guizhou medicine plants catalogue》With《Simao, Yunnan Chinese herbal medicine is selected》Record, it has cough-relieving
Relieving asthma, the effect such as blood circulation promoting and blood stasis dispelling, for treating pneumonia, trachea and bronchitis, gastric and duodenal ulcers, fracture and traumatic injury labor
Wound etc..Herba Thuniae Albae has very high medical value, there is huge market prospect.Because Herba Thuniae Albae seed has, the tiny amount of seed is big, nothing
Endosperm, needs to be sprouted with suitable mycorrhizal fungi under natural conditions, natural environment destruction and artificially excessively excavation, is adding
Germination Condition is harsh, causes wild resource extremely difficult in original Habitat pattern.
At present, also without a kind of method with regard to Herba Thuniae Albae tissue cultivating and seedling, going out for artificial growth and artificial breeding is had no
It is existing, scale breeding and the plantation of Herba Thuniae Albae are constrained to a certain extent.
The content of the invention
The technical problem to be solved in the present invention is：A kind of Herba Thuniae Albae tissue cultivating and seedling method is provided, Herba Thuniae Albae tissue cultured seedling is improved
The survival rate of cultivation, it is cost-effective, provide key technology for the production of Herba Thuniae Albae scale breeding.
The purpose of the present invention is achieved through the following technical solutions：
A kind of Herba Thuniae Albae tissue cultivating and seedling method, comprises the following steps：
（1）It is prepared by culture medium：Culture medium includes induction germination medium, proliferated culture medium and root media；Wherein, induction is sprouted
Send out culture medium and spend precious No. 1 0.7g/L+ banana puree 30g/L+ mashed potatoes 20g/L+ sucrose for MS+NAA0.2mg/L+KT0.5mg/L+
30g/L+ agar 5g/L+PH5.9；Proliferated culture medium is MS+IBA0.5mg/L+6-BA0.5mg/L+ sucrose 30g/L+ agar 5g/L
+PH5.9；Root media is 2/3MS+NAA0.8mg/L+IAA0.5mg/L+ sucrose 30g/L+ activated carbon 0.5g/L+ agar 5g/
L+ph5.9；By the induction germination medium for preparing with every bottle of 60ml, every bottle of 120ml of root media is fitted in culture bottle, adopts
Carried out with 121 DEG C, 0.5Mpa standby after cooled and solidified after sterilizing 25min；
（2）It is prepared by explant：Autumn plucks full outstanding achievement as explant on ripe plant, and is cleaned, sterilized；
（3）Inoculated and cultured：Inoculation utensil is sterilized, after both hands sterilization, by step（2）In the explant for preparing ultra-clean
In workbench, outstanding achievement is cut by sterile working's requirement, seed therein is uniformly sprinkled in induction germination medium, covered
Being positioned over after bottle cap in the standard culture room after sterilization carries out induction sprouting culture, accesses after emerging and is given birth in root media
Root culture 2 ~ 3 months；
（4）Domestication：Root culture is obtained into seedling to take out, soaked with 1000 times of carbendazim plant after flowing water is rinsed after 10min in
In peat soil, perlite, wood sawdust mixed-matrix, the domestication initial stage is sheltered from heat or light 10 days with 3 pin shade nets, and domestication can transplant big for 60 days
Field.
Preferably, step（2）The step of cleaning of middle explant, sterilization is：Rinse well after being steeped 30 minutes with soap water enchroachment (invasion)
Superficial stain, then flowing water flushing at least 30min, with pocket knife by the scar removal in outstanding achievement, after surface moisture evaporation, is surpassing
5 ~ 10min of bubble is invaded with 75% ethanol on net workbench, outstanding achievement is constantly rocked halfway, then with using nothing after aseptic water washing 3 ~ 5 times
Bacterium tweezers take out and are placed on concentration to invade 10 ~ 15min of bubble in 0.1 ~ 0.2% mercuric chloride, and outstanding achievement is constantly rocked in midway, then uses sterilized water
Taken out with aseptic nipper after rinsing 5 ~ 8 times and be placed on the inoculation disk for being lined with aseptic absorbent paper, you can.
Preferably, step（3）In, induction sprouting culture, the concrete operations of root culture are carried out in culturing room is：In training
Foster interior carries out induction sprouts culture includes light culture and optical culture, and temperature control is in 18 ~ 22 DEG C, humidity 40- during light culture
50%, light culture proceeds to afterwards optical culture for 7 ~ 10 days, and optical culture illumination 3500LUX is daily 8 hours；Treat that culture height of seedling 2 is sprouted in induction
~ 3cm, Seedling of the stalk with diameter greater than 0.3cm is transferred by sterility requirements in superclean bench and taken root in root media
Culture, root culture temperature is 25 DEG C, and illumination 3500LUX is daily 8 hours；Other seedling can go in proliferated culture medium and expand numerous
Grow.
Preferably, step（4）In, peat soil, perlite, the volume ratio of wood sawdust three are 4：1：1, it is mixed to obtain
Planting matrix PH6.5 ~ 6.8.
Beneficial effects of the present invention：Herba Thuniae Albae tissue cultivating and seedling is carried out with the present invention, germination rate reaches more than 90%, numerous
Grow coefficient and reach 3 ~ 5, with breeding coefficient it is high, reproduction speed is fast the characteristics of, and the seedling for breeding out has and parent phase
Same plant characteristics, better resistance.The difficult problem of Herba Thuniae Albae breeding is significantly solved, is conducive to Herba Thuniae Albae to carry out scale, batch production
Production.
Specific embodiment
Invention is described further with reference to specific embodiment：
A kind of Herba Thuniae Albae tissue cultivating and seedling method, comprises the following steps：
（1）It is prepared by culture medium：Culture medium includes induction germination medium, proliferated culture medium and root media；Wherein, induction is sprouted
Send out culture medium and spend precious No. 1 0.7g/L+ banana puree 30g/L+ mashed potatoes 20g/L+ sucrose for MS+NAA0.2mg/L+KT0.5mg/L+
30g/L+ agar 5g/L+PH5.9；Proliferated culture medium is MS+IBA0.5mg/L+6-BA0.5mg/L+ sucrose 30g/L+ agar 5g/L
+PH5.9；Root media is 2/3MS+NAA0.8mg/L+IAA0.5mg/L+ sucrose 30g/L+ activated carbon 0.5g/L+ agar 5g/
L+ph5.9；By the induction germination medium for preparing with every bottle of 60ml, every bottle of 120ml of root media is fitted in culture bottle, adopts
Carried out with 121 DEG C, 0.5Mpa standby after cooled and solidified after sterilizing 25min.
（2）It is prepared by explant：Autumn plucks full outstanding achievement as explant on ripe plant, and is cleaned, disappeared
Poison, step is：Superficial stain is rinsed well after being steeped 30 minutes with soap water enchroachment (invasion), then flowing water rinses at least 30min, will with pocket knife
Scar removal in outstanding achievement, after surface moisture evaporation, 5 ~ 10min of bubble is invaded on superclean bench with 75% ethanol, and midway is not
It is disconnected to rock outstanding achievement, then concentration is placed on to invade in 0.1 ~ 0.2% mercuric chloride with being taken out with aseptic nipper after aseptic water washing 3 ~ 5 times
Outstanding achievement is constantly rocked in 10 ~ 15min of bubble, midway, then with taken out with aseptic nipper after aseptic water washing 5 ~ 8 times be placed on be lined with it is aseptic
On the inoculation disk of absorbent paper, you can.
（3）Inoculated and cultured：Inoculation utensil is sterilized, after both hands sterilization, by step（2）In the explant for preparing exist
In superclean bench, outstanding achievement is cut by sterile working's requirement, seed therein is uniformly sprinkled in induction germination medium,
Covering carries out induction sprouting culture, root culture in the standard culture room being positioned over after bottle cap after sterilization, concrete operations are as follows：
Induction sprouting culture, the concrete operations of root culture are carried out in culturing room is：Carrying out induction sprouting culture in culture interior includes
Light culture and optical culture, in 18 ~ 22 DEG C, humidity 40-50%, light culture proceeds to afterwards light training for 7 ~ 10 days to temperature control during light culture
Support, optical culture illumination 3500LUX is daily 8 hours；Treat that culture 2 ~ 3cm of height of seedling is sprouted in induction, Seedling of the stalk with diameter greater than 0.3cm exists
Transfer by sterility requirements in superclean bench and carry out root culture in root media, root culture temperature is 25 DEG C, illumination
3500LUX is daily 8 hours；Other seedling can go to expanding propagation in proliferated culture medium.
（4）Domestication：Root culture is obtained into seedling to take out, is soaked after flowing water is rinsed after 10min with 1000 times of carbendazim and is planted
Plant in peat soil, perlite, wood sawdust mixed-matrix, peat soil, perlite, the volume ratio of wood sawdust three are 4：1：1, institute
Planting matrix PH6.5 ~ 6.8 being mixed to get, the domestication initial stage is sheltered from heat or light 10 days with 3 pin shade nets, and domestication can transplant land for growing field crops in 60 days.
Above content is to combine specific preferred implementation further description made for the present invention, it is impossible to assert
The present invention be embodied as be confined to these explanations.For general technical staff of the technical field of the invention,
On the premise of without departing from present inventive concept, some simple deduction or replace can also be made, should all be considered as belonging to the present invention's
Protection domain.
Claims (4)
1. a kind of Herba Thuniae Albae tissue cultivating and seedling method, it is characterised in that comprise the following steps：
（1）It is prepared by culture medium：Culture medium includes induction germination medium, proliferated culture medium and root media；Wherein, induction is sprouted
Send out culture medium and spend precious No. 1 0.7g/L+ banana puree 30g/L+ mashed potatoes 20g/L+ sucrose for MS+NAA0.2mg/L+KT0.5mg/L+
30g/L+ agar 5g/L+PH5.9；Proliferated culture medium is MS+IBA0.5mg/L+6-BA0.5mg/L+ sucrose 30g/L+ agar 5g/L
+PH5.9；Root media is 2/3MS+NAA0.8mg/L+IAA0.5mg/L+ sucrose 30g/L+ activated carbon 0.5g/L+ agar 5g/
L+ph5.9；By the induction germination medium for preparing with every bottle of 60ml, every bottle of 120ml of root media is fitted in culture bottle, adopts
Carried out with 121 DEG C, 0.5Mpa standby after cooled and solidified after sterilizing 25min；
（2）It is prepared by explant：Autumn plucks full outstanding achievement as explant on ripe plant, and is cleaned, sterilized；
（3）Inoculated and cultured：Inoculation utensil is sterilized, after both hands sterilization, by step（2）In the explant for preparing ultra-clean
In workbench, outstanding achievement is cut by sterile working's requirement, seed therein is uniformly sprinkled in induction germination medium, covered
Being positioned over after bottle cap in the standard culture room after sterilization carries out induction sprouting culture, accesses after emerging and is given birth in root media
Root culture 2 ~ 3 months；
（4）Domestication：Root culture is obtained into seedling to take out, soaked with 1000 times of carbendazim plant after flowing water is rinsed after 10min in
In peat soil, perlite, wood sawdust mixed-matrix, the domestication initial stage is sheltered from heat or light 10 days with 3 pin shade nets, and domestication can transplant big for 60 days
Field.
2. a kind of Herba Thuniae Albae tissue cultivating and seedling method according to claim 1, it is characterised in that：Step（2）Middle explant
Cleaning, sterilization the step of be：Superficial stain is rinsed well after being steeped 30 minutes with soap water enchroachment (invasion), then flowing water rinses at least 30min,
With pocket knife by the scar removal in outstanding achievement, after surface moisture evaporation, on superclean bench with 75% ethanol invade bubble 5 ~
Outstanding achievement is constantly rocked in 10min, midway, then with taken out with aseptic nipper after aseptic water washing 3 ~ 5 times be placed on concentration for 0.1 ~
Invade 10 ~ 15min of bubble in 0.2% mercuric chloride, outstanding achievement is constantly rocked in midway, then put with being taken out with aseptic nipper after aseptic water washing 5 ~ 8 times
Put and be lined with the inoculation disk of aseptic absorbent paper, you can.
3. a kind of Herba Thuniae Albae tissue cultivating and seedling method according to claim 1, it is characterised in that：Step（3）In, in culture
Induction sprouting culture, the concrete operations of root culture are carried out in room is：Induction is carried out in culture interior sprout culture including dark training
Support and optical culture, in 18 ~ 22 DEG C, humidity 40-50%, light culture proceeds to afterwards optical culture, light in 7 ~ 10 days to temperature control during light culture
Illumination 3500LUX is daily 8 hours for culture；Treat that culture 2 ~ 3cm of height of seedling is sprouted in induction, Seedling of the stalk with diameter greater than 0.3cm is ultra-clean
Transfer by sterility requirements in workbench and carry out root culture in root media, root culture temperature is 25 DEG C, illumination
3500LUX is daily 8 hours；Other seedling can go to expanding propagation in proliferated culture medium.
4. a kind of Herba Thuniae Albae tissue cultivating and seedling method according to claim 1, it is characterised in that：Step（4）In, peat
Soil, perlite, the volume ratio of wood sawdust three are 4：1：1, mixed planting matrix PH6.5 ~ 6.8 for obtaining.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611157500.1A CN106665355B (en) | 2016-12-15 | 2016-12-15 | A kind of rock bamboo shoot tissue cultivating and seedling method |
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|---|---|---|---|
| CN201611157500.1A CN106665355B (en) | 2016-12-15 | 2016-12-15 | A kind of rock bamboo shoot tissue cultivating and seedling method |
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| CN106665355B CN106665355B (en) | 2018-10-02 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107466858A (en) * | 2017-09-25 | 2017-12-15 | 贵州大学 | A kind of bamboo shoot orchid species seedling rapid propagation method |
| CN115644059A (en) * | 2022-10-08 | 2023-01-31 | 贵州省植物园（贵州省园林科学研究所、贵州省植物研究所） | Tissue culture and rapid propagation method for common spider moss |
-
2016
- 2016-12-15 CN CN201611157500.1A patent/CN106665355B/en active Active
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107466858A (en) * | 2017-09-25 | 2017-12-15 | 贵州大学 | A kind of bamboo shoot orchid species seedling rapid propagation method |
| CN107466858B (en) * | 2017-09-25 | 2019-12-03 | 贵州大学 | A kind of bamboo shoot orchid species seedling rapid propagation method |
| CN115644059A (en) * | 2022-10-08 | 2023-01-31 | 贵州省植物园（贵州省园林科学研究所、贵州省植物研究所） | Tissue culture and rapid propagation method for common spider moss |
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Effective date of registration: 20210317 Address after: 211111 No. 12, Mazhou East Road, Mau Ling Street, Jiangning District, Nanjing, Jiangsu Patentee after: JIANGSU ZHIJU INTELLECTUAL PROPERTY SERVICE Co.,Ltd. Address before: No. 705, No. 2 Building, Guantang R&D Center, No. 10 Shuangren Road, Liudong New District, Liuzhou City, Guangxi Zhuang Autonomous Region (Trusted by Liuzhou Gaochuang Business Secretary Co., Ltd.) Patentee before: Liuzhou Shuangcheng Technology Co.,Ltd. |