CN105941156B - A kind of oil tissue culture medium (TCM) and its cultural method of tree peony - Google Patents
A kind of oil tissue culture medium (TCM) and its cultural method of tree peony Download PDFInfo
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- CN105941156B CN105941156B CN201610385679.XA CN201610385679A CN105941156B CN 105941156 B CN105941156 B CN 105941156B CN 201610385679 A CN201610385679 A CN 201610385679A CN 105941156 B CN105941156 B CN 105941156B
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- explant
- tree peony
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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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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
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Abstract
The invention discloses it is a kind of oil use tree peony tissue culture medium (TCM), including：Improve MS+0.5~0.8mgL‑1IAA+0.3~0.5mgL‑1IBA+0.5~0.6mgL‑1The gL of NAA+0.2~0.3‑1Activated carbon+2~3gL‑1Sucrose, pH=5.6~5.8.The present invention discloses its cultural methods.Step of the present invention is simple, it is easily operated, by improveing the supplementary measures such as minimal medium and ingredient, adjustment condition of culture, so that the bud ratio of test tube seedling has been more than 98%, and transplanting shoot survival percent is high, growth is fast, substantially reduces the cycle that oil is cultivated with tree peony, it reduces production and rises this, improve the economic value with tree peony.
Description
Technical field
The present invention relates to the tissue culture medium (TCM)s and its cultural method that a kind of oil uses tree peony, belong to phytobiology technology neck
Domain.
Background technology
Oil is a kind of emerging woody oleiferous plants crop with tree peony, possesses the characteristics of prominent " three high and one low ":High production (five
Year raw per mu yield is up to 300 kilograms, and mu comprehensive benefit is up to ten thousand yuan), high oil content (seed oil content 22%), high-quality (unsaturated lipid
Fat acid content 92%), (oil is suitble to green barren hill afforestation, underwood planting with the drought-enduring impoverishment tolerant of tree peony to low cost;1 year kind a century
It receives, at low cost).The oil being currently used in commodity production only has two kinds of Feng Dan and purple plague purpura with tree peony, and is to be suitble to China absolutely
Most of ecological region planting, high yield seed the oil original seed of tree peony, wherein, phoenix pellet seed oil-containing more than 22%, higher than domestic soybean 17.5%
Oil content, while peony seed oil also contain numerous medicinal tree peony active ingredient.
Tree peony can be bred by the methods of sowing, plant division, grafting, cuttage, press strip and tissue culture.Oil is generally adopted with tree peony
It is carried out with the method for seed propagation, seed tree seedling generally only has terminal bud growth then, and current increment is small, only 5-10cm.Oil
It is fruit with the economic position of tree peony, and tree peony is Dan Shengzhi tops and as a result, therefore oil uses the hair branch quantity of tree peony to yield
Formation is of great significance.From rootstock, terminal bud is grown 1 year raw tree peony seedling there are one general upwards, and current growth length
Only 5-10cm, to there is a pair of of kryptoblast at rootstock, if will not be sprouted without manual intervention.
The content of the invention
To solve the above-mentioned problems, it is an object of the invention to provide a kind of step is simple, germination rate is high and grows up soon
The oil tissue culture medium (TCM) and its cultural method of tree peony.
In order to solve the above technical problems, the technical scheme is that：
A kind of oil tissue culture medium (TCM) of tree peony, including：Improve MS+0.5~0.8mgL-1IAA+0.3~0.5mgL-1IBA+0.5~0.6mgL-1The gL of NAA+0.2~0.3-1Activated carbon+2~3gL-1Sucrose, pH=5.6~5.8.
Further, above-mentioned improvement MS includes grand nutrition element, micronutrient element and organic reagent.
Wherein, the component of grand nutrition element concentration corresponding with its is：1800 mg/L of potassium nitrate；Ammonium nitrate 1650
mg/L；Calcium chloride dihydrate 350mg/L；Anhydrous potassium dihydrogenphosphate 300mg/L；Epsom salt：360 mg/L；Ethylenediamine tetra-acetic acid
Disodium 20.5mg/L.
The component of micronutrient element concentration corresponding with its is：Four water manganese sulfates, 18.6 mg/L；White vitriol
7.5mg/L；4.8 mg/L of boric acid；Potassium iodide 1.5mg/L；Sodium molybdate 0.5mg/L；0.1 mg/L of CoCL2 6H2O.
The component of organic reagent concentration corresponding with its is：0.5 mg/L of thiamine hydrochloride；0.5 mg/L of niacin；Sweet ammonia
Acid：0.5 mg/L；0.5 mg/L of puridoxine hydrochloride；100.0 mg/L of inositol.
Further, a kind of oil method for tissue culture of tree peony, comprises the following steps：
（1）Oil is chosen by the use of the buff fruit of tree peony as explant；
（2）It carries out disinfection to explant：First, explant 3-5 times, each 5-8min is rinsed with clear water, then using pure
Explant 2-3 times, each 8-10min is rinsed in water purification, again after, explant is moved into superclean bench and is carried out disinfection place
Reason；
（3）Sowing：The moisture on explant surface is blotted using aseptic paper, is then cut off explant with disinfection cutter, it will
The seed of explant is put into oil with carrying out tissue cultures in the tissue culture medium (TCM) of tree peony, and condition of culture is：Temperature is 20-25 DEG C,
Light application time is 10-12h/d, intensity of illumination 1000lx, and culture 3-4 months is to growing seedling, during height of seedling 4-5cm, is moved
It plants；
（4）Transplanting：Bottle cap is opened, after standing 7-10d, test tube seedling is taken out, the culture medium of base portion is washed away with clear water, keep
Indoor temperature is 20-25 DEG C, is placed in culturing room, and passes through bag film moisturizing, and bag film is removed after 15d, is watered once daily,
Trial zone is transplanted to after 30d, carries out Routine Management.
And step（2）The disinfection treatment method is：It is carried out disinfection first with 75% alcohol to explant 30-45s,
Then aseptic water washing explant is used 2-3 times, washing time is each 3-5min, finally sterilizes 6- with the mercuric chloride that concentration is 0.1%
8min, then with aseptic water washing explant 2-3 times, washing time is each 3-5min.
Beneficial effects of the present invention are：Step of the present invention is simple, easily operated, by improve minimal medium and ingredient,
Adjust the supplementary measures such as condition of culture so that the bud ratio of test tube seedling has been more than 98%, and transplants shoot survival percent height, and growth is fast, greatly
It in the big cycle for shortening oil and being cultivated with tree peony, reduces production and rises this, improve the economic value with tree peony.
Specific embodiment
With reference to specific embodiment, the present invention will be described in detail.
The improvement MS that all examples below of the present invention uses includes grand nutrition element, micronutrient element and organic
Reagent.
And the component of grand nutrition element concentration corresponding with its is：1800 mg/L of potassium nitrate；1650 mg/L of ammonium nitrate；
Calcium chloride dihydrate 350mg/L；Anhydrous potassium dihydrogenphosphate 300mg/L；Epsom salt：360 mg/L；Disodium ethylene diamine tetraacetate
20.5mg/L。
The component of micronutrient element concentration corresponding with its is：Four water manganese sulfates, 18.6 mg/L；White vitriol
7.5mg/L；4.8 mg/L of boric acid；Potassium iodide 1.5mg/L；Sodium molybdate 0.5mg/L；0.1 mg/L of CoCL2 6H2O.
The component of organic reagent concentration corresponding with its is：0.5 mg/L of thiamine hydrochloride；0.5 mg/L of niacin；Sweet ammonia
Acid：0.5 mg/L；0.5 mg/L of puridoxine hydrochloride；100.0 mg/L of inositol.
Embodiment 1：
（1）Oil is chosen by the use of the buff fruit of tree peony as explant；
（2）It carries out disinfection to explant：First, explant is rinsed 4 times with clear water, then each 6min utilizes pure water
Rinse explant 2 times, each 9min, again after, explant is moved into superclean bench and is carried out disinfection processing, specific steps
For：It is carried out disinfection first with 75% alcohol to explant 40s, then with aseptic water washing explant 3 times, washing time is every
Secondary 4min finally sterilizes 7min with the mercuric chloride that concentration is 0.1%, then with aseptic water washing explant 3 times, washing time is each
4min；
（3）Sowing：The moisture on explant surface is blotted using aseptic paper, is then cut off explant with disinfection cutter, it will
The seed of explant is put into oil includes improvement MS with tissue cultures, the tissue culture medium (TCM) is carried out in the tissue culture medium (TCM) of tree peony
+0.6mg·L-1IAA +0.4mgL-1 IBA +0.6mgL-1NAA +0.2gL-1Activated carbon+2.5gL-1Sucrose, pH=5.7, and train
Foster condition is：Temperature is 22 DEG C, light application time 11h/d, intensity of illumination 1000lx, is sprouted after cultivating 27d, during height of seedling 5cm,
It is transplanted；
（4）Transplanting：Bottle cap is opened, after standing 8d, test tube seedling is taken out, the culture medium of base portion is washed away with clear water, keeps indoor
Temperature is 22 DEG C, is placed in culturing room, and passes through bag film moisturizing, and bag film is removed after 15d, is watered once daily, after 30d
Trial zone is transplanted to, carries out Routine Management.
Embodiment 2：
（1）Oil is chosen by the use of the buff fruit of tree peony as explant；
（2）It carries out disinfection to explant：First, explant is rinsed 3 times with clear water, then each 8min utilizes pure water
Rinse explant 2 times, each 10min, again after, explant is moved into superclean bench and is carried out disinfection processing, specific step
Suddenly it is：It is carried out disinfection first with 75% alcohol to explant 30s, then with aseptic water washing explant 2 times, washing time is
Each 5min finally sterilizes 6min with the mercuric chloride that concentration is 0.1%, then with aseptic water washing explant 2 times, washing time is every
Secondary 5min；
（3）Sowing：The moisture on explant surface is blotted using aseptic paper, is then cut off explant with disinfection cutter, it will
The seed of explant is put into oil includes improvement MS with tissue cultures, the tissue culture medium (TCM) is carried out in the tissue culture medium (TCM) of tree peony
+0.5mg·L-1IAA +0.3mgL-1 IBA +0.5mgL-1NAA +0.2gL-1Activated carbon+2gL-1Sucrose, pH=5.6, and cultivate
Condition is：Temperature is 20 DEG C, light application time 10h/d, intensity of illumination 1000lx, is sprouted after cultivating 30d, during height of seedling 4cm, into
Row transplanting；
（4）Transplanting：Bottle cap is opened, after standing 7d, test tube seedling is taken out, the culture medium of base portion is washed away with clear water, keeps indoor
Temperature is 20 DEG C, is placed in culturing room, and passes through bag film moisturizing, and bag film is removed after 15d, is watered once daily, after 30d
Trial zone is transplanted to, carries out Routine Management.
Embodiment 3：
（1）Oil is chosen by the use of the buff fruit of tree peony as explant；
（2）It carries out disinfection to explant：First, explant is rinsed 5 times with clear water, then each 5min utilizes pure water
Rinse explant 3 times, each 8min, again after, explant is moved into superclean bench and is carried out disinfection processing, specific steps
For：It is carried out disinfection first with 75% alcohol to explant 45s, then with aseptic water washing explant 3 times, washing time is every
Secondary 3min finally sterilizes 8min with the mercuric chloride that concentration is 0.1%, then with aseptic water washing explant 3 times, washing time is each
3min；
（3）Sowing：The moisture on explant surface is blotted using aseptic paper, is then cut off explant with disinfection cutter, it will
The seed of explant is put into oil includes improvement MS with tissue cultures, the tissue culture medium (TCM) is carried out in the tissue culture medium (TCM) of tree peony
+0.8mg·L-1IAA +0.5mgL-1 IBA +0.6mgL-1NAA +0.3 gL-1Activated carbon+3gL-1Sucrose, pH=5.8, and train
Foster condition is：Temperature is 25 DEG C, light application time 12h/d, intensity of illumination 1000lx, is sprouted after cultivating 29d, during height of seedling 5cm,
It is transplanted；
（4）Transplanting：Bottle cap is opened, after standing 10d, test tube seedling is taken out, the culture medium of base portion is washed away with clear water, keep room
Interior temperature is 25 DEG C, is placed in culturing room, and passes through bag film moisturizing, and bag film is removed after 15d, is watered once daily, 30d
After be transplanted to trial zone, carry out Routine Management.
Tissue is carried out with tree peony explant to 160 oil respectively using the method described in above-described embodiment 1-3 and culture medium
Cultivate and transplant nursery, while nursery is carried out using conventional seed type of seeding, and in its bud ratio, the budding time, transplanting into
The statistical form of motility rate is as shown in table 1：
Table 1：
As shown in Table 1：By culture medium of the present invention carry out tissue cultures oil with tree peony bud ratio, budding when
Between, transplanting survival rate etc. the method that is significantly better than traditional seed seeding and seedling raising, substantially reduce oil and cultivated with tree peony
Cycle, reduce production rise this, improve the economic value with tree peony.
The invention is not limited in any way for more than specific embodiment, every in a manner of equivalent substitution or equivalent transformation
The technical solution obtained, all falls in protection scope of the present invention.
Claims (3)
1. a kind of oil tissue culture medium (TCM) of tree peony, which is characterized in that including：Improve MS+0.5~0.8mgL-1IAA +0.3
~0.5mgL-1IBA+0.5~0.6mgL-1The gL of NAA+0.2~0.3-1Activated carbon+2~3gL-1Sucrose, pH=5.6~
5.8, the improvement MS include grand nutrition element, micronutrient element and organic reagent, the grand nutrition element
Component concentration corresponding with its is：1800 mg/L of potassium nitrate；1650 mg/L of ammonium nitrate；Calcium chloride dihydrate 350mg/L；Anhydrous phosphorus
Acid dihydride potassium 300mg/L；Epsom salt：360 mg/L；Disodium ethylene diamine tetraacetate 20.5mg/L, micronutrient member
Element component concentration corresponding with its be：Four water manganese sulfates, 18.6 mg/L；White vitriol 7.5mg/L；4.8 mg/L of boric acid；
Potassium iodide 1.5mg/L；Sodium molybdate 0.5mg/L；0.1 mg/L of CoCL2 6H2O, the component of the organic reagent are corresponding with its
Concentration is：0.5 mg/L of thiamine hydrochloride；0.5 mg/L of niacin；Glycine：0.5 mg/L；0.5 mg/L of puridoxine hydrochloride；Flesh
100.0 mg/L of alcohol.
2. a kind of oil method for tissue culture of tree peony, which is characterized in that comprise the following steps：
（1）Oil is chosen by the use of the buff fruit of tree peony as explant；
（2）It carries out disinfection to explant：First, explant is rinsed 3-5 times with clear water, then each 5-8min utilizes pure water
Rinse explant 2-3 times, each 8-10min, again after, explant is moved into superclean bench and is carried out disinfection processing；
（3）Sowing：The moisture on explant surface is blotted using aseptic paper, is then cut off explant with disinfection cutter, by explant
The seed of body is put into oil with carrying out tissue cultures in the tissue culture medium (TCM) of tree peony, and condition of culture is：Temperature is 20-25 DEG C, illumination
Time is 10-12h/d, intensity of illumination 1000lx, and culture 3-4 months is to growing seedling, during height of seedling 4-5cm, is transplanted；
（4）Transplanting：Bottle cap is opened, after standing 7-10d, test tube seedling is taken out, the culture medium of base portion is washed away with clear water, keeps indoor
Temperature is 20-25 DEG C, is placed in culturing room, and passes through bag film moisturizing, and bag film is removed after 15d, is watered once daily, 30d
After be transplanted to trial zone, carry out Routine Management.
A kind of 3. oil according to claim 2 method for tissue culture of tree peony, which is characterized in that step（2）Described
Disinfection treatment method is：It is carried out disinfection first with 75% alcohol to explant 30-45s, then with aseptic water washing explant 2-
3 times, washing time is each 3-5min, finally sterilizes 6-8min with the mercuric chloride that concentration is 0.1%, then with aseptic water washing explant
Body 2-3 times, washing time are each 3-5min.
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| CN108094197A (en) * | 2017-11-30 | 2018-06-01 | 安徽心缘康生物科技有限公司 | A kind of oil tree peony phoenix pellet asexual multiplication seedling method |
| CN110476807B (en) * | 2019-07-31 | 2021-06-08 | 河南科技大学 | Method for establishing sterile culture system of mature seed embryo of peony 'paeonia ostii' for oil |
Citations (3)
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|---|---|---|---|---|
| CN103355165A (en) * | 2012-04-09 | 2013-10-23 | 上海植物园 | Culture method of peony embryonic callus as well as culture medium |
| CN104206417A (en) * | 2013-06-04 | 2014-12-17 | 中国科学院上海生命科学研究院 | Method for promoting paeonia suffruticosa seedling growth and application thereof |
| CN104770103A (en) * | 2015-04-15 | 2015-07-15 | 芜湖市丹洋现代农业科技发展有限公司 | Seedling raising method for oil-used peony seeds |
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2016
- 2016-06-03 CN CN201610385679.XA patent/CN105941156B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103355165A (en) * | 2012-04-09 | 2013-10-23 | 上海植物园 | Culture method of peony embryonic callus as well as culture medium |
| CN104206417A (en) * | 2013-06-04 | 2014-12-17 | 中国科学院上海生命科学研究院 | Method for promoting paeonia suffruticosa seedling growth and application thereof |
| CN104770103A (en) * | 2015-04-15 | 2015-07-15 | 芜湖市丹洋现代农业科技发展有限公司 | Seedling raising method for oil-used peony seeds |
Non-Patent Citations (1)
| Title |
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| 三种牡丹组织培养比较研究;范小峰等;《北方园艺》;20101231(第4期);摘要，1.1 试验材料，1.3 培养基及培养条件 * |
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