CN106106185B - A kind of blue or green money willow rooting of vitro seedling culture medium and culture of rootage method - Google Patents
A kind of blue or green money willow rooting of vitro seedling culture medium and culture of rootage method Download PDFInfo
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- CN106106185B CN106106185B CN201610691975.2A CN201610691975A CN106106185B CN 106106185 B CN106106185 B CN 106106185B CN 201610691975 A CN201610691975 A CN 201610691975A CN 106106185 B CN106106185 B CN 106106185B
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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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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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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Abstract
The invention discloses a kind of blue or green money willow rooting of vitro seedling culture medium and culture of rootage method.The formula of the root media is：Improve MS+IBA0.5~2.0mg/L+IAA0~2.0mg/L+ diethyl aminoethyl hexanoate 0.02~0.1mg/L+, 15~20g/L of sucrose, pH5.5~6.0；Wherein, the formula for improveing MS is：A great number of elements：Potassium nitrate 700mg/L, ammonium nitrate 600mg/L, epsom salt 185mg/L, potassium dihydrogen phosphate 60mg/L, calcium chloride dihydrate 165mg/L；Organic compound：Inositol 100mg/L, nicotinic acid 1mg/L, puridoxine hydrochloride 1mg/L, thiamine hydrochloride 0.2mg/L, glycine 2mg/L；Trace element is the same as MS culture mediums.Culture of rootage, rooting rate >=70% are carried out to blue or green money willow test tube seedling with above-mentioned culture medium.
Description
Technical field
The present invention relates to the tissue culture technique of plant, and in particular to a kind of blue or green money willow rooting of vitro seedling culture medium and takes root
Cultural method.
Background technology
It is blue or green that blue or green money willow [Cyclocarya paliurus (Batal.) Iljinskaja] belongs to Juglandaceae (Juglandaceae)
Money sallow, alias money tree, numb willow, blue or green money Lee, mountain fiber crops willow, Shan Huashu.Blue or green money willow is known as " plant giant panda ", " doctor
3rd tree of educational circles ", is the rare tree species that the discipline of glacier four survives, exists only in China, be the in imminent danger of special-protection-by-the-State
Plant.Blue or green money willow is distributed mainly on the ground such as Jiangxi, Zhejiang, Jiangsu, Anhui, Fujian, Taiwan, Hubei, Sichuan, Guizhou, Yunnan, more
It is fragmentary to be distributed in remote, thickly forested mountains and some nature reserve areas.
Blue or green money willow is with more unique health care and medical value.It is among the people to do tea with Qingqian Willow leaf for a long time, tea gas delicate fragrance,
It is sweet.《Chinese medicine resource will will》(1994) record, Qingqian Willow leaf tool heat-clearing is quenched one's thirst the effect of removing toxic substances.《National Chinese herbal medicine name mirror》
(1996) record, blue or green money willow bark, leaf, root have the effect of desinsection is antipruritic, anti-inflammatory analgetic wind-dispelling.Modern pharmacology research proves,
Blue or green money willow has lowering blood pressure and blood fat and anti-ageing medicinal efficacy of waiting for a long time.Research finds, Qingqian Willow leaf contain substantial amounts of potassium, calcium, magnesium,
The macroelements such as phosphorus, also containing trace elements such as manganese, iron, copper, chromium, zinc, selenium, vanadium, germanium.Wherein, make with glycometabolism and insulin
Content with closely related elemental chromium, nickel, lithium, selenium, vanadium is higher, can assist in insulin and plays hypoglycemic effect, and can change
Kind sugar tolerance.Qingqian Willow leaf also contains the organic compounds such as abundant polysaccharide, flavones, triterpene, steroid, saponin(e, coumarin, this
A little compounds can effectively reduce triglycerides and cholesterol, blue or green money willow is had the function of blood pressure lowering, hypoglycemic, reducing blood lipid, together
When to enhancing body immunity, anti-oxidant, anti-ageing wait for a long time with unique effect.
At present, the blue or green money willow nursery stock of in the market is mostly the seedling from seed of wild resource transplanting or artificial culture.Due to green grass or young crops
Money willow introduces a collection is limited, and the full rate of seed is low, also with deep dormancy characteristic, is grown using conventional seed planting method for culturing seedlings breeding cycle, effect
Rate is low, it is difficult to produces a large amount of seedlings in a short time, it is impossible to meet the needs of market.In recent years, domestic and international researcher with
In vitro Embryo, blade, tender shoots, stem section etc. carry out the research of the tissue culture and rapid proliferation of blue or green money willow for explant in succession, such as thank
The third of the twelve Earthly Branches (the Primary Study northwests Botany Gazette of the blue or green money willow tissue culture rapid propagation systems such as Xie Yinfeng, 2009,29 (11) such as peak：2331-
2338) rapid propagation system of blue or green money willow is tentatively established using In vitro Embryo, using WPM+0.2mg/L IBA+20g/L sucrose culture mediums
Induced, rooting rate 16.67%, by 15d, secretly induction is handled, and rooting rate can bring up to 23.33%.Qiao Qingmei etc. is (tall
Minister in ancient times plum, Cheng Maogao, influence north gardening 2009 (7) of the Wei Zhi China differences exogenous hormone to blue or green money willow stem section tissue cultures：
125-127) research is found, in blue or green money willow stem section tissue cultures, the appropriate media of root induction is improvement MS+5.0mg/
L IBA, inductivity 40.00%.Lu Meng etc. (the Techniques of in Vitro Culture Nanjing Forestry University of blue or green money willow stem segment with axillary buds such as sprouts in Shandong
Journal (natural science edition), 2013,37 (6)：6-10) using WPM as minimal medium, IBA, GGR-6, ABT-1 are to blue or green money for research
The influence of Liu Shenggen, the results showed that, 10%GGR+1.0mg/L IBA combined treatment effects are best, rooting rate 25%.It is existing
Result of study shows, blue or green money willow test tube seedling can successfully expand numerous, but its rooting rate is low, quality of rooting is poor, it is difficult to meets batch production
The requirement of nursery.It can be seen from the above that the rooting technique of blue or green money willow tissue-cultured seedling has become restricts the pass that blue or green money willow seedling is factory produced
Key link, the breakthrough of tissue-cultured seedling rooting technique, to establishing the efficient tissue culture rapid propagation system of blue or green money willow, expanding planting scale with important
Meaning.
The content of the invention
The technical problem to be solved in the present invention is to provide a kind of blue or green money willow rooting of vitro seedling culture medium and culture of rootage method.
Culture of rootage is carried out to conventional blue or green money willow test tube seedling using root media of the present invention, more than 70% life can be obtained
Root rate, and root system development is good, plant strain growth is healthy and strong, and transplanting survival rate is high.
The formula of blue or green money willow rooting of vitro seedling culture medium of the present invention is：
Improve MS+IBA0.5~2.0mg/L+IAA0~15~20g/ of 2.0mg/L+ diethyl aminoethyl hexanoate 0.02~0.1mg/L+ sucrose
L, pH5.5~6.0；
Wherein, the formula for improveing MS is：
The component and concentration of a great number of elements be：Potassium nitrate 700mg/L, ammonium nitrate 600mg/L, epsom salt 185mg/L,
Potassium dihydrogen phosphate 60mg/L, calcium chloride dihydrate 165mg/L；
The component and concentration of organic compound be：Inositol 100mg/L, nicotinic acid 1mg/L, puridoxine hydrochloride 1mg/L, hydrochloric acid sulphur
Amine element 0.2mg/L, glycine 2mg/L；
The component and concentration of trace element are the same as MS culture mediums.
In above-mentioned technical proposal, the formula of the blue or green money willow root media is preferably：Improve MS+IBA0.5~1.0mg/
L+IAA 0.8~1.5mg/L+ diethyl aminoethyl hexanoate 0.05~0.08mg/L+, 18~20g/L of sucrose, pH5.5~6.0；Most preferably improve
MS+IBA 0.5mg/L+IAA 1.0mg/L+ diethyl aminoethyl hexanoate 0.05mg/L+ sucrose 20g/L, pH5.5~6.0.The blue or green money willow is taken root
The pH value of culture medium is preferably 5.8~6.0.
Present invention additionally comprises the culture of rootage method of blue or green money willow test tube seedling, is specially：Single plant test tube seedling is cut, access is taken root
At least 25d is cultivated in culture medium；Wherein：
The formula of the root media is：
Improve MS+IBA0.5~2.0mg/L+IAA0~15~20g/ of 2.0mg/L+ diethyl aminoethyl hexanoate 0.02~0.1mg/L+ sucrose
L, pH5.5~6.0；
Wherein, the formula for improveing MS is：
The component and concentration of a great number of elements be：Potassium nitrate 700mg/L, ammonium nitrate 600mg/L, epsom salt 185mg/L,
Potassium dihydrogen phosphate 60mg/L, calcium chloride dihydrate 165mg/L；
The component and concentration of organic compound be：Inositol 100mg/L, nicotinic acid 1mg/L, puridoxine hydrochloride 1mg/L, hydrochloric acid sulphur
Amine element 0.2mg/L, glycine 2mg/L；
The component and concentration of trace element are the same as MS culture mediums.
In the above method, the culture of rootage condition of blue or green money willow test tube seedling can be same as the prior art.It is preferably as follows：Culture
Temperature is 20~30 DEG C, and periodicity of illumination is 10~14h/d, and intensity of illumination is 2000~3000Lx；Further preferred culture bar
Part is：Cultivation temperature is 23~27 DEG C, and periodicity of illumination is 10~12h/d, and intensity of illumination is 2000~3000Lx.
In the above method, blue or green money willow test tube seedling in root media under above-mentioned condition of culture cultivate 25d when, rooting rate
More than 70% is can reach, and root system development is good, and plant strain growth is healthy and strong.In the method for the invention, preferably test tube seedling is in life
Incubation time in root culture medium is 25~35d.
In the above method, the formula of the blue or green money willow root media is preferably：Improve MS+IBA0.5~1.0mg/L+
IAA 0.8~1.5mg/L+ diethyl aminoethyl hexanoate 0.05~0.08mg/L+, 18~20g/L of sucrose, pH5.5~6.0；Most preferably improve MS
+ IBA 0.5mg/L+IAA 1.0mg/L+ diethyl aminoethyl hexanoate 0.05mg/L+ sucrose 20g/L, pH5.5~6.0.The blue or green money willow is taken root training
The pH value for supporting base is preferably 5.8~6.0.
In the method for the invention, the test tube seedling for carrying out culture of rootage is obtained by existing conventional method cultured in vitro,
Preferably with blue or green money willow current-year branch, according to a conventional method obtained by cultured in vitro.Preferably choose the single plant test tube of height >=2cm
Seedling carries out culture of rootage, more preferably chooses height >=3cm and the single plant test tube seedling with 3~4 compound leaves carries out culture of rootage,
The transplanting survival rate of plant can so be further improved.
Compared with prior art, the method have the characteristics that：By improveing minimal medium concentration of component, spy being used in combination
Determine diethyl aminoethyl hexanoate, IBA the and IAA plant growth regulator of concentration, gained root media is effectively facilitated blue or green money willow test tube
Seedling is taken root.Culture of rootage is carried out to blue or green money willow test tube seedling using the root media, rooting rate can be made to reach more than 70% (excellent
Embodiment is selected up to 86%), and root system development is good, plant strain growth is healthy and strong, and transplanting survival rate is high, to realize that blue or green money willow tissue culture is produced
Industry and popularization and application provide technical support.
Brief description of the drawings
Fig. 1 is the picture of the blue or green money willow test tube seedling obtained according to a conventional method in embodiment 1；
The picture of rooted seedling when Fig. 2 is using root media culture 25d described in the embodiment of the present invention 1.
Embodiment
With reference to specific embodiment, the present invention is described in further detail, to more fully understand present disclosure, but
The present invention is not limited to following embodiments.
In following embodiment, involved improvement MS includes a great number of elements, trace element and organic compound, specifically
Formula is as follows：
The component and concentration of a great number of elements be：Potassium nitrate 700mg/L, ammonium nitrate 600mg/L, epsom salt 185mg/L,
Potassium dihydrogen phosphate 60mg/L, calcium chloride dihydrate 165mg/L；
Trace element component and concentration be：Potassium iodide 0.83mg/L, boric acid 6.2mg/L, four water manganese sulfate 22.3mg/L,
White vitriol 8.6mg/L, Sodium Molybdate Dihydrate 0.25mg/L, cupric sulfate pentahydrate 0.025mg/L, CoCL2 6H2O 0.025mg/L,
Ferrous sulfate heptahydrate 27.8mg/L.Wherein, ferrous sulfate heptahydrate is chelated with disodium ethylene diamine tetraacetate, disodium ethylene diamine tetraacetate
Concentration is 37.3mg/L.
The component and concentration of organic compound be：Inositol 100mg/L, nicotinic acid 1mg/L, puridoxine hydrochloride 1mg/L, hydrochloric acid sulphur
Amine element 0.2mg/L, glycine 2mg/L.
Embodiment 1
1) blue or green money willow current-year branch is taken, cultured in vitro obtains test tube seedling according to a conventional method, as shown in Figure 1；
2) culture of rootage：Cut height and be more than 3.5cm and with the single plant test tube seedling of 3~4 compound leaves, access culture of rootage
25d (as shown in Figure 2 when cultivating 25d) is cultivated in base；Wherein：
The formula of the root media is：Improve MS+IBA0.5mg/L+IAA1.0mg/L+ diethyl aminoethyl hexanoate 0.05mg/L+ sugarcanes
Sugared 20g/L, pH5.8, use filter paper bridge as supporter.
Culture of rootage condition is：Temperature is 23~25 DEG C, periodicity of illumination 12h/d, intensity of illumination 2000Lx.
Rooting rate (same as below), rooting rate 86% are calculated by strain number of taking root/inoculation strain number × 100%.
Comparative example 1
Prescription of rooting medium, is simply revised as by the method for repeating embodiment 1：Improve MS+IBA0.5mg/L+IAA
1.0mg/L+ sucrose 20g/L, pH5.8.
It is computed, rooting rate 25%.
Comparative example 2
Prescription of rooting medium, is simply revised as by the method for repeating embodiment 1：Improve MS+IBA 0.5mg/L+IAA
1.0mg/L+ diethyl aminoethyl hexanoate 0.01mg/L+ sucrose 20g/L, pH5.8.
It is computed, rooting rate 40%.
Comparative example 3
Prescription of rooting medium, is simply revised as by the method for repeating embodiment 1：Improve MS+IBA0.5mg/L+IAA
1.0mg/L+ diethyl aminoethyl hexanoate 0.15mg/L+ sucrose 20g/L, pH5.8.
It is computed, rooting rate 5%.
Embodiment 2
1) blue or green money willow current-year branch is taken, cultured in vitro obtains test tube seedling according to a conventional method；
2) culture of rootage：Cut higher than 3.5cm and with the single plant test tube seedling of 3~4 compound leaves, access in root media
Cultivate 25d；Wherein：
The formula of the root media is：MS+IBA1.5mg/L+ diethyl aminoethyl hexanoate 0.05mg/L+ sucrose 20g/L are improved,
PH5.8, uses filter paper bridge as supporter.
Condition of culture in root media is：Temperature is 23~25 DEG C, periodicity of illumination 12h/d, and intensity of illumination is
2000Lx。
It is computed, rooting rate 80%.
Embodiment 3
1) blue or green money willow current-year branch is taken, cultured in vitro obtains test tube seedling according to a conventional method；
2) culture of rootage：Cut higher than 2.5cm and with the single plant test tube seedling of 2~3 compound leaves, access in root media
Cultivate 30d；Wherein：
The formula of the root media is：Improve MS+IBA0.5mg/L+IAA 2.0mg/L+ diethyl aminoethyl hexanoates 0.02mg/L+
Sucrose 15g/L, pH6.0, use filter paper bridge as supporter.
Condition of culture in root media is：Temperature is 25 DEG C, periodicity of illumination 10h/d, and intensity of illumination is
3000Lx。
It is computed, rooting rate 75%.
Embodiment 4
1) blue or green money willow current-year branch is taken, cultured in vitro obtains test tube seedling according to a conventional method；
2) culture of rootage：Cut higher than 2.5cm and with the single plant test tube seedling of 1~2 compound leaf, access in root media
Cultivate 27d；Wherein：
The formula of the root media is：Improve MS+IBA1.5mg/L+IAA0.5mg/L+ diethyl aminoethyl hexanoate 0.1mg/L+ sugarcanes
Sugared 18g/L, pH5.5, use filter paper bridge as supporter.
Condition of culture in root media is：Temperature is 20 DEG C, periodicity of illumination 14h/d, and intensity of illumination is
2500Lx。
It is computed, rooting rate 70%.
Embodiment 5
1) blue or green money willow current-year branch is taken, cultured in vitro obtains test tube seedling according to a conventional method；
2) culture of rootage：Choose higher than 3.5cm and with the single plant test tube seedling of 3~4 compound leaves, access in root media
Cultivate 28d；Wherein：
The formula of the root media is：Improve MS+IBA1.0mg/L+IAA1.5mg/L+ diethyl aminoethyl hexanoate 0.08mg/L+ sugarcanes
Sugared 16g/L, pH5.9, use filter paper bridge as supporter.
Condition of culture in root media is：Temperature is 30 DEG C, periodicity of illumination 12h/d, intensity of illumination 2000
~2200Lx.
It is computed, rooting rate 82%.
Claims (7)
- A kind of 1. blue or green money willow rooting of vitro seedling culture medium, it is characterised in that：The formula of the root media is：MS+IBA0.5~2.0mg/L+IAA0~2.0mg/L+ diethyl aminoethyl hexanoate 0.02~0.1mg/L+, 15~20g/L of sucrose are improved, PH5.5~6.0；Wherein, the formula for improveing MS is：The component and concentration of a great number of elements be：Potassium nitrate 700mg/L, ammonium nitrate 600mg/L, epsom salt 185mg/L, phosphoric acid Potassium dihydrogen 60mg/L, calcium chloride dihydrate 165mg/L；The component and concentration of organic compound be：Inositol 100mg/L, nicotinic acid 1mg/L, puridoxine hydrochloride 1mg/L, thiamine hydrochloride 0.2mg/L, glycine 2mg/L；The component and concentration of trace element are the same as MS culture mediums.
- 2. blue or green money willow rooting of vitro seedling culture medium according to claim 1, it is characterised in that：The root media is matched somebody with somebody Fang Wei：Improve MS+IBA0.5mg/L+IAA1.0mg/L+ diethyl aminoethyl hexanoate 0.05mg/L+ sucrose 20g/L, pH5.5~6.0.
- A kind of 3. culture of rootage method of blue or green money willow test tube seedling, it is characterised in that：Single plant test tube seedling is cut, accesses root media Middle culture at least 25d；Wherein：The formula of the root media is：MS+IBA0.5~2.0mg/L+IAA0~2.0mg/L+ diethyl aminoethyl hexanoate 0.02~0.1mg/L+, 15~20g/L of sucrose are improved, PH5.5~6.0；Wherein, the formula for improveing MS is：The component and concentration of a great number of elements be：Potassium nitrate 700mg/L, ammonium nitrate 600mg/L, epsom salt 185mg/L, phosphoric acid Potassium dihydrogen 60mg/L, calcium chloride dihydrate 165mg/L；The component and concentration of organic compound be：Inositol 100mg/L, nicotinic acid 1mg/L, puridoxine hydrochloride 1mg/L, thiamine hydrochloride 0.2mg/L, glycine 2mg/L；The component and concentration of trace element are the same as MS culture mediums.
- 4. the culture of rootage method of blue or green money willow test tube seedling according to claim 3, it is characterised in that：The training of taking root of test tube seedling Foster condition is：Cultivation temperature is 20~30 DEG C, and periodicity of illumination is 10~14h/d, and intensity of illumination is 2000~3000Lx.
- 5. the culture of rootage method of blue or green money willow test tube seedling according to claim 4, it is characterised in that：The training of taking root of test tube seedling Foster condition is：Cultivation temperature is 23~27 DEG C, and periodicity of illumination is 10~12h/d, and intensity of illumination is 2000~3000Lx.
- 6. the culture of rootage method of the blue or green money willow test tube seedling according to any one of claim 3~5, it is characterised in that：Examination Pipe seedling rooting incubation time is 25~35d.
- 7. the culture of rootage method of the blue or green money willow test tube seedling according to any one of claim 3~5, it is characterised in that：Institute The formula for stating root media is：Improve MS+IBA0.5mg/L+IAA1.0mg/L+ diethyl aminoethyl hexanoate 0.05mg/L+ sucrose 20g/L, pH5.5~6.0.
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