CN105875405A - Culture medium inducing proliferation of lamiophlomis rotata and preparation method thereof - Google Patents
Culture medium inducing proliferation of lamiophlomis rotata and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a culture medium inducing proliferation of lamiophlomis rotata and a preparation method thereof, and is characterized in that the culture medium comprises MS, 20-40 g/L of white sugar, 6.5-11 g/L of carrageenan, 0.2-3 mg/L of BA, 0.1-1.5 mg/L of NAA, 0.1-1.0 mg/L of IBA, and 0.1-4 mg/L of ZT. According to the lamiophlomis rotata culture medium, with joint coordination regulation action of a specific content of the MS culture medium, BA, NAA, IBA and ZT, rich nutrition elements are provided for tissue culture of the lamiophlomis rotate, the production cost is greatly reduced, the culture medium is not affected by natural conditions, the propagation coefficient is improved, and thus the economic value of the lamiophlomis rotata is greatly improved.
Description
Technical field
The invention belongs to biological technical field, be specifically related to a kind of group training proliferated culture medium and preparation method thereof.
Background technology
Labiatae lamiophlomis rotata belongs to unique species, and perennial herb is high 2.5-10 centimetre；Rhizome extends, slightly
Thickness, footpath reaches 1 centimetre.Normal 4 pieces of blade, spoke shape is relative two-by-two, water chestnut shape circle, rhombus, sector, horizontal kidney
Shape is so that triangle, bitter, is slightly cold.There is promoting blood circulation, effect of swelling and pain relieving.Lamiophlomis rotata is Qinghai-Tibet Platean
The important medicinal plant of distinctive one, is distributed widely in Tibet, and in Qinghai, Gansu, Sichuan and Yunnan only
There is fragmentary distribution, be often born in stone matter mesophorbium, coryphile, flood land or the intensity weathering of height above sea level 3500-5100 rice
On rubble beach.In existing tissue culture technology, the cultivation to lamiophlomis rotata is reported seldom, the most externally implant protocorm
Carry out Fiber differentiation, breed, break up, biochemical culture, obtain seedling, habitat conditions is required severe by lamiophlomis rotata
Carving, difficulty opens low altitude area cultivation, and under the conditions of artificial pseudo-wild cultivating, seed germination rate is at a fairly low and wild
The extinction in imminent danger of production-goods source；Therefore, in the training of lamiophlomis rotata group, proliferation-inducing is cultivated improving its breeding coefficient and resource
Protection has very important significance；And though the breeding of existing application prior seed can obtain certain germination rate
Rate, but germination rate is low, and cost is high, and resistance is poor, and easy infection disease.
Further, in group training under normal circumstances, the general mode using hot filling in the preparation process of culture medium,
I.e. white sugar and carragheen is the most heated boil after pour in bottle placer filling, culture medium that the method is prepared battalion
Support element and plant hormone and run off serious, and due to time canned temperature be gradually lowered carragheen and present semigel shape
Stir uneven cause canned uneven to tissue culture bottle Middle nutrition liquid, be unfavorable for inoculation, switching and the growth of seedling.
Summary of the invention
It is an object of the invention to provide a kind of induction lamiophlomis rotata proliferated culture medium and preparation method thereof, this cultivation
Base overcomes in existing traditional cultivation seeding technique that the seed nutritional of lamiophlomis rotata is bad causes breeding coefficient low
Problem, induction lamiophlomis rotata is Proliferation, Differentiation in group training, improves the breeding coefficient of lamiophlomis rotata.
In order to reach foregoing invention purpose, the technical solution used in the present invention is: provide one induction lamiophlomis rotata
The culture medium of propagation, it is characterised in that: include MS+ white sugar 20～40g/L+ carragheen 6.5～11g/L+BA
0.2～3mg/L+NAA0.1～1.5mg/L+IBA 0.1～1.0mg/L+ZT0.1～4mg/L.Described lamiophlomis rotata lures
Leading in proliferated culture medium, in every liter of MS culture medium, the content of each component is as follows: NH4NO3 165mg、KNO3
1900mg、CaCl2·2H2O 440mg、MgSO4·7H2O 370mg、KH2PO4 170mg、KI 0.83mg、
H3BO3 6.2mg、MnSO4·H2O 16.9mg、ZnSO4·7H2O 8.6mg、Na2MoO4·2H2O
0.25mg、CuSO4·5H2O 0.025mg、CoCl2·6H2O 0.025mg、FeSO4·7H2O 27.8mg、
Na2-EDTA·2H2O 37.3mg, inositol 100mg, nicotinic acid 0.5mg, hydrochloric acid pyrrole tremble zinc 0.5mg,
Thiamine hydrochloride 0.1mg, glycine 2mg.
Lamiophlomis rotata proliferative induction culture medium of the present invention, it is characterised in that: include MS+ white sugar 30g/L+
Carragheen 6.5g/L+BA 1.0mg/L+NAA 0.2mg/L+IBA 0.30mg/L+ZT0.2mg/L.
The preparation method of above-mentioned lamiophlomis rotata proliferative induction culture medium comprises the following steps:
A, first white sugar and carragheen are pre-processed: respectively white sugar and carragheen are proportionally weighed
The distilled water adding 15 times of quality dissolves, and stands 3～4 hours；
The preparation of b, MS culture medium mother liquor, including mother liquor 1, mother liquor 2, mother liquor 3, mother liquor 4, mother liquor 5
Mother liquor 6, mother liquor 7 and the preparation of mother liquor 8, wherein
The preparation of mother liquor 1: weigh 170g KH2PO4It is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 2: weigh 1900g KNO3It is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 3: weigh 1650g NH4NO3It is placed in beaker, adds distilled water stirring and dissolving,
It is settled to 10000ml, is saved in brown break-in bottle；
The preparation of mother liquor 4: 370g MgSO4·7H2O is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 5: weigh 440g CaCl2·2H2O is placed in beaker, adds distilled water stirring and dissolving,
It is settled to 10000ml, is saved in brown break-in bottle；
The preparation of mother liquor 6: weigh 1.66g KI, 12.4g H respectively3BO3、33.8g MnSO4·H2O、17.2g
ZnSO4·7H2O、0.5g Na2MoO4·2H2O、0.05g CuSO4·5H2O and 0.05g CoCl2·6H2O,
Add in beaker and dissolve, be settled to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 7: weigh 200g inositol respectively, 1g nicotinic acid, 1g hydrochloric acid pyrrole tremble zinc, 0.2g salt
Allithiamine element and 4g glycine, add in beaker and dissolve, be settled to 10000ml, be saved in brown break-in
In Ping；
The preparation of mother liquor 8: weigh 55.6g FeSO4·7H2O、74.6g Na2-EDTA·2H2O adds respectively
Entering 3000ml distilled water to dissolve, then by two kinds of solution mixing, heating is boiled 15 minutes, and is stirred continuously,
Fully chelating, then it is settled to 10000ml, it is saved in brown break-in bottle；
C, by pretreatment white sugar and the carragheen aqueous solution, 10ml/L mother liquor 1,10ml/L mother liquor 2,10ml/L
Mother liquor 3,10ml/L mother liquor 4,10ml/L mother liquor 5.5ml/L mother liquor 6,5ml/L mother liquor 7,5ml/L
Mother liquor 8 is poured in bottle placer respectively, adds BA, NAA, IBA, ZT.Add water constant volume, and stirs
15min, regulation pH value is to 5.8, rear filling, 121 DEG C, sterilizing 23 minutes under conditions of 0.16MPa,
Prepare after condensation.
The described filling time is 3s, and the intermittent time is 0.7s.
Lamiophlomis rotata proliferative induction culture medium of the present invention is by using the MS culture medium of certain content, this lamiophlomis rotata
Culture medium is by using certain content MS culture medium, the common synergic adjustment of BA, NAA, IBA and ZT
The group training of effect, the most only lamiophlomis rotata provides abundant nutrient, greatly reduces production cost low, and
And do not significantly improved its breeding coefficient by effect of natural conditions, thus it is greatly improved the economic worth of lamiophlomis rotata.
The preparation method of the lamiophlomis rotata proliferative induction culture medium that the present invention provides is by carrying out pre-by carragheen and white sugar
Process and use cold filling mode, the nutrients such as trace element, vitamin and plant hormone can be greatly reduced
Loss during preparing culture medium, and the culture medium hardness prepared is moderate, bright in white color, for
Lamiophlomis rotata proliferative induction provides the culture medium condition of key；After this proliferative induction medium culture only one
The breeding potential of taste brings up to 90%, and plant aberration rate is low, and plant structural integrity robustness is good.
Detailed description of the invention
Embodiment 1
The formula of this lamiophlomis rotata proliferative induction culture medium is: MS+ white sugar 30g/L+ carragheen 6.5g/L+BA
1.0mg/L+NAA 0.2mg/L+IBA 0.30mg/L+ZT0.2mg/L.Wherein, MS minimal medium
Formula is: in every liter of culture medium, the content of component is as follows:
NH4NO3 165mg、KNO3 1900mg、CaCl2·2H2O 440mg、MgSO4·7H2O 370mg、
KH2PO4 170mg、KI 0.83mg、H3BO3 6.2mg、MnSO4·H2O 16.9mg、ZnSO4·7H2O
8.6mg、Na2MoO4·2H2O 0.25mg、CuSO4·5H2O 0.025mg、CoCl2·6H2O 0.025mg、
FeSO4·7H2O 27.8mg、Na2-EDTA·2H2O 37.3mg, inositol 100mg, nicotinic acid 0.5mg,
Puridoxine hydrochloride 0.5mg, thiamine hydrochloride 0.1mg, glycine 2mg.
The process for preparation of above-mentioned lamiophlomis rotata proliferative induction culture medium is:
(1) preparation of MS minimal medium mother liquor: include mother liquor 1, mother liquor 2, mother liquor 3, mother liquor 4,
Mother liquor 5, mother liquor 6, mother liquor 7 and the preparation of mother liquor 8, wherein
The preparation of mother liquor 1: weigh 170g KH2PO4It is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 2: weigh 1900g KNO3It is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 3: weigh 1650g NH4NO3It is placed in beaker, adds distilled water stirring and dissolving,
It is settled to 10000ml, is saved in brown break-in bottle；
The preparation of mother liquor 4: 370g MgSO4·7H2O is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 5: weigh 440g CaCl2·2H2O is placed in beaker, adds distilled water stirring and dissolving,
It is settled to 10000ml, is saved in brown break-in bottle；
The preparation of mother liquor 6: weigh 1.66g KI, 12.4g H respectively3BO3、33.8g MnSO4·H2O、17.2g
ZnSO4·7H2O、0.5g Na2MoO4·2H2O、0.05g CuSO4·5H2O and 0.05g CoCl2·6H2O,
Add in beaker and dissolve, be settled to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 7: weigh 200g inositol respectively, 1g nicotinic acid, 1g hydrochloric acid pyrrole tremble zinc, 0.2g salt
Allithiamine element and 4g glycine, add in beaker and dissolve, be settled to 10000ml, be saved in brown break-in
In Ping；
The preparation of mother liquor 8: weigh 55.6g FeSO4·7H2O、74.6g Na2-EDTA·2H2O adds respectively
Entering 3000ml distilled water to dissolve, then by two kinds of solution mixing, heating is boiled 15 minutes, and is stirred continuously,
Fully chelating, then it is settled to 10000ml, it is saved in brown break-in bottle；
(2) white sugar and carragheen pre-process: white sugar and carragheen are proportionally weighed respectively and add
The distilled water entering 15 times of quality dissolves, and stands 4 hours；
(3) by pretreatment white sugar and the carragheen aqueous solution, 10ml/L mother liquor 1,10ml/L mother liquor 2,
10ml/L mother liquor 3,510ml/L mother liquor 4,10ml/L mother liquor 5.5ml/L mother liquor 6,5ml/L mother liquor 7,
5ml/L mother liquor 8 is poured in bottle placer respectively, adds BA 1.0mg/L, NAA 0.2mg/L, IBA
0.30mg/L、ZT0.2mg/L.Add water constant volume, and stirs 15min, and regulation pH value is to 5.8, rear filling,
121 DEG C, sterilizing 23 minutes under conditions of 0.16MPa, prepare after condensation；Wherein, the filling time is
3s, the intermittent time is 0.7s.
Embodiment 2
The formula of this lamiophlomis rotata proliferation-inducing culture medium is: MS+ white sugar 20g/L+ carragheen 8g/L+BA
3.0mg/L+NAA 0.5mg/L+IBA 0.50mg/L+ZT0.8mg/L.Wherein, MS minimal medium
Formula is: in every liter of culture medium, the content of component is as follows:
NH4NO3 165mg、KNO3 1900mg、CaCl2·2H2O 440mg、MgSO4·7H2O 370mg、
KH2PO4 170mg、KI 0.83mg、H3BO3 6.2mg、MnSO4·H2O 16.9mg、ZnSO4·7H2O
8.6mg、Na2MoO4·2H2O 0.25mg、CuSO4·5H2O 0.025mg、CoCl2·6H2O 0.025mg、
FeSO4·7H2O 27.8mg、Na2-EDTA·2H2O 37.3mg, inositol 100mg, nicotinic acid 0.5mg,
Puridoxine hydrochloride 0.5mg, thiamine hydrochloride 0.1mg, glycine 2mg.
The process for preparation of above-mentioned lamiophlomis rotata proliferative induction culture medium is:
(1) preparation of MS minimal medium mother liquor: include mother liquor 1, mother liquor 2, mother liquor 3, mother liquor 4,
Mother liquor 5, mother liquor 6, mother liquor 7 and the preparation of mother liquor 8, wherein
The preparation of mother liquor 1: weigh 170g KH2PO4It is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 2: weigh 1900g KNO3It is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 3: weigh 1650g NH4NO3It is placed in beaker, adds distilled water stirring and dissolving,
It is settled to 10000ml, is saved in brown break-in bottle；
The preparation of mother liquor 4: 370g MgSO4·7H2O is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 5: weigh 440g CaCl2·2H2O is placed in beaker, adds distilled water stirring and dissolving,
It is settled to 10000ml, is saved in brown break-in bottle；
The preparation of mother liquor 6: weigh 1.66g KI, 12.4g H respectively3BO3、33.8g MnSO4·H2O、17.2g
ZnSO4·7H2O、0.5g Na2MoO4·2H2O、0.05g CuSO4·5H2O and 0.05g CoCl2·6H2O,
Add in beaker and dissolve, be settled to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 7: weigh 200g inositol respectively, 1g nicotinic acid, 1g hydrochloric acid pyrrole tremble zinc, 0.2g salt
Allithiamine element and 4g glycine, add in beaker and dissolve, be settled to 10000ml, be saved in brown break-in
In Ping；
The preparation of mother liquor 8: weigh 55.6g FeSO4·7H2O、74.6g Na2-EDTA·2H2O adds respectively
Entering 3000ml distilled water to dissolve, then by two kinds of solution mixing, heating is boiled 15 minutes, and is stirred continuously,
Fully chelating, then it is settled to 10000ml, it is saved in brown break-in bottle；
(2) white sugar and carragheen pre-process: white sugar and carragheen are proportionally weighed respectively and add
The distilled water entering 15 times of quality dissolves, and stands 4 hours；
(3) by pretreatment white sugar and the carragheen aqueous solution, 10ml/L mother liquor 1,10ml/L mother liquor 2,
10ml/L mother liquor 3,10ml/L mother liquor 4,10ml/L mother liquor 5.5ml/L mother liquor 6,5ml/L mother liquor 7,
5ml/L mother liquor 8 is poured in bottle placer respectively, adds BA 3.0mg/L, NAA 0.5mg/L, IBA
0.50mg/L、ZT0.8mg/L.Add water constant volume, and stirs 15min, and regulation pH value is to 5.8, rear filling,
121 DEG C, sterilizing 23 minutes under conditions of 0.16MPa, prepare after condensation；Wherein, the filling time is
3s, the intermittent time is 0.7s.
Embodiment 3
The formula of this dendrobium candidum flower induction culture medium is: 1/2MS+ white sugar 40g/L+ carragheen
10g/L+BA 2.0mg/L+NAA 1.5mg/L+IBA 0.80mg/L+ZT0.4mg/L.Wherein, 1/2MS base
The formula of basal culture medium is: in every liter of culture medium, the content of component is as follows:
NH4NO3 82.5mg、KNO3 950mg、CaCl2·2H2O 220mg、MgSO4·7H2O 185mg、
KH2PO4 85mg、KI 0.415mg、H3BO3 3.1mg、MnSO4·H2O 8.45mg、ZnSO4·7H2O
4.3mg、Na2MoO4·2H2O 0.125mg、CuSO4·5H2O 0.0125mg、CoCl2·6H2O
0.0125mg、FeSO4·7H2O 13.9mg、Na2-EDTA·2H2O 18.65mg, inositol 50mg,
Nicotinic acid 0.25mg, puridoxine hydrochloride 0.25mg, thiamine hydrochloride 0.05mg, glycine 1mg.
The process for preparation of above-mentioned lamiophlomis rotata proliferative induction culture medium is:
(1) preparation of MS minimal medium mother liquor: include mother liquor 1, mother liquor 2, mother liquor 3, mother liquor 4,
Mother liquor 5, mother liquor 6, mother liquor 7 and the preparation of mother liquor 8, wherein
The preparation of mother liquor 1: weigh 170g KH2PO4It is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 2: weigh 1900g KNO3It is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 3: weigh 1650g NH4NO3It is placed in beaker, adds distilled water stirring and dissolving,
It is settled to 10000ml, is saved in brown break-in bottle；
The preparation of mother liquor 4: 370g MgSO4·7H2O is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 5: weigh 440g CaCl2·2H2O is placed in beaker, adds distilled water stirring and dissolving,
It is settled to 10000ml, is saved in brown break-in bottle；
The preparation of mother liquor 6: weigh 1.66g KI, 12.4g H respectively3BO3、33.8g MnSO4·H2O、17.2g
ZnSO4·7H2O、0.5g Na2MoO4·2H2O、0.05g CuSO4·5H2O and 0.05g CoCl2·6H2O,
Add in beaker and dissolve, be settled to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 7: weigh 200g inositol respectively, 1g nicotinic acid, 1g hydrochloric acid pyrrole tremble zinc, 0.2g salt
Allithiamine element and 4g glycine, add in beaker and dissolve, be settled to 10000ml, be saved in brown break-in
In Ping；
The preparation of mother liquor 8: weigh 55.6g FeSO4·7H2O、74.6g Na2-EDTA·2H2O adds respectively
Entering 3000ml distilled water to dissolve, then by two kinds of solution mixing, heating is boiled 15 minutes, and is stirred continuously,
Fully chelating, then it is settled to 10000ml, it is saved in brown break-in bottle；
(2) white sugar and carragheen pre-process: white sugar and carragheen are proportionally weighed respectively and add
The distilled water entering 15 times of quality dissolves, and stands 4 hours；
(3) by white sugar and the carragheen aqueous solution, 5ml/L mother liquor 1,5ml/L mother liquor 2, the 5ml/L of pretreatment
Mother liquor 3,5ml/L mother liquor 4,5ml/L mother liquor 5.2.5ml/L mother liquor 6,2.5ml/L mother liquor 7,2.5ml/L
Mother liquor 8 is poured in bottle placer respectively, add BA2.0mg/L, NAA 1.5mg/L, IBA 0.80mg/L,
ZT0.4mg/L adds water constant volume, and stirs 15min, and regulation pH value is to 5.8, rear filling, 121 DEG C,
Sterilizing 23 minutes under conditions of 0.16MPa, prepare after condensation；Wherein, the filling time is 3s, time intermittently
Between be 0.7s.
Test example
The plant leaf blade choosing the wild lamiophlomis rotata of county, Aba state of Sichuan province cultivates evoked callus, warp as outer implant
After Fiber differentiation and Multiplying culture, obtain Multiple Buds, this Multiple Buds is inoculated in the embodiment of the present invention 1 and prepares
Lamiophlomis rotata proliferative induction culture medium in, use 28 watts of T5 incandescent lamp fluorescent tubes as light source, intensity of illumination to be
3000lx, day temperature be 24 DEG C night temperature 22 DEG C, culturing room's air humidity is 55%, light application time 13 hours
Under conditions of/sky, cultivate 50 days, carry out proliferative induction；Wherein, table 1 is for using proliferative induction culture medium
Carry out cultivating and the compareing of the Reducing sugar using simple culture media to carry out cultivating:
Table 1
As can be seen from the above table, after using proliferative induction medium culture, the breeding coefficient ratio of Multiple Buds obtains
It is greatly improved.
Although in conjunction with specific embodiments the detailed description of the invention of the present invention is described in detail, but not
It it is the restriction to this patent protection domain.In claims limited range, those skilled in the art
Various amendments or adjustment that member can make without creative work are still protected by this patent.
Claims (4)
1. an induction lamiophlomis rotata proliferated culture medium, it is characterised in that: MS+ white sugar 20～40g/L+ carragheen
6.5～11g/L+BA 0.2～3mg/L+NAA0.1～1.5mg/L+IBA 0.1～1.0mg/L+ZT0.1～4mg/L.
In described lamiophlomis rotata proliferative induction culture medium, in every liter of MS culture medium, the content of each component is as follows: NH4NO3
165mg、KNO3 1900mg、CaCl2·2H2O 440mg、MgSO4·7H2O 370mg、KH2PO4
170mg、KI 0.83mg、H3BO36.2mg、MnSO4·H2O 16.9mg、ZnSO4·7H2O 8.6mg、
Na2MoO4·2H2O 0.25mg、CuSO4·5H2O 0.025mg、CoCl2·6H2O 0.025mg、
FeSO4·7H2O 27.8mg、Na2-EDTA·2H2O 37.3mg, inositol 100mg, nicotinic acid 0.5mg,
Hydrochloric acid pyrrole is trembled zinc 0.5mg, thiamine hydrochloride 0.1mg, glycine 2mg.
Lamiophlomis rotata group the most according to claim 1 training culture medium, it is characterised in that: include that MS+ is white
Sugar 30g/L+ carragheen 6.5g/L+BA 1.0mg/L+NAA 0.2mg/L+IBA 0.30mg/L
+ZT0.2mg/L。
3. a preparation method for the lamiophlomis rotata proliferative induction culture medium described in claim 1, its feature exists
In, including:
A, first white sugar and carragheen are pre-processed: respectively white sugar and carragheen are proportionally weighed
The distilled water adding 15 times of quality dissolves, and stands 3～4 hours；
The preparation of b, MS culture medium mother liquor, including mother liquor 1, mother liquor 2, mother liquor 3, mother liquor 4, mother liquor 5
Mother liquor 6, mother liquor 7 and the preparation of mother liquor 8, wherein
The preparation of mother liquor 1: weigh 170g KH2PO4It is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 2: weigh 1900g KNO3It is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 3: weigh 1650g NH4NO3It is placed in beaker, adds distilled water stirring and dissolving,
It is settled to 10000ml, is saved in brown break-in bottle；
The preparation of mother liquor 4: 370g MgSO4·7H2O is placed in beaker, adds distilled water stirring and dissolving, fixed
Hold to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 5: weigh 440g CaCl2·2H2O is placed in beaker, adds distilled water stirring and dissolving,
It is settled to 10000ml, is saved in brown break-in bottle；
The preparation of mother liquor 6: weigh 1.66g KI, 12.4g H respectively3BO3、33.8g MnSO4·H2O、17.2g
ZnSO4·7H2O、0.5g Na2MoO4·2H2O、0.05g CuSO4·5H2O and 0.05g CoCl2·6H2O,
Add in beaker and dissolve, be settled to 10000ml, be saved in brown break-in bottle；
The preparation of mother liquor 7: weigh 200g inositol respectively, 1g nicotinic acid, 1g hydrochloric acid pyrrole tremble zinc, 0.2g salt
Allithiamine element and 4g glycine, add in beaker and dissolve, be settled to 10000ml, be saved in brown break-in
In Ping；
The preparation of mother liquor 8: weigh 55.6g FeSO4·7H2O、74.6g Na2-EDTA·2H2O adds respectively
Entering 3000ml distilled water to dissolve, then by two kinds of solution mixing, heating is boiled 15 minutes, and is stirred continuously,
Fully chelating, then it is settled to 10000ml, it is saved in brown break-in bottle；
C, by pretreatment white sugar and the carragheen aqueous solution, 10ml/L mother liquor 1,10ml/L mother liquor 2,10ml/L
Mother liquor 3,10ml/L mother liquor 4,10ml/L mother liquor 5.5ml/L mother liquor 6,5ml/L mother liquor 7,5ml/L
Mother liquor 8 is poured in bottle placer respectively, adds BA, NAA, IBA, ZT.Add water constant volume, and stirs
15min, regulation pH value is to 5.8, rear filling, 121 DEG C, sterilizing 23 minutes under conditions of 0.16MPa,
Prepare after condensation.
The preparation method of lamiophlomis rotata proliferative induction culture medium the most according to claim 3, its feature exists
In: the described filling time is 3s, and the intermittent time is 0.7s.
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