CN104845908B - A kind of richness strontium hair weeds cells and its cultural method - Google Patents
A kind of richness strontium hair weeds cells and its cultural method Download PDFInfo
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Abstract
The invention discloses a kind of rich strontium hair weeds cells and its cultural methods, belong to biological food beneficiation technologies field.The richness strontium hair weeds cells are placed in cultivate in rich strontium culture medium by hair weeds cells and be obtained, and strontium chloride are added in rich strontium culture medium, and FeSO is added4·7H2O、Na2SiO3Equal substances, collective effect, it is organic strontium by cell transformation to promote absorption of the hair weeds cells to strontium, the inorganic strontium that hair weeds cells are absorbed from culture medium, to improve content of strontium in hair weeds cells, while improving the biomass of hair weeds cells.The preparation method of richness strontium hair weeds cells of the invention, is soaked in using that will deliver vegetables containing FeSO4·7H2O、Na2SiO3Culture obtains hair weeds cells in the culture medium of equal substances, then the method that hair weeds cells are cultivated in rich strontium culture medium, improves the content of strontium in hair weeds cells, and easy to operate, process is easily controllable, is suitable for industrial application.
Description
Technical field
The invention belongs to biological food Beneficiation technical fields, and in particular to a kind of richness strontium hair weeds cells and its preparation side
Method.
Background technology
Strontium is one of the trace element needed for tissue progress normal activities.Strontium to the function of human body mainly with
The formation of bone is closely related, is the normal component part of skeleton and tooth.Its function with blood vessel and it is configured with relationship,
Strontium competes absorption site with sodium in the intestine, can reduce absorption of the human body to sodium, increase the excretion of sodium.Internal sodium is excessive, easily causes
Hypertension, angiocardiopathy, and strontium can reduce absorption of the human body to sodium, play the role of preventing disease.
It delivers vegetables, also known as hair-like nostoc, because its color is black and elongated, gains the name such as the hair of people, and it is humorous with " making a good deal of money " to deliver vegetables
Sound, very popular over the past thousands of years, nutritive value of delivering vegetables is abundant, and 22.8g containing protein in every 100g natural lavers, is chicken
1.5 times of egg, fat content is low, rich in trace elements such as calcium, iron, iodine, magnesium, zinc, selenium, manganese.In recent years, excessive due to delivering vegetables
Harvesting has had resulted in serious environmental problem, and great harm is caused to ecological environment and social stability.Currently, from delivering vegetables
Hair weeds cells are detached in frond carrying out liquid suspension culture and be gradually valued by people, hair weeds cells culture technique is from delivering vegetables
The hair weeds cells for providing vigorous splitting ability are isolated and purified in frond, keep hair weeds cells largely numerous by liquid suspension culture
It grows, is handled by reprocessing, realize that hair weeds cells are factory produced, the growth rate of hair weeds cells is wild hair in incubation
30 times of dish growth rate realize the fast-growth breeding of hair-like seaweed resource.
There are two types of existing forms in hair weeds cells for strontium：One is the ionic state form of strontium, the second is being combined with organic matter
Organic strontium form.Ionic state strontium can be dissolved in water, be absorbed into the cell, a portion is combined further with protein
It is converted to organic strontium, this is the premise that hair weeds cells are enriched with organic strontium.And ionic state strontium soluble in water, it is special in process of production
It Rong Yi not be lost in, cause utilization rate not high, economic benefit unobvious.If ingesting high-content ionic state strontium for a long time, not only make
At waste, strontium intoxicating phenomenon there is also；On the contrary, organic strontium toxicity is low, bioactivity is high, is easily absorbed and utilized by humans and animals,
It is safe and efficient strontium preparation.Being enriched with organic strontium hair weeds cells has the characteristics of high Commercial cultivation rate, edible safety, and organic strontium is delivered vegetables
Cell can be used for a variety of diseases because caused by lacking strontium.Therefore, biological concentration and conversion are carried out as carrier using hair weeds cells
Inorganic strontium is that organic strontium is the approach for having at present development prospect.
Chinese patent CN103976411A discloses a kind of manufacturing method for product of delivering vegetables, by the way that soil is added in the medium
The mode of earth extracting solution and strontium solution promotes absorption of the hair weeds cells to strontium, while playing and prevent cardiovascular disease, but the public affairs
Content of strontium is still relatively low in the hair weeds cells of the preparation method culture for the product of delivering vegetables opened, how by adjusting culture medium composition
Ingredient improves the content of strontium of hair weeds cells, and the utilization of enrichment and hair weeds cells to strontium have great importance.
Invention content
In order to overcome the deficiencies of existing technologies, one of the objects of the present invention is to provide a kind of biomass height, content of strontium are high
Hair weeds cells.
Meanwhile the present invention also resides in the cultural method for providing a kind of rich strontium hair weeds cells.
To achieve the goals above, the technical solution adopted by the present invention is as follows：
A kind of richness strontium hair weeds cells, hair weeds cells are placed in rich strontium culture medium and cultivate acquisition, in the richness strontium culture medium
SrCl2Concentration and Na2SiO3Concentration ratio be（1~8）: 116.
A kind of cultural method of richness strontium hair weeds cells, including following operation：
1）Dry deliver vegetables is soaked in minimal medium, hair weeds cells inoculation liquid is obtained after homogenate；
2）Take step 1）Hair weeds cells inoculation liquid streak inoculation on solid medium of preparation, purifying culture are delivered vegetables
Cell culture, the single algae for therefrom selecting purifying falls, spare；
3）Take step 2）Single algae of middle purifying, which falls to be placed in minimal medium, expands culture, obtains hair weeds cells；
4）By step 3）The hair weeds cells of preparation, which are seeded in rich strontium, delivers vegetables and cultivates in culture medium, and filtering is stood after the completion of culture
Hair weeds cells are isolated to get the rich strontium hair weeds cells；
Wherein solid medium is that agar is added in minimal medium to be prepared；
The richness strontium culture medium is by SrCl2It is added in minimal medium and is mixed to prepare, SrCl2A concentration of 0.5 ~
4mg/L；Wherein mainly it is formulated by the raw material of following quality per 1000mL minimal mediums：NaNO3250mg, MgSO4·
7H2O 75mg, CaCl2·2H2O 36mg, H3BO40.286mg, MnCl2·4H2O 0.18mg, ZnSO4·7H2O 0.02mg,
CuSO4·5H2O 0.08mg, NaMoO4·2H2O 0.04mg, CoCl2·6H2O 0.001mg, EDTA-Na2（Ethanedioic acid tetrem
Acid disodium）0.lmg, FeSO4·7H2O 0.48mg, citric acid 0.06mg, Na2SiO358mg, K2HPO440mg, surplus are water.
Further, step 1）In it is dry deliver vegetables be soaked in minimal medium before pre-processed, specific method is：By dry hair
Dish sterilizes after totally drying wash with distilled water, then is rinsed with water clean.
Further, the disinfection is using 70% ~ 75% 0.5 ~ 1.0min of alcohol disinfecting.
Further, step 1）In deliver vegetables in basic medium soaking time be 8 ~ 11h.
Further, step 3）Middle expansion culture is to exponential phase up to hair weeds cells.
Further, step 4）Described in the condition of culture cultivated be：Temperature is 25 DEG C；Periodicity of illumination is 12h illumination 12h
Dark, intensity of illumination 2000Lux；Incubation time is 15 ~ 20 days.
Further, the cultural method of above-mentioned rich strontium hair weeds cells, further includes by step 4）In the hair weeds cells isolated
It is freeze-dried after being washed with water, the water catching bin temperature in freezing dry process is -50 ~ -55 DEG C, and the time is 12 ~ 20h to get institute
The rich strontium hair weeds cells product stated.
Further, the minimal medium is prepared by the method including following operating procedure：
Ⅰ：Raw material predissolve：
a：Take CaCl2、MgSO4·7H2O obtains solution A with water dissolution；
b：Take NaMoO4·2H2O、ZnSO4·7H2O、H3BO4、MnCl2·4H2O、EDTA-Na2、FeSO4·7H2O、
CuSO4·5H2O、CoCl2·6H2O, citric acid obtains solution B with water dissolution；
c：Take K2HPO4·3H2O obtains solution C with water dissolution；
d：Take Na2SiO3·9H2O obtains solution D with water dissolution；
Ⅱ：Sterilizing obtains solution E after solution A and solution B are mixed；After solution C and solution D are mixed, NaNO is added3,
Sterilize to obtain solution F；
Ⅲ：Solution E and solution F are mixed to get the minimal medium.
4 DEG C of the solution B is stored refrigerated spare.
The solution A, C, D room temperature save backup.
Sterilizing described in step II is that wet heating sterilizes, and temperature is 121 DEG C, time 20min.
The richness strontium culture medium is prepared by following methods：Take SrCl2After water dissolution, SrCl is obtained2Mother liquor, will
SrCl2It is added in minimal medium to get the rich strontium hair weeds cells culture medium after mother liquor sterilizing.The rich strontium hair weeds cells of adjustment
The pH of culture medium is 7.5.
The present invention also protects a kind of rich strontium hair weeds cells prepared using above-mentioned cultural method.
Advantageous effect
Richness strontium hair weeds cells of the invention, hair weeds cells are placed in rich strontium culture medium and cultivate acquisition, are added in rich strontium culture medium
Enter strontium chloride, and FeSO is added4·7H2O、Na2SiO3Equal substances, collective effect promote absorption of the hair weeds cells to strontium, deliver vegetables
The inorganic strontium that cell is absorbed from culture medium is organic strontium by cell transformation, to improve content of strontium in hair weeds cells, together
Biomass in Shi Tigao hair weeds cells.
The preparation method of richness strontium hair weeds cells of the invention is obtained using that will deliver vegetables to be soaked in basic culture solution to extract to be inoculated with
The hair weeds cells list algae of purifying falls, and obtains hair weeds cells by the expansion culture of minimal medium, and hair weeds cells are placed in richness
Culture obtains rich strontium hair weeds cells in strontium culture medium, improves absorption of the hair weeds cells to inorganic strontium in culture medium, and then improve
Content of strontium in hair weeds cells, while improving the biomass in hair weeds cells.Preparation method of the present invention, easy to operate, process is easy
In control, it is suitable for industrial application.
Specific implementation mode
With reference to specific embodiment, invention is further described in detail, but does not constitute any limit to the present invention
System.
Embodiment 1
The present embodiment richness strontium hair weeds cells are that hair weeds cells are cultivated to acquisition in rich strontium culture medium, wherein rich strontium culture medium
For by SrCl2It is added in minimal medium and is mixed to prepare, SrCl2A concentration of 2mg/L；Wherein per 1000mL minimal mediums
Mainly it is formulated by the raw material of following quality：NaNO3250mg, MgSO4·7H2O 75mg, CaCl2·2H2O 36mg,
H3BO40.286mg, MnCl2·4H2O 0.18mg, ZnSO4·7H2O 0.02mg, CuSO4·5H2O 0.08mg, NaMoO4·
2H2O 0.04mg, CoCl2·6H2O 0.001mg, EDTA-Na2（Ethanedioic acid tetraacethyl disodium）0.lmg, FeSO4·7H2O
0.48mg, citric acid 0.06mg, Na2SiO358mg, K2HPO4 40mg。
The preparation method of the present embodiment richness strontium hair weeds cells, concrete operation step are as follows：
1）Prepare culture medium：
A：Prepare minimal medium：
Ⅰ：Raw material predissolve：
a：Weigh CaCl2·2H2O 2.05g、MgSO4·7H2O 3.75g are settled to 500mL, are obtained with distillation water dissolution
To solution A, room temperature saves backup；
b：Weigh NaMoO4·2H2O 20mg、ZnSO4·7H2O 10mg、H3BO4 143mg、MnCl2·4H2O 90mg、
EDTA-Na2 50mg、FeSO4·7H2O 240mg、CuSO4·5H2O 40mg、CoCl2·6H2O 0.5mg, citric acid 30mg,
After distillation water dissolution, it is settled to 500mL, obtains solution B, 4 DEG C of refrigerations are spare；
c：Weigh K2HPO4·3H2O 2g are settled to 500mL after distillation water dissolution, obtain solution C, and room temperature preserves standby
With；
d：Weigh Na2SiO3·9H2O 2.9g are settled to 500mL with distillation water dissolution, obtain solution D, and room temperature preserves standby
With；
Ⅱ：With the accurate draw solution A 20mL of pipette, solution B 2mL mixing adds distilled water to be settled to 1000mL, dispenses
Into 10 250mL conical flasks, every bottle of 100mL is wrapped up, and 121 DEG C of wet methods sterilizing 20min obtain solution E；It is accurate with pipette
NaNO is added after solution D 20mL mixing in draw solution C 20mL32.5g adds distilled water to be settled to 1000mL, is dispensed into 10
In 250mL conical flasks, every bottle of 100mL is wrapped up, and 121 DEG C of wet methods sterilizing 20min obtain solution F；
Ⅲ：Solution E and solution F are placed under superclean bench aseptic condition and are cooled to room temperature, after mixing to get institute
The minimal medium stated.
B：Prepare rich strontium culture medium：
Weigh SiCl250mg is settled to 50mL, the mother liquor of 1mg/mL is configured to, using filtration method pair after distilling water dissolution
Mother liquor carries out degerming, and the mother liquor 2mL after sterilizing is taken to be added in 998mL minimal mediums, is uniformly mixed, adjusts the pH value of solution
It is 7.5, obtains rich strontium culture medium, SiCl in the culture medium2A concentration of 2mg/L.
C：Prepare solid medium：It takes 100g minimal mediums that 1.7g agar is added, solid medium is made.
2）Hair weeds cells strain isolates and purifies
A：Deliver vegetables dry 3 times wash with distilled water, after drying with 75% alcohol disinfecting 0.8min, then rushed with distilled water
Wash clean obtains the dry of cleaning and delivers vegetables；
B：Clean dry deliver vegetables prepared by step A is soaked in 10h in minimal medium, then uses glass pressure-even pulp crusher equal
Slurry, obtains hair weeds cells inoculation liquid；
C：The step B hair weeds cells prepared are seeded in streak inoculation on solid medium, are cultivated through 5 ~ 6 times purifying
To hair weeds cells culture；
D：The single algae purified in picking hair weeds cells culture falls, and is placed in inflation in minimal medium and expands culture, harvest
The cell to exponential phase is cultivated to get hair weeds cells.
3）The culture of rich strontium hair weeds cells
Aseptically, by step 2）The hair weeds cells of preparation are seeded in rich strontium culture medium, are put into 25 DEG C of constant temperature
It is cultivated in biochemical cultivation case case, the periodicity of illumination of culture is 12h illumination 12h dark, and intensity of illumination 2000Lux is cultivated 20 days,
Stationary culture liquid, filters to isolate hair weeds cells, after the hair weeds cells isolated are washed with distilled water three times, pours into tablet
In, it is freeze-dried, the water catching bin temperature in freezing dry process is -53 DEG C, and the time is 18h to get rich strontium hair weeds cells.
Embodiment 2
The present embodiment richness strontium hair weeds cells are that hair weeds cells are cultivated to acquisition in rich strontium culture medium, wherein rich strontium culture medium
For by SrCl2It is added in minimal medium and is mixed to prepare, SrCl2A concentration of 4mg/L；Wherein per 1000mL minimal mediums
Mainly it is formulated by the raw material of following quality：NaNO3250mg, MgSO4·7H2O 75mg, CaCl2·2H2O 36mg,
H3BO40.286mg, MnCl2·4H2O 0.18mg, ZnSO4·7H2O 0.02mg, CuSO4·5H2O 0.08mg, NaMoO4·
2H2O 0.04mg, CoCl2·6H2O 0.001mg, EDTA-Na2（Ethanedioic acid tetraacethyl disodium）0.lmg, FeSO4·7H2O
0.48mg, citric acid 0.06mg, Na2SiO358mg, K2HPO4 40mg。
The preparation method of the present embodiment richness strontium hair weeds cells, concrete operation step are as follows：
1）Prepare culture medium：
A：Prepare minimal medium：
Ⅰ：Raw material predissolve：
a：Weigh CaCl2·2H2O 2.05g、MgSO4·7H2O 3.75g are settled to 500mL, are obtained with distillation water dissolution
To solution A, room temperature saves backup；
b：Weigh NaMoO4·2H2O 20mg、ZnSO4·7H2O 10mg、H3BO4 143mg、MnCl2·4H2O 90mg、
EDTA-Na2 50mg、FeSO4·7H2O 240mg、CuSO4·5H2O 40mg、CoCl2·6H2O 0.5mg, citric acid 30mg,
After distillation water dissolution, it is settled to 500mL, obtains solution B, 4 DEG C of refrigerations are spare；
c：Weigh K2HPO4·3H2O 2g are settled to 500mL after distillation water dissolution, obtain solution C, and room temperature preserves standby
With；
d：Weigh Na2SiO3·9H2O 2.9g are settled to 500mL with distillation water dissolution, obtain solution D, and room temperature preserves standby
With；
Ⅱ：With the accurate draw solution A 20mL of pipette, solution B 2mL mixing adds distilled water to be settled to 1000mL, dispenses
Into 10 250mL conical flasks, every bottle of 100mL is wrapped up, and 121 DEG C of wet methods sterilizing 20min obtain solution E；It is accurate with pipette
NaNO is added after solution D 20mL mixing in draw solution C 20mL32.5g adds distilled water to be settled to 1000mL, is dispensed into 10
In 250mL conical flasks, every bottle of 100mL is wrapped up, and 121 DEG C of wet methods sterilizing 20min obtain solution F；
Ⅲ：Solution E and solution F are placed under superclean bench aseptic condition and are cooled to room temperature, after mixing to get institute
The minimal medium stated.
B：Prepare rich strontium culture medium：
Weigh SiCl250mg is settled to 50mL, the mother liquor of 1mg/mL is configured to, using filtration method pair after distilling water dissolution
Mother liquor carries out degerming, and the mother liquor 4mL after sterilizing is taken to be added in 996mL minimal mediums, is uniformly mixed, adjusts the pH value of solution
It is 7.5, obtains rich strontium culture medium, SiCl in the culture medium2A concentration of 4mg/L.
C：Prepare solid medium：It takes 100g minimal mediums that 1.7g agar is added, solid medium is made.
2）Hair weeds cells strain isolates and purifies
A：Deliver vegetables dry 4 times wash with distilled water, after drying with 75% alcohol disinfecting 1.0min, then rushed with distilled water
Wash clean obtains the dry of cleaning and delivers vegetables；
B：Clean dry deliver vegetables prepared by step A is soaked in 8h in minimal medium, then uses glass pressure-even pulp crusher homogenate,
Obtain hair weeds cells inoculation liquid；
C：The step B hair weeds cells prepared are seeded in streak inoculation on solid medium, are obtained through 5 ~ 6 purifying cultures
Hair weeds cells culture；
D：The single algae purified in picking hair weeds cells culture falls, and is placed in inflation in minimal medium and expands culture, harvest
The cell to exponential phase is cultivated to get hair weeds cells.
3）The culture of rich strontium hair weeds cells
Aseptically, by step 2）The hair weeds cells of preparation are seeded in rich strontium culture medium, are put into 25 DEG C of constant temperature
It is cultivated in biochemical cultivation case case, the periodicity of illumination of culture is 12h illumination 12h dark, and intensity of illumination 2000Lux is cultivated 15 days,
Stationary culture liquid, filters to isolate hair weeds cells, after the hair weeds cells isolated are washed with distilled water three times, pours into tablet
In, it is freeze-dried, the water catching bin temperature in freezing dry process is -53 DEG C, and the time is 18h to get rich strontium hair weeds cells.
Embodiment 3
The present embodiment richness strontium hair weeds cells are that hair weeds cells are cultivated to acquisition in rich strontium culture medium, wherein rich strontium culture medium
For by SrCl2It is added in minimal medium and is mixed to prepare, SrCl2A concentration of 0.5mg/L；Wherein cultivated substantially per 1000mL
Base is mainly formulated by the raw material of following quality：NaNO3250mg, MgSO4·7H2O 75mg, CaCl2·2H2O 36mg,
H3BO40.286mg, MnCl2·4H2O 0.18mg, ZnSO4·7H2O 0.02mg, CuSO4·5H2O 0.08mg, NaMoO4·
2H2O 0.04mg, CoCl2·6H2O 0.001mg, EDTA-Na2（Ethanedioic acid tetraacethyl disodium）0.lmg, FeSO4·7H2O
0.48mg, citric acid 0.06mg, Na2SiO358mg, K2HPO4 40mg。
The preparation method of the present embodiment richness strontium hair weeds cells, concrete operation step are as follows：
1）Prepare culture medium：
A：Prepare minimal medium：
Ⅰ：Raw material predissolve：
a：Weigh CaCl2·2H2O 2.05g、MgSO4·7H2O 3.75g are settled to 500mL, are obtained with distillation water dissolution
To solution A, room temperature saves backup；
b：Weigh NaMoO4·2H2O 20mg、ZnSO4·7H2O 10mg、H3BO4 143mg、MnCl2·4H2O 90mg、
EDTA-Na2 50mg、FeSO4·7H2O 240mg、CuSO4·5H2O 40mg、CoCl2·6H2O 0.5mg, citric acid 30mg,
After distillation water dissolution, it is settled to 500mL, obtains solution B, 4 DEG C of refrigerations are spare；
c：Weigh K2HPO4·3H2O 2g are settled to 500mL after distillation water dissolution, obtain solution C, and room temperature preserves standby
With；
d：Weigh Na2SiO3·9H2O 2.9g are settled to 500mL with distillation water dissolution, obtain solution D, and room temperature preserves standby
With；
Ⅱ：With the accurate draw solution A 20mL of pipette, solution B 2mL mixing adds distilled water to be settled to 1000mL, dispenses
Into 10 250mL conical flasks, every bottle of 100mL is wrapped up, and 121 DEG C of wet methods sterilizing 20min obtain solution E；It is accurate with pipette
NaNO is added after solution D 20mL mixing in draw solution C 20mL32.5g adds distilled water to be settled to 1000mL, is dispensed into 10
In a 250mL conical flasks, every bottle of 100mL is wrapped up, and 121 DEG C of wet methods sterilizing 20min obtain solution F；Solution A and solution B are mixed
Sterilizing obtains solution E after conjunction；After solution C and solution D are mixed, NaNO is added3, sterilize to obtain solution F；
Ⅲ：Solution E and solution F are placed under superclean bench aseptic condition and are cooled to room temperature, after mixing to get institute
The minimal medium stated.
B：Prepare rich strontium culture medium：
Weigh SiCl250mg is settled to 50mL, the mother liquor of 1mg/mL is configured to, using filtration method pair after distilling water dissolution
Mother liquor carries out degerming, and the mother liquor 0.5L after sterilizing is taken to be added in 999.5mL minimal mediums, is uniformly mixed, adjusts solution
PH value is 7.5, obtains rich strontium culture medium, SiCl in the culture medium2A concentration of 0.5mg/L.
C：Prepare solid medium：It takes 100g minimal mediums that 1.7g agar is added, solid medium is made.
2）Hair weeds cells strain isolates and purifies
A：Deliver vegetables dry 3 times wash with distilled water, after drying with 75% alcohol disinfecting 0.8min, then rushed with distilled water
Wash clean obtains the dry of cleaning and delivers vegetables；
B：Clean dry deliver vegetables prepared by step A is soaked in 11h in minimal medium, then uses glass pressure-even pulp crusher equal
Slurry, obtains hair weeds cells inoculation liquid；
C：The step B hair weeds cells prepared are seeded in streak inoculation on solid medium, are obtained through 5-6 purifying culture
Hair weeds cells culture；
D：The single algae purified in picking hair weeds cells culture falls, and is placed in inflation in minimal medium and expands culture, harvest
The cell to exponential phase is cultivated to get hair weeds cells.
3）The culture of rich strontium hair weeds cells
Aseptically, by step 2）The hair weeds cells of preparation are seeded in rich strontium culture medium, are put into 25 DEG C of constant temperature
It is cultivated in biochemical cultivation case case, the periodicity of illumination of culture is 12h illumination 12h dark, and intensity of illumination 2000Lux is cultivated 18 days,
Stationary culture liquid, filters to isolate hair weeds cells, after the hair weeds cells isolated are washed with distilled water three times, pours into tablet
In, it is freeze-dried, control water catching bin temperature is -53 DEG C in freezing dry process, and the time is 18h to get rich strontium hair weeds cells.
Test example
Test method：Change the formula of rich strontium culture medium, biology in the hair weeds cells of more different richness strontium medium cultures
Content and content of strontium；
It is specific as follows：
A groups：FeSO is not added in culture medium4·7H2O、Na2SiO3、SrCl2With strontium solution, other components are the same as embodiment 1.
B groups：FeSO is not added in culture medium4·7H2O and Na2SiO3, add SrCl2, strontium chloride is final dense in culture medium
Degree is 2mg/L, and other components are the same as embodiment 1.
C groups：FeSO is not added in culture medium4·7H2O and Na2SiO3, strontium solution is added, the ultimate density of strontium in culture medium
For 2mg/L, other components are the same as embodiment 1.
D groups：The embodiment of the present invention 1
E groups：FeSO is added in culture medium4·7H2O and Na2SiO3, strontium solution is added, the ultimate density of strontium is in culture solution
2mg/L, other components are the same as embodiment 1.
Strontium solution is to be obtained from rich strontium mineral water in this test example.
The above each group respectively does 3 groups of parallel tests, and cultural method measures hair weeds cells biomass respectively with embodiment 1, uses
Micro-wave digestion, ICP-OMS methods measure the content of strontium element in cell, and results are averaged, and test result is as shown in table 1 below：
Influence of the more different rich strontium culture medium prescription of table 1 to hair weeds cells biomass and content of strontium
From table 1, compares A, B, C group and D, E group biomass of delivering vegetables can be seen that and adds FeSO in culture solution4·7H2O and
Na2SiO3The biomass of hair weeds cells can be significantly improved；Compare D groups and E groups obtain, compare strontium solution, adds SrCl2It can be slightly
The biomass of hair weeds cells is improved, and the content of strontium element in hair weeds cells can be significantly improved.
Show to be added in the culture medium in rich strontium hair weeds cells incubation of the invention by above-mentioned test result
FeSO4·7H2O and Na2SiO3And use SrCl2For inorganic barium source, each ingredient in culture medium is made mutually to act synergistically, improves hair
Strontium element content in dish cellular biomass and cell makes the biomass of hair weeds cells reach 232mg/L, strontium element in hair weeds cells
Content reach 236 μ g/g.
Claims (8)
1. a kind of cultural method of richness strontium hair weeds cells, which is characterized in that including following operation：
1）It will deliver vegetables and be soaked in minimal medium, hair weeds cells inoculation liquid is obtained after homogenate；
2）Take step 1）Hair weeds cells inoculation liquid streak inoculation on solid medium of preparation, purifying culture obtain hair weeds cells
Culture, the single algae for therefrom selecting purifying falls, spare；
3）By step 2）Single algae of middle purifying, which falls to be placed in minimal medium, expands culture, obtains hair weeds cells；
4）By step 3）The hair weeds cells of preparation are seeded in rich strontium culture medium and cultivate, and stand to be separated by filtration after the completion of culture and set out
Dish cell, the as rich strontium hair weeds cells；
Wherein solid medium is to be prepared after agar is added in minimal medium；
The richness strontium culture medium is by SrCl2It is added in minimal medium and is mixed to prepare, SrCl in rich strontium culture medium2Concentration
For 0.5 ~ 4mg/L；Wherein it is formulated by the raw material of following quality per 1000mL minimal mediums：NaNO3250mg, MgSO4·
7H2O 75mg, CaCl2·2H2O 36mg, H3BO40.286mg, MnCl2·4H2O 0.18mg, ZnSO4·7H2O 0.02mg,
CuSO4·5H2O 0.08mg, NaMoO4·2H2O 0.04mg, CoCl2·6H2O 0.001mg, EDTA-Na20.lmg,
FeSO4·7H2O 0.48mg, citric acid 0.06mg, Na2SiO358mg, K2HPO440mg, surplus are water.
2. the cultural method of richness strontium hair weeds cells as described in claim 1, which is characterized in that step 1）In deliver vegetables and be soaked in base
It is pre-processed before basal culture medium, specific method is：It will deliver vegetables and be sterilized after totally drying wash with distilled water, then be rinsed with water dry
Only.
3. the cultural method of richness strontium hair weeds cells as claimed in claim 2, which is characterized in that the disinfection for using 70% ~
75% 0.5 ~ 1.0min of alcohol disinfecting.
4. the cultural method of richness strontium hair weeds cells as described in claim 1, which is characterized in that step 1）In deliver vegetables and training substantially
It is 8 ~ 11h to support soaking time in base.
5. the cultural method of richness strontium hair weeds cells as described in claim 1, which is characterized in that step 3）Middle expansion is cultivated to right
Number growth period is up to hair weeds cells.
6. the cultural method of richness strontium hair weeds cells as described in claim 1, which is characterized in that step 4）Described in the training cultivated
Foster condition is：Temperature is 25 DEG C；Periodicity of illumination is 12h illumination 12h dark, intensity of illumination 2000Lux；Incubation time be 15 ~
20 days.
7. the cultural method of richness strontium hair weeds cells as described in claim 1, which is characterized in that further include by step 4）Middle separation
Freeze-drying is to get rich strontium hair weeds cells product after the hair weeds cells gone out are washed with water.
8. the cultural method of richness strontium hair weeds cells as described in claim 1, which is characterized in that the preparation of the minimal medium
Method includes the following steps：
Ⅰ：Raw material predissolve：
a：Take CaCl2·2H2O、MgSO4·7H2O obtains solution A with water dissolution；
b：Take NaMoO4·2H2O、ZnSO4·7H2O、H3BO4、MnCl2·4H2O、EDTA-Na2、FeSO4·7H2O、CuSO4·
5H2O、CoCl2·6H2O, citric acid obtains solution B with water dissolution；
c：Take K2HPO4·3H2O obtains solution C with water dissolution；
d：Take Na2SiO3·9H2O obtains solution D with water dissolution；
Ⅱ：Sterilizing obtains solution E after solution A and solution B are mixed；After solution C and solution D are mixed, NaNO is added3, sterilize
Solution F；
Ⅲ：Solution E and solution F are mixed to get the minimal medium.
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| CN1392245A (en) * | 2002-07-26 | 2003-01-22 | 中国科学院水生生物研究所 | Method for cultivating hair weeds cell |
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| CN103976411A (en) * | 2014-04-30 | 2014-08-13 | 河南科技大学 | Manufacturing method of nostoc flagelliforme product |
| CN104403985A (en) * | 2014-12-26 | 2015-03-11 | 天津科技大学 | Culture method of iodine-rich Nostoc flagelliforme |
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| CN103976411A (en) * | 2014-04-30 | 2014-08-13 | 河南科技大学 | Manufacturing method of nostoc flagelliforme product |
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