CN104845908A - Strontium-rich hair weed cell and culture method thereof - Google Patents

Strontium-rich hair weed cell and culture method thereof Download PDF

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CN104845908A
CN104845908A CN201510206703.4A CN201510206703A CN104845908A CN 104845908 A CN104845908 A CN 104845908A CN 201510206703 A CN201510206703 A CN 201510206703A CN 104845908 A CN104845908 A CN 104845908A
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strontium
hair weeds
weeds cells
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hair
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CN104845908B (en
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郭金英
朱蓓茹
任国艳
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Guo Jinying
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Henan University of Science and Technology
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Abstract

The invention discloses a strontium-rich hair weed cell and a culture method thereof, belonging to the technical field of food bio-concentration. The strontium-rich hair weed cell is prepared by culturing a hair weed cell in a strontium-rich medium; strontium chloride and substances like FeSO4.7H2O and Na2SiO3 are added into the strontium-rich medium and cooperatively promote strontium absorption of the hair weed cell; and the inorganic strontium absorbed from the medium by the hair weed cell is transformed into organic strontium through cell transformation, so the content of strontium in the hair weed cell is increased, and the biomass of the hair weed cell is improved. According to the culture method for the strontium-rich hair weed cell, hair weed is soaked and cultured in the medium containing substances like FeSO4.7H2O and Na2SiO3 so as to obtain the hair weed cell, and the hair weed cell is cultured in the strontium-rich medium, so the content of strontium in the hair weed cell is increased; moreover, operation is simple and convenient, process is easy to control, and the method is suitable for industrial popularization and application.

Description

A kind of rich strontium hair weeds cells and cultural method thereof
Technical field
The invention belongs to biological food Beneficiation technical field, be specifically related to a kind of rich strontium hair weeds cells and preparation method thereof.
Background technology
Strontium is one of tissue trace element of carrying out needed for normal activities.Strontium is mainly closely related with the formation of bone to the function of human body, is the normal integral part of skeleton and tooth.The function of it and blood vessel and be configured with relation, strontium competes absorption site with sodium in intestines, can reduce the absorption of human body to sodium, increases the excretion of sodium.In body, sodium is too much, easily cause hypertension, cardiovascular disorder, and strontium can reduce the absorption of human body to sodium, have prophylactic effect.
Deliver vegetables, also known as hair-like nostoc, Yin Qisehei and elongated, as people hair and gain the name, and deliver vegetables and " making a good deal of money " homophonic, very popular over the past thousands of years, delivering vegetables rich in nutritive value, containing protein 22.8g in every 100g natural laver, is 1.5 times of egg, lipid content is low, is rich in the trace elements such as calcium, iron, iodine, magnesium, zinc, selenium, manganese.In recent years, owing to delivering vegetables, excessive gathering brings serious environmental problem, causes great harm to ecotope and social stability.At present, from frond of delivering vegetables, be separated hair weeds cells carry out fluid suspension culture and be subject to people's attention gradually, hair weeds cells culture technique is the hair weeds cells that separation and purification goes out to have vigorous splitting ability from frond of delivering vegetables, hair weeds cells amount reproduction is made by fluid suspension culture, by reprocessing process, realize hair weeds cells factorial praluction, in culturing process, the growth velocity of hair weeds cells is 30 times of wild growth velocity of delivering vegetables, and achieves the quick growth and breeding of hair-like seaweed resource.
Strontium has two kinds of existing forms in hair weeds cells: the ionic state form of first strontium, and it two is the organic strontium forms be combined with organism.Ionic state strontium can be dissolved in water, is absorbed in cell, and wherein a part and protein bound change into organic strontium further, and this is the prerequisite of the organic strontium of hair weeds cells enrichment.And ionic state strontium soluble in water, especially easily run off in process of production, cause utilization ratio not high, economic benefit is not obvious.If ingested for a long time, high-content ionic state strontium, not only causes waste, also there will be strontium intoxicating phenomenon; On the contrary, organic strontium toxicity is low, and biological activity is high, is easily absorbed by humans and animals, is strontium preparation safely and efficiently.Enrichment organic strontium hair weeds cells has that Commercial cultivation rate is high, the feature of edible safety, and organic strontium hair weeds cells can be used for the various diseases caused because of scarce strontium.Therefore, hair weeds cells is utilized to carry out biomagnification and transform inorganic strontium be organic strontium being the approach at present with development prospect as carrier.
Chinese patent CN103976411A discloses a kind of manufacture method of product of delivering vegetables, by adding the mode of soil extract and strontium solution in the medium, promote that hair weeds cells is to the absorption of strontium, serve simultaneously and prevent cardiovascular diseases, but content of strontium is still lower in the hair weeds cells that the preparation method of the product of delivering vegetables of the disclosure cultivates, how to pass through the moiety of adjustment substratum, improve the content of strontium of hair weeds cells, the enrichment of strontium and the exploitation of hair weeds cells are had great importance.
Summary of the invention
In order to overcome the defect of prior art, an object of the present invention is to provide the hair weeds cells that a kind of biomass is high, content of strontium is high.
Meanwhile, the present invention is also the cultural method providing a kind of rich strontium hair weeds cells.
To achieve these goals, the technical solution used in the present invention is as follows:
A kind of rich strontium hair weeds cells, hair weeds cells is placed in rich strontium substratum and cultivates acquisition, described rich strontium substratum is by SrCl 2be added into mixing in minimum medium obtained, SrCl 2concentration be 0.5 ~ 4mg/L; Wherein every 1000mL minimum medium forms primarily of the preparation of raw material of following quality: NaNO 3250mg, MgSO 47H 2o 75mg, CaCl 22H 2o 36mg, H 3bO 40.286mg, MnCl 24H 2o 0.18mg, ZnSO 47H 2o 0.02mg, CuSO 45H 2o 0.08mg, NaMoO 42H 2o 0.04mg, CoCl 26H 2o 0.001mg, EDTA-Na 2(oxalic acid tetraacethyl disodium) 0.lmg, FeSO 47H 2o 0.48mg, citric acid 0.06mg, Na 2siO 358mg, K 2hPO 440mg, surplus is water.
SrCl in described rich strontium substratum 2concentration and Na 2siO 3concentration ratio be (1 ~ 8): 116.
The cultural method of above-mentioned rich strontium hair weeds cells, comprises following operation:
1) dry delivering vegetables is soaked in minimum medium, after homogenate, obtains hair weeds cells inoculation liquid;
2) get hair weeds cells inoculation liquid streak inoculation on solid medium prepared by step 1), purifying is cultivated and is obtained hair weeds cells culture, and the single algae therefrom selecting purifying falls, for subsequent use;
3) get step 2) in single algae of purifying fall to being placed in minimum medium enlarged culturing, obtain hair weeds cells;
4) hair weeds cells prepared by step 3) is seeded in rich strontium to deliver vegetables in substratum and cultivate, has cultivated rear leaving standstill and filtered to isolate hair weeds cells, obtained described rich strontium hair weeds cells;
Wherein solid medium is add agar preparation in minimum medium to obtain.
In described solid culture, the mass ratio of minimum medium and agar is 100:1.7.
In step 1) dry deliver vegetables be soaked in minimum medium before carry out pre-treatment, concrete grammar is: by dry deliver vegetables to clean up with distilled water dry rear sterilization, then to rinse well with water.
Described sterilization is the alcohol disinfecting 0.5 ~ 1.0min of employing 70% ~ 75%.
Soak time in minimum medium of delivering vegetables in step 1) is 8 ~ 11h.
In step 3), namely enlarged culturing obtains hair weeds cells to logarithmic phase.
The culture condition cultivated described in step 4) is: temperature is 25 DEG C; Periodicity of illumination is that 12h illumination 12h is dark, and intensity of illumination is 2000Lux; Incubation time is 15 ~ 20 days.
The cultural method of above-mentioned rich strontium hair weeds cells, also comprises and washes hair weeds cells isolated in step 4) with water postlyophilization, and the water catching bin temperature in freezing dry process is-50 ~-55 DEG C, and the time is 12 ~ 20h, obtains described rich strontium hair weeds cells product.
Described minimum medium is prepared by the method comprising following operation steps and obtains:
I: raw material predissolve:
A: get CaCl 2, MgSO 47H 2o, by water dissolution, obtains solution A;
B: get NaMoO 42H 2o, ZnSO 47H 2o, H 3bO 4, MnCl 24H 2o, EDTA-Na 2, FeSO 47H 2o, CuSO 45H 2o, CoCl 26H 2o, citric acid, by water dissolution, obtain solution B;
C: get K 2hPO 43H 2o, by water dissolution, obtains solution C;
D: get Na 2siO 39H 2o, by water dissolution, obtains solution D;
II: sterilizing after solution A and solution B mixing is obtained solution E; After solution C and solution D mixing, add NaNO 3, sterilizing obtains solution F;
III: solution E and solution F are mixed, obtains described minimum medium.
Described solution B 4 DEG C of stored refrigerated are for subsequent use.
Described solution A, C, D normal temperature save backup.
Sterilizing described in step II is wet-heating sterilizing, and temperature is 121 DEG C, and the time is 20min.
Described rich strontium substratum is prepared by following methods and obtains: get SrCl 2after water dissolution, obtain SrCl 2mother liquor, by SrCl 2add after mother liquor sterilizing in minimum medium, obtain described rich strontium hair weeds cells substratum.The pH adjusting rich strontium hair weeds cells substratum is 7.5.
beneficial effect
The rich strontium hair weeds cells of the present invention, is placed in rich strontium substratum and cultivates acquisition, add strontium chloride, and add FeSO in rich strontium substratum by hair weeds cells 47H 2o, Na 2siO 3deng material, acting in conjunction, promote that hair weeds cells is to the absorption of strontium, the inorganic strontium that hair weeds cells absorbs from substratum is organic strontium by cell transformation, thus improves content of strontium in hair weeds cells, improves the biomass in hair weeds cells simultaneously.
The preparation method of the rich strontium hair weeds cells of the present invention, the hair weeds cells list algae being soaked in extraction inoculation acquisition purifying in basic culture solution by delivering vegetables is adopted to fall, and obtain hair weeds cells by the enlarged culturing of minimum medium, hair weeds cells is placed in rich strontium substratum and cultivates the rich strontium hair weeds cells of acquisition, improve the absorption of hair weeds cells to strontium inorganic in substratum, and then improve content of strontium in hair weeds cells, improve the biomass in hair weeds cells simultaneously.Preparation method of the present invention, easy and simple to handle, process is easy to control, and is suitable for industrial application.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in further detail, but does not form any limitation of the invention.
Embodiment 1
The rich strontium hair weeds cells of the present embodiment is that hair weeds cells is cultivated acquisition in rich strontium substratum, and wherein rich strontium substratum is by SrCl 2be added into mixing in minimum medium obtained, SrCl 2concentration be 2mg/L; Wherein every 1000mL minimum medium forms primarily of the preparation of raw material of following quality: NaNO 3250mg, MgSO 47H 2o 75mg, CaCl 22H 2o 36mg, H 3bO 40.286mg, MnCl 24H 2o 0.18mg, ZnSO 47H 2o 0.02mg, CuSO 45H 2o 0.08mg, NaMoO 42H 2o 0.04mg, CoCl 26H 2o 0.001mg, EDTA-Na 2(oxalic acid tetraacethyl disodium) 0.lmg, FeSO 47H 2o 0.48mg, citric acid 0.06mg, Na 2siO 358mg, K 2hPO 440mg.
The preparation method of the rich strontium hair weeds cells of the present embodiment, concrete operation step is as follows:
1) substratum is prepared:
A: preparation minimum medium:
I: raw material predissolve:
A: take CaCl 22H 2o 2.05g, MgSO 47H 2o 3.75g, dissolve with distilled water, be settled to 500mL, obtain solution A, normal temperature saves backup;
B: take NaMoO 42H 2o 20mg, ZnSO 47H 2o 10mg, H 3bO 4143mg, MnCl 24H 2o 90mg, EDTA-Na 250mg, FeSO 47H 2o 240mg, CuSO 45H 2o 40mg, CoCl 26H 2o 0.5mg, citric acid 30mg, after dissolving with distilled water, be settled to 500mL, obtain solution B, 4 DEG C of refrigerations are for subsequent use;
C: take K 2hPO 43H 2o 2g, after dissolving with distilled water, be settled to 500mL, obtain solution C, normal temperature saves backup;
D: take Na 2siO 39H 2o 2.9g, dissolve with distilled water, be settled to 500mL, obtain solution D, normal temperature saves backup;
II: with accurate draw solution A 20mL, the solution B 2mL mixing of transfer pipet, adding distil water is settled to 1000mL, is dispensed in 10 250mL Erlenmeyer flasks, every bottle of 100mL, wrapping, and 121 DEG C of wet method sterilizing 20min, obtain solution E; After accurate draw solution C 20mL, the solution D 20mL mixing of transfer pipet, add NaNO 32.5g, adding distil water is settled to 1000mL, is dispensed in 10 250mL Erlenmeyer flasks, every bottle of 100mL, wrapping, and 121 DEG C of wet method sterilizing 20min, obtain solution F;
III: be cooled to room temperature under solution E and solution F are placed on Bechtop aseptic condition, after mixing, obtain described minimum medium.
B: prepare rich strontium substratum:
Take SiCl 250mg, distilled water is settled to 50mL, is mixed with the mother liquor of 1mg/mL after dissolving, adopt filtration method to carry out degerming to mother liquor, get the mother liquor 2mL after sterilizing and join in 998mL minimum medium, mix, the pH value of adjustment solution is 7.5, obtains rich strontium substratum, SiCl in this substratum 2concentration is 2mg/L.
C: preparation solid medium: get 100g minimum medium and add 1.7g agar, obtained solid medium.
2) separation and purification of hair weeds cells bacterial classification
A: dry delivering vegetables is cleaned 3 times with distilled water, dries the alcohol disinfecting 0.8min of rear use 75%, then clean with distilled water flushing, obtain clean dryly to deliver vegetables;
B: dry the delivering vegetables of cleaning steps A prepared is soaked in 10h in minimum medium, then uses glass pressure-even pulp crusher homogenate, obtains hair weeds cells inoculation liquid;
C: the hair weeds cells prepared by step B is seeded in streak inoculation on solid medium, cultivates through 5 ~ 6 times purifying and obtains hair weeds cells culture;
D: in picking hair weeds cells culture, single algae of purifying falls, and is placed in minimum medium and inflates enlarged culturing, results are cultured to the cell of logarithmic phase, obtain hair weeds cells.
3) cultivation of rich strontium hair weeds cells
Aseptically, by step 2) hair weeds cells prepared is seeded in rich strontium substratum, and the constant temperature biochemical cultivation case case putting into 25 DEG C is cultivated, the periodicity of illumination cultivated is that 12h illumination 12h is dark, and intensity of illumination is 2000Lux, cultivates 20 days, quiescent culture liquid, filter to isolate hair weeds cells, after isolated hair weeds cells distilled water wash three times, pour in flat board, lyophilize, water catching bin temperature in freezing dry process is-53 DEG C, and the time is 18h, obtains rich strontium hair weeds cells.
Embodiment 2
The rich strontium hair weeds cells of the present embodiment is that hair weeds cells is cultivated acquisition in rich strontium substratum, and wherein rich strontium substratum is by SrCl 2be added into mixing in minimum medium obtained, SrCl 2concentration be 4mg/L; Wherein every 1000mL minimum medium forms primarily of the preparation of raw material of following quality: NaNO 3250mg, MgSO 47H 2o 75mg, CaCl 22H 2o 36mg, H 3bO 40.286mg, MnCl 24H 2o 0.18mg, ZnSO 47H 2o 0.02mg, CuSO 45H 2o 0.08mg, NaMoO 42H 2o 0.04mg, CoCl 26H 2o 0.001mg, EDTA-Na 2(oxalic acid tetraacethyl disodium) 0.lmg, FeSO 47H 2o 0.48mg, citric acid 0.06mg, Na 2siO 358mg, K 2hPO 440mg.
The preparation method of the rich strontium hair weeds cells of the present embodiment, concrete operation step is as follows:
1) substratum is prepared:
A: preparation minimum medium:
I: raw material predissolve:
A: take CaCl 22H 2o 2.05g, MgSO 47H 2o 3.75g, dissolve with distilled water, be settled to 500mL, obtain solution A, normal temperature saves backup;
B: take NaMoO 42H 2o 20mg, ZnSO 47H 2o 10mg, H 3bO 4143mg, MnCl 24H 2o 90mg, EDTA-Na 250mg, FeSO 47H 2o 240mg, CuSO 45H 2o 40mg, CoCl 26H 2o 0.5mg, citric acid 30mg, after dissolving with distilled water, be settled to 500mL, obtain solution B, 4 DEG C of refrigerations are for subsequent use;
C: take K 2hPO 43H 2o 2g, after dissolving with distilled water, be settled to 500mL, obtain solution C, normal temperature saves backup;
D: take Na 2siO 39H 2o 2.9g, dissolve with distilled water, be settled to 500mL, obtain solution D, normal temperature saves backup;
II: with accurate draw solution A 20mL, the solution B 2mL mixing of transfer pipet, adding distil water is settled to 1000mL, is dispensed in 10 250mL Erlenmeyer flasks, every bottle of 100mL, wrapping, and 121 DEG C of wet method sterilizing 20min, obtain solution E; After accurate draw solution C 20mL, the solution D 20mL mixing of transfer pipet, add NaNO 32.5g, adding distil water is settled to 1000mL, is dispensed in 10 250mL Erlenmeyer flasks, every bottle of 100mL, wrapping, and 121 DEG C of wet method sterilizing 20min, obtain solution F;
III: be cooled to room temperature under solution E and solution F are placed on Bechtop aseptic condition, after mixing, obtain described minimum medium.
B: prepare rich strontium substratum:
Take SiCl 250mg, distilled water is settled to 50mL, is mixed with the mother liquor of 1mg/mL after dissolving, adopt filtration method to carry out degerming to mother liquor, get the mother liquor 4mL after sterilizing and join in 996mL minimum medium, mix, the pH value of adjustment solution is 7.5, obtains rich strontium substratum, SiCl in this substratum 2concentration is 4mg/L.
C: preparation solid medium: get 100g minimum medium and add 1.7g agar, obtained solid medium.
2) separation and purification of hair weeds cells bacterial classification
A: dry delivering vegetables is cleaned 4 times with distilled water, dries the alcohol disinfecting 1.0min of rear use 75%, then clean with distilled water flushing, obtain clean dryly to deliver vegetables;
B: dry the delivering vegetables of cleaning steps A prepared is soaked in 8h in minimum medium, then uses glass pressure-even pulp crusher homogenate, obtains hair weeds cells inoculation liquid;
C: the hair weeds cells prepared by step B is seeded in streak inoculation on solid medium, cultivates through 5 ~ 6 purifying and obtains hair weeds cells culture;
D: in picking hair weeds cells culture, single algae of purifying falls, and is placed in minimum medium and inflates enlarged culturing, results are cultured to the cell of logarithmic phase, obtain hair weeds cells.
3) cultivation of rich strontium hair weeds cells
Aseptically, by step 2) hair weeds cells prepared is seeded in rich strontium substratum, and the constant temperature biochemical cultivation case case putting into 25 DEG C is cultivated, the periodicity of illumination cultivated is that 12h illumination 12h is dark, and intensity of illumination is 2000Lux, cultivates 15 days, quiescent culture liquid, filter to isolate hair weeds cells, after isolated hair weeds cells distilled water wash three times, pour in flat board, lyophilize, water catching bin temperature in freezing dry process is-53 DEG C, and the time is 18h, obtains rich strontium hair weeds cells.
Embodiment 3
The rich strontium hair weeds cells of the present embodiment is that hair weeds cells is cultivated acquisition in rich strontium substratum, and wherein rich strontium substratum is by SrCl 2be added into mixing in minimum medium obtained, SrCl 2concentration be 0.5mg/L; Wherein every 1000mL minimum medium forms primarily of the preparation of raw material of following quality: NaNO 3250mg, MgSO 47H 2o 75mg, CaCl 22H 2o 36mg, H 3bO 40.286mg, MnCl 24H 2o 0.18mg, ZnSO 47H 2o 0.02mg, CuSO 45H 2o 0.08mg, NaMoO 42H 2o 0.04mg, CoCl 26H 2o 0.001mg, EDTA-Na 2(oxalic acid tetraacethyl disodium) 0.lmg, FeSO 47H 2o 0.48mg, citric acid 0.06mg, Na 2siO 358mg, K 2hPO 440mg.
The preparation method of the rich strontium hair weeds cells of the present embodiment, concrete operation step is as follows:
1) substratum is prepared:
A: preparation minimum medium:
I: raw material predissolve:
A: take CaCl 22H 2o 2.05g, MgSO 47H 2o 3.75g, dissolve with distilled water, be settled to 500mL, obtain solution A, normal temperature saves backup;
B: take NaMoO 42H 2o 20mg, ZnSO 47H 2o 10mg, H 3bO 4143mg, MnCl 24H 2o 90mg, EDTA-Na 250mg, FeSO 47H 2o 240mg, CuSO 45H 2o 40mg, CoCl 26H 2o 0.5mg, citric acid 30mg, after dissolving with distilled water, be settled to 500mL, obtain solution B, 4 DEG C of refrigerations are for subsequent use;
C: take K 2hPO 43H 2o 2g, after dissolving with distilled water, be settled to 500mL, obtain solution C, normal temperature saves backup;
D: take Na 2siO 39H 2o 2.9g, dissolve with distilled water, be settled to 500mL, obtain solution D, normal temperature saves backup;
II: with accurate draw solution A 20mL, the solution B 2mL mixing of transfer pipet, adding distil water is settled to 1000mL, is dispensed in 10 250mL Erlenmeyer flasks, every bottle of 100mL, wrapping, and 121 DEG C of wet method sterilizing 20min, obtain solution E; After accurate draw solution C 20mL, the solution D 20mL mixing of transfer pipet, add NaNO 32.5g, adding distil water is settled to 1000mL, is dispensed in 10 250mL Erlenmeyer flasks, every bottle of 100mL, wrapping, and 121 DEG C of wet method sterilizing 20min, obtain solution F; Sterilizing after solution A and solution B mixing is obtained solution E; After solution C and solution D mixing, add NaNO 3, sterilizing obtains solution F;
III: be cooled to room temperature under solution E and solution F are placed on Bechtop aseptic condition, after mixing, obtain described minimum medium.
B: prepare rich strontium substratum:
Take SiCl 250mg, distilled water is settled to 50mL, is mixed with the mother liquor of 1mg/mL after dissolving, adopt filtration method to carry out degerming to mother liquor, get the mother liquor 0.5L after sterilizing and join in 999.5mL minimum medium, mix, the pH value of adjustment solution is 7.5, obtains rich strontium substratum, SiCl in this substratum 2concentration is 0.5mg/L.
C: preparation solid medium: get 100g minimum medium and add 1.7g agar, obtained solid medium.
2) separation and purification of hair weeds cells bacterial classification
A: dry delivering vegetables is cleaned 3 times with distilled water, dries the alcohol disinfecting 0.8min of rear use 75%, then clean with distilled water flushing, obtain clean dryly to deliver vegetables;
B: dry the delivering vegetables of cleaning steps A prepared is soaked in 11h in minimum medium, then uses glass pressure-even pulp crusher homogenate, obtains hair weeds cells inoculation liquid;
C: the hair weeds cells prepared by step B is seeded in streak inoculation on solid medium, cultivates through 5-6 purifying and obtains hair weeds cells culture;
D: in picking hair weeds cells culture, single algae of purifying falls, and is placed in minimum medium and inflates enlarged culturing, results are cultured to the cell of logarithmic phase, obtain hair weeds cells.
3) cultivation of rich strontium hair weeds cells
Aseptically, by step 2) hair weeds cells prepared is seeded in rich strontium substratum, and the constant temperature biochemical cultivation case case putting into 25 DEG C is cultivated, the periodicity of illumination cultivated is that 12h illumination 12h is dark, and intensity of illumination is 2000Lux, cultivates 18 days, quiescent culture liquid, filter to isolate hair weeds cells, after isolated hair weeds cells distilled water wash three times, pour in flat board, lyophilize, control water catching bin temperature in freezing dry process and be-53 DEG C, the time is 18h, obtains rich strontium hair weeds cells.
Test example
Test method: the formula changing rich strontium substratum, biological content and content of strontium in the hair weeds cells of more different rich strontium culture medium culturing;
Specific as follows:
A group: do not add FeSO in substratum 47H 2o, Na 2siO 3, SrCl 2with strontium solution, other components are with embodiment 1.
B group: do not add FeSO in substratum 47H 2o and Na 2siO 3, add SrCl 2, in substratum, the ultimate density of strontium chloride is 2mg/L, and other components are with embodiment 1.
C group: do not add FeSO in substratum 47H 2o and Na 2siO 3, add strontium solution, in substratum, the ultimate density of strontium is 2mg/L, and other components are with embodiment 1.
D group: the embodiment of the present invention 1
E group: add FeSO in substratum 47H 2o and Na 2siO 3, add strontium solution, in nutrient solution, the ultimate density of strontium is 2mg/L, and other components are with embodiment 1.
In this test example, strontium solution is for obtain from rich strontium mineral water.
Each group is respectively done 3 groups of parallel tests above, and cultural method, with embodiment 1, measures hair weeds cells biomass respectively, adopts micro-wave digestion, and ICP-OMS method measures the content of strontium element in cell, results averaged, and test-results is as shown in table 1 below:
The more different rich strontium culture medium prescription of table 1 is on the impact of hair weeds cells biomass and content of strontium
From table 1, comparing A, B, C group and D, E group biomass of delivering vegetables can find out, adds FeSO in nutrient solution 47H 2o and Na 2siO 3the biomass of hair weeds cells can be significantly improved; Relatively D group and E group draw, compare strontium solution, add SrCl 2slightly can improve the biomass of hair weeds cells, and the content of strontium element in hair weeds cells can be significantly improved.
Shown by above-mentioned test-results, in the substratum in the present invention's rich strontium hair weeds cells culturing process, be added with FeSO 47H 2o and Na 2siO 3, and adopt SrCl 2for inorganic strontium source, each composition in substratum is acted synergistically mutually, improve the biomass in hair weeds cells and content of strontium, make the biomass of hair weeds cells reach 232mg/L, in hair weeds cells, the content of strontium element reaches 236 μ g/g.

Claims (9)

1. a rich strontium hair weeds cells, is characterized in that, hair weeds cells is placed in rich strontium substratum and cultivates acquisition, described rich strontium substratum is by SrCl 2be added into mixing in minimum medium obtained, SrCl in rich strontium substratum 2concentration be 0.5 ~ 4mg/L; Wherein every 1000mL minimum medium is formed by the preparation of raw material of following quality: NaNO 3250mg, MgSO 47H 2o 75mg, CaCl 22H 2o 36mg, H 3bO 40.286mg, MnCl 24H 2o 0.18mg, ZnSO 47H 2o 0.02mg, CuSO 45H 2o 0.08mg, NaMoO 42H 2o 0.04mg, CoCl 26H 2o 0.001mg, EDTA-Na 20.lmg, FeSO 47H 2o 0.48mg, citric acid 0.06mg, Na 2siO 358mg, K 2hPO 440mg, surplus is water.
2. the cultural method of rich strontium hair weeds cells as claimed in claim 1, is characterized in that, comprise following operation:
1) will deliver vegetables and be soaked in minimum medium, after homogenate, obtain hair weeds cells inoculation liquid;
2) get hair weeds cells inoculation liquid streak inoculation on solid medium prepared by step 1), purifying is cultivated and is obtained hair weeds cells culture, and the single algae therefrom selecting purifying falls, for subsequent use;
3) by step 2) in single algae of purifying fall to being placed in minimum medium enlarged culturing, obtain hair weeds cells;
4) hair weeds cells prepared by step 3) is seeded in rich strontium substratum cultivates, cultivated rear leaving standstill and filtered to isolate hair weeds cells, be described rich strontium hair weeds cells;
Wherein solid medium is prepare after adding agar in minimum medium.
3. the cultural method of rich strontium hair weeds cells as claimed in claim 2, is characterized in that, deliver vegetables in step 1) before being soaked in minimum medium and carry out pre-treatment, concrete grammar is: will deliver vegetables to clean up with distilled water and dry rear sterilization, then rinse well with water.
4. the cultural method of rich strontium hair weeds cells as claimed in claim 3, is characterized in that, described sterilization is the alcohol disinfecting 0.5 ~ 1.0min of employing 70% ~ 75%.
5. the cultural method of rich strontium hair weeds cells as claimed in claim 2, is characterized in that, soak time in minimum medium of delivering vegetables in step 1) is 8 ~ 11h.
6. the cultural method of rich strontium hair weeds cells as claimed in claim 2, it is characterized in that, in step 3), namely enlarged culturing obtains hair weeds cells to logarithmic phase.
7. the cultural method of rich strontium hair weeds cells as claimed in claim 2, it is characterized in that, the culture condition cultivated described in step 4) is: temperature is 25 DEG C; Periodicity of illumination is that 12h illumination 12h is dark, and intensity of illumination is 2000Lux; Incubation time is 15 ~ 20 days.
8. the cultural method of rich strontium hair weeds cells as claimed in claim 2, is characterized in that, also comprises and washes hair weeds cells isolated in step 4) with water postlyophilization, obtains rich strontium hair weeds cells product.
9. the cultural method of rich strontium hair weeds cells as claimed in claim 2, it is characterized in that, the preparation method of described minimum medium comprises the following steps:
I: raw material predissolve:
A: get CaCl 22H 2o, MgSO 47H 2o, by water dissolution, obtains solution A;
B: get NaMoO 42H 2o, ZnSO 47H 2o, H 3bO 4, MnCl 24H 2o, EDTA-Na 2, FeSO 47H 2o, CuSO 45H 2o, CoCl 26H 2o, citric acid, by water dissolution, obtain solution B;
C: get K 2hPO 43H 2o, by water dissolution, obtains solution C;
D: get Na 2siO 39H 2o, by water dissolution, obtains solution D;
II: sterilizing after solution A and solution B mixing is obtained solution E; After solution C and solution D mixing, add NaNO 3, sterilizing obtains solution F;
III: solution E and solution F are mixed, obtains described minimum medium.
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