CN102827916B - Method for quantifying inorganic carbon approach for microalgae utilization - Google Patents

Method for quantifying inorganic carbon approach for microalgae utilization Download PDF

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CN102827916B
CN102827916B CN201210348447.9A CN201210348447A CN102827916B CN 102827916 B CN102827916 B CN 102827916B CN 201210348447 A CN201210348447 A CN 201210348447A CN 102827916 B CN102827916 B CN 102827916B
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inorganic carbon
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CN102827916A (en
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吴沿友
李海涛
刘丛强
王宝利
梁小兵
徐莹
谢腾祥
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Institute of Geochemistry of CAS
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Abstract

The invention discloses a method for quantifying a microalgae using an inorganic carbon approach. The prior art can not quantify microalgae using an inorganic carbon approach. The solving method comprises the following steps of: respectively adding two sodium bicarbonate of which the delta 13C value difference values are larger than 0.8% as an isotope label 1 and an isotope label 2 to cultivate microalgae to be tested in a nutrient solution; and respectively cultivating the microalgae to be tested for 4 days under the above condition and the cultivating condition of 16mM of sodium bicarbonate and 10mM of MAZ, testing the delta 13C values, computing the portion fB of the used and added inorganic carbon source under various microalgae cultivating conditions and the delta 13C value delta a of a microalgae body when a carbon dioxide approach in the atmosphere is completely used, and obtaining the portions of the two approaches in which the microalgae uses the inorganic carbon according to the obtained data. The method can quickly quantify the microalgae using inorganic carbon approach, and can develop a cultivating test under the complete same experiment condition to obtain the reliable data of the portion of the microalgae using inorganic carbon approach, which can not be achieved in the prior art.

Description

Micro-algae utilizes the quantivative approach of inorganic carbon approach
Technical field
The present invention relates to the quantivative approach that a kind of micro-algae utilizes inorganic carbon approach, belong to ecological environment treatment and marine biotechnology field.
Background technology
Micro-algae (microalgae) comprises the swim microphyte of mode of life of all Shui Zhongying of living in, and conventionally just refers to planktonic algae.Micro-algal structure is simple, and its physiological process is also relatively simple, and some kind is the model plant of scientific research, as: Chlamydomonas reinhardtii, chlorella, all right artificial culture of many types, this research that is us is provided convenience.Micro-algae has two kinds of modes to the utilization of water body inorganic carbon, and (1) utilizes the carbonic acid gas in atmosphere.CO 2as linear non-polar molecule, be electric neutrality, it can enter cell bilayer lipid membrane by free diffusing, enters the CO in cell 2by the photosynthesis of microalgae cell is utilized; (2) utilize bicarbonate ion in solution.Bicarbonate ion both can cross-docking also indirectly in transporte to cells by microalgae cell is utilized.The cross-docking of bicarbonate ion refers to through cytoplasmic membrane surface carrier proteins or anion-exchange protein, directly, in bicarbonate ion transporte to cells, in born of the same parents, through carbonic anhydrase, is converted into CO 2or directly with the form of bicarbonate ion by chloroplast membranes albumen active transport in chloroplast(id), through carbonic anhydrase, change into CO 2for ribulose-1,5-bisphosphate, 5 bisphosphate carboxylation/oxygenases (Rubisco) are fixing; The indirect transhipment of bicarbonate ion refers to the indirect transhipment of the bicarbonate ion that depends on the outer carbonic anhydrase of born of the same parents.Carbonic anhydrase (EC 4.2.1.1) is a kind of metalloenzyme containing zinc, catalysis CO 2with HCO 3 -reversible conversion: CO 2+ H 2o H ++ HCO 3 -.Carbonic anhydrase is divided in plasma membrane and outer two kinds of plasma membrane.The outer carbonic anhydrase of plasma membrane (claiming again born of the same parents outer carbonic anhydrase) is connected with cell walls internal surface by metal ion, and in catalysis cellular invasion layer, bicarbonate ion is hydrolyzed into rapidly free CO 2thereby, guaranteed the CO of cell 2fast supply.
Some algae only utilizes CO 2; Some algae can not only utilize CO 2, can also or directly utilize bicarbonate ion, or by the outer carbonic anhydrase indirect utilization bicarbonate ion of born of the same parents, or both have both at the same time.But prior art cannot be utilized inorganic carbon approach by quantitative micro-algae.Understand micro-algae and utilize mode and share to contribute to the development of Microalgae biotechnology, meanwhile, for the improvement of wawter bloom red tide provides scientific basis.
Summary of the invention
The technical problem to be solved in the present invention is, provides a kind of micro-algae to utilize the recognition methods of inorganic carbon approach, cannot quantitative micro-algae utilizes the deficiency of inorganic carbon approach to overcome prior art.
The present invention takes following technical scheme: it comprises the following steps: the first, select two kinds of δ 13c value difference value is greater than 8 ‰ sodium bicarbonate and adds to respectively in nutrient solution and cultivate and treat micrometer algae as isotopic labeling 1 and isotopic labeling 2; The δ of the sodium bicarbonate of isotopic labeling 1 13c value is δ c1, the δ of the sodium bicarbonate of isotopic labeling 2 13c value is δ c2; Measure simultaneously and do not add sodium bicarbonate, complete in inorganic carbon δ in the original culture of air 13c value is δ c0;
Second, until micrometer algae, under the culture condition of being investigated and under the culture condition of 16 mM sodium bicarbonates and 10mM AZ, cultivate after 4 days simultaneously, measure respectively stable carbon isotope under corresponding each culture condition that two kinds of isotope-labeled nutrient solutions cultivate, that investigated micro-algae and form δ 13the value δ of C t1, δ t2, δ t1-AZand δ t2-AZ;
The 3rd, pass through equation , calculate the share f that utilizes the inorganic carbon source of adding under the each culture condition of micro-algae b;
The 4th, according to the stable carbon isotope value δ that treats micrometer algae under the culture condition of 16 mM sodium bicarbonates and 10mM AZ t1-AZor δ t2-AZ, according to equation: δ a=δ ti-f bd i, the δ of frond when calculating micro-algae and utilizing the carbonic acid gas approach in atmosphere completely 13c value is δ a;
The 5th, according to the stable carbon isotope value δ that treats micrometer algae under each culture condition t1and δ t2; The δ of frond when micro-algae utilizes the carbonic acid gas approach in atmosphere completely 13c value δ a; Under the each culture condition of micro-algae, utilize the share f of the inorganic carbon source of adding band the δ of corresponding isotope-labeled sodium bicarbonate 13c value δ ciwith do not add sodium bicarbonate, completely from inorganic carbon δ in the original culture of air 13c value δ c0poor D iinformation, substitution equation: f b=1000 (δ ti-δ a-f bd iin)/9, calculate micro-algae and utilize the share f of bicarbonate ion approach b, 1-f bfor micro-algae utilizes the share of carbonic acid gas approach;
advantage of the present invention is as follows:
Occurring in nature carbon has two kinds of stable isotopes: 12c and 13c, their natural average abundance is respectively 98.89% and 1.11%.Stable carbon isotope composition is used δ conventionally 13c(‰) represent occurring in nature δ 13c is changed to-90 ‰ ~+20 ‰.The strong fractionation feature of stable carbon isotope is the basis in the micro-algae inorganic carbon of identification source.Mass balance principle and isotropic substance mixture model and chemometrics method are the bases in the micro-algae inorganic carbon of quantitatively identification source.
Two kinds of inorganic carbons of micro-algae utilize the isotope fractionation difference that approach causes to have obvious difference.Utilize the carbonic acid gas approach in atmosphere can cause maximum isotope fractionation (δ a).The cross-docking of bicarbonate ion causes isotope fractionation little by 9 ‰ (PDB) with the isotope fractionation Billy that the indirect transporting pathway that depends on the bicarbonate ion of carbonic anhydrase outside born of the same parents causes by the carbonic acid gas approach in atmosphere.
Acetazolamide (acetazolamide, AZ) is the heterocyclic sulfonamide carbonic anhydrase extracellular enzyme inhibitor containing 1,3,4-Thiadiazole.Utilize carbonic anhydrase extracellular enzyme inhibitor can suppress the feature of carbonic anhydrase extracellular enzyme single-mindedly, micro-algae isotropic substance under research carbonic anhydrase Extracellular enzyme activity is suppressed changes, and can reversal of identification depend on the born of the same parents indirect transporting pathway of the bicarbonate ion of carbonic anhydrase utilizes inorganic carbon contribution and share outward.Depending on the suppressed while of the indirect transporting pathway of the bicarbonate ion of carbonic anhydrase outside born of the same parents, the cross-docking approach of bicarbonate ion is also simultaneously suppressed.High density carbonic acid hydrogen sodium also has restraining effect to carbonic anhydrase extracellular enzyme, under the effect of high density carbonic acid hydrogen sodium and 10mM AZ, the indirect transporting pathway of bicarbonate ion and the cross-docking approach of bicarbonate ion that depend on the outer carbonic anhydrase of born of the same parents are suppressed the while completely.
The present invention takes following thinking: add respectively two kinds of δ 13the sodium bicarbonate of the different great disparity of C value difference is cultivated simultaneously and is treated micrometer algae, measures frond δ 13c value, utilizes the isotropic substance mixture model of two end members to obtain the share that micro-algae utilization comes from the inorganic carbon source of air and utilizes the inorganic carbon source of adding, and adds respectively subsequently two kinds of δ 13c value difference sodium bicarbonate different great disparity, high density is cultivated and is treated micrometer algae under the condition of 10mM AZ, measures frond δ 13c value, obtains the isotope fractionation value (δ that micro-algae utilizes the carbonic acid gas approach in atmosphere to cause completely a), last, according to above-mentioned information, utilize the isotropic substance mixture model of two end members, obtain respectively the share of two kinds of approach.
Obtain the principle of inorganic carbon source share:
In water body there are four kinds of form: CO in total inorganic carbon (DIC) dissolving 2, HCO 3 -, H 2cO 3and CO 3 2-, and they exist following chemical equilibrium: CO 2+ H 2o → H 2cO 3→ HCO 3 -+ H +→ CO 3 2-+ 2H +.Derive from the inorganic carbon of air or the inorganic carbon of interpolation and all have two kinds of inorganic carbons---CO 2and HCO 3 -for micro-algae, utilize.Therefore, can utilize the isotropic substance mixture model of two end members to obtain the share that micro-algae utilization comes from the inorganic carbon source of air and utilizes the inorganic carbon source of adding.
The isotropic substance mixture model of two end members can be expressed as:
δ Ti=δ Ai- f Biδ Ai +f Biδ Bi （i=1，2，3，------）（1）
Here δ tifor the δ of micro-algae 13c value, δ aithe δ of frond while utilizing the inorganic carbon source of air completely for being assumed to micro-algae 13c value, δ bithe δ of frond while utilizing the inorganic carbon source of interpolation completely for being assumed to micro-algae 13c value, f bifor the micro-algae of this investigation utilizes the shared share of inorganic carbon source of adding.
Obviously, only know δ tibe difficult to obtain f bi, therefore, the present invention adopts the δ with larger difference 13c value sodium bicarbonate is cultivated respectively micro-algae simultaneously, identifies the share of the inorganic carbon source of micro-algae utilization interpolation with stable carbon isotope double-tagging.
For isotopic labeling 1(i=1), equation (1) is expressed as follows formula:
δ T1=δ A1- f B1δ A1 +f B1δ B1 （2）
Here δ t1for using the known δ of the first 13the δ of micro-algae frond that the sodium bicarbonate of C value is cultivated 13c value, δ a1the δ of frond while utilizing the inorganic carbon source of air completely for being assumed to micro-algae 13c value, δ b1the δ of frond while utilizing the inorganic carbon source of interpolation completely for being assumed to micro-algae 13c value, f b1for the micro-algae of this investigation utilizes add inorganic, it is the shared share of carbon source.
For isotopic labeling 2(i=2), equation (1) is expressed as follows formula:
δ T2=δ A2 - f B2δ A2 +f B2δ B2 （3）
Here δ t2for using the known δ of the first 13the δ of micro-algae frond that the sodium bicarbonate of C value is cultivated 13c value, δ a2the δ of frond while utilizing the inorganic carbon source of air completely for being assumed to micro-algae 13c value, δ b2the δ of frond while utilizing the inorganic carbon source of interpolation completely for being assumed to micro-algae 13c value, f b2for the micro-algae of this investigation utilizes the shared share of inorganic carbon source of adding.
(2) δ and in (3) two equations a1=δ a2, f b=f bi=f b1=f b2, simultaneous solution
Figure 838356DEST_PATH_IMAGE002
（4）
(4) δ in formula b1-δ b2can be converted into the δ of the sodium bicarbonate of isotopic labeling 1 13c value δ c1δ with the sodium bicarbonate of isotopic labeling 2 13c value δ c2poor:
Figure 431142DEST_PATH_IMAGE001
（5）
Therefore, can be by the δ of the sodium bicarbonate of mensuration isotopic labeling 1 13c value δ c1δ with the sodium bicarbonate of isotopic labeling 2 13c value δ c2, the δ of micro-algae that mensuration is cultivated with the sodium bicarbonate of corresponding mark simultaneously 13c value, determines δ t1and δ t2value, calculates the micro-algae of this investigation according to (5) formula and utilizes the shared share of inorganic carbon source of adding.
Obtain the principle that micro-algae inorganic carbon utilizes approach share:
Derive from the inorganic carbon of air or the inorganic carbon of interpolation and all have two kinds of inorganic carbons---CO 2and HCO 3 -for micro-algae, utilize.Therefore, can utilize the isotropic substance mixture model of two end members to obtain two kinds, micro-algae to utilize the share information of inorganic carbon approach.
The isotropic substance mixture model of two end members can be expressed as:
δ Ti=(1-f B)[(1-f b)δ a+f b(δ a +9‰)]+f B [(1-f b)δ bi+f b(δ bi +9‰)]（i=1，2）（6）
Here δ tifor the δ of micro-algae 13c value, δ athe δ of frond while utilizing the carbonic acid gas approach in atmosphere completely for micro-algae 13c value, δ bibe assumed to that micro-algae utilizes the inorganic carbon source of adding, the δ of frond while carrying out carbonic acid gas approach completely 13c value, f bfor the micro-algae of this investigation utilizes the shared share of bicarbonate ion approach.
Here, δ bi-δ acan be converted into the δ of isotope-labeled sodium bicarbonate 13c value δ ci(or the δ of the sodium bicarbonate of isotopic labeling 1 13c value δ c1or the δ of the sodium bicarbonate of isotopic labeling 2 13c value δ c2) and do not add sodium bicarbonate, complete in inorganic carbon δ in the original culture of air 13c value δ c0poor D i.
Therefore, (6) formula can be simplified to:
f b= 1000 (δ Ti-δ a- f BD i)/9 （i=1，2） （7）
The δ of frond when micro-algae utilizes the carbonic acid gas approach in atmosphere completely 13c value is also δ aassignment principle:
Under the effect of high density carbonic acid hydrogen sodium and 10mM AZ, (7) formula f b=0
Also be δ ti-δ a-f bd i=0,
δ a=δ Ti- f BD i （i=1，2） （8）
Fully utilize above-mentioned principle, can utilize inorganic carbon approach by the micro-algae of fast quantification; Under identical experiment condition, carry out culture experiment, obtain micro-algae and utilize the data of inorganic carbon approach share reliable, this is that prior art all cannot be accomplished.
Embodiment
Embodiments of the invention: first step, measure the sodium bicarbonate that different manufacturers is produced, select two kinds of δ 13c value difference value is greater than 8 ‰ sodium bicarbonate and is added to respectively in nutrient solution as isotopic labeling 1 and isotopic labeling 2.The δ of isotope-labeled sodium bicarbonate 13c value is designated as δ ci, the wherein δ of the sodium bicarbonate of isotopic labeling 1 13c value is δ c1, the δ of the sodium bicarbonate of isotopic labeling 2 13c value is δ c2.The δ of isotope-labeled sodium bicarbonate 13c value δ cwith do not add sodium bicarbonate, completely from inorganic carbon δ in the original culture of air 13c value δ c0difference be D i, wherein D 1for the δ of the sodium bicarbonate of isotopic labeling 1 13c value δ c1with do not add sodium bicarbonate, completely from inorganic carbon δ in the original culture of air 13c value δ c0poor, D 2for the δ of the sodium bicarbonate of isotopic labeling 2 13c value δ c2with do not add sodium bicarbonate, completely from inorganic carbon δ in the original culture of air 13c value δ c0poor.
Second step, simultaneously with the nutrient solution of two kinds of isotopic labeling sodium bicarbonates respectively simultaneously under the culture condition of being investigated and under the culture condition of 16 mM sodium bicarbonates and 10mM AZ, micro-algae that cultivation is investigated, after 4 days, measure respectively stable carbon isotope under corresponding each culture condition that two kinds of isotope-labeled nutrient solutions cultivate, that investigated micro-algae simultaneously and form δ 13the value δ of C t1, δ t2, δ t1-AZand δ t2-AZ; .
Third step, by the δ of the sodium bicarbonate of isotopic labeling 1 13c value is as δ c1, by δ under isotopic labeling 1 culture condition that cultivate, each, that investigated micro-algae 13c value is as δ t1, the δ of the sodium bicarbonate of isotopic labeling 2 13c value is as δ c2, by δ under that cultivate, the corresponding culture condition of isotopic labeling 2, that investigated micro-algae 13c value is as δ t2, bring into
Figure 826351DEST_PATH_IMAGE001
, calculate the share f that is investigated the inorganic carbon source of micro-algae utilization interpolation under each culture condition b, it is 1-f that micro-algae utilizes the inorganic carbon source share of air b.
The 4th step, according to the stable carbon isotope value δ that is investigated micro-algae cultivating under the culture condition of 16 mM sodium bicarbonates and 10mM AZ ti(δ t1-AZor δ t2-AZ), according to equation: δ a=δ ti-f bd i, the δ of frond when calculating micro-algae and utilizing the carbonic acid gas approach in atmosphere completely 13c value δ a.
The 5th step, according to the stable carbon isotope value δ that treats micrometer algae under each culture condition ti, the wherein stable carbon isotope value δ that treats micrometer algae under isotopic labeling 1 sodium bicarbonate is cultivated t1, the stable carbon isotope value δ that treats micrometer algae under isotopic labeling 2 sodium bicarbonates are cultivated t2; The δ of frond when micro-algae utilizes the carbonic acid gas approach in atmosphere completely 13c value δ a, utilize the share f of the inorganic carbon source of adding under the each culture condition of micro-algae band the δ of corresponding isotope-labeled sodium bicarbonate 13c value δ ci(the δ of the sodium bicarbonate of isotopic labeling 1 13c value δ c1or the δ of the sodium bicarbonate of isotopic labeling 2 13c value δ c2) and do not add sodium bicarbonate, complete in inorganic carbon δ in the original culture of air 13c value δ c0poor D iinformation, substitution equation: f b=1000 (δ ti-δ a-f bd iin)/9, calculate micro-algae and utilize the share f of bicarbonate ion approach b, 1-f bfor micro-algae utilizes the share of carbonic acid gas approach.
Implementation result of the present invention is as follows:
Cultivated material is: chlamydomonas and chlorella.Basic culture solution adopts SE substratum, and basic culture condition is: photoperiod L/D:12h/12h; 25 ℃ of temperature; Intensity of illumination is 100 μ molm -2s -1, pH value 8.0 or 8.2(regulate with hydrochloric acid and sodium hydroxide).Other culture condition is as table 1, the δ of the sodium bicarbonate wherein adding 13c is respectively-26.3 ‰ (PDB) (δ c1) and-16.5 ‰ (PDB) (δ c2), the culture condition of 16 mM sodium bicarbonates and 10mM AZ is essential.Do not add sodium bicarbonate, complete in inorganic carbon δ in the original culture of air 13c value δ c0for-11.8 ‰ (PDB).After cultivating 4 days, measure respectively micro-algae δ of each culture condition and each experiment 13c value.By the inventive method, draw the share f that utilizes the inorganic carbon source of adding under each culture condition of chlamydomonas and chlorella band utilize the inorganic carbon source share 1-f of air b, as table 2.Subsequently, utilize the present invention, draw under each culture condition of chlamydomonas and chlorella and utilize bicarbonate ion approach and the share of utilizing carbonic acid gas approach, as table 3.
The culture condition of table 1 chlamydomonas and the each culture experiment of chlorella
Under each culture condition of table 2 chlamydomonas and chlorella, utilize the share of the inorganic carbon source of adding
Figure 525503DEST_PATH_IMAGE005
Under each culture condition of table 3 chlamydomonas and chlorella, utilize bicarbonate ion approach and the share of utilizing carbonic acid gas approach
Cultivation number f b 1-f b Cultivation number f b 1-f b
YB1 0.97 0.03 XB1 0.71 0.29
YB2 0.99 0.01 XB2 0.75 0.25
YB3 1.07 -0.07 XB3 0.87 0.13
YB4 1.03 -0.03 XB4 1.04 -0.04
YB5 1.11 -0.11 XB5 1.22 -0.22
YB6 0.46 0.54 XB6 0.46 0.54
YBA1 0.68 0.32 XBA1 0.64 0.36
YBA2 0.65 0.35 XBA2 0.53 0.47
YBA3 0.66 0.34 XBA3 0.62 0.38
YBA4 0.40 0.60 XBA4 0.34 0.66
YBA5 0.26 0.74 XBA5 0.13 0.87
YBA6 0.00 1.00 XBA6 0.07 0.93
YBAT1 -0.27 1.27 XBAT1 -0.48 1.48
YBAT2 -0.03 1.03 XBAT2 -0.30 1.30
YBAT3 0.12 0.88 XBAT3 -0.04 1.04
YBAT4 0.25 0.75 XBAT4 -0.02 1.02
YBAT5 0.18 0.82 XBAT5 0.01 0.99
YBAT6 0.00 1.00 XBAT6 0.00 1.00
YD1 1.08 -0.08 XD1 0.81 0.19
YD2 1.07 -0.07 XD2 0.86 0.14
YD3 1.09 -0.09 XD3 0.90 0.10
YD4 1.14 -0.14 XD4 0.69 0.31
YD5 -0.04 1.04 XD5 -0.30 1.30
As can be seen from Table 2, under different culture condition, utilize the share of the inorganic carbon source of adding obviously different, present interpolation sodium bicarbonate more, utilize the larger trend (except YB6 and XB6) of share of the inorganic carbon source of adding.Utilizing the share of inorganic carbon source of adding to be less than 0 YBA2, YBA3 and XBA3 is that error at measurment causes, but also approaches very much 0.These and practical situation are consistent, and also in certain concentration range, add sodium bicarbonate more, utilize the share of the inorganic carbon source of adding larger, when sodium bicarbonate concentration exceedes certain limit, some physiological process is suppressed, causes utilizing the share of the inorganic carbon source of adding to reduce.Show that micro-algae that the present invention draws utilizes the share of the inorganic carbon source of adding to have reliability.
As can be seen from Table 3, under different culture condition, micro-algae utilizes the share of bicarbonate ion approach obviously different, presents interpolation AZ concentration larger, and micro-algae utilizes the trend that the share of bicarbonate ion approach is less.This is because AZ is the inhibitor of the outer carbonic anhydrase of born of the same parents.See on the whole, chlamydomonas utilizes the share of bicarbonate ion approach to be greater than chlorella to utilize the share of bicarbonate ion approach.DIDS(4,4'-diisothiocyano-stilbene-2,2'-disulfonate) can suppress bicarbonate ion and enter cell, the feature of utilizing DIDS to suppress bicarbonate ion to enter cell single-mindedly, research bicarbonate ion enters and is blocked lower micro-algae isotropic substance and changes, contribution and the share of cross-docking approach that can reversal of identification bicarbonate ion to inorganic carbon utilization.When bicarbonate ion cross-docking approach is suppressed completely, the indirect transporting pathway that depends on the bicarbonate ion of carbonic anhydrase outside born of the same parents is also simultaneously completely suppressed.The DIDS of lower concentration utilizes the impact of approach less on bicarbonate ion.The DIDS(2mM of high density) on utilizing the approach impact of bicarbonate ion larger, almost suppress to utilize the approach of bicarbonate ion completely.This is consistent with practical situation.Due to error at measurment, occur that some are greater than 1 or be less than 0 situation, this all recoverable be 1 or 0, the share of utilizing bicarbonate ion approach of for example YB4 and XB4 is respectively 1.03 and 1.04, and all recoverable is 1, and the share of utilizing carbonic acid gas approach all recoverable is 0, show when adding the sodium bicarbonate of 4mM, two kinds of micro-algaes all do not utilize carbonic acid gas approach, only utilize bicarbonate ion approach, and this is also consistent with practical situation.It is upper that some large deviations appear at XD5, YBAT1, XBAT1 and XBAT2, this be because, compare with micro-algae of cultivating under 16 mM sodium bicarbonates and 10mMAZ, now micro algae growth speed is very little, cell volume is less, the δ of micro-algae actual measurement 13c value is than partially negative 1 to 2 ‰ (PDB) of theoretical value, therefore, make micro-algae utilize the share of carbonic acid gas approach to increase on apparent, but this situation (inorganic carbon is supplied with few) seldom occurs at occurring in nature.Also there is larger deviation in YB5 and XB5, this be because, compare with micro-algae of cultivating under 16 mM sodium bicarbonates and 10mMAZ, now micro algae growth speed is large, cell volume is large, the δ of micro-algae actual measurement 13c value is than theoretical value polarization 0.5 to 1 ‰ PDB, but this situation (sodium bicarbonate is up to 8mM) also seldom occurs at occurring in nature equally.The normal situation about occurring of nature, the present invention can estimate micro-algae and utilize the share of bicarbonate ion approach well.

Claims (1)

1. micro-algae utilizes a quantivative approach for inorganic carbon approach, and its feature comprises following steps:
The first, select two kinds of δ 13c value difference value is greater than 8 ‰ sodium bicarbonate and adds to respectively in nutrient solution and cultivate and treat micrometer algae as isotopic labeling 1 and isotopic labeling 2; The δ of the sodium bicarbonate of isotopic labeling 1 13c value is δ c1, the δ of the sodium bicarbonate of isotopic labeling 2 13c value is δ c2; Measure simultaneously and do not add sodium bicarbonate, complete in inorganic carbon δ in the original culture of air 13c value is δ c0;
Second, until micrometer algae, under the culture condition of being investigated and under the culture condition of 16 mM sodium bicarbonates and 10mM AZ, cultivate after 4 days simultaneously, measure respectively stable carbon isotope under corresponding each culture condition that two kinds of isotope-labeled nutrient solutions cultivate, that investigated micro-algae and form δ 13the value δ of C t1, δ t2, δ t1-AZand δ t2-AZ;
The 3rd, pass through equation
Figure 2012103484479100001DEST_PATH_IMAGE002
, calculate the share f that utilizes the inorganic carbon source of adding under the each culture condition of micro-algae b;
The 4th, according to the stable carbon isotope value δ that treats micrometer algae under the culture condition of 16 mM sodium bicarbonates and 10mM AZ t1-AZor δ t2-AZ, according to equation: δ a=δ ti-AZ-f bd i, the δ of frond when calculating micro-algae and utilizing the carbonic acid gas approach in atmosphere completely 13c value is δ a;
The 5th, according to the stable carbon isotope value δ that treats micrometer algae under each culture condition t1and δ t2; The δ of frond when micro-algae utilizes the carbonic acid gas approach in atmosphere completely 13c value δ a; Under the each culture condition of micro-algae, utilize the share f of the inorganic carbon source of adding band the δ of corresponding isotope-labeled sodium bicarbonate 13c value δ ciwith do not add sodium bicarbonate, completely from inorganic carbon δ in the original culture of air 13c value δ c0poor D iinformation, substitution equation: f b=1000 (δ ti-δ a-f bd iin)/9, calculate micro-algae and utilize the share f of bicarbonate ion approach b, 1-f bfor micro-algae utilizes the share of carbonic acid gas approach.
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CN103074411B (en) * 2013-01-05 2014-06-04 中国科学院地球化学研究所 Method for detecting and quantifying utilization of carbon source in calcium carbonate by microalgae
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CN106092944B (en) * 2016-06-12 2018-12-07 中国科学院地球化学研究所 A method of quantitative determination plant 1,5- ribulose diphosphate power of regeneration
CN107219349B (en) * 2017-05-24 2019-05-07 中国科学院地球化学研究所 A kind of quantitative microalgae utilizes the inorganic carbon capability approach derived from Silicate Rocks weathering
CN108319820B (en) * 2018-04-09 2020-04-14 中国科学院地球化学研究所 Method for obtaining utilization share of plant bicarbonate in field habitat
CN109444349B (en) * 2018-12-26 2020-07-07 中国科学院地球化学研究所 Method for measuring metabolic water utilization share and actual water demand of indoor plant

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5112740A (en) * 1989-12-18 1992-05-12 Eastman Kodak Company Carbon dioxide assay for body fluids comprising carbonic anhydrase
CN100390528C (en) * 2006-05-31 2008-05-28 山东大学 Quantitative detection method for algae in water
CN101926267B (en) * 2010-08-09 2012-01-11 中国科学院地球化学研究所 Method for measuring bicarbonate ion utilizing capability of plant
CN102365925B (en) * 2010-12-29 2013-02-27 中国科学院地球化学研究所 Method for determining ability of plant on using nitrate
CN102511362B (en) * 2011-10-27 2013-05-15 中国科学院地球化学研究所 Method for utilizing double markers to acquire share of inorganic carbon source utilized by plants
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