CN101926267B - Method for measuring bicarbonate ion utilizing capability of plant - Google Patents
Method for measuring bicarbonate ion utilizing capability of plant Download PDFInfo
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- CN101926267B CN101926267B CN 201010247881 CN201010247881A CN101926267B CN 101926267 B CN101926267 B CN 101926267B CN 201010247881 CN201010247881 CN 201010247881 CN 201010247881 A CN201010247881 A CN 201010247881A CN 101926267 B CN101926267 B CN 101926267B
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Abstract
The invention discloses a method for measuring the bicarbonate ion utilizing capability of a plant. The method comprises the following steps of: (1) respectively measuring stable carbon isotope compositions, i.e. delta 13C values of leaves of a plant to be studied and a reference plant; (2) measuring the stable carbon isotope composition, i.e. delta 13C value of microalgae; (3) substituting the delta 13C value of the leaves of the plant to be studied, the delta 13C value of the leaves of the reference plant and the delta 13C value of the microalgae into a two-end-member model and calculating the proportional share of bicarbonate ions utilized by the plant to be studied in an inorganic carbon source; (4) measuring the net photosynthetic rate of the leaves of the plant to be studied; and (5) deriving the bicarbonate ion utilizing capability of the plant to be studied according to the net photosynthetic rate of the leaves of the plant to be studied and the proportional share of the bicarbonate ions utilized by the plant to be studied in the inorganic carbon source. The invention can be used for quantitatively measuring the bicarbonate ion utilizing capability of the plant with less plant materials and less steps and simple calculation.
Description
Technical field
The present invention relates to a kind of method of measuring plant utilization bicarbonate ion ability, belong to the ecological environment treatment field.
Background technology
Plant can not only utilize the carbon dioxide of air to carry out photosynthesis for raw material, and can utilize the bicarbonate ion that stores to carry out photosynthesis for raw material through the effect of carbonic anhydrase.Concerning the karst, the ability of plant utilization bicarbonate ion is particularly important.The karst adaptive plant is after suffering karst adverse circumstance (karst arid, high calcium, pH, heavy carbonic radical ion and low inorganic nutrients etc.); Carbonic anhydrase activity in the blade raises; One side causes stomatal conductance to reduce or closes; Reduce transpiration to prevent that plant from further dewatering, on the other hand intracellular bicarbonate ion is changed into water and CO
2, reduce because of stomatal conductance or close moisture and the CO that causes with reply
2Deficiency, under the karst adverse circumstance, carry out photosynthetic carbon reduction, utilize inorganic carbon.The ability of plant utilization bicarbonate ion can become an evaluation criterion of karst adaptive plant.To screening karst adaptive plant, utilize biological method to administer and recover fragile karst ecologic environment to have important effect.
At present, measuring the photosynthetic instrument such as the portable photosynthetic appearance of Li-6400 of plant leaf blade more exactly, is to adopt the gas exchange method to measure photosynthesis of plant, through the measurement CO before and after the leaf chamber that flows through
2The variation of concentration and humidity change Net Photosynthetic Rate and the transpiration rate that calculates plant, and calculate stomatal conductance and intercellular CO
2Concentration.Can not be measured but plant leaf blade utilizes bicarbonate ion to carry out photosynthesis for the portable photosynthetic appearance of Li-6400; Because the inorganic carbon source of this part is without the leaf chamber, so the photosynthetic utilization that can't measure this part carbon source with the instrument as the portable photosynthetic appearance of Li-6400.Therefore, must seek the information that a kind of method is obtained the plant utilization bicarbonate ion.
The strong fractionation characteristic of stable carbon isotope is the basis in identification plant inorganic carbon source.The occurring in nature carbon has two kinds of stable isotopes:
12C with
13C, their natural average abundance is respectively 98.89% and 1.11%.The stable carbon isotope composition is used δ usually
13C (‰) expression, occurring in nature δ
13C is changed to-90 ‰ ~+20 ‰.The strong fractionation characteristic of stable carbon isotope helps discerning plant inorganic carbon source.Mass balance principle and isotope mixture model and chemometrics method; It is the basis of quantitatively discerning inorganic carbon source in the plant; Therefore, the present invention utilizes isotope technology to combine the mensuration of conventional Net Photosynthetic Rate to obtain the information of plant utilization bicarbonate ion.
Summary of the invention
The technical matters that the present invention will solve is, a kind of method of measuring plant utilization bicarbonate ion ability is provided, and utilizes bicarbonate ion to carry out photosynthetic deficiency to overcome the plant leaf blade of can not measuring that exists in the prior art.
The present invention takes following technical scheme: it may further comprise the steps, and the first, measure respectively and investigated plant and form δ with stable carbon isotope with reference to the blade of plant
13The value of C; The second, measure the stable carbon isotope of little algae and form δ
13The C value; The 3rd, will be by the δ of investigation plant leaf blade
13The C value, with reference to the δ of the blade of species
13The δ of C value, little algae
13The C value is brought two end member models into, calculates the bicarbonate ion of being investigated plant utilization and accounts for inorganic carbon source ratio share; The 4th, measure the Net Photosynthetic Rate of being investigated plant leaf blade; The 5th, account for inorganic carbon source ratio share according to the Net Photosynthetic Rate of being investigated plant leaf blade with by the bicarbonate ion of investigation plant utilization, obtain the ability of being investigated the plant utilization bicarbonate ion.
In first step; Select the extremely low plant of carbonic anhydrase activity to do with reference to plant; The plant that will investigate with reference to the planting seed of plant in the environment that will investigate; After treating that plant strain growth has 4 above true leaves, measure respectively and investigated plant and form δ with reference to first stable carbon isotope of launching leaf fully of plant
13The C value.
In second step, utilize the soil liquid preparing nutrient solution of being investigated environment of being investigated the vegetable seeds sowing, culturing microalgae is measured the δ of little algae
13The C value.
In third step, calculate the bicarbonate ion of being investigated plant utilization and account for inorganic carbon source ratio share, be with the δ that is investigated plant leaf blade
13The C value is as δ
T, with reference to the δ of the blade of plant
13The C value is δ
A, the δ of little algae
13The C value is δ
B, bring two end member model δ into
T=δ
A-f
Bδ
A+ f
Bδ
B, calculate the bicarbonate ion of being investigated plant utilization and account for inorganic carbon source ratio share f
B
In the 5th step, obtain the ability of being investigated the plant utilization bicarbonate ion.To be investigated second of plant to launch the Net Photosynthetic Rate Pn and the f of leaf fully
BBring formula BBUC=f into
BPn/ (1-f
B) in, obtain the ability of this plant utilization bicarbonate ion.
In second step, be cultivate to investigate plant with reference to plant in culturing microalgae, incubation time was two thoughtful five weeks.
Principle of the present invention is: utilize two end member model δ
T=δ
A-f
Bδ
A+ f
Bδ
BCalculate f
BHere δ
TFor being investigated the δ of plant leaf blade
13The C value, δ
ADo not make the δ of the blade of the extremely low plant of inorganic carbon source, carbonic anhydrase activity for not utilizing bicarbonate ion basically
13The C value, δ
BFor seldom utilizing carbon dioxide to make carbon source, but be the δ of little algae of main inorganic carbon source with bicarbonate ion
13The C value, f
BFor the bicarbonate ion of plant utilization accounts for inorganic carbon source ratio share.Through calculating, can obtain f
BPhotosynthetic rate according to photosynthetic appearance such as the portable photosynthetic appearance mensuration of Li-6400 is Pn, utilizes formula BBUC=f
BPn/ (1-f
B) can record the ability of this plant utilization bicarbonate ion, BBUC is the ability of plant utilization bicarbonate ion here.
Advantage of the present invention is following:
1) ability of this method ability quantitative measurement plant utilization bicarbonate ion.
2) the required vegetable material of this method is few, therefore takes up an area of little.
3) step of this method employing is few, calculates simple.
Embodiment
Embodiments of the invention: it may further comprise the steps, and the first, measure respectively and investigated plant and form δ with stable carbon isotope with reference to the blade of plant
13The value of C; The second, measure the stable carbon isotope of little algae and form δ
13The C value; The 3rd, will be by the δ of investigation plant leaf blade
13The C value, with reference to the δ of the blade of species
13The δ of C value, little algae
13The C value is brought two end member models into, calculates the bicarbonate ion of being investigated plant utilization and accounts for inorganic carbon source ratio share; The 4th, measure the Net Photosynthetic Rate of being investigated plant leaf blade; The 5th, account for inorganic carbon source ratio share according to the Net Photosynthetic Rate of being investigated plant leaf blade with by the bicarbonate ion of investigation plant utilization, obtain the ability of being investigated the plant utilization bicarbonate ion.
Detailed implementation process and theing contents are as follows:
Select the extremely low plant species of carbonic anhydrase activity to do reference; The plant species that will investigate with reference to the planting seed of plant species in the environment that will investigate; After treating that plant strain growth has 4 above true leaves, utilize the portable photosynthetic appearance of photosynthetic appearance such as Li-6400 to measure and investigated second Net Photosynthetic Rate Pn that launches leaf fully of plant.Take off respectively subsequently and investigated plant and launch leaf fully with reference to first of species; Place 60 ℃ of thermostatic drying chambers to dry; After the sample grinding with above-mentioned oven dry; Cross 0.1 mm sieve, after conventional the processing, last isotope mass spectrometer such as isotope mass spectrometer MAT252 carry out stable carbon isotope and form δ
13C measures.Utilize the soil liquid preparing nutrient solution of being investigated environment simultaneously, cultivate chlorella after three weeks, centrifugal collection frond, oven dry is ground, and after conventional the processing, last isotope mass spectrometer such as isotope mass spectrometer MAT252 carry out the chlorella stable carbon isotope and form δ
13The mensuration of C.
According to two end member model δ
T=δ
A-f
Bδ
A+ f
Bδ
B, calculate the bicarbonate ion of being investigated plant utilization and account for inorganic carbon source ratio share, δ here
TFor being investigated the δ of plant leaf blade
13The C value, δ
ADo not make the δ of the blade of the extremely low plant of inorganic carbon source, carbonic anhydrase activity for not utilizing bicarbonate ion basically
13The C value, δ
BFor seldom utilizing carbon dioxide to make carbon source is the δ of little algae of main inorganic carbon source with bicarbonate ion
13The C value, f
BFor the bicarbonate ion of plant utilization accounts for inorganic carbon source ratio share.With the top δ that is investigated plant leaf blade
13The C value is as δ
T, with reference to the δ of the blade of species
13The C value is δ
A, the δ of chlorella
13The C value is δ
B, bring two end member model δ into
T=δ
A-f
Bδ
A+ f
Bδ
B, calculate the bicarbonate ion of being investigated plant utilization and account for inorganic carbon source ratio share f
B
According to formula BBUC=f
BPn/ (1-f
B), calculate the ability of being investigated the plant utilization bicarbonate ion, here, BBUC is an ability of being investigated the plant utilization bicarbonate ion, Pn is for being investigated second Net Photosynthetic Rate of launching leaf fully of plant, f
BBe similarly the bicarbonate ion of being investigated plant utilization and account for inorganic carbon source ratio share.The bicarbonate ion that to be investigated Net Photosynthetic Rate Pn that second of plant launch leaf fully and its utilization accounts for inorganic carbon source ratio share f
BBring formula BBUC=f into
BPn/ (1-f
B) in, calculate the ability BBUC of this plant utilization bicarbonate ion.
In the present embodiment, the planting seed of getting rape, Orychophragmus violaceus, paper mulberry and mulberry tree and plane tree is to the soil of karst.Rape, Orychophragmus violaceus, paper mulberry and mulberry tree are investigated species, and the carbonic anhydrase activity of plane tree can't be measured with conventional pH meter method, prove that its carbonic anhydrase activity is minimum, can be used as with reference to species.With the soil liquid preparing nutrient solution of this area, cultivate chlorella after three weeks, use the inventive method, draw the ability of various plant utilization bicarbonate ions, like table 1.
Table 1 several plant utilizes the comparison of the ability of bicarbonate ion
Can find out that from table 1 BBUC of paper mulberry and Orychophragmus violaceus is significantly greater than the BBUC of mulberry tree and rape, this and paper mulberry and Orychophragmus violaceus are that the fact of karst adaptive plant is coincide.Especially paper mulberry though its ability of utilizing carbon dioxide is added to BBUC together less than mulberry tree, can be found out, the whole carbon assimilation ability of paper mulberry is apparently higher than the whole carbon assimilation ability of mulberry tree, and this is the information that present photosynthetic appearance can't obtain.
Claims (4)
1. method of measuring plant utilization bicarbonate ion ability, it is characterized in that: it may further comprise the steps, and the first, measure respectively and investigated plant and form δ with stable carbon isotope with reference to the blade of plant
13The value of C; The second, measure the stable carbon isotope of little algae and form δ
13The C value; The 3rd, will be by the δ of investigation plant leaf blade
13The C value, with reference to the δ of the blade of species
13The δ of C value, little algae
13The C value is brought two end member models into, calculates the bicarbonate ion of being investigated plant utilization and accounts for inorganic carbon source ratio share; The 4th, measure the Net Photosynthetic Rate of being investigated plant leaf blade; The 5th, account for inorganic carbon source ratio share according to the Net Photosynthetic Rate of being investigated plant leaf blade with by the bicarbonate ion of investigation plant utilization, obtain the ability of being investigated the plant utilization bicarbonate ion; In third step, calculate the bicarbonate ion of being investigated plant utilization and account for inorganic carbon source ratio share, be with the δ that is investigated plant leaf blade
13The C value is as δ
T, with reference to the δ of the blade of plant
13The C value is δ
A, the δ of little algae
13The C value is δ
B, bring two end member model δ into
T=δ
A-f
Bδ
A+ f
Bδ
B, calculate the bicarbonate ion of being investigated plant utilization and account for inorganic carbon source ratio share f
BIn the 5th step, obtain the ability of being investigated the plant utilization bicarbonate ion, be to be launched the Net Photosynthetic Rate Pn and the f of leaf fully with investigating second of plant
BBring formula BBUC=f into
BPn/ (1-f
B) in, obtain the ability of this plant utilization bicarbonate ion.
2. the method for mensuration plant utilization bicarbonate ion ability according to claim 1; It is characterized in that: in first step; Select the extremely low plant of carbonic anhydrase activity to do with reference to plant; In the plant that will investigate and the environment that will investigate to institute with reference to the planting seed of plant, treat that plant strain growth has 4 above true leaves after, measure respectively and investigated plant and first stable carbon isotope composition δ that launches leaf fully with reference to plant
13The C value.
3. the method for mensuration plant utilization bicarbonate ion ability according to claim 1 is characterized in that: in second step, utilize the soil liquid preparing nutrient solution of being investigated environment of being investigated the vegetable seeds sowing, culturing microalgae is measured the δ of little algae
13The C value.
4. the method for mensuration plant utilization bicarbonate ion ability according to claim 3 is characterized in that: in second step, be cultivate to investigate plant with reference to plant in culturing microalgae, incubation time was two thoughtful five weeks.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010247881 CN101926267B (en) | 2010-08-09 | 2010-08-09 | Method for measuring bicarbonate ion utilizing capability of plant |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010247881 CN101926267B (en) | 2010-08-09 | 2010-08-09 | Method for measuring bicarbonate ion utilizing capability of plant |
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| CN101926267A CN101926267A (en) | 2010-12-29 |
| CN101926267B true CN101926267B (en) | 2012-01-11 |
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Families Citing this family (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN102792891A (en) * | 2012-08-09 | 2012-11-28 | 中国科学院地球化学研究所 | Method for determining tissue culture seedling autotrophic portion |
| CN102827916B (en) * | 2012-08-09 | 2014-04-16 | 中国科学院地球化学研究所 | Method for quantifying inorganic carbon approach for microalgae utilization |
| CN103125281B (en) * | 2013-02-20 | 2014-04-16 | 中国科学院地球化学研究所 | Method for screening plants using bicarbonate radical ions efficiently by using photosynthetic carbon dioxide response curve |
| CN103616477B (en) * | 2013-11-13 | 2015-05-20 | 中国科学院地球化学研究所 | Method for measuring daily mean stable carbon isotope composition of atmospheric carbon dioxide |
| CN104142382B (en) * | 2014-07-17 | 2016-02-17 | 中国农业大学 | A kind of isotope stem stalk double-tagging tracing method |
| CN105067772B (en) * | 2015-08-10 | 2016-09-21 | 中国科学院地球化学研究所 | A kind of method measuring plant gross photosynthesis carbon assimilation ability |
| CN105973851B (en) * | 2016-06-12 | 2018-10-23 | 中国科学院地球化学研究所 | A kind of bathmic method of quantitative determination plant photosynthesis |
| 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 |
| CN107027626B (en) * | 2017-04-12 | 2019-06-11 | 中国科学院地球化学研究所 | A method of quantitative tissue-cultured seedling nitrate utilization efficiency |
| CN108333311B (en) * | 2018-02-12 | 2019-12-03 | 中国科学院地球化学研究所 | A kind of method of quick obtaining plant inorganic carbon supplement reuse share |
| 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 |
| CN112379054B (en) * | 2020-11-12 | 2021-10-15 | 中国科学院地球化学研究所 | Method for judging proper amplitude and optimal concentration of plant growth bicarbonate |
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| CN101926267A (en) | 2010-12-29 |
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