CN110132868A - A kind of water plant root activity measuring method - Google Patents
A kind of water plant root activity measuring method Download PDFInfo
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- CN110132868A CN110132868A CN201910497038.7A CN201910497038A CN110132868A CN 110132868 A CN110132868 A CN 110132868A CN 201910497038 A CN201910497038 A CN 201910497038A CN 110132868 A CN110132868 A CN 110132868A
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- methylene blue
- blue solution
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- soak
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
Abstract
The invention discloses a kind of water plant root activity measuring methods, including the following steps being connected in order: 1) compound concentration is 0.01 ± 0.002mgmL‑1Methylene blue solution C0；2) methylene blue solution C is used0Series standard solution is prepared, OD is measured at 672nm, using methylene blue solution concentration as abscissa, OD is ordinate, draws standard curve；It 3) is V with volume0Methylene blue solution C0The root system of plant to be measured that quality is m is impregnated, soak is obtained；4) OD of soak is measured at 672nm；5) calibration curve equation is brought into using the OD of soak to calculate, obtain the concentration C of soak0', the total absorption area TAA of root system of plant unit mass to be measured is calculated using following formula:Water plant root activity measuring method of the present invention, realizes the simple, accurate of water plant root activity, rapid survey.
Description
Technical field
The present invention relates to a kind of water plant root activity measuring methods, belong to botany method determination techniques field.
Background technique
Root system plant is active absorption organ, and the upgrowth situation and vigor of root system determine the nutrient absorption and light of plant
Close ability to function.
Water plant, which refers to, physiologically depends on water environment, and photosynthesis organ is for good and all or in 1 year in several months sinking water
Or the plant kept afloat.Water plant root system and terrestrial plant root system are due to the difference of growing environment, and there are many differences.By
Low in terrestial enviornment water content, nutrition is relatively low, and terrestrial plant has prosperity to absorb moisture and nutrition from soil
Root system, and water content is high in water environment, nutrition is dissolved in the water, is easier to be absorbed by plants, and root system of plant water content is high, less sends out
It reaches, some water plant root systems only serve fixed function；Water plant rhizome space between cells is especially flourishing, and often also development has spy
Different tubulose aerating tissue, this structure guarantee can there is enough oxygen in the underwater portion of plant, and plant can make to have under water
Oxygen breathing；Most of terrestrial plant space between cells is more undeveloped, and root system is immersed in the water for a long time, will do it anaerobic under anoxia condition
Breathing generates alcohol or lactic acid, causes to poison to plant, or even dead；Compared to terrestrial plant, water plant root system cellulosic ratio
It is more relatively thin, moisture and nutrition can be absorbed directly from water.
To the measuring method of terrestrial plant root activity at present there is several methods that, as TTC reduction method, alpha-naphthylamine aoxidize
Method, root system bleeding method etc., especially with TTC reduction method using relatively broad, but these types of method operation is relatively complicated, and part is tried
There are toxicity for agent.Patent application if publication No. is CN108562471A measures root activity using TTC, uses strong corrosive examination
Agent sulfuric acid and toxic methanol；The waste liquid amount that the patent application that publication No. is CN104649455A thinks that TTC method generates is big, right
Environmental pollution is serious.In addition water plant root system and terrestrial plant root system significant difference, can not be by terrestrial plant root activity
Measuring method be directly used in the measurement of water plant root activity, although there is many researchers to be related to water plant root
It is the research of vitality test, but so far, there is no simple, accurately and rapidly water plant root activity measuring method.
Summary of the invention
The present invention provides a kind of water plant root activity measuring method, realize water plant root activity it is simple,
Accurately, rapid survey.
In order to solve the above technical problems, the technical solution adopted in the present invention is as follows:
A kind of water plant root activity measuring method, including the following steps being connected in order:
1) compound concentration is 0.010 ± 0.002mgmL-1Methylene blue solution C0；
2) methylene blue solution C is used0Series standard solution is prepared, OD is measured at 672nm, is cross with methylene blue solution concentration
Coordinate, OD are ordinate, draw standard curve；
It 3) is V with volume0Methylene blue solution C0The root system of plant to be measured that quality is m is impregnated, soak is obtained；
4) OD of soak is measured at 672nm；
5) calibration curve equation is brought into using the OD of soak to calculate, obtain the concentration C of soak0', utilize following formula
Calculate the total absorption area TAA of root system of plant unit mass to be measured:
Coloring agent methylene blue (molecular formula: C16H18ClN3S) toxicity is very low, is easy to remove, cheap.Methylene blue can be
Root system of plant adsorption forms uniform monolayer, when root system reaches saturation to the absorption of methylene blue, can will adsorb
Methylene blue be further diverted into and continue to adsorb in root cell, and the amount of methylene blue absorption can pass through dyeing front and back methylene blue
The variation of concentration calculates；Due to water plant root system and terrestrial plant root system significant difference, the survey of terrestrial plant root activity
The method of determining can not be directly used in the measurement of water plant root activity.When using methylene blue determining adsorption, the two is to methylene blue
There is also differences for the requirement of solution concentration, and wavelength including absorption peak etc. is also not fully different, this is also those skilled in the art
Member cannot get always the water plant root activity measurement reason for it of efficiently and accurately.
In step 1), the selection of methylene blue solution concentration is extremely important, too high or too low, and the accuracy not only detected can drop
It is low, and standard curve can be detached from.
In step 2), the selection of wavelength 672nm is extremely important, and in addition to this wavelength, other wave bands include 660nm, is used for water
Group difference is big when plant root activity measures, and accuracy is low.Applicant it has been investigated that, water plant root system dyeing front and back
Absorption peak it is different, by water plant eel grass after scanning all band (400-800nm) methylene blue reagent dyeing, black algae and
The root systems such as Haloragidaceae Myriophyllum spicatum are found that while that the pure solution of methylene blue has absorption peak on 660nm, but contaminate for water plant
Color is crossed root system of plant absorption peak and is both present at 672nm, on this wavelength and under specific methylene blue solution concentration, standard
Curve be it is linear, root activity can be characterized well, this is because the differences such as water plant and terrestrial plant structure, water content
Caused by, reason is that the moisture of terrestrial plant root system adsorption and root system growing model are low, and dyestuff is easy uniformly penetrating to plant
In object root system, water plant root system growing model is high, and dyestuff penetration is poor, is generally gathered in root system surface, can not penetrate into root
System is internal.
In order to improve the accuracy of solution concentration preparation, in step 1), the methylene blue of 100 times of concentration of aimed concn is first configured
Then solution obtains aimed concn by diluting the method for constant volume again as 0.010 ± 0.002mgmL-1Methylene blue solution C0。
In order to improve the accuracy of measurement, in step 2), the concentration of series standard solution is respectively methylene blue solution C0It is dense
0~0.6 times of degree.
In order to further increase the accuracy of measurement, in step 3), the quality of root system of plant to be measured uses one thousandth gram
Balance weighs.
In order to reduce interference, takes into account the accuracy and efficiency of measurement, in step 3), root system of plant to be measured is cleaned, with suction
After water paper suck dry moisture and weighing, methylene blue solution C is immersed0In, it is taken out after impregnating 1 ± 0.1min, obtains soak.Applicant
The measurement of water plant root activity can be realized within 1 minute or so accurately it has been investigated that impregnating, it is high-efficient.
To make the first adhered on root system to further increase the accuracy of measurement after root system of plant to be measured impregnates taking-up
Alkene indigo plant solution is flow back into soak.
Above-mentioned steps 4) in 672nm determination method, include the following steps:
4.1) compound concentration is 0.010 ± 0.002mgmL-1Methylene blue solution C0；
4.2) methylene blue solution C is used0Water plant root system is impregnated, soak is obtained；
4.3) OD that soak is measured using ultrapure water as blank control, was dyed water plant root system absorption peak
For 672nm.
As a kind of preferred concrete scheme of the application, water plant root activity measuring method, including what is be connected in order
Following steps:
1) 0.1000g methylene blue reagent is accurately weighed, is dissolved in 100mL volumetric flask, 1.000mgmL is made-1Methylene
Blue solutionTake 1mL methylene blue solutionIt is dissolved in 100mL volumetric flask, 0.01mgmL is made-1Methylene blue solutionIt is dense
Degree is denoted as C0；
2) methylene blue solution is usedSeries standard solution is prepared, methylene blue standard curve is made, OD is measured at 672nm,
Using methylene blue solution concentration as abscissa, OD is ordinate, draws standard curve；
3) 8mL methylene blue solution is drawnIt is placed in 10mL test tube, root system of plant to be measured is cleaned, is inhaled with blotting paper
Solid carbon dioxide point, immerses in aforementioned test tube, takes out after impregnating 1min, flow back into the methylene blue solution adhered on root system in test tube, contaminates
Methylene blue solution after color is labeled as methylene blue solution
4) taking wall thickness is the quartz colorimetric utensil of 1mm, first uses methylene blue solutionRinse, then by methylene blue solutionIt pours into
At 2/3 volume of cuvette, cuvette outer wall is dried with lens wiping paper, measures OD at 672nm using ultrapure water as blank control；
5) calibration curve equation calculating is brought into using the OD of soak find out methylene blue solutionConcentration, be denoted as C0'。
The total absorption area TAA of water plant root system (total absorption area) calculates (single according to following formula
Position: cm2):
In formula, C0For 0.010mgmL-1；C0' it is methylene blue solutionConcentration；V0=8mL.
The unmentioned technology of the present invention is referring to the prior art.
Water plant root activity measuring method of the present invention is designed for water plant feature, and test method is easy, measurement
As a result group difference is small, and repeatability is high, and test speed is fast, small toxicity.
Detailed description of the invention
Fig. 1 is that three kinds of water plant root systems of the present invention dye 400-800nm absorption value figure；
Fig. 2 is the canonical plotting in embodiment 2；
Fig. 3 is the canonical plotting in embodiment 3.
Specific embodiment
For a better understanding of the present invention, below with reference to the embodiment content that the present invention is furture elucidated, but it is of the invention
Content is not limited solely to the following examples.
Test equipment is Shimadzu UV-2450 ultraviolet-visible spectrophotometer in each example.
Embodiment 1
The determination of absorption peak:
1) 0.1000g methylene blue reagent is accurately weighed, is placed in 100mL volumetric flask, is settled to 100mL with ultrapure water,
1.000mgmL is made-1Methylene blue solution is 1.；Take 1mL methylene blue solution 1. in 100mL volumetric flask with liquid-transfering gun, use is ultrapure
Water is settled to 100mL, and 0.01mgmL is made-1Methylene blue solution is 2.；
2) it takes 8mL methylene blue solution to be 2. placed in 10mL test tube, root system of plant is cleaned, with blotting paper suck dry moisture,
It immerses in above-mentioned test tube solution, is taken out after impregnating 1min, flow back into the methylene blue solution adhered on root system in former test tube, impregnate
3. the methylene blue solution crossed is labeled as methylene blue solution；
3) take wall thickness be 1mm quartz colorimetric utensil, first with methylene blue solution 3. rinse three times, then by methylene blue solution
3. pouring at about 2/3 volume of cuvette, cuvette outer wall is dried with lens wiping paper, OD is measured using ultrapure water as blank control
(optical density, optical density), each OD is repeated three times；
Water plant root system absorption peak was dyed present in 672nm by verifying discovery, as shown in Figure 1.
Embodiment 2
The measurement of water caltrop root activity, specifically follows the steps below, and synchronous request for utilization publication No.
The TTC method measurement of CN108562471A is compared (specific method is referring to the embodiment 2 in CN108562471A):
1) 0.1000g methylene blue reagent really is weighed, is dissolved in 100mL volumetric flask, 1.000mgmL is made-1Methylene blue
SolutionTake 1mL methylene blue solutionIt is dissolved in 100mL volumetric flask, 0.01mgmL is made-1Methylene blue solutionIt is denoted as
C0；
2) by system methylene blue solution shown in table 1Series standard solution is prepared, methylene blue standard curve is made, in
OD is measured at 672nm, using methylene blue solution concentration as abscissa, OD is ordinate, draws standard curve such as Fig. 2；
1 methylene blue solution standard curve of table
3) 8mL methylene blue solution is takenIt is placed in 10mL test tube, water caltrop root system is cleaned, with blotting paper suck dry moisture,
Each water caltrop root system quality m is weighed using one thousandth gram balance, is immersed in above-mentioned test tube, takes out, makes on root system after impregnating 1min
The methylene blue solution of attachment is flow back into former test tube, and the methylene blue solution after dyeing is labeled as methylene blue solution
4) taking wall thickness is the quartz colorimetric utensil of 1mm, first with a little methylene blue solutionRinse, then by methylene blue solutionIt pours at about 2/3 volume of cuvette, cuvette outer wall is dried with lens wiping paper, using ultrapure water as blank control in 672nm
Place's measurement OD；
5) the dense of methylene blue after each test tube root immersion can be found out by bringing calibration curve equation into using the OD of soak and calculating
Degree, is denoted as C0', the total absorption area TAA of water plant root system unit mass calculates (unit: cm according to following formula2):
C in formula0For 0.010mgmL-1；C0' it is methylene blue solutionConcentration；V0=8mL.Step 3) -5) it uses respectively
5 plants of water caltrop root systems have made 5 groups of parallel laboratory tests, and test result is as shown in table 2.
2 water caltrop root activity measurement result of table
| Serial number | Root system quality m (g) | Concentration C after immersion0'(mg/mL) | TAA(cm2) |
| 1 | 0.177 | 0.0024 | 3.772 |
| 2 | 0.183 | 0.0021 | 3.791 |
| 3 | 0.172 | 0.0026 | 3.787 |
| 4 | 0.143 | 0.0039 | 3.743 |
| 5 | 0.182 | 0.0027 | 3.521 |
The average value of TAA is 3.723 ± 0.114cm in embodiment 22, error 3.1%, comparison measured using TTC method,
TTC also commercial weight is 0.124 ± 0.013mg/gh, error 10.5%.
Embodiment 3
Water quality grass roots system vitality test, specifically follows the steps below, and synchronous request for utilization publication No.
The TTC method measurement of CN108562471A is compared (specific method is referring to the embodiment 2 in CN108562471A):
1) 0.1000g methylene blue reagent is accurately weighed, is dissolved in 100mL volumetric flask, 1.000mgmL is made-1Methylene
Blue solutionTake 1mL methylene blue solutionIt is dissolved in 100mL volumetric flask, 0.01mgmL is made-1Methylene blue solutionIt is denoted as
C0；
2) by system methylene blue solution shown in table 3Series standard solution is prepared, methylene blue standard curve is made, in
OD is measured at 672nm, using methylene blue solution concentration as abscissa, OD is ordinate, draws standard curve such as Fig. 3；
3 methylene blue solution standard curve of table
3) 8mL methylene blue solution is drawnIt is placed in 10mL test tube, water quality grass roots system is cleaned, blots water with blotting paper
Point, each water quality grass root system quality m is weighed using one thousandth gram balance, is immersed in above-mentioned test tube, is taken out after impregnating 1min, makes root
The methylene blue solution for fastening attachment is flow back into former test tube, and the methylene blue solution after dyeing is labeled as methylene blue solution
4) taking wall thickness is the quartz colorimetric utensil of 1mm, first with a little methylene blue solutionRinse, then by methylene blue solutionIt pours at about 2/3 volume of cuvette, cuvette outer wall is dried with lens wiping paper, using ultrapure water as blank control in 672nm
Place's measurement OD；
5) the dense of methylene blue after each test tube root immersion can be found out by bringing calibration curve equation into using the OD of soak and calculating
Degree, is denoted as C0, the total absorption area TAA of water quality grass roots system unit mass is according to following formula calculating (unit: cm2):
C in formula0For 0.01mgmL-1；C0' it is methylene blue solutionConcentration；V0=8mL, step 3) -5) respectively with 5
Strain water quality grass roots system has made 5 groups of parallel laboratory tests, and test result is as shown in table 4.
4 water quality grass roots system vitality test result of table
| Serial number | Root system quality m (g) | Concentration C after immersion0'(mg/mL) | TAA(cm2) |
| 1 | 0.045 | 0.0021 | 10.658 |
| 2 | 0.054 | 0.0026 | 11.993 |
| 3 | 0.055 | 0.0026 | 11.776 |
| 4 | 0.043 | 0.0041 | 12.074 |
| 5 | 0.061 | 0.0022 | 11.216 |
The average value of TAA is 11.543 ± 0.598cm in embodiment 22, error 5.2%, comparison measured using TTC method,
TTC also commercial weight is 0.391 ± 0.029mg/gh, error 7.4%.
Claims (8)
1. a kind of water plant root activity measuring method, it is characterised in that: including the following steps being connected in order:
1) compound concentration is 0.010 ± 0.002mgmL-1Methylene blue solution C0；
2) methylene blue solution C is used0Series standard solution is prepared, OD is measured at 672nm, using methylene blue solution concentration as abscissa,
OD is ordinate, draws standard curve；
It 3) is V with volume0Methylene blue solution C0The root system of plant to be measured that quality is m is impregnated, soak is obtained；
4) OD of soak is measured at 672nm；
5) calibration curve equation is brought into using the OD of soak to calculate, obtain the concentration C of soak0', it is calculated using following formula
The total absorption area TAA of root system of plant unit mass to be measured:
2. water plant root activity measuring method as described in claim 1, it is characterised in that: in step 1), first configure mesh
Mark the methylene blue solution of 100 times of concentration of concentration, then again by the method for dilution constant volume obtain aimed concn for 0.010 ±
0.002mg·mL-1Methylene blue solution C0。
3. water plant root activity measuring method as described in claim 1, it is characterised in that: in step 2), series standard
The concentration of solution is respectively methylene blue solution C00~0.6 times of concentration.
4. water plant root activity measuring method as claimed in any one of claims 1-3, it is characterised in that: step 3)
In, the quality of root system of plant to be measured is weighed using one thousandth gram balance.
5. water plant root activity measuring method as claimed in claim 4, it is characterised in that: in step 3), by plant to be measured
Object root system is cleaned, and with blotting paper suck dry moisture and after weighing, immerses methylene blue solution C0In, it takes out, obtains after impregnating 1 ± 0.1min
Soak.
6. water plant root activity measuring method as claimed in claim 5, it is characterised in that: root system of plant immersion to be measured takes
After out, it flow back into the methylene blue solution adhered on root system in soak.
7. water plant root activity measuring method as claimed in any one of claims 1-3, it is characterised in that: in step 4)
The determination method of 672nm, includes the following steps:
4.1) compound concentration is 0.010 ± 0.002mgmL-1Methylene blue solution C0；
4.2) methylene blue solution C is used0Water plant root system is impregnated, soak is obtained；
4.3) OD of soak is measured using ultrapure water as blank control, being dyed water plant root system absorption peak is
672nm。
8. water plant root activity measuring method as claimed in any one of claims 1-3, it is characterised in that: including sequence
The following steps to connect:
1) 0.1000g methylene blue reagent is accurately weighed, is dissolved in 100mL volumetric flask, 1.000mgmL is made-1Methylene blue is molten
LiquidTake 1mL methylene blue solutionIt is dissolved in 100mL volumetric flask, 0.010mgmL is made-1Methylene blue solutionConcentration note
For C0；
2) methylene blue solution is usedSeries standard solution is prepared, methylene blue standard curve is made, measures OD, at 672nm with methylene
Blue solution concentration is abscissa, and OD is ordinate, draws standard curve；
3) 8mL methylene blue solution is drawnIt is placed in 10mL test tube, root system of plant to be measured is cleaned, blot water with blotting paper
Point, it immerses in aforementioned test tube, is taken out after impregnating 1min, flow back into the methylene blue solution adhered on root system in test tube, after dyeing
Methylene blue solution be labeled as methylene blue solution
4) taking wall thickness is the quartz colorimetric utensil of 1mm, first uses methylene blue solutionRinse, then by methylene blue solutionPour into colorimetric
At 2/3 volume of ware, cuvette outer wall is dried with lens wiping paper, measures OD at 672nm using ultrapure water as blank control；
5) calibration curve equation calculating is brought into using the OD of soak find out methylene blue solutionConcentration, be denoted as C0', aquatic plant
The total absorption area TAA of object root system calculates (unit: cm according to following formula2):
In formula, C0For 0.010mgmL-1；C0' it is methylene blue solutionConcentration；V0=8mL.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113740329A (en) * | 2021-09-08 | 2021-12-03 | 河北省农林科学院旱作农业研究所 | Winter wheat water conservation identification method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104649455A (en) * | 2015-03-17 | 2015-05-27 | 内蒙古民族大学 | Method for recycling ethyl acetate applied to determination of root activity by virtue of TTC method |
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2019
- 2019-06-10 CN CN201910497038.7A patent/CN110132868A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104649455A (en) * | 2015-03-17 | 2015-05-27 | 内蒙古民族大学 | Method for recycling ethyl acetate applied to determination of root activity by virtue of TTC method |
Non-Patent Citations (2)
| Title |
|---|
| 丁华侨 等: "不同栽培基质下水生植物的适应性", 《湖南农业大学学报(自然科学版)》 * |
| 王晶英等编: "《植物生理生化实验技术与原理》", 31 December 2003 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113740329A (en) * | 2021-09-08 | 2021-12-03 | 河北省农林科学院旱作农业研究所 | Winter wheat water conservation identification method |
| CN113740329B (en) * | 2021-09-08 | 2023-06-20 | 河北省农林科学院旱作农业研究所 | Winter wheat water-saving property identification method |
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Application publication date: 20190816 |