CN108254254B - Method for quantitatively detecting water retention capacity of plant leaves - Google Patents

Method for quantitatively detecting water retention capacity of plant leaves Download PDF

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CN108254254B
CN108254254B CN201810220649.2A CN201810220649A CN108254254B CN 108254254 B CN108254254 B CN 108254254B CN 201810220649 A CN201810220649 A CN 201810220649A CN 108254254 B CN108254254 B CN 108254254B
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邢德科
束宇
彭健
李志祥
李玉龙
吴沿友
陈晓乐
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Jiangsu University
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Abstract

The invention discloses a method for quantitatively detecting the water retention capacity of plant leaves, which comprises the following steps: cleaning plant leaves to be detected, soaking the plant leaves in water, taking out the leaves after a period of time, and sucking surface water to be dry; respectively measuring the maximum compressive internal force of the leaves after t hours of water loss by using a texture analyzer, and calculating the ultimate stress of the leaves at each water loss moment; simultaneously, the water potential and physiological capacitance of the leaves at each dehydration moment are respectively measured, and the tensity of the leaves is calculated; calculating the rigidity of the plant leaves at each dehydration moment according to the ultimate stress and the tension of the leaves; and taking the rigidity of the plant leaves at the 0 moment as a reference, calculating the relative leaf rigidity of the plant at each water loss moment, and respectively adding the relative leaf rigidity to obtain the accumulated relative leaf rigidity at each water loss moment. Comparing the accumulated relative leaf stiffness among different plants, and quantifying the water retention capacity of the leaves of different plants. The method has simple and quick determination process and high result accuracy, and can quantitatively compare the drought resistance of different plants according to the water retention capacity of the leaves.

Description

Method for quantitatively detecting water retention capacity of plant leaves
Technical Field
The invention belongs to the technical field of drought-resistant breeding, crop cultivation and crop information detection, and particularly relates to a method for quantitatively detecting water retention capacity of plant leaves.
Background
The growth and development of plants are often affected by various biotic and abiotic stresses, drought being one of the most important abiotic stresses. The drought resistances of different plants are different, and the detection of the drought resistance of the plants is very important in order to implement a reasonable irrigation system for crops with different drought resistances. Meanwhile, a variety which can adapt to the environment which tends to be drought is found, drought can be effectively resisted, the survival rate of plants is improved, the ecological environment is improved, desertification is restrained, and the method has important significance for promoting the agricultural sustainable development and the ecological construction synchronous development in China.
The leaves are the main organs of plants for assimilation and transpiration, are closely related to the surrounding environment, and are one of the most obvious indexes for judging the drought resistance degree of a plant variety. The water retention capacity of leaves is generally used to indicate the ability of tree tissues to resist dehydration, and the stronger the water retention capacity of leaves, the stronger the drought resistance of plants. The plant leaf is composed of cells, and the mechanical property of the leaf is closely related to the mechanical behavior of single cells. Plant cells consist of a cell wall, a thick, tough and somewhat elastic structure located outside the cell membrane, and luminal material (primarily protoplasts). The cell wall acts as a boundary of the life form of the plant cell, prevents the unrestricted flow of water and nutrients, maintains the internal pressure of the cell, and the like, and also bears the external load to provide the function of maintaining the mechanical strength of the plant body.
The lignin is an essential element of all vascular plant cell walls, and the lignin is filled in a cellulose skeleton in the cell walls, so that the hardness of the cell walls is increased, and the mechanical supporting force and the compressive strength of cells are enhanced. Generally, plants with strong drought resistance can enhance the mechanical strength of cell walls by rapidly promoting the synthesis of lignin under drought conditions. The liquid pressure formed by the free water wrapped by the semipermeable plasma membrane acts on the cell wall to cause the cell wall to generate tensile deformation, and the yield of the cell wall inevitably causes the reduction of turgor pressure, thereby causing the reduction of the power for driving the expansion of the cells. When the cells are dehydrated and shrunk, the cell walls have an outward drawing effect, the higher the mechanical strength of the cell walls is, the higher the rigidity is, the larger the negative pressure potential generated by the cell walls is, so that the cell water potential can be greatly reduced, the continuous dehydration of the cells is effectively limited, and the water retention capacity of the leaves is improved.
At present, the drying method is still the main means for measuring the water retention capacity of plant leaves. However, the above method is time-consuming, the measurement process is complicated, the qualitative analysis is biased, and the specific quantitative technique is lacked. Compared with the methods, the method can acquire related data information more simply, quickly and reliably by measuring the mechanical index and the electrophysiological property of the leaves, has short experimental period, low requirement on experimental conditions and easy observation, and has important application value in the aspects of future drought-resistant breeding and water-saving irrigation research.
Disclosure of Invention
The invention aims to solve the technical problem of providing a method for quantitatively detecting the water retention capacity of plant leaves, which can accurately detect the water retention capacity of the plant leaves through mechanical indexes and physiological and electrical characteristics, is simple, convenient and quick, has high accuracy and provides scientific support for drought-resistant breeding and water-saving irrigation technologies.
The technical scheme adopted by the invention comprises the following steps:
taking fresh branches of plants to be detected with leaves, and taking measures to slow down water diffusion;
step two, cleaning the leaves, picking a plurality of leaves with consistent growth vigor, and soaking the leaves in water;
step three, taking out the soaked water-saturated leaves after the leaves are in a water-saturated state, drying and dehydrating the leaves after treatment, and measuring the maximum compression internal force F of the left half part of the main veins of the dry and dehydrated leaves by using a texture analyzer at the tth hour after the leaves are dried and dehydratedmaxMeasuring the water potential W and the physiological capacitance CP of the right half part of the main leaf vein of the leaf at the same time period; maximum compressive internal force FmaxIn the measuring process, the enough pressing pressure of the texture analyzer probe needs to be ensured, so that the blade can be crushed;
step four, according to the maximum compression internal force F of the blademaxAnd the diameter d of the circular cross section of the push down probePCalculating the ultimate stress sigma of the blademax，
Figure BDA0001599725820000021
FmaxThe unit is N, pi represents the circumference ratio, dPIn units of mm, σmaxUnit is N/mm2；
Step five, calculating the leaf tensity LT according to the leaf water potential W and the physiological capacitance CP,
Figure BDA0001599725820000022
wherein LT is in cm2In/cm, i is the dissociation coefficient, R is the gas constant, T is the thermodynamic temperature,0is a vacuum dielectric constant, a is a relative dielectric constant of cytosol, M is a relative molecular mass of the cytosol, CP is plant leaf physiological capacitance, and W is plant leaf water potential;
step six, utilizing the ultimate stress sigmamaxAnd leaf stress LT to calculate plant leaf stiffness LCS,
Figure BDA0001599725820000023
LCS unit is N/mm, sigmamaxRepresenting the maximum compressive internal force per unit area of the blade in contact with the circular cross-section of the pressing probe, dLARepresents the effective thickness of the blade per unit area, and has a unit of cm/cm2；
Seventhly, the rigidity LCS of the plant leaves at the time 0 is measured0As a reference, the relative leaf stiffness value RLCS of the plant at each water loss time is calculatedi，RLCSi＝LCSi/LCS0In which the LCSiThe stiffness of the plant leaves at the ith water loss moment is respectively 0, 1, 2 and 3 …;
step eight, adding the relative leaf stiffness values of the plants at all the water loss moments to obtain an accumulated relative leaf stiffness value TRLCS of the plants, namely TRLCS ═ Sigma RLCSi＝RLCS0+RLCS1+RLCS2+RLCS3…；
And step nine, comparing the accumulated relative leaf stiffness values TRLCS of different plants respectively, thereby quantitatively calculating the water retention capacities of the leaves of the different plants.
The invention has the following beneficial effects:
1) the method measures the maximum compression internal force of the blade by using the texture analyzer, the result is not limited by the surrounding environment factors, and the method is accurate in result, high in reliability, simple, convenient and quick.
2) The method combines the comprehensive measurement of the mechanical index and the electrophysiological property of the leaves, can represent the change condition of the cell walls of the leaves, can also represent the influence of the water change of the leaves on the cell volume, and can more accurately reflect the drought and water loss condition of the plant leaves.
3) Based on the determination of the ultimate stress and the stress degree of the leaves of the plants, the leaf rigidity of the plants is rapidly obtained, the relative leaf rigidity under different drought stresses is calculated, the accumulated relative leaf rigidity of different plants is obtained, the water retention capacities of the leaves among different plants are compared, and further the drought resistance of the plants can be judged; the relative leaf rigidity of the plants obtained by the method can better and more directly reflect the water retention capacity of the leaves, and can be directly used for quantitatively comparing the drought resistance of different plants.
Detailed Description
The invention will be further explained below, but the scope of protection of the invention is not limited thereto.
The basic principle of the invention is as follows:
the plant leaf is composed of cells, and the mechanical property of the leaf is closely related to the mechanical behavior of single cells. Plant cells are composed of cell walls and luminal material (mainly protoplasm), and the cell walls not only serve as the boundary of the life forms of plant cells, prevent the unrestricted flow of water and nutrients, and maintain the internal pressure of the cells, but also bear external loads and provide the function of maintaining the mechanical strength of plant bodies.
When the cells are dehydrated and shrunk, the cell walls have an outward drawing effect, the greater the rigidity of the cell walls is, the greater the negative pressure potential generated by the cell walls is, so that the cell water potential can be greatly reduced, the continuous dehydration of the cells is effectively limited, and the water retention capacity of the leaves is improved. Therefore, the water retention capacity of leaves can be quantitatively reflected by the rigidity of the cell walls of their constituent cells.
The stiffness calculation formula is as follows:
Figure BDA0001599725820000031
wherein CS is stiffness in N/m; p is the force acting on the object in N; is the deformation caused by the force acting on the object, in m.
And at the moment of crushing the blade by the external force, the moving distance of the external force is the effective thickness of the blade. The effective thickness per unit area of the blade is denoted dLAThe calculation can be carried out through the blade tensity, and the blade tensity calculation formula is as follows:
Figure BDA0001599725820000032
wherein LT is in cm2I/cm, i dissociationA coefficient having a value of 1; r is a gas constant, 0.0083 L.MPa/mol.K; t is thermodynamic temperature K, T is 273+ T ℃, and T is environmental temperature (20 ℃ in the experiment);0dielectric constant in vacuum, 8.854 × 10-12F/m; a is the relative dielectric constant of cytosol, M is the relative molecular mass of cytosol, and the cytosol of the leaf is assumed to be sucrose, where a is 3.3F/M and M is 342; CP is the physiological capacitance of the plant leaf, and W is the water potential of the plant leaf.
The calculation formula of the effective thickness per unit area of the blade is as follows:
Figure BDA0001599725820000041
wherein d isLAHas the unit of cm/cm2。
At the moment of crushing the blade by an external force, the maximum compressive internal force borne by the blade per unit area can be defined by the ultimate stress sigmamaxIs expressed as σmaxUnit is N/mm2The ultimate stress calculation formula is as follows:
Figure BDA0001599725820000042
wherein, FmaxThe maximum compression internal force of the blade is N; dPThe diameter of the circular cross section of the pressing probe is in mm; and pi represents the circumferential ratio.
Stiffness represents the force required to cause a unit displacement, so at the instant the blade is crushed by an external force, the blade stiffness can then be calculated from the ratio between the maximum compressive internal force experienced per unit area of blade and the effective thickness per unit area of blade, i.e.:
Figure BDA0001599725820000043
where LCS is the blade stiffness in N/mm.
In order to eliminate the difference among different plants, the leaf stiffness measured at each water loss time of each plant is divided by the leaf stiffness of each plant in a water-saturated state, relative values are taken, the relative leaf stiffness is accumulated and added to obtain the accumulated relative leaf stiffness of the different plants, and the water retention capacity of the leaves among the different plants can be quantitatively compared by comparing the accumulated relative leaf stiffness values among the different plants.
The specific implementation process of the invention is as follows:
step one, taking a fresh branch of a plant to be detected with leaves, and wrapping the base of the plant branch by using wet cloth to slow down water diffusion.
And step two, rapidly returning to a laboratory, picking 10 leaves with consistent growth vigor on the fresh branches after cleaning dust on the surfaces of the leaves, and soaking in a container filled with water for 30 minutes.
Step three, after the leaves are soaked for 30 minutes and become a water-saturated state, taking out 10 soaked water-saturated leaves, quickly and lightly sucking the surface water of all the leaves with a paper towel, placing the leaves on a dry and ventilated table top to dry and dehydrate the leaves, respectively taking out 1 piece of the dry and dehydrated leaves in 0, 1, 2 and 3 … hours after the leaves are dried and dehydrated, and measuring the maximum compression internal force F of the left half part of the main leaf vein of the dry and dehydrated leaves by using a texture analyzermax(ii) a Simultaneously measuring the water potential W and the physiological capacitance CP of the right half part of the main leaf vein of the leaf at the same time period; maximum compressive internal force FmaxThe measurement process of (2) needs to ensure that the pressure for pressing down the texture analyzer probe is enough to crush the blade.
Step four, according to the maximum compression internal force F of the blademaxAnd the diameter d of the circular cross section of the push down probePCalculating the ultimate stress sigma of the blademaxWherein
Figure BDA0001599725820000051
FmaxThe unit is N, pi represents the circumference ratio, dPIn units of mm, σmaxUnit is N/mm2。
Step five, calculating the leaf tensity LT (formula (2)) according to the leaf water potential W and the physiological capacitance CP, wherein the unit of LT is cm2/cm。
Step six, utilizing ultimate stressσmaxAnd leaf stress LT calculating the leaf stiffness LCS of the plant, wherein
Figure BDA0001599725820000052
LCS unit is N/mm, sigmamaxRepresents the maximum compressive internal force per unit area of the blade in contact with the circular cross section of the hold-down probe; dLARepresents the effective thickness of the blade per unit area, and has a unit of cm/cm2。
Seventhly, the rigidity LCS of the plant leaves at the time 0 is measured0As a reference, the relative leaf stiffness value RLCS of the plant at each water loss time is calculatediThe calculation formula of the relative leaf stiffness value of the plant at each water loss time is RLCSi＝LCSi/LCS0In which the LCSiThe stiffness of the plant leaves at the ith water loss moment is respectively 0, 1, 2 and 3 ….
Step eight, adding the relative leaf stiffness values of the plants at all the water loss moments to obtain an accumulated relative leaf stiffness value TRLCS of the plants, namely TRCS ═ Sigma RLCSi＝RLCS0+RLCS1+RLCS2+RLCS3…。
And step nine, comparing the accumulated relative leaf stiffness values TRLCS of different plants respectively, thereby quantitatively calculating the water retention capacities of the leaves of the different plants.
The specific implementation process of the steps is as follows:
example 1:
taking a fresh branch of a paper mulberry with leaves, and wrapping the base of the branch of the plant with wet cloth to slow down the water diffusion; rapidly returning to a laboratory, after cleaning dust on the surfaces of the paper mulberry leaves, picking 10 leaves with relatively consistent growth vigor on the fresh branches, and soaking in a container filled with water for 30 minutes; after the leaves are soaked for 30 minutes and become a water-saturated state, taking out 10 soaked water-saturated leaves, quickly and lightly sucking the surface water of all the leaves with a paper towel, placing the leaves on a dry and ventilated table top to dry and dehydrate the leaves, respectively taking out 1 piece of the dry and dehydrated leaves at 0, 1, 2, 3, 4 and 5 hours after the leaves are dried and dehydrated, and measuring the maximum left half part of the main leaf vein of the dry and dehydrated leaves with a texture analyzerCompressive internal force Fmax(see table 1); the water potential W (water potential meter) and the physiological capacitance CP (LCR tester) of the right half of the main leaf vein of the leaf were measured at the same time in the same period (see Table 1).
TABLE 1 maximum compressive internal force F of Broussonetia papyrifera leaves at different moments of water lossmaxLeaf water potential W and physiological capacitance CP
Figure BDA0001599725820000061
According to maximum compressive internal force F of blademaxAnd the diameter d of the circular cross section of the push down probePCalculating the ultimate stress sigma of the blademax(using equation (4), the results are shown in Table 2), and the circular cross-sectional diameter d of the depressor probe used in this exampleP2mm, pi-3.14; calculating the leaf tensity LT according to the leaf water potential W and the physiological capacitance CP (by adopting a formula (2), the result is shown in a table 2); at the same time, the ultimate stress sigma is utilizedmaxAnd leaf stress LT (using equation (5), the results are shown in Table 2).
TABLE 2 ultimate stress σ of Broussonetia papyrifera leaves at different moments of water lossmaxBlade tension LT and blade stiffness LCS
Figure BDA0001599725820000071
The plant leaf stiffness at time 0 LCS0As a reference, the relative leaf stiffness value RLCS of the plant at each water loss time is calculatedi(see Table 3), the relative leaf stiffness value of the plant at each water loss time is calculated as RLCSi＝LCSi/LCS0In which the LCSiThe stiffness of the plant leaves at the ith water loss moment is respectively 0, 1, 2, 3, 4 and 5; adding the relative leaf stiffness values of the plants at the respective water loss moments to obtain the cumulative relative leaf stiffness value TRLCS (see Table 3), i.e. TRLCS ═ Sigma RLCSi＝RLCS0+RLCS1+RLCS2+RLCS3…, respectively; and respectively comparing the accumulated relative leaf stiffness values TRLCS of different plants, thereby quantitatively calculating the water retention capacity of the leaves of different plants.
TABLE 3 mean blade stiffness LCS, relative blade stiffness value RLCS and cumulative relative blade stiffness value TRLCS of the Broussonetia papyrifera at different moments of water loss
Figure BDA0001599725820000072
Figure BDA0001599725820000081
Therefore, the water retention capacity of the leaves of the paper mulberry is 9.16.
Example 2:
taking mulberry as an example, all the procedures are the same as example 1.
TABLE 4 maximum compressive internal force F of mulberry leaves at different moments of water lossmaxLeaf water potential W and physiological capacitance CP
Figure BDA0001599725820000082
TABLE 5 ultimate stress σ of Mulberry leaves at different moments of water lossmaxBlade tension LT and blade stiffness LCS
Figure BDA0001599725820000083
Figure BDA0001599725820000091
TABLE 6 mean leaf stiffness LCS, relative leaf stiffness value RLCS and cumulative relative leaf stiffness value TRLCS of mulberry trees at different moments of water loss
Figure BDA0001599725820000092
Therefore, the water retention capacity of the leaves of the mulberry was 5.33.
The implementation effect of the invention is as follows:
as can be seen from tables 3 and 6, the water retention capacity (9.16) of the leaves of the paper mulberry is higher than that of the mulberry (5.33), which indicates that the drought resistance of the paper mulberry is higher than that of the mulberry, and the true situation is also met, and the cumulative relative leaf stiffness value TRLCS of the plants can represent the water retention capacity of the leaves of the plants, and the drought resistance of different plants is quantitatively compared.
The above description is only a preferred embodiment of the present invention, and the present invention is not limited to the above embodiment, and it should be noted that any equivalent substitution, obvious modification made by those skilled in the art under the teaching of the present specification are within the spirit scope of the present specification, and the present invention should be protected.

Claims (5)

1. A method for quantitatively detecting the water retention capacity of plant leaves is characterized by comprising the following steps:
taking fresh branches of plants to be detected with leaves, and taking measures to slow down water diffusion;
step two, cleaning the leaves, picking a plurality of leaves with consistent growth vigor, and soaking the leaves in water;
step three, taking out the soaked water-saturated leaves after the leaves are in a water-saturated state, drying and dehydrating the leaves after treatment, and measuring the maximum compression internal force F of the left half part of the main veins of the dried and dehydrated leaves at the tth hour after the leaves are dried and dehydratedmaxMeasuring the water potential W and the physiological capacitance CP of the right half part of the main leaf vein of the leaf at the same time period;
step four, according to the maximum compression internal force F of the blademaxAnd the diameter d of the circular cross section of the push down probePCalculating the ultimate stress sigma of the blademax；
Ultimate stress of blade
Figure FDA0002446103940000011
FmaxThe unit is N; pi represents a circumferential ratio; dPThe unit is mm; sigmamaxUnit is N/mm2；
Step five, calculating the tension LT of the leaves according to the water potential W of the leaves and the physiological capacitance CP;
said calculation of blade tension LTIs given by the formula
Figure FDA0002446103940000012
Wherein LT is in cm2In/cm, i is the dissociation coefficient, R is the gas constant, T is the thermodynamic temperature,0is a vacuum dielectric constant, a is a relative dielectric constant of cytosol, M is a relative molecular mass of the cytosol, CP is plant leaf physiological capacitance, and W is plant leaf water potential;
step six, utilizing the ultimate stress sigmamaxCalculating the leaf stiffness LCS of the plant according to the leaf tension LT;
plant leaf stiffness
Figure FDA0002446103940000013
Wherein LCS unit is N/mm, and ultimate stress sigmamaxRepresenting the maximum compressive internal force per unit area of the blade in contact with the circular cross-section of the pressing probe, dLARepresents the effective thickness of the blade per unit area, and has a unit of cm/cm2；
Seventhly, the rigidity LCS of the plant leaves at the time 0 is measured0As a reference, the relative leaf stiffness value RLCS of the plant at each water loss time is calculatedi；
Adding the relative leaf stiffness values of the plants at each water loss moment to obtain an accumulated relative leaf stiffness value TRLCS of the plants;
and step nine, comparing the accumulated relative leaf stiffness values TRLCS of different plants respectively, thereby quantitatively calculating the water retention capacities of the leaves of the different plants.
2. The method for quantitatively detecting the water retention capacity of plant leaves as claimed in claim 1, wherein the instrument for measuring the maximum compressive internal force in the third step is a texture analyzer.
3. The method for quantitatively detecting the water retention capacity of plant leaves as claimed in claim 1 or 2, wherein the maximum compressive internal force F in the third stepmaxThe pressure of the texture analyzer probe pressing is enough to be ensured in the measuring processLarge, it can crush the blade.
4. The method for quantitatively detecting the water retention capacity of plant leaves as claimed in claim 1, wherein the calculation formula of the relative leaf stiffness value of the plant at each dehydration time in the seventh step is RLCSi＝LCSi/LCS0In which the LCSiThe stiffness of the plant leaves at the ith water loss moment is respectively 0, 1, 2 and 3 ….
5. The method for quantitatively determining the water retention capacity of plant leaves as claimed in claim 1, wherein the cumulative relative leaf stiffness value TRLCS ∑ RLCS of the plants in the eighth stepi＝RLCS0+RLCS1+RLCS2+RLCS3…。
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CN109633092A (en) * 2018-12-25 2019-04-16 北京林业大学 A method of detection poplar drought stress critical value
CN110133185B (en) * 2019-04-17 2022-03-22 江苏大学 Method for judging water retention capacity of plant stem based on stress relaxation time
CN110108632B (en) * 2019-04-24 2021-08-03 江苏大学 Method for detecting dehydration resistance of plant tissue based on energy change
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CN103630656A (en) * 2013-12-18 2014-03-12 江苏大学 Method for quantitatively detecting drought resistance of plants
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