阅读说明：本技术 一种测定多花梾木耐涝能力的方法 (method for measuring waterlogging tolerance of cornus floribunda ) 是由 张文煊 周余华 王莹 孙杰 周雨 王红梅 于 2019-09-12 设计创作，主要内容包括：本发明公开了一种测定多花梾木耐涝能力的方法,试验经过淹水延续天数的变化,记录以及测定多花梾木叶片颜色形态、色差、叶绿素、荧光参数,发掘多花梾木耐水湿能力的临界值,有利于充分了解淹水对多花梾木光合作用的影响过程和作用机理,并且该研究结果为多花梾木耐涝机理研究奠定了理论基础,对科学合理地制定苗期抗涝对策具有一定的现实意义,也为多花梾木实地推广应用提供理论依据。(The invention discloses a method for determining waterlogging tolerance of cornus floribunda, which is characterized in that the change of the number of days of the test after the waterlogging lasts, the color form, the color difference, the chlorophyll and the fluorescence parameters of leaves of the cornus floribunda are recorded and determined, the critical value of the water-moisture tolerance of the cornus floribunda is found, the influence process and the action mechanism of the waterlogging on the photosynthesis of the cornus floribunda are favorably and fully known, the research result lays a theoretical foundation for the waterlogging tolerance mechanism research of the cornus floribunda, a certain practical significance is realized for scientifically and reasonably formulating waterlogging tolerance strategies in the seedling stage, and a theoretical basis is provided for the field popularization and application of the cornus floribunda.)

1. A method for measuring waterlogging tolerance of cornus floribunda is characterized by comprising the following steps:

(1) Performing flooding treatment on the cornus floribunda, wherein the water level exceeds the soil surface by 5cm, and continuing until the leaves of the plants are bare;

(2) Observing the color of leaves by naked eyes, measuring the color difference of the leaves of the cornus floribunda by using a spectrocolorimeter, measuring the chlorophyll content of the cornus floribunda by using a spectroscopic analyzer or measuring the chlorophyll fluorescence of the cornus floribunda by using a fluorometer;

(3) Grading and grading according to 4 indexes of the leaf color, leaf color difference, chlorophyll content and chlorophyll fluorescence of the cornus floribunda and scoring:

(4) and calculating the total score of the 4 indexes, and judging the flooding degree of the cornus floribunda according to the score so as to judge the flooding tolerance of the cornus floribunda.

2. the method of claim 1, wherein determining the leaf chromatic aberration using a spectrocolorimeter comprises the steps of:

(1) obtaining a fresh leaf sample;

(2) and (3) measuring the leaf color of the fixed leaves in the middle of the plant by using a spectrocolorimeter, taking the data measured for the first time as a standard color, taking the later data as a contrast color, comparing the color difference value of the leaves tested each time, and researching the change of the leaf color.

3. The method of claim 1, wherein the step of determining the chlorophyll content using a spectroscopic analyzer comprises the steps of:

(1) Obtaining a fresh leaf sample;

(2) And selecting the middle part of the leaf, and detecting the chlorophyll content by using a spectrum analyzer so as to calculate the photosynthetic rate.

4. the method of claim 1, wherein the step of measuring chlorophyll fluorescence using a fluorometer comprises the steps of:

(1) Obtaining a fresh leaf sample;

(2) chlorophyll of the leaves is measured by a fluorimeter, the leaves to be measured are clamped by a leaf clamp for dark treatment, and then the measurement is carried out.

5. the method according to claim 1, wherein the step (3) of assigning the grade and the score criteria are as follows:

(I) leaf color

A level: normally growing green or dark green leaves, to obtain 0 point;

b stage: the edge of the leaf is slightly reddish and is divided into 1 point;

c level: the leaves are orange, and 2 points are obtained;

d stage: leaves are reddish brown, and 3 points are obtained;

e, grade: withering and yellow leaves and withering, and obtaining 4 points;

(II) color difference of blade

A level: the total color difference delta Eab is 0 to obtain 0 point;

b stage: the total color difference delta Eab is more than 0 and less than or equal to 5, and 1 is obtained;

C level: the total color difference delta Eab is more than 22 and less than or equal to 25 or the total color difference delta Eab is more than 5 and less than or equal to 15, and 2 points are obtained;

D stage: the total color difference delta Eab is less than or equal to 20 and is more than 15, and 3 points are obtained;

E, grade: the total color difference delta Eab is less than or equal to 22 and is more than 20, and 4 points are obtained;

(III) chlorophyll content

A level: R800/R700 is more than or equal to 5.9 and less than or equal to 6.2 or R800/R640 is more than or equal to 2.7 and less than or equal to 2.8, and 0 point is obtained;

B stage: R800/R700 is more than or equal to 5.6 and less than 5.9, or R800/R640 is more than 2.8 and less than or equal to 3.0, and 1 point is obtained;

c level: R800/R700 is more than or equal to 4.9 and less than 5.6 or R800/R640 is more than or equal to 2.6 and less than 2.7, and 2 points are obtained;

d stage: R800/R700 is more than or equal to 3.7 and less than 4.9 or R800/R640 is more than or equal to 2.1 and less than 2.6, and 3 points are obtained;

E, grade: R800/R700 is more than or equal to 1.0 and less than 3.7 or R800/R640 is more than or equal to 1.0 and less than 2.1, and 4 points are obtained;

(IV) chlorophyll fluorescence

A level: Fv/Fm is more than or equal to 0.7 and less than or equal to 0.8, and 0 point is obtained;

B stage: Fv/Fm is more than or equal to 0.6 and less than 0.7, and 1 is obtained;

c level: Fv/Fm is more than or equal to 0.5 and less than 0.6, and 2 points are obtained;

D stage: Fv/Fm is more than or equal to 0.3 and less than 0.5, and 3 points are obtained;

e, grade: Fv/Fm is more than or equal to 0.2 and less than 0.3, and 4 points are obtained;

Calculating the total scores of (I) to (IV), wherein the total score is 13-16, which is the serious flooding degree and the flooding tolerance is poor; 9-12 is moderate flooding degree, moderate flooding tolerance and the like; 1-8 is slight flooding degree, and has better flooding resistance; 0 is not flooded, and the flooding resistance is good.

Technical Field

the invention relates to a method in the agricultural field, in particular to a method for measuring waterlogging tolerance of cornus floribundus.

background

The influence of water stress and rehydration on physiological characteristics and spectral characteristics of flue-cured tobacco is researched by Betaer waves and the like in 'influence of water stress on physiological characteristics and spectral characteristics of flue-cured tobacco', the chlorophyll and soluble protein of leaves, the photosynthetic rate and the transpiration rate of the leaves, the accumulation amount of dry matters, the membrane peroxidation degree of the leaves and the spectral reflectivity are investigated, and the spectral characteristics are used as important means for diagnosing the water stress degree of the flue-cured tobacco.

According to Zhuzhuo and the like, in research on growth and photosynthetic characteristics of Paeonia ostii in the south of the Yangtze river under flooding stress, waterlogging stress is carried out on Paeonia ostii in a waterlogging stress simulating mode, physiological and biochemical change conditions of Paeonia ostii under stress are researched by measuring changes of indexes such as relative conductivity, chlorophyll, soluble protein, Malondialdehyde (MDA), superoxide dismutase (SOD), peroxidase (CAT) and catalase (POD) activity, and main component analysis is carried out by taking waterlogging tolerance coefficients of various indexes as measurement indexes.

the method comprises the steps of conducting flooding stress on the countryside wind box fruits and the introduced purple leaf wind box fruits in 'the influence of the flooding stress on the photosynthesis characteristics of the countryside wind box fruits and the introduced purple leaf wind box fruits', measuring the influence on the photosynthesis characteristics of leaves, and measuring the net photosynthesis rate (Pn), the air pore conductance (Gs) and the transpiration rate (Tr) of the leaves, the non-photochemical quenching (NPQ) and the excess light energy (1-qP)/NPQ.

wangyongqiang et al, in the "Change in major Biochemical indicators of Ginkgo biloba under flooding stress", point out that the change in the content of Malondialdehyde (MDA), free Proline (PRO), soluble sugar and superoxide dismutase (SOD), which are major biochemical indicators in ginkgo biloba leaves of 3 years, is measured at different times after flooding treatment.

Therefore, the current prior art contains many disadvantages: 1. the degree of flooding of the plant cannot be immediately judged from the plant morphology; 2. the determination efficiency is not high; 3. the procedure is complicated; 4. the measuring cost is very high; 5. some medicaments are extremely toxic during measurement, belong to control medicaments and are difficult to obtain during measurement.

disclosure of Invention

the purpose of the invention is as follows: the invention aims to disclose a method for measuring waterlogging tolerance of cornus floribunda, which can be used for quickly measuring the flooding condition of the cornus floribunda and exploring a critical value of the cornus floribunda for water-moisture resistance.

The technical scheme is as follows: the invention provides a method for measuring waterlogging tolerance of cornus floribunda, which is characterized by comprising the following steps of:

(1) performing flooding treatment on the cornus floribunda, wherein the water level exceeds the soil surface by 5cm, and continuing until the leaves of the plants are bare;

(2) observing the color of leaves by naked eyes, measuring the color difference of the leaves of the cornus floribunda by using a spectrocolorimeter, measuring the chlorophyll content of the cornus floribunda by using a spectroscopic analyzer or measuring the chlorophyll fluorescence of the cornus floribunda by using a fluorometer;

(3) Grading and grading according to 4 indexes of the leaf color, leaf color difference, chlorophyll content and chlorophyll fluorescence of the cornus floribunda and scoring:

(4) And calculating the total score of the 4 indexes, and judging the flooding degree of the cornus floribunda according to the score so as to judge the flooding tolerance of the cornus floribunda.

the method for measuring the color difference of the blade by using the spectrocolorimeter comprises the following steps:

(1) Obtaining a fresh leaf sample;

(2) And (3) measuring the leaf color of the fixed leaves in the middle of the plant by using a spectrocolorimeter, taking the data measured for the first time as a standard color, taking the later data as a contrast color, comparing the color difference value of the leaves tested each time, and researching the change of the leaf color.

The method for measuring the chlorophyll content by using a spectrum analyzer comprises the following steps:

(1) obtaining a fresh leaf sample;

(2) And selecting the middle part of the leaf, and detecting the chlorophyll content by using a spectrum analyzer so as to calculate the photosynthetic rate.

The measurement of chlorophyll fluorescence using a fluorometer comprises the steps of:

(1) obtaining a fresh leaf sample;

(2) chlorophyll of the leaves is measured by a fluorimeter, the leaves to be measured are clamped by a leaf clamp for dark treatment, and then the measurement is carried out.

The calibration grade and score criteria in step (3) are as follows:

(I) leaf color

a level: normally growing green or dark green leaves, to obtain 0 point;

b stage: the edge of the leaf is slightly reddish and is divided into 1 point;

C level: the leaves are orange, and 2 points are obtained;

D stage: leaves are reddish brown, and 3 points are obtained;

e, grade: withering and yellow leaves and withering, and obtaining 4 points;

(II) color difference of blade

A level: the total color difference delta Eab is 0 to obtain 0 point;

b stage: the total color difference delta Eab is more than 0 and less than or equal to 5, and 1 is obtained;

c level: the total color difference delta Eab is more than 22 and less than or equal to 25 or the total color difference delta Eab is more than 5 and less than or equal to 15, and 2 points are obtained;

d stage: the total color difference delta Eab is less than or equal to 20 and is more than 15, and 3 points are obtained;

E, grade: the total color difference delta Eab is less than or equal to 22 and is more than 20, and 4 points are obtained;

(III) chlorophyll content

a level: R800/R700 is more than or equal to 5.9 and less than or equal to 6.2 or R800/R640 is more than or equal to 2.7 and less than or equal to 2.8, and 0 point is obtained;

b stage: R800/R700 is more than or equal to 5.6 and less than 5.9, or R800/R640 is more than 2.8 and less than or equal to 3.0, and 1 point is obtained;

C level: R800/R700 is more than or equal to 4.9 and less than 5.6 or R800/R640 is more than or equal to 2.6 and less than 2.7, and 2 points are obtained;

D stage: R800/R700 is more than or equal to 3.7 and less than 4.9 or R800/R640 is more than or equal to 2.1 and less than 2.6, and 3 points are obtained;

e, grade: R800/R700 is more than or equal to 1.0 and less than 3.7 or R800/R640 is more than or equal to 1.0 and less than 2.1, and 4 points are obtained;

(IV) chlorophyll fluorescence

A level: Fv/Fm is more than or equal to 0.7 and less than or equal to 0.8, and 0 point is obtained;

B stage: Fv/Fm is more than or equal to 0.6 and less than 0.7, and 1 is obtained;

C level: Fv/Fm is more than or equal to 0.5 and less than 0.6, and 2 points are obtained;

d stage: Fv/Fm is more than or equal to 0.3 and less than 0.5, and 3 points are obtained;

e, grade: Fv/Fm is more than or equal to 0.2 and less than 0.3, and 4 points are obtained;

calculating the total scores of (I) to (IV), wherein the total score is 13-16, which is the serious flooding degree and the flooding tolerance is poor; 9-12 is moderate flooding degree, moderate flooding tolerance and the like; 1-8 is slight flooding degree, and has better flooding resistance; 0 is not flooded, and the flooding resistance is good.

has the advantages that: compared with the prior art, the method of the invention has the following advantages: 1. the flooding degree of the plant can be immediately judged from the plant morphology; 2. the determination efficiency is high; 3. the procedure is simple; 4. the determination cost is reduced; 5. leaves containing toxic agents can be assayed.

Drawings

FIG. 1 is a graph of the color change of a leaf under flooding;

FIG. 2 is a color difference change diagram of a blade under flooding (wherein, DeltaL is black-white color difference, DeltaL is positive, which indicates that a measured sample is lighter (white) than a standard sample and is negative, which indicates that the measured sample is darker (black) than the standard sample, Deltaa is red-green color difference, Deltaa is positive, which indicates that the measured sample is negative than the red (red) of the standard sample, which indicates that the measured sample is greener (green) than the standard sample, and Deltab is yellow-blue color difference, which indicates that the measured sample is negative than the yellow (yellow) of the standard sample, which indicates that the measured sample is bluer (blue) than the standard sample, and DeltaEab is total color difference);

FIG. 3 is a graph showing changes in chlorophyll content measured by a spectrum under flooding (R800/R700 and R800/R640 are parameters positively correlated with chlorophyll content);

FIG. 4 is a graph of chlorophyll fluorescence change under flooding (F0 is initial fluorescence, Fm is maximal fluorescence, Fv is variable fluorescence);

FIG. 5 is a graph of the fluorescence change of chlorophyll under flooding (Fo/Fm is the potential activity of PSI, Fv/Fm is the maximum photochemical efficiency at which the reaction center of PS II is fully opened under dark adaptation, and Fv/Fo is the potential activity of PS II).

Detailed Description