阅读说明：本技术 一种基于电阻值快速评价柳树品种耐盐性的方法 (Method for rapidly evaluating salt tolerance of willow variety based on resistance value ) 是由 张健 刘国元 陈艳红 江钰娜 余春梅 钟非 连博琳 柯勇超 郭安芳 朱星兆 刘昱 于 2020-08-14 设计创作，主要内容包括：本发明公开了一种基于电阻值快速评价柳树品种耐盐性的方法,取每种待测柳树品种一年生粗度3-5mm的枝条若干根,每根长度相同；测量每根枝条的中间粗度、枝条的初始含水量以及初始电阻值；然后将枝条浸入NaCl溶液中,在浸入2h、4h、6h、8h时拿出测定电阻值,并在浸入192h时拿出测定枝条的含水量、皮孔黑孔数和电导率值；采用基于初始电阻值的柳树耐盐分级评价模型进行分级。本方法操作方便,评价柳树品种耐盐性快捷且不受外界环境影响,可在10日内评价出待测样品的耐盐能力。(The invention discloses a method for rapidly evaluating salt tolerance of willow varieties based on resistance values, which comprises the steps of taking a plurality of branches with annual thickness of 3-5mm of each willow variety to be tested, wherein the lengths of the branches are the same; measuring the intermediate thickness of each branch, the initial water content of the branch and the initial resistance value; then immersing the branches in NaCl solution, taking out the branches for measuring the resistance value when the branches are immersed for 2h, 4h, 6h and 8h, and taking out the branches for measuring the water content, the number of skin holes and black holes and the conductivity value when the branches are immersed for 192 h; and (4) grading by adopting a willow salt tolerance grading evaluation model based on an initial resistance value. The method is convenient to operate, the salt tolerance of the willow variety can be evaluated quickly and is not influenced by the external environment, and the salt tolerance of the sample to be tested can be evaluated within 10 days.)

1. A method for rapidly evaluating the salt tolerance of willow varieties based on resistance values is characterized by comprising the following steps:

taking a plurality of branches with annual thickness of 3-5mm of each willow variety to be detected, wherein the length of each branch is the same; measuring the intermediate thickness of each branch, the initial water content of the branch and the initial resistance value; then immersing the branches in NaCl solution, taking out the branches for measuring the resistance value when the branches are immersed for 2h, 4h, 6h and 8h, and taking out the branches for measuring the water content, the number of skin holes and black holes and the conductivity value when the branches are immersed for 192 h; for each variety to be tested, after the measurement data of each branch is averaged, a willow salt tolerance grading evaluation model based on an initial resistance value is adopted for grading:

when R is₀<At 200, the grading evaluation model is as follows:

Y＝3.869[-0.124(X₁-4.273)/0.396+0.229(X₂-79.05)/14.214+0.258(X₃-69.95)/7.338+0.238(X₄-61.4)/5.236+0.176(X₅-52.15)/4.87-0.207(X₆-51.412)/3.196-0.043(X₈-13.275)/7.347-0.072(X₉-440.084)/240.969]²

the salt tolerance was divided according to the evaluation value Y as:

(a) y is more than 0 and less than or equal to 1, and the willow variety to be detected is extremely salt-tolerant;

(b) 1< Y < 2, the willow variety to be tested is salt-tolerant;

(c) 2< Y is less than or equal to 3, and the willow variety to be detected is moderately salt-tolerant;

(d) y is more than 3, and the sensitive salt of the willow variety to be detected.

When R is more than or equal to 200₀<At 250, the grading evaluation model is:

Y＝[-0.276(X₁-3.69)/0.271+0.231(X₂-94.65)/7.795+0.485(X₃-76.5)/7.578+0.422(X₄-68.45)/8.062+0.456(X₅-64.95)/5.246-0.371(X₆-47.413)/3.146+0.119(X₇-93.444)/0.408+0.159(X₈-14.42)/4.383+0.268(X₉-163.745)/97.068+1]²+[0.336(X₁-3.69)/0.271-0.316(X₂-94.65)/7.795+0.046(X₃-76.5)/7.578+0.195(X₄-68.45)/8.062+0.004(X₅-64.95)/5.246-0.187(X₆-47.413)/3.146+0.048(X₇-93.444)/0.408+0.357(X₈-14.42)/4.383-0.274(X₉-163.745)/97.068+1]²

the salt tolerance was divided according to the evaluation value Y as:

(a) y is more than 0 and less than or equal to 1, and the willow variety to be detected is extremely salt-tolerant;

(b) 1< Y < 5, the willow variety to be tested is salt-tolerant;

(c) y is less than or equal to 10 when 5, and the willow variety to be tested has moderate salt tolerance;

(d) y is more than 10, and the sensitive salt of the willow variety to be detected.

When R is₀When the evaluation value is more than or equal to 250, the grading evaluation model is as follows:

Y＝[-0.243(X₁-3.565)/0.352+0.259(X₂-108.15)/11.695+0.218(X₃-81.55)/11.941+0.151(X₄-70.5)/8.281+0.269(X₅-71.55)/6.143-0.049(X₆-44.88)/4.727-0.071(X₇-93.093)/0.623+0.083(X₈-12.39)/2.648-0.016(X₉-164.428)/65.417]²+[0.141(X₁-3.565)/0.352+0.01(X₂-108.15)/11.695-0.22(X₃-81.55)/11.941+0.577(X₄-70.5)/8.281-0.081(X₅-71.55)/6.143-0.272(X₆-44.88)/4.727+0.339(X₇-93.093)/0.623+0.405(X₈-12.39)/2.648+0.49(X₉-164.428)/65.417+2]²

the salt tolerance was divided according to the evaluation value Y as:

(a) y is more than 0 and less than or equal to 1, and the willow variety to be detected is extremely salt-tolerant;

(b) 1< Y < 5, the willow variety to be tested is salt-tolerant;

(c) y is less than or equal to 10 when 5, and the willow variety to be tested has moderate salt tolerance;

(d) y is more than 10, and the sensitive salt of the willow variety to be detected.

Wherein: r₀Is an initial resistance value; x₁Is the intermediate thickness; x₂Resistance value of 2 h; x₃Resistance value of 4 h; x₄Resistance value of 6 h; x₅Resistance value of 8 h; x₆Is the initial moisture content; x₇The water content is 192 h; x₈The number of the black holes is 192 h; x₉Was 192h conductivity.

2. The method for rapidly evaluating the salt tolerance of the willow variety according to claim 1, wherein the concentration of the NaCl solution is 100 mM/L.

3. The method for rapidly evaluating the salt tolerance of the willow variety based on the resistance value according to claim 1, wherein the conductivity value measuring method comprises the following steps: rinsing the branches with ultrapure water for 2 times, then sucking dry surface water, putting the branches into a centrifuge tube, adding 50mL of ultrapure water into the centrifuge tube, and measuring the conductivity value of water in the centrifuge tube by using a digital conductivity meter after the branches are completely soaked for 2 hours.

Technical Field

The invention belongs to the field of plant stress resistance, and particularly relates to a method for rapidly identifying and evaluating salt tolerance of willow varieties.

Background

The salt tolerance is the main character of plant stress resistance, the traditional salt tolerance identification method is to plant plants on saline soil, the survival rate and the growth condition of the plants are measured to evaluate, the salt tolerance of the variety to be tested can be generally evaluated within more than 1 year, and the salt tolerance is greatly influenced by the external environment. Willow is a general term of salix and salix rhynchophylla of salicaceae, is an important stress-resistant tree species, plays an important role in the aspects of soil heavy metal remediation, saline-alkali soil greening, biofuel and the like, and has good salt tolerance of many varieties, and the salt tolerance of some varieties is over 4 per mill (extremely salt tolerance).

Disclosure of Invention

The purpose of the invention is as follows: aiming at the prior art, a method for rapidly evaluating the salt tolerance of willow varieties based on resistance values is provided, and the salt tolerance of willows can be rapidly identified.

The technical scheme is as follows: a method for rapidly evaluating the salt tolerance of willow varieties based on resistance values comprises the following steps:

taking a plurality of branches with annual thickness of 3-5mm of each willow variety to be detected, wherein the length of each branch is the same; measuring the intermediate thickness of each branch, the initial water content of the branch and the initial resistance value; then immersing the branches in NaCl solution, taking out the branches for measuring the resistance value when the branches are immersed for 2h, 4h, 6h and 8h, and taking out the branches for measuring the water content, the number of skin holes and black holes and the conductivity value when the branches are immersed for 192 h; for each variety to be tested, after the measurement data of each branch is averaged, a willow salt tolerance grading evaluation model based on an initial resistance value is adopted for grading:

when R is₀<At 200, the grading evaluation model is as follows:

Y＝3.869[-0.124(X₁-4.273)/0.396+0.229(X₂-79.05)/14.214+0.258(X₃-69.95)/7.338+0.238(X₄-61.4)/5.236+0.176(X₅-52.15)/4.87-0.207(X₆-51.412)/3.196-0.043(X₈-13.275)/7.347-0.072(X₉-440.084)/240.969]²

the salt tolerance was divided according to the evaluation value Y as:

(a) y is more than 0 and less than or equal to 1, and the willow variety to be detected is extremely salt-tolerant;

(b) 1< Y < 2, the willow variety to be tested is salt-tolerant;

(c) 2< Y is less than or equal to 3, and the willow variety to be detected is moderately salt-tolerant;

(d) y is more than 3, and the sensitive salt of the willow variety to be detected.

When R is more than or equal to 200₀<At 250, the grading evaluation model is:

Y＝[-0.276(X₁-3.69)/0.271+0.231(X₂-94.65)/7.795+0.485(X₃-76.5)/7.578+0.422(X₄-68.45)/8.062+0.456(X₅-64.95)/5.246-0.371(X₆-47.413)/3.146+0.119(X₇-93.444)/0.408+0.159(X₈-14.42)/4.383+0.268(X₉-163.745)/97.068+1]²+[0.336(X₁-3.69)/0.271-0.316(X₂-94.65)/7.795+0.046(X₃-76.5)/7.578+0.195(X₄-68.45)/8.062+0.004(X₅-64.95)/5.246-0.187(X₆-47.413)/3.146+0.048(X₇-93.444)/0.408+0.357(X₈-14.42)/4.383-0.274(X₉-163.745)/97.068+1]²

the salt tolerance was divided according to the evaluation value Y as:

(a) y is more than 0 and less than or equal to 1, and the willow variety to be detected is extremely salt-tolerant;

(b) 1< Y < 5, the willow variety to be tested is salt-tolerant;

(c) y is less than or equal to 10 when 5, and the willow variety to be tested has moderate salt tolerance;

(d) y is more than 10, and the sensitive salt of the willow variety to be detected.

When R is₀When the evaluation value is more than or equal to 250, the grading evaluation model is as follows:

Y＝[-0.243(X₁-3.565)/0.352+0.259(X₂-108.15)/11.695+0.218(X₃-81.55)/11.941+0.151(X₄-70.5)/8.281+0.269(X₅-71.55)/6.143-0.049(X₆-44.88)/4.727-0.071(X₇-93.093)/0.623+0.083(X₈-12.39)/2.648-0.016(X₉-164.428)/65.417]²+[0.141(X₁-3.565)/0.352+0.01(X₂-108.15)/11.695-0.22(X₃-81.55)/11.941+0.577(X₄-70.5)/8.281-0.081(X₅-71.55)/6.143-0.272(X₆-44.88)/4.727+0.339(X₇-93.093)/0.623+0.405(X₈-12.39)/2.648+0.49(X₉-164.428)/65.417+2]²

the salt tolerance was divided according to the evaluation value Y as:

(a) y is more than 0 and less than or equal to 1, and the willow variety to be detected is extremely salt-tolerant;

(b) 1< Y < 5, the willow variety to be tested is salt-tolerant;

(c) y is less than or equal to 10 when 5, and the willow variety to be tested has moderate salt tolerance;

(d) y is more than 10, and the sensitive salt of the willow variety to be detected.

Wherein: r₀Is an initial resistance value; x₁Is the intermediate thickness; x₂Resistance value of 2 h; x₃Resistance value of 4 h; x₄Resistance value of 6 h; x₅Resistance value of 8 h; x₆Is the initial moisture content; x₇The water content is 192 h; x₈The number of the black holes is 192 h; x₉Was 192h conductivity.

Further, the concentration of the NaCl solution is 100 mM/L.

Further, the conductivity value measuring method comprises the following steps: rinsing the branches with ultrapure water for 2 times, then sucking dry surface water, putting the branches into a centrifuge tube, adding 50mL of ultrapure water into the centrifuge tube, and measuring the conductivity value of water in the centrifuge tube by using a digital conductivity meter after the branches are completely soaked for 2 hours.

Has the advantages that: the method is convenient to operate, the salt tolerance of the willow variety can be evaluated quickly and is not influenced by the external environment, and the salt tolerance of the sample to be tested can be evaluated within 10 days.

Detailed Description

The present invention is explained further below.

(1) Taking materials and treating

Taking branches of the willow variety to be detected with annual thickness of 3-5mm, taking 10 branches for each treatment, wherein each branch is 8cm long, and measuring the middle thickness of each branch, the initial water content and the initial resistance value of each branch by using a vernier caliper; the initial values were determined in triplicate and averaged. Then, the branches were immersed in 100mM/L NaCl solution and taken out at intervals of 2, 4, 6, and 8 hours to measure the primary resistance. The moisture content, the number of black skin pores and the conductivity value of the shoots were measured at 192h (8 d).

(2) Determination of water content

Two probes were inserted into the ends of the branches in a 1mm reading mode in the White Poplat kerring test mode by using the YHT01 moisture tester manufactured by Shenzhen Source Hengtong technology Co.

(3) Determination of resistance value

Resistance shelves of MF-47A internal magnetic pointer type mechanical multimeters manufactured by Nanjing Jinyaoyou instruments Limited are adopted, and two detecting heads are inserted into two ends of branches for reading by 1 mm.

(4) And (4) measuring the number of black skin pores.

The skin pores are small openings formed on the periderm of the branches and are the material exchange tissues of the cortex. The quantity of the black skin pores under salt stress means that the quantity of skin pore tissues can exchange substances such as gas and water with the outside, and the number of the black skin pores is selected as one of important salt tolerance indexes. The specific method for measuring the number of the black pores comprises the following steps: the number of dark spots in the skin of each shoot produced following salt stress was visually assessed.

(5) Conductivity value

Rinsing the branches with ultrapure water for 2 times, draining surface water, putting the branches into a centrifuge tube, adding 50mL of ultrapure water into the centrifuge tube, completely soaking the branches for 2 hours, and measuring the conductivity value of water in the centrifuge tube by using a Control 320C-06A digital conductivity meter produced by Saimer Feishell science and technology.

(6) For each variety to be measured, the measured data of each branch are averaged, and the method is based on the initial resistance value (R)₀) The willow salt-tolerant grading evaluation model is graded, and the evaluation model is established according to principal component clustering analysis:

when R is₀<When 200, the process:

Y＝3.869[-0.124(X₁-4.273)/0.396+0.229(X₂-79.05)/14.214+0.258(X₃-69.95)/7.338+0.238(X₄-61.4)/5.236+0.176(X₅-52.15)/4.87-0.207(X₆-51.412)/3.196-0.043(X₈-13.275)/7.347-0.072(X₉-440.084)/240.969]²(X₇coefficient 0, not set forth in equation)

(a) Y is more than 0 and less than or equal to 1, and the willow variety to be tested is extremely salt-tolerant (the salt tolerance is more than 4 per mill of mass concentration);

(b) 1< Y < 2, the willow variety to be detected is salt-tolerant (salt-tolerant degree is 3-4-thousandth mass concentration);

(c) 2< Y is less than or equal to 3, and the willow variety to be detected has moderate salt tolerance (the salt tolerance degree is 2-3 per mill mass concentration);

(d) y is more than 3, and the salt tolerance of the willow variety to be tested is less than 2 per mill of mass concentration.

When R is more than or equal to 200₀<At 250 time:

Y＝[-0.276(X₁-3.69)/0.271+0.231(X₂-94.65)/7.795+0.485(X₃-76.5)/7.578+0.422(X₄-68.45)/8.062+0.456(X₅-64.95)/5.246-0.371(X₆-47.413)/3.146+0.119(X₇-93.444)/0.408+0.159(X₈-14.42)/4.383+0.268(X₉-163.745)/97.068+1]²+[0.336(X₁-3.69)/0.271-0.316(X₂-94.65)/7.795+0.046(X₃-76.5)/7.578+0.195(X₄-68.45)/8.062+0.004(X₅-64.95)/5.246-0.187(X₆-47.413)/3.146+0.048(X₇-93.444)/0.408+0.357(X₈-14.42)/4.383-0.274(X₉-163.745)/97.068+1]²

(a) y is more than 0 and less than or equal to 1, and the willow variety to be tested is extremely salt-tolerant (the salt tolerance is more than 4 per mill of mass concentration);

(b) 1< Y < 5 > and the willow variety to be detected is salt-tolerant (salt-tolerant degree is 3-4 mass percent);

(c) 5< Y < 10, moderate salt tolerance (salt tolerance degree 2-3 mass per mill) of willow variety to be detected;

(d) y is more than 10, and the salt tolerance of the willow variety to be tested is less than 2 per mill of mass concentration.

When R is₀When the ratio is more than or equal to 250:

Y＝[-0.243(X₁-3.565)/0.352+0.259(X₂-108.15)/11.695+0.218(X₃-81.55)/11.941+0.151(X₄-70.5)/8.281+0.269(X₅-71.55)/6.143-0.049(X₆-44.88)/4.727-0.071(X₇-93.093)/0.623+0.083(X₈-12.39)/2.648-0.016(X₉-164.428)/65.417]²+[0.141(X₁-3.565)/0.352+0.01(X₂-108.15)/11.695-0.22(X₃-81.55)/11.941+0.577(X₄-70.5)/8.281-0.081(X₅-71.55)/6.143-0.272(X₆-44.88)/4.727+0.339(X₇-93.093)/0.623+0.405(X₈-12.39)/2.648+0.49(X₉-164.428)/65.417+2]²

(a) y is more than 0 and less than or equal to 1, and the willow variety to be tested is extremely salt-tolerant (the salt tolerance is more than 4 per mill of mass concentration);

(b) 1< Y < 5 > and the willow variety to be detected is salt-tolerant (salt-tolerant degree is 3-4 mass percent);

(c) 5< Y < 10, moderate salt tolerance (salt tolerance degree 2-3 mass per mill) of willow variety to be detected;

(d) y is more than 10, and the salt tolerance of the willow variety to be tested is less than 2 per mill of mass concentration.

Wherein:

R₀: an initial resistance value (k Ω); x₁: intermediate thickness (mm); x₂: 2h resistance value (k Ω); x₃: 4h resistance value (k Ω); x₄: a resistance value (k Ω) of 6 h; x₅: resistance value (k Ω) of 8 h; x₆: initial water content (%); x₇: 192h water content (%); x₈: 192h of black hole number (number); x₉: 192h conductivity (. mu.S/cm).

The salt tolerance of a batch of willow varieties is randomly evaluated by using the method, the evaluation result is shown in table 1, and the result is basically consistent with the statistical evaluation result of more than one year of willow varieties planted in saline soil.

TABLE 1

The method is convenient to operate, the salt tolerance of the willow variety can be evaluated quickly and is not influenced by the external environment, and the salt tolerance of the sample to be tested can be evaluated within 10 days.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.