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A kind of method for measuring the tolerance of plants to sub-low temperature or sub-high temperature

A sub-low temperature and sub-high temperature technology, applied in the field of plant temperature tolerance measurement, can solve the problems of inability to effectively distinguish physiological activity and decrease of physiological activity, and achieve the effect of novel technical principle, rapid method and universal applicability

Inactive Publication Date: 2017-06-23
HAINAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in sub-low temperature or sub-high temperature environment, the integrity of plant cell membrane has not been affected, but the physiological activity is reduced
The application of conductivity method cannot effectively distinguish the change of physiological activity

Method used

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  • A kind of method for measuring the tolerance of plants to sub-low temperature or sub-high temperature
  • A kind of method for measuring the tolerance of plants to sub-low temperature or sub-high temperature
  • A kind of method for measuring the tolerance of plants to sub-low temperature or sub-high temperature

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Embodiment 1 The method for preparing protoplasts with different enzyme combinations and different enzymolysis times

[0053] (1) Get the new leaf that the watermelon seedling that grows about 30d fully expands, weighs 1g, cuts into the block that size is about 2 * 2mm;

[0054] (2) Add 5mL of enzymatic hydrolysis solution to the material and mix well, place at room temperature 25°C, oscillate for enzymolysis on a shaker at 80r / min, and enzymolysis for 150min;

[0055] The enzymatic solution combination has 2 kinds of enzyme solution combinations:

[0056] ①2.7% (w / v, that is, 100mL mannitol aqueous solution contains 2.7g cellulase) cellulase + 0.78% (w / v) isolated enzyme R-10, dissolved in 0.4mol / L mannitol in aqueous solution;

[0057] ②2.7% (w / v) cellulase + 0.78% (w / v) isolated enzyme R-10 + 0.155% (w / v) pectinase, dissolved in mannitol with an initial concentration of 0.4mol / L ;

[0058] (3) The solution after the enzymolysis is filtered with a 160-mesh nylon m...

Embodiment 2

[0062] The preparation method of protoplast under the different osmotic potentials of embodiment 2

[0063] (1) Get the fully expanded new leaves of watermelon seedlings that grow about 30d, weigh 1g, and cut into chunks of about 2 × 3mm;

[0064] (2) Add 5mL of enzymatic hydrolysis solution to the material, mix well, place at room temperature 25°C, and oscillate for enzymatic hydrolysis on a shaker at 80r / min;

[0065] The formula of the enzymatic hydrolysis solution is: 2.7% cellulase + 0.78% isolated enzyme R-10; dissolved in 0.3M, 0.35M, 0.4M, 0.45M, 0.5M aqueous mannitol solutions to optimize the concentration of mannitol aqueous solutions Effect on protoplast yield;

[0066] (3) After 120 minutes of enzymolysis, the enzymolysis solution was filtered with a 160-mesh nylon mesh, and the filtrate was collected;

[0067] (4) Then wash with 0.4mol / L mannitol, dilute to 6mL, centrifuge at a centrifugal force of 24×g for 5min, discard the supernatant, and repeat once;

[006...

Embodiment 3

[0069] The effect of embodiment 3 low temperature on the expansion of dendrobium protoplasts

[0070] (1) Cut 0.7g of new leaves of Dendrobium dendrobii into pieces of 3×3mm;

[0071] (2) Transfer to 2mL enzymatic hydrolysis solution;

[0072] The formula of the enzymatic solution is: 0.16% (w / v) pectinase + 2.8% (w / v) cellulase + 0.85% (w / v) isolated enzyme R-10, dissolved in 0.35M mannitol aqueous solution middle;

[0073] (3) 25°C, 100 minutes of enzymatic hydrolysis on a shaker at 80r / min;

[0074] (4) Filter the solution after enzymolysis with a 160-mesh nylon mesh, collect the filtrate, wash with 0.35mol / L mannitol, dilute to 5mL, centrifuge at a centrifugal force of 24×g for 5min, discard the supernatant, and repeat once;

[0075] (5) Add hydrolytic enzyme in the prepared protoplast suspension solution, the composition of hydrolytic enzyme is as follows: the cellulase of 1% (w / v), the isolated enzyme R10 of 0.3% (w / v), be dissolved in concentration In the mannitol a...

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Abstract

The invention discloses a method for measuring the sub-low temperature or sub-high temperature tolerance of plants, which comprises the following steps: (1) selecting tender plant tissues, after crushing, adding enzymatic hydrolysis solution and mixing them evenly, and enzymolyzing their cell walls; (2) After the product is washed and purified, a protoplast suspension is obtained, and hydrolytic enzymes are added to the protoplast suspension to maintain low-level regeneration of cell wall cellulose; Warm bath, after the warm bath is over, add ultrapure water to reduce the osmotic potential of the solution, and regularly measure the diameter of the protoplast; (4) use the obtained diameter value of the protoplast to calculate the unit time, unit osmotic potential, and cell membrane per unit area of ​​multiple protoplasts The average value of the net velocity Lp of water entering the cell from the outside of the cell, the greater the difference between the average value of Lp and the Lp value at the appropriate temperature of the selected plant, the worse its sub-low temperature or sub-high temperature tolerance. This method has the characteristics of wide applicability and high resolution.

Description

technical field [0001] The invention belongs to the technical field of measuring plant temperature tolerance, in particular to a method for measuring the tolerance of plants to sub-low temperature or sub-high temperature. Background technique [0002] Temperature is the most important environmental factor for plant growth. High temperature or low temperature will cause the growth and development of plants to be hindered or even damaged. [0003] Different plants have different temperature requirements, and even the same plant has different temperature requirements in different growth stages. The so-called sub-low temperature or sub-high temperature refers to the temperature condition that only slows down the growth of crops but does not cause major damage to the crops and cause their growth to stagnate. [0004] The tolerance of plants to sub-low temperature and sub-high temperature is called sub-low temperature tolerance or sub-high temperature tolerance. different from e...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/02
Inventor 江雪飞乔飞穆大伟党选民丛汉卿王静
Owner HAINAN UNIV
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