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Method for measuring sub-low temperature or sub-high temperature tolerance of plant

A sub-low temperature and sub-high temperature technology is applied in the field of plant temperature tolerance measurement, which can solve the problems of reduced physiological activity and inability to effectively distinguish physiological activities, and achieves the effects of rapid method, novel technical principle and good stability.

Inactive Publication Date: 2015-08-12
HAINAN UNIVERSITY
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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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  • Method for measuring sub-low temperature or sub-high temperature tolerance of plant
  • Method for measuring sub-low temperature or sub-high temperature tolerance of plant
  • Method for measuring sub-low temperature or sub-high temperature tolerance of plant

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 a plant. The method comprises the following steps: (1) selecting the tender tissue of the plant, crushing, adding an enzymatic hydrolysate and mixing evenly to realize the enzymatic hydrolysis of the cell walls of the tissue; (2) washing and purifying the product to obtain a protoplast suspension, and adding a hydrolase to the protoplast suspension to keep low-level regeneration of the cell wall cellulose; (3) putting the protoplast added with the hydrolase in a warm bath under a sub-low temperature or sub-high temperature condition; after the warm bath is finished, adding ultrapure water to reduce the osmotic potential of the solution and regularly measuring the diameter of the protoplast; (4) calculating unit times and the unit osmotic potential of a plurality of protoplast and the average value of the net rates Lp of moisture on the cell membrane in unit area going into cells from the outside of the cells according to the obtained diameter value of the protoplast, wherein if the larger the difference between the Lp average value and the Lp value of the selected plant at an appropriate temperature is, the worse the sub-low temperature or sub-high temperature tolerance of the plant is. The method has the characteristics of wide adaptability 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 Applications(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 UNIVERSITY
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