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Method for screening salt-tolerant day lily varieties based on glyoxalase activity

A technology of glyoxalase and Hemerocallis, which is applied in the field of screening salt-tolerant Hemerocallis varieties based on glyoxalase activity, can solve the problems of costing a lot of manpower, polluting soil, etc., and achieve the effect of low cost, short cycle and accurate screening results

Pending Publication Date: 2020-04-03
SHANGHAI INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Existing methods for screening salt-tolerant Hemerocallis varieties require a lot of manpower, and large-scale screening may pollute the soil

Method used

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  • Method for screening salt-tolerant day lily varieties based on glyoxalase activity
  • Method for screening salt-tolerant day lily varieties based on glyoxalase activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The method for screening hemerocallis tolerant varieties based on glyoxalase activity of this embodiment includes the following steps:

[0023] (1) Take 0.1g of the young leaves of the Hemerocallis variety ‘Tranquil Morgan’, crush them into fine powder in liquid nitrogen, and add 1 mL of 0.1 mol·L -1 Phosphate buffer solution, shake well; at 4℃, 1000 revolutions·min -1 Centrifuge for 10 min, take the supernatant, which is the protein solution of the young leaves of Hemerocallis, and divide it into two equal parts, one for the determination of protein content and the other for the determination of glyoxalase activity; the phosphate buffer is pH= 7.2, also contains 1mmol·L -1 Phenylmethylsulfonyl fluoride and 1% polyvinylpyrrolidone;

[0024] (2) The method for determining protein content: Coomassie brilliant blue method.

[0025] Take 60 μL of the leaf protein solution in a 10 mL centrifuge tube, add the reaction solution [Coomassie Brilliant Blue 4 mL, distilled water 940 μL] ...

Embodiment 2

[0029] The method for screening hemerocallis tolerant varieties based on glyoxalase activity of this embodiment includes the following steps:

[0030] (1) Take 0.1g of the young leaves of the Hemerocallis variety "Duplex", crush them into fine powder in liquid nitrogen, and add 1mL of 0.1mol·L -1 Phosphate buffer solution, shake well; at 4℃, 1000 revolutions·min -1 Centrifuge for 10 min, take the supernatant, which is the protein solution of the young leaves of Hemerocallis, and divide it into two equal parts, one for the determination of protein content and the other for the determination of glyoxalase activity; the phosphate buffer is pH= 7.2, also contains 1mmol·L -1 Phenylmethylsulfonyl fluoride and 1% polyvinylpyrrolidone;

[0031] (2) The method for determining protein content: Coomassie brilliant blue method.

[0032] Take 60 μL of the leaf protein solution in a 10 mL centrifuge tube, add the reaction solution [Coomassie Brilliant Blue 4 mL, 940 μL of distilled water] to the s...

Embodiment 3

[0036] The method for screening hemerocallis tolerant varieties based on glyoxalase activity of this embodiment includes the following steps:

[0037] (1) Take 0.1g of the young leaves of the Hemerocallis variety ‘Golden Doll’, pulverize them into fine powder in liquid nitrogen, and add 1 mL of 0.1 mol·L -1 Phosphate buffer, shake well; at 4℃, 1000 revolutions.min -1 Centrifuge for 10 min, take the supernatant, which is the protein solution of the young leaves of Hemerocallis, and divide it into two equal parts, one for the determination of protein content and the other for the determination of glyoxalase activity; the phosphate buffer is pH= 7.2, also contains 1mmol·L -1 Phenylmethylsulfonyl fluoride and 1% polyvinylpyrrolidone;

[0038] (2) The method for determining protein content: Coomassie brilliant blue method.

[0039] Take 60 μL of the leaf protein solution in a 10 mL centrifuge tube, add the reaction solution [Coomassie Brilliant Blue 4 mL, 940 μL of distilled water] to the...

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Abstract

The invention discloses a method for screening salt-tolerant day lily varieties based on glyoxalase activity. The method comprises the following steps: taking tender day lily leaves; putting into liquid nitrogen and crushing into fine powder; adding phosphate buffer, after oscillating and shaking up, centrifuging at low temperature, then taking the supernatant, dividing into two equal parts, one part is used for determining the protein content, the other part is used for determining the activity of glyoxalase, the activity of glyoxalase is calculated according to the consumption of the substrate methyl glyoxalase and the protein content per minute, and the plant with high glyoxalase activity is salt-tolerant. The salt-tolerant day lily variety is screened by measuring the activity of glyoxalase in vivo. Meanwhile, in order to avoid errors caused by fresh weight, the protein content in vivo is firstly measured, and the salt tolerance of the day lily is screened according to the glyoxalase activity of unit protein. Therefore, the method provided by the invention has the advantages of short period, accurate screening result, low cost and the like, and is of great significance to day lily salt resistance research and breeding.

Description

Technical field [0001] The invention relates to a method for screening salt-tolerant plant varieties, in particular to a method for screening salt-tolerant hemerocallis varieties based on glyoxalase activity. Background technique [0002] Hemerocallis is a perennial perennial herb with a wide range of varieties, rich and colorful flowers, and strong adaptability. It can be planted once and can be viewed for many years. It is often introduced and cultivated as the first choice ornamental plant in urban greening. However, it has an obvious defect that it is not adapted to the environment of saline-alkali soil. Once planted in saline-alkali soil, it usually ends up in large areas of dead seedlings. Therefore, the selection of salt-tolerant hemerocallis varieties is particularly important. [0003] Existing methods for screening salina-tolerant varieties require a lot of manpower, and large-scale screening may pollute soil. We found that methylglyoxal is at a normal level under norma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 倪迪安梁锦钟荣谭雨馨吴寒杨华英
Owner SHANGHAI INST OF TECH
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