Molecular detection method for rapidly verifying salt-tolerant fraxinus velutina

A technology of fluffy white wax and molecular detection, which is applied in the detection field of molecular biology in the field of biology, and achieves the effects of short detection time, high sensitivity, and increased ecology

Inactive Publication Date: 2012-06-27
SHANDONG FOREST SCI RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are no relevant reports on molecular rapid detection of salt

Method used

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  • Molecular detection method for rapidly verifying salt-tolerant fraxinus velutina
  • Molecular detection method for rapidly verifying salt-tolerant fraxinus velutina
  • Molecular detection method for rapidly verifying salt-tolerant fraxinus velutina

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] 1. Select 300 seeds of the F1 generation of Luwa No. 3 variety that is plump and free from diseases and insect pests (a new salt-tolerant variety of fluffy white wax bred by Liu Dexi of Shandong Academy of Forestry Sciences), collected from Shandong Shouguang Experimental Station. Put them in 1.5ml centrifuge tubes, rinse with tap water, soak for 10 minutes, sterilize with 70% ethanol for 30 seconds, then sterilize with 0.1% mercuric chloride for 8 minutes, and then wash with sterile water for 5 times. Inoculate into the germination medium of MS+5g / L agar+30g / L sucrose (pH value 5.8), 10 seeds in each bottle of medium, 30 bottles in total, the culture conditions are daytime temperature 25°C, night temperature 18°C , light intensity 40μmol·m -2 ·s -1 , light time 14h·d -1 (The same under the culture condition), obtain 3 leaf 1 heart seedlings.

[0024] Among them, the components of MS medium are: KNO 3 1900mg·L -1 , NH 4 NO 3 1650mg·L -1 , MgSO 4 ·7H 2 O370mg...

Embodiment 2

[0037] 1. Select 400 seeds of the F1 generation of Plump No. Luwa No. 3 (a new salt-tolerant variety of fluffy white wax selected by Liu Dexi of Shandong Academy of Forestry Sciences), which is free from diseases and insect pests, and collected from Shandong Shouguang Experimental Station. Soak in water at a temperature of 45°C, change the water once a day, and sow the seeds in peat, perlite, and frog stone (volume ratio 1:1:1) substrate after 4 weeks of whitening, at a temperature of 25-30°C and relative humidity In 50-70% of the greenhouses, when the seedlings grow to 4 true leaves, clean the peat, perlite and frog stones at the roots of the seedlings, and then transplant them into Hoagland nutrient solution with a concentration of 1 / 2. 10 plants per plastic container, 250 plants were transplanted in 25 plastic containers, and the evaporated water was replenished to the original volume with deionized water every two days.

[0038] Wherein Hoagland's (Hoagland's) nutrient sol...

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Abstract

The invention discloses a molecular detection method for rapidly verifying salt-tolerant fraxinus velutina. The method comprises the following steps of: performing PCR (Polymerase Chain Reaction) amplification with two primers of a nucleotide sequence which is shown as SEQ ID No.1 and serves as an SCAR (Sequence Characterized Amplified Region) mark by taking DNA (Deoxyribonucleic Acid) of a fraxinus velutina plant as a template at the annealing temperature of 54-56 DEG C, performing electrophoretic separation on an amplified product on 1.0 percent agarose gel, observing a PCR result, and verifying whether the plant is a salt-tolerant fraxinus velutina plant; if a DNA stripe of 592bp is amplified, determining that the plant is a salt-tolerant fraxinus velutina plant; and if a NDA stripe of592bp cannot be amplified, determining that the plant is a salt-sensitive plant. Due to the adoption of the molecular detection method, the salt tolerance of the fraxinus velutina plant can be detected at any time without influencing normal growth; fraxinus velutina plantes can be classified as a salt-tolerant material and a salt-sensitive material for being subjected to molecular detection and judgment; and the detection is rapid and cost is low.

Description

technical field [0001] The invention relates to a molecular detection method for rapidly identifying salt-tolerant fluffy white wax, which belongs to the detection method of molecular biology in the field of biology. Background technique [0002] Salinity is one of the main environmental factors limiting plant growth and yield, and soil salinization is a worldwide problem. Breeding of new salt-tolerant plant varieties has always been the focus of research at home and abroad. Fraxinus Velutina Torr. is a deciduous tree of the genus Fraxinus Linn. in the family Oleaceae. It is native to the United States. It grows fast, has straight trunks, beautiful textures, and is resistant to stress. It is the only large tree species used for afforestation in saline-alkali land. However, most white waxes cannot grow and reproduce normally on saline-alkali soils with a salt content of more than 0.3%. Among them, Fraxinus Velutina Torr. Very strong salt tolerance, able to grow and develop...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 夏阳燕丽萍李双云刘翠兰束靖
Owner SHANDONG FOREST SCI RES INST
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