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Method for rapid identification of ploidy of salicaceae plant

A willow, rapid technology, applied in the biological field, to achieve the effect of simple preparation method, high resolution and reliable technical means

Active Publication Date: 2016-05-25
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the experimental system of using flow cytometry to detect the ploidy of Salicaceae plants at home and abroad

Method used

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  • Method for rapid identification of ploidy of salicaceae plant
  • Method for rapid identification of ploidy of salicaceae plant
  • Method for rapid identification of ploidy of salicaceae plant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1 weeping willow ( Salixbabylonica ) ploidy identification

[0034] The young leaves of the weeping willow to be tested were collected from Xuanwu Lake Park in Nanjing, and the diploid dustpan willow ( S. suchowensis ) for the control. Before the experiment, collect fresh young leaves, wrap them in wet filter paper and bring them back, store them in a refrigerator at 4°C, and use them within 3 days.

[0035] (1) Preparation of nucleus extraction buffer

[0036] The nucleus extraction buffer was prepared according to the following components and ratios: 0.5mM spermine tetrahydrochloride, 30mM sodium citrate, 200mM Tris, 80mM potassium chloride, 0.5% (v / v) TritonX-100. Adjust the pH to 7.5 with 1M hydrochloric acid or 1M sodium hydroxide, and store at 4°C.

[0037] (2) Preparation of nuclei staining buffer

[0038] Dissolve 1 mg of propidium iodide (PI) powder in 1 mL of deionized water to prepare a 1 mg / mL PI stock solution and store at 4°C.

[0039] (...

Embodiment 2

[0047] Embodiment 2 Chinese wolfberry ( S. integra ) ploidy identification

[0048] The sample to be tested willow willow is a hydroponic seedling, which comes from the germplasm resource nursery of Nanjing Forestry University in Chenxu Forest Farm, Sihong, Jiangsu. The diploid glandular willow ( S. chaenomeloides ) for the control.

[0049] (1) Preparation of nucleus extraction buffer

[0050] The nucleus extraction buffer was prepared according to the following components and ratios: 0.5mM spermine tetrahydrochloride, 30mM sodium citrate, 200mM Tris, 80mM potassium chloride, 0.5% (v / v) TritonX-100. Adjust the pH to 7.5 with 1M hydrochloric acid or 1M sodium hydroxide, and store at 4°C.

[0051] (2) Preparation of nuclei staining buffer

[0052] Dissolve 1 mg of propidium iodide (PI) powder in 1 mL of deionized water to prepare a 1 mg / mL PI stock solution and store at 4°C.

[0053] (3) Preparation of cell nucleus suspension

[0054] Take 30~50 mg of fresh young leaves...

Embodiment 363

[0061] Example 363 poplar ( Populus deltides ‘Harvord’) ploidy determination

[0062] Sample 63 poplar to be tested came from the campus of Nanjing Forestry University, and the young leaves of its branches were collected as the test material. The diploid Populus tomentosa ( P. tomentosa ) for the control.

[0063] (1) Preparation of nucleus extraction buffer

[0064] The nucleus extraction buffer was prepared according to the following components and ratios: 0.5mM spermine tetrahydrochloride, 30mM sodium citrate, 200mM Tris, 80mM potassium chloride, 0.5% (v / v) TritonX-100. Adjust the pH to 7.5 with 1M hydrochloric acid or 1M sodium hydroxide, and store at 4°C.

[0065] (2) Preparation of nuclei staining buffer

[0066] Dissolve 1 mg of propidium iodide (PI) powder in 1 mL of deionized water to prepare a 1 mg / mL PI stock solution and store at 4°C.

[0067] (3) Preparation of cell nucleus suspension

[0068] Take 30~50 mg of fresh young leaves, wash the dust on the surfa...

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Abstract

The invention discloses a method for rapid identification of the ploidy of a salicaceae plant. The method comprises the steps of cell nucleus extraction buffer solution preparation, cell nucleus dyeing buffer solution preparation, cell nucleus suspension preparation, cell nucleus DNA specific dyeing, flow cytometry detection and ploidy judgment. The method has the advantages of being wide in application range, low in sample consumption and simple in preparing method when used for determining the ploidy of the salicaceae plant. Extracted cell nucleus DNA is high in average flourescence intensity and small in variable coefficient. The determination result can be obtained quickly and is accurate, and resolution ratio is high. The method provides a reliable technical means for salicaceae plant polyploid breeding and has broad application prospects.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for rapidly identifying ploidy of Salicaceae plants. Background technique [0002] Salicaceae plants belong to the phylum Magnoliophyta, the class Magnoliopsida, the subclass Dilleniidae, the order Salicales, and the family Salicaceae. As an important industrial timber tree species, Salixaceae plants occupy a very important position in my country's forestry production due to their excellent characteristics such as rapid growth, strong adaptability and easy vegetative reproduction. In addition, willow plays an important role in phytoremediation and ecosystem restoration, and is also an important ornamental tree species in gardens (Tu Zhongyu, 1989; Ball et al., 2005; Tang Guimei et al., 2007). Salicaceae plants have high biomass production and have also been used as important biomass energy tree species in recent years (Adegbidi et al., 2001; Perlack et al., 2011; Zalesny et a...

Claims

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Application Information

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IPC IPC(8): G01N15/14
CPCG01N15/14
Inventor 李淑娴郭炜胡南李小平尹佟明
Owner NANJING FORESTRY UNIV
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