Vitrification ultralow-temperature preservation method of flax seeds

A technology for vitrifying ultra-low temperature and flax seeds, which is applied in the fields of plant preservation, botanical equipment and methods, and applications, and achieves the effects of simple process, simple and convenient operation, and reliable stability.

Active Publication Date: 2014-12-24
INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The vitrification ultra-low temperature method has simple process, convenient operation, good effect and reproducibility. It is an effective and safe method for long-term safe preservation of plant germplasm. It has been applied to the protocorm of Dendrobium wardianum and the tip of chrysanthemum (Chrysanthemum). , Pota

Method used

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  • Vitrification ultralow-temperature preservation method of flax seeds
  • Vitrification ultralow-temperature preservation method of flax seeds
  • Vitrification ultralow-temperature preservation method of flax seeds

Examples

Experimental program
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Effect test

Example Embodiment

[0026] Example 1: Ultra-low temperature preservation and thawing process of seeds

[0027] Take the flax "Zhongya No. 1" seeds harvested that year (provided by the Flax Breeding Research Group of the Flax Research Institute of the Chinese Academy of Agricultural Sciences), dry them naturally, put the flax seeds with a moisture content of about 9% into the loading liquid, and load them for a certain period of time. The flax seeds are taken out and placed in a centrifuge tube containing a glass protectant. The centrifuge tube is placed in a gradient cooling box (manufactured by Nalgene, USA, model 5100-0001) with 100% isopropanol, and then the gradient cooling box is placed in- 80°C low-temperature refrigerator performs gradient cooling, and the isopropanol in the gradient cooling box is renewed after repeated use 4-5 times. It is lowered to a certain temperature and stored in liquid nitrogen. The flax seeds after ultra-low temperature preservation are naturally thawed, and the th...

Example Embodiment

[0030] Example 2: Determine the relationship between the loading time (minutes), the temperature of the gradient drop before liquid nitrogen is added and the germination rate of flax seeds

[0031] This example is based on Example 1 and the design loading time is 0min, 5min, 10min, 15min, 20min, and the gradient drop temperature is -10℃, -20℃, -20℃, -30℃, -40℃, -50°C, -60°C, -70°C, see Table 1 for details. A total of 40 groups of experiments were designed, with 50 flax seeds in each group, repeated three times. After the flax seeds are stored at ultra-low temperature, they are evenly placed on moist filter paper (the filter paper is placed in a petri dish, and a thin layer of absorbent cotton is placed under the filter paper, which is moistened), and then placed in an incubator at 25°C for 3 days. Take out the petri dish and count the number of sprouts.

[0032] According to the results of the germination rate, the effects of loading time, gradient cooling and their interaction on...

Example Embodiment

[0039] Example 3: The relationship between different storage and treatment methods of flax seeds and seed germination rate

[0040] In this embodiment, the different ultra-low temperature storage methods and treatments designed on the basis of embodiment 1 are:

[0041] a Fresh flax seeds are put into the freezing tube without any treatment, and directly put into liquid nitrogen for storage;

[0042] b Flax seeds are not loaded, directly add glass protective agent and put them in the cryotube, and put them in liquid nitrogen for ultra-low temperature storage;

[0043] c. Load the flax seeds for 20 minutes, add vitrification protectant, drop the temperature to -40℃ gradually, and put in liquid nitrogen for ultra-low temperature storage;

[0044] d. Flax seeds are loaded for 20 minutes, and vitrification protectant is added. Without gradient cooling, they are stored in liquid nitrogen at ultra-low temperature.

[0045] In this example, 50 full flax seeds were selected for each test, and ea...

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Abstract

The invention discloses a vitrification ultralow-temperature preservation method of flax seeds. The vitrification ultralow-temperature preservation method specifically comprises the following steps: collecting sundried fresh flax seeds, loading in a loading solution for 20min under the normal temperature condition, then transferring into a freeze-preservation pipe filled with a vitrification protecting agent, putting in the freeze-preservation pipe in a gradient cooling box, reducing the temperature to -30 DEG C to -60 DEG C, and charging liquid nitrogen for ultralow-temperature preservation; and putting the freeze-preservation pipe out from the liquid nitrogen, naturally unfreezing the flax seeds when the room temperature is greater than or equal to 20 DEG C, rapidly unfreezing the flax seeds in a 30-DEG C water bath box for 1 minute when the room temperature is less than 20 DEG C, and washing for 2-3 times so as to obtain the flax seeds capable of being directly cultured for growth. The vitrification ultralow-temperature preservation method is simple, and convenient to operate; the preserved flax seeds are stable in inheritable characters, the germination rate of 98.67 percent can be obtained in germination tests of the seeds, and the flax seeds can be preserved for a long time.

Description

technical field [0001] The invention belongs to the technical field of flax germplasm resource preservation, and in particular relates to a vitrification ultra-low temperature preservation method of flax seeds. Background technique [0002] Flax (Linum usitatissimum) is an ancient bast fiber crop and oil crop in the world, which originated in the Near Middle East and the Mediterranean coast. Flax has a long history of cultivation in China and is a characteristic economic crop. Flax fiber has the best effect among the five hemp spinning industrial raw material crops, and is known as the queen of hemp. Flax germplasm resources are the basic work of flax breeding, scientific research, production and teaching. The primary task of germplasm resources research is to collect germplasm and preserve germplasm safely. At present, the preservation methods of flax germplasm resources are normal temperature and low temperature preservation of flax seeds. Flax seeds stored at room tempe...

Claims

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Application Information

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IPC IPC(8): A01N3/00
Inventor 许英陈建华王晓飞王玉富朱爱国栾明宝孙志民
Owner INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI
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