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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). , Potato (Solanum tuberosum L) stem tips, etc., different species, and different parts have very different specific requirements and preservation effects for vitrification ultra-low temperature preservation. The present invention uses flax seeds as materials, and vitrification ultra-low temperature preservation Not yet reported

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
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: Seed cryopreservation and thawing process

[0027] Take the flax "Zhongya No. 1" seeds harvested in the same year (provided by the flax breeding research group of the Institute of Hemp Research, Chinese Academy of Agricultural Sciences), dry them naturally, put the flax seeds with a water content of about 9% into the loading solution, load them for a certain period of time, The flax seeds are taken out and put into a centrifuge tube equipped with a glass protectant, and the centrifuge tube is placed in a gradient cooling box (produced by Nalgene Company of the United States, model 5100-0001) with 100% isopropanol, and then the gradient cooling box is put into- The 80°C low-temperature refrigerator was used for gradient cooling, and the isopropanol in the gradient cooling box was reused 4-5 times before being renewed. Lowered to a certain temperature, put into liquid nitrogen for storage. The flax seeds preserved in ultra-low temperature were thawed natura...

Embodiment 2

[0030] Embodiment 2: Determine the relationship between loading time (minutes), gradient drop to temperature and flax seed germination rate before dropping into liquid nitrogen

[0031] In this example, on the basis of Example 1, the loading time is designed to be 0min, 5min, 10min, 15min, and 20min, and the gradient drops to temperatures of -10°C, -20°C, -20°C, -30°C, -40°C, For -50°C, -60°C, -70°C, see Table 1 for details. A total of 40 groups of experiments were designed, and the number of flax seeds in each group was 50, 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, and the absorbent cotton is moist), and then placed in an incubator at 25°C for 3 days. Remove the Petri dish and count the number of sprouts.

[0032] According to the results of germination rate, the effects of loadi...

Embodiment 3

[0039] Embodiment 3: The relationship between the different preservation and treatment methods of flax seeds and the germination rate of flax seeds

[0040] In the present embodiment, different cryopreservation modes and processes designed on the basis of Example 1 are:

[0041] a. Fresh flax seeds are put into cryopreservation tubes without any treatment, and are directly stored in liquid nitrogen;

[0042] b The flax seeds are not loaded, directly add glass protective agent and put them in the cryopreservation tube, and store them in liquid nitrogen at ultra-low temperature;

[0043] c. Load the flax seeds for 20 minutes, add vitrification protective agent, gradually cool down to -40°C, and put them into liquid nitrogen for ultra-low temperature storage;

[0044] d Flaxseeds were loaded for 20 minutes, added vitrification protective agent, and put into liquid nitrogen for ultra-low temperature storage without gradient cooling.

[0045] In this embodiment, 50 plump flax see...

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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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