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A method for ultra-low temperature in vitro preservation of peach dormant bud stem tips

A technology of in vitro preservation and dormant buds, applied in the fields of plant preservation, botanical equipment and methods, and applications, can solve problems such as difficulty in regeneration rate and tissue browning, and achieve optimized ultra-low temperature preservation steps, less damage, and simplified loading. The effect of liquid treatment

Active Publication Date: 2021-10-26
ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Peach is rich in phenolic substances. The cutting of dormant buds in actual operation, the stress pressure caused by hypertonic solution, and physical damage in low temperature environment during the processing will make it show strong phenolic substances. These phenolic compounds May cause tissue browning, which inhibits bud reproduction and recovery after liquid nitrogen storage, thus making it more difficult to achieve high regeneration rates after freezing
In the prior art, there is no research report on ultra-low temperature in vitro preservation of peach dormant bud stem tips as materials

Method used

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  • A method for ultra-low temperature in vitro preservation of peach dormant bud stem tips
  • A method for ultra-low temperature in vitro preservation of peach dormant bud stem tips
  • A method for ultra-low temperature in vitro preservation of peach dormant bud stem tips

Examples

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Effect test

Embodiment 1

[0051] The method for ultra-low temperature in vitro preservation of peach dormant shoot tips provided in this example, the specific steps are as follows:

[0052] (1) Dormant branches of peaches in winter (variety: Zhongtao Hongyu) are collected outdoors, the flower buds on the long branches are removed, and the branches are processed into small sections of 2 to 3 cm each containing only one dormant bud;

[0053] (2) Peel off the scales: Use tweezers to peel off 4 to 5 layers of scales on the surface of each dormant bud until you get a light green wrapped bud body;

[0054] (3) In the ultra-clean workbench, first sterilize with 75% alcohol (v / v) for 1 min, then sterilize with 10% (w / w) sodium hypochlorite for 6 min, rinse with sterile water for 5 times, and strip the stem tip under a dissecting microscope ;

[0055] (4) The shoot tip is placed on a plate containing pre-medium, wrapped in parafilm and pre-cultivated in the dark at 4°C for 4 days. The composition of the pre-me...

Embodiment 2

[0061] The method for ultra-low temperature in vitro preservation of peach dormant shoot tips provided in this example, the specific steps are as follows:

[0062] (1) Dormant branches of peaches in winter (variety: Zhongtao Hongyu) are collected outdoors, the flower buds on the long branches are removed, and the branches are processed into small sections of 2 to 3 cm each containing only one dormant bud;

[0063] (2) Peel off the scales: Use tweezers to peel off 4 to 5 layers of scales on the surface of each dormant bud until you get a light green wrapped bud body;

[0064] (3) In the ultra-clean workbench, first sterilize with 75% alcohol (v / v) for 1 min, then sterilize with 10% (w / w) sodium hypochlorite for 6 min, rinse with sterile water for 5 times, and strip the stem tip under a dissecting microscope ;

[0065] (4) The shoot tip is placed on a plate containing pre-medium, wrapped in parafilm and pre-cultivated in the dark at 4°C for 6 days. The composition of the pre-me...

Embodiment 3

[0073] The method for ultra-low temperature in vitro preservation of peach dormant shoot tips provided in this example, the specific steps are as follows:

[0074] (1) Dormant branches of peaches in winter (variety: Zhongtao Hongyu) are collected outdoors, the flower buds on the long branches are removed, and the branches are processed into small sections of 2 to 3 cm each containing only one dormant bud;

[0075] (2) Peel off the scales: Use tweezers to peel off 4 to 5 layers of scales on the surface of each dormant bud until you get a light green wrapped bud body;

[0076] (3) In the ultra-clean workbench, first disinfect with 75% alcohol (v / v) for 30 seconds, then disinfect with 8% (w / w) sodium hypochlorite for 7 minutes, rinse with sterile water for 5 times, and strip the stem tip under a dissecting microscope ;

[0077] (4) The shoot tip is placed on the plate containing the pre-medium, wrapped in parafilm and pre-cultivated in the dark at 2°C for 2 days, the composition...

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Abstract

The invention discloses a method for ultra-low temperature in vitro preservation of peach dormant bud stem tips, comprising the following steps: pre-cultivation: taking peach dormant bud stem tips and placing them in a pre-culture medium for pre-cultivation; vitrification and dehydration treatment: using PVS3 solution to carry out Dehydration treatment for 80-120 minutes; cryopreservation in liquid nitrogen; thawing, unloading; recovery culture: the unloaded peach dormant shoot tips were placed in the recovery medium for recovery culture; a certain amount of Activated Charcoal and Ascorbic Acid. The present invention uses peach dormant bud stem tip as material, reduces low-temperature domestication steps, simplifies loading solution treatment, vitrification solution dehydration treatment steps, improves pre-culture medium and recovery medium, and causes damage to the stem tip during the treatment of each step. The damage is small, and the stem tip regeneration rate after preservation is high, which provides a new way for the long-term and stable preservation of peach germplasm resources.

Description

【Technical field】 [0001] The invention belongs to the technical field of low-temperature preservation of plant germplasm, and in particular relates to a method for ultra-low temperature in vitro preservation of peach dormant bud stem tips using a vitrification method. 【Background technique】 [0002] Peach is a deciduous woody perennial plant that is native to China and then spread all over the world. Edible peaches are the mainstay of the development of my country's peach industry, with rich types and varieties. With the development of breeding methods, my country has formed thousands of peach commercial varieties by 2019. It provides a greater possibility, so it is particularly important to collect and preserve peach germplasm resources reasonably. Cryogenic preservation of plant germplasm resources refers to a set of biotechnology for long-term preservation of plant cells, tissues or organs under ultra-low temperature conditions of liquid nitrogen (-196°C), and certain met...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N3/00
CPCA01N3/00
Inventor 方伟超陈璋王力荣曹珂王新卫陈昌文
Owner ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI