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A kind of litsea cubeba tissue culture method

A technique for tissue culture, Radix chinensis, applied in horticultural methods, microorganism-based methods, botanical equipment and methods, etc., can solve the problem of death, restricting the efficiency of rapid propagation of Radix chinensis in vitro, abnormal growth of explants, etc. problems, to achieve accurate test results, reduce manpower, and promote development.

Active Publication Date: 2022-02-18
HUNAN NUOZ BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the prior art, in the process of using the bud stem section of Litsea cubeba as explants for tissue culture, the explants often grow abnormally or even die, which seriously restricts the efficiency of Litsea cubeba rapid propagation in vitro

Method used

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  • A kind of litsea cubeba tissue culture method
  • A kind of litsea cubeba tissue culture method
  • A kind of litsea cubeba tissue culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] In early April 2019, wild Litsea Cubeba spring budding stems were collected in Shending Mountain, Miluo City, Hunan Province. The collected budding stems were washed with detergent or washing powder, rinsed under running water for 8 hours, and then placed in Pretreatment at a low temperature of 4°C for 8 hours.

[0037] In the aseptic operating table, put the budded stem section after low-temperature pretreatment in 75% ethanol for 10 seconds, rinse it with sterile water for 4 times, and put it in 0.1% mercuric chloride solution for 5 minutes to sterilize , followed by rinsing with sterile water for 4 times, and after the rinsing, the sterile budded stem section is placed in sterile water for subsequent use.

[0038] Randomly select 1-2 stem segments with buds from the same spring apex, use sterilized surgical scissors to cut out multiple stem tissue fragments with a thickness of no more than 1mm and a total weight of 0.6g, and place them in 1ml of DNA extraction soluti...

Embodiment 2

[0045] At the beginning of April 2019, the wild Litsea cubeba spring with slightly budded stems were collected in Taohualing Park, Changsha City, Hunan Province. The collected budded stems were washed with detergent or washing powder, rinsed under running water for 10 hours, and then placed Pretreatment at a low temperature of 6°C for 12 hours.

[0046] In the aseptic operating table, put the budded stem section after low temperature pretreatment in 75% ethanol for 15 seconds, rinse it with sterile water for 5 times, and put it in 0.1% mercuric chloride solution for 7 minutes to sterilize , followed by 5 times of rinsing with sterile water as well. After rinsing, the aseptic budded stem section is placed in sterilized water for subsequent use.

[0047] Randomly select 1-2 stem segments with buds from the same spring apex, use sterilized surgical scissors to cut out multiple stem tissue fragments with a thickness of no more than 1mm and a total weight of 1g, and place them in 1...

Embodiment 3

[0054] In early May 2019, wild Litsea Cubeba spring budding stems were collected in Huju Town, Zhuzhou City, Hunan Province. The collected budding stems were washed with detergent or washing powder, rinsed under running water for 9 hours, and then placed in Pretreatment at a low temperature of 5°C for 10 hours.

[0055] In the aseptic operating table, put the budded stem section after low temperature pretreatment in 75% ethanol for 12 seconds, rinse it with sterile water for 5 times, and put it in 0.1% mercuric chloride solution for 6 minutes to sterilize , followed by 5 times of rinsing with sterile water as well. After rinsing, the aseptic budded stem section is placed in sterilized water for subsequent use.

[0056] Randomly select 1-2 stem segments with buds from the same spring apex, use sterilized surgical scissors to cut out multiple stem tissue fragments with a thickness of no more than 1mm and a total weight of 0.8g, and place them in 1ml of DNA extraction solution ,...

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Abstract

A Litsea Cubeba tissue culture method disclosed by the invention comprises the following steps: taking Litsea Cubeba L. bud stem section as an explant, after pretreatment and disinfection, detecting Arthrobacter endophytes, and screening out the explants The explants of the endophyte of the genus Arthrobacter in the explant tissue were cultured subsequently. Litsea cubeba tissue culture method provided by the present invention detects Arthrobacter endophytes in explants before culturing, so that explants without endophytes are selected for subsequent culture, thereby greatly reducing manpower, The waste of material resources has improved the efficiency of Litsea cubeba rapid propagation in vitro and promoted the development of Litsea cubeba industry.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture and in vitro rapid propagation, and more specifically relates to a method for culturing litsea cubeba tissue. Background technique [0002] Litsea cubeba (Litsea cubeba) is a unique industrial raw material and spice tree species in southern my country. The essential oil extracted from its fruit has a high content of citral and is widely used in medicine, food, cosmetics and other fields. The establishment of the tissue culture and rapid propagation system of Litsea cubeba can not only promote the development of Litsea cubeba industry, but also lay the foundation for the basic research of Litsea cubeba. [0003] In the prior art, in the process of using the bud stem section of Litsea cubeba as explants for tissue culture, the explants often grow abnormally or even die, which seriously restricts the efficiency of Litsea cubeba rapid propagation in vitro . Contents of the invention ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00C12Q1/689C12Q1/6806C12Q1/04C12R1/06
CPCA01H4/00C12Q1/689C12Q1/6806
Inventor 陈昊王阳裴莹张付豪
Owner HUNAN NUOZ BIOLOGICAL TECH CO LTD
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