A kind of seed embryo disinfection method for bamboo tissue culture

A disinfection method and tissue culture technology, applied in the field of plant cultivation, can solve the problems of difficult disinfection of moso bamboo seed embryos, difficult control of pollution rate, high toxicity, etc., and achieve the effects of solving endophyte pollution, promoting tissue culture work, and simple operation

Active Publication Date: 2021-10-22
INT CENT FOR BAMBOO & RATTAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the tissue culture of Moso bamboo, the most important economic bamboo species in my country, has not been successful yet, and germ germ disinfection of Moso bamboo is one of the major problems.
Previous disinfection methods were either difficult to control the contamination rate or had high toxicity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The explants are full-grown, bright-colored, and healthy seeds of bamboo that are free from diseases and insect pests. The disinfection method is the aseptic treatment method for the naked seeds that have been stripped off their shells before tissue culture, including shelling, soaking, and washing. , disinfection, dissection and other steps.

[0027] 1. Peel off the lemma of the moso bamboo seed embryo and put it into a glass bottle with a cover.

[0028] 2. Soak the seed embryos with tap water, put them into a 30°C constant temperature water bath, and treat them for 36 hours, changing the water every 6 hours on average.

[0029] 3. Put the seed embryo in a beaker, cover it with gauze and wash it with running water for 1 hour to remove the remaining paddles; add an appropriate amount of detergent or a few drops of Tween 40 to the beaker, cover it with gauze, and continue to wash it with running water for 1 hour. The flow speed should be such that the seed embryos are ...

Embodiment 2

[0034] In this embodiment, step 2 is to soak the seed embryos with tap water, put them into a constant temperature water bath at 30°C, and treat them for 48 hours, changing the water every 6 hours on average.

[0035] Step 4. In the ultra-clean workbench, first soak the seed embryos with 70% alcohol for 1 min, rinse them with sterile water for 5 times, and transfer them to a sterile bottle with a cover during the process; then disinfect them with 1.5% formaldehyde solution for 4 h, and make 1 time / h manual shock treatment; finally rinsed with sterile water 5 times, during which time the embryos were transferred to a sterile bottle with a lid again.

[0036] Other operations are the same as in Example 1. In the present embodiment, the initial contamination rate is controlled below 5%, and the final contamination rate is also below 5%, but the embryo germination rate and callus induction rate are at extremely low levels in this example. It shows that the concentration of formal...

Embodiment 3

[0038] In this embodiment, step 4 is, in the ultra-clean workbench, first soak the seed embryo with 70% alcohol for 1 min, rinse it with sterile water 5 times, and transfer it to a sterile bottle with a cover during the period; then sterilize it with 0.5% formaldehyde solution The treatment was performed for 3 hours, during which manual shaking was performed once per hour; finally, the embryos were rinsed with sterile water for 5 times, during which time the seed embryos were transferred to a sterile bottle with a lid again.

[0039] Other operations are the same as in Example 1. In this embodiment, the initial pollution rate can be controlled below 20%, and the final pollution rate can be controlled below 30%.

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PUM

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Abstract

The present invention proposes a method for disinfecting seed embryos used in bamboo tissue culture, comprising the following steps: (1) peeling off the lemma of bamboo seed embryos; (2) soaking the seed embryos at a constant temperature of 28 to 32°C for 30 to 48 hours, and changing the water during 6 hours 1 time; (3) Rinse with running water for 1-3 hours; (4) In the ultra-clean workbench, first soak the seed embryos with alcohol, then rinse with sterile water 4-6 times, and then disinfect with 0.8-1.6% formaldehyde solution for 3 ~5h, (5) Dissection: in the ultra-clean workbench, cut off the tail of the bamboo germ embryo with awn. The disinfection method proposed by the present invention is simple to operate, the core step uses formaldehyde solution with relatively low toxicity, and solves the difficult problem of endophytic bacteria pollution in the process of bamboo embryo tissue culture, greatly reduces the pollution rate, and greatly promotes the Tissue culture of bamboo seed embryos. The invention lays a technical foundation for raising seedlings of Phyllostachys pubescens, establishing a callus induction regeneration system of molecular breeding, and the like.

Description

technical field [0001] The invention belongs to the technical field of plant cultivation, and in particular relates to a seed embryo disinfection method in the tissue culture process. Background technique [0002] Bamboo belongs to Poaceae ( Gramineae ) Bamboo subfamily ( Bambusoideae ) Perennial evergreen plants are extremely important non-woody renewable forestry resources, and are also important landscaping plants. There are more than 70 genera and more than 1,200 species of bamboo plants in the world, mainly distributed in tropical and subtropical regions, and a few species are distributed in temperate and frigid regions. my country is very rich in bamboo resources. It is the most important bamboo cultivation and production country in the world. There are more than 500 species of bamboo plants in 39 genera. The bamboo forest area is 6.01 million hectares, accounting for more than 1 / 4 of the world's bamboo forest area and about the total forest area of ​​the country. 4...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/00
Inventor 马艳军高健程占超
Owner INT CENT FOR BAMBOO & RATTAN
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