Embryo disinfecting method for bamboo tissue culture

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

Active Publication Date: 2018-10-16
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 in tap water, put them in a constant temperature water bath at 30°C, 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 residual lemma; 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 turn...

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 minute, 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 hours, and do 1 time during the process / 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 this 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 conce...

Embodiment 3

[0038] In this embodiment, step 4 is, in the ultra-clean workbench, first soak the seed embryo with 70% alcohol for 1 minute, rinse it with sterile water for 5 times, and transfer it to a sterile bottle with a cover during the period; then use 0.5% formaldehyde solution for disinfection treatment 3h, during which manual shaking was performed once / h; finally, rinsed with sterile water for 5 times, during which time the 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 invention provides an embryo disinfecting method for bamboo tissue culture. The method comprises the following steps: (1) stripping off inferior palea of bamboo embryos; (2) soaking the embryos for 30 to 48 hours at the constant temperature of 28 DEG C to 32 DEG C, and changing water once every 6 hours during soaking; (3) carrying out flushing with flowing water for 1 to 3 hours; (4) soaking the embryos with spirit firstly in a superclean bench, then, flushing the embryos with sterile water for 4 to 6 times, and then, carrying out disinfecting treatment for 3 to 5 hours with a 0.8%-1.6% formaldehyde solution; and (5) carrying out dissection: cutting tails of bamboo embryos aristate in a superclean bench. According to the disinfecting method provided by the invention, the operation is simple, a core step employs the formaldehyde solution with relatively low toxicity, the problem, i.e., endophyte pollution in a bamboo embryo tissue culture process is solved, the pollution rate is greatly lowered, and the tissue culture work of phyllostachys pubescens embryos is greatly promoted. The method settles a technical foundation for the seedling raising of phyllostachys pubescens seedlingsprouts, the establishment of a caluus induced 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 is a perennial evergreen plant of the Gramineae subfamily Bambusoideae. It is an extremely important non-woody renewable forestry resource and an important landscaping plant. 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 and is the most important bamboo cultivation and production country in the world. There are more than 500 species of bamboo plants in 39 genera, and the bamboo forest area is 6.01 million ha 2 , accounting for more than 1 / 4 of the world's bamboo forest area, accounting for about 4% (3.8%) of the country's total forest area; the annual...

Claims

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Application Information

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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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