Method for preparing paper mulberry somatic embryo and application thereof to paper mulberry expanding propagation

A somatic embryo and paper mulberry technology, applied in the biological field, can solve the problems of low efficiency, high labor cost and high cost, and achieve the effect of solving the complicated preparation and solving the market shortage.

Active Publication Date: 2015-10-21
大连中植环境生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cuttings have problems such as seasonality, large site occupation, and low efficiency.
Conventi

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1, preparation somatic embryo

[0050] 1. Preparation of embryogenic callus

[0051] (1) Get the leaves of the aseptic seedlings of the tree, cut into 0.5 × 0.5cm 2, inoculated on Medium A, and cultivated in the dark at 23±2°C for 25 days. At this time, it can be observed that light yellow callus grows from the leaf cut, that is, embryogenic callus.

[0052] (2) The embryogenic callus tissue obtained in step (1) was taken, inoculated on medium A, and cultured in the dark at 23±2° C. for 25 days.

[0053] (3) The embryogenic callus tissue obtained in step (2) was taken, inoculated on medium A, and cultured in the dark at 23±2° C. for 25 days.

[0054] In actual operation, the subculture can be continuously carried out according to the method of step (2).

[0055] Medium A (solid): MS medium containing 0.5mg / L 2,4-dichlorophenoxyacetic acid, 30g / L sucrose and 5g / L agar powder.

[0056] 2. Liquid suspension culture

[0057] (1) Add 1 g of the embryogenic c...

Embodiment 2

[0067] Embodiment 2, somatic embryo plant regeneration

[0068] Dissolve sodium alginate in 1 / 2 MS medium to obtain sodium alginate solution.

[0069] 1. Put the somatic embryo obtained in Example 1 into 3.0g / 100mL sodium alginate solution and soak at room temperature for 3-4min, then transfer to 0.5g / 100mL CaCl 2 Soak in an aqueous solution for 3 minutes at room temperature to form artificial seeds with a certain hardness, then rinse with sterile deionized water for 3-4 times to terminate the reaction, and place them on sterile paper to absorb the surface moisture of the artificial seeds.

[0070] 2. Place the artificial seeds on the culture medium, light at 24°C for 12 hours (the light intensity in this example is 1500 lx; ​​in actual operation, the light intensity is 1500-2000 lx) / 12 hours dark, static culture 30d.

[0071] Medium D (solid): containing 0.5mg / L NAA, 0.5mg / L 6-BA, 1mg / L GA 3 (Gibberellin), 1 / 2MS medium of 30g / L sucrose and 7g / L agar.

[0072] Experiments ...

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PUM

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Abstract

The invention discloses a method for preparing a paper mulberry somatic embryo and application thereof to paper mulberry expanding propagation. The provided method for preparing the paper mulberry somatic embryo includes the following steps that 1, embryonic callus of paper mulberry is added into a second culture medium to be cultured, and the second culture medium is a 1/2MS culture medium containing 0.5 mg/L 2,4-dichlorphenoxyacetic acid and 30 g/L saccharose; 2, after the step 1 is finished, the culture system is transferred to a third culture medium to be cultured, and the third culture medium is a 1/2MS culture medium containing 0.5 mg/L NAA, 0.2 mM TDZ, 0.4 mg/L IBA, 0.4 mg/L 6-BA and 30 g/L saccharose. The whole technological system of efficiently inducing somatic cells of the paper mulberry from explants and culturing the somatic cells in fluid in a suspension mode is completed for the first time, the technology is provided for large-scale industrialized production, and the problems that it is tedious to manufacture paper mulberry germchit, and the paper mulberry germchit is highly demanded in the market are solved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing somatic embryos of mulberry tree and its application in the propagation of mulberry tree. Background technique [0002] Broussoneti Papyrifera (L) Vent, also known as mulberry tree, is a deciduous tree belonging to Moraceae. It can grow in almost all types of soil, has few pests and diseases, is resistant to dry and cold, damp and hot, and has developed root system. An excellent tree species for water and soil conservation. The mulberry tree is planted in one year and harvested in many years. The whole body of the mulberry tree can be used: the bark fiber is white and slender, and it is a high-grade paper and textile raw material with high value and large market demand; the wooden pole is a good raw material for the production of paper and fiberboard; the leaves are rich in crude protein and amino acids , trace elements, is a high-quality feed raw...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 张宗申张朝晖窦旖君金朝霞李毅熊伟
Owner 大连中植环境生物科技有限公司
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