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Method for establishing small pine nut tissue culture regeneration system

A tissue culture and construction method technology, applied in the field of forestry economic plants, achieves the effects of easy operation, solving the scarcity of seedlings, and simple process route

Inactive Publication Date: 2016-11-23
李军
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, tidewater power generation, wind power generation, etc. have been developed in the world, while the field of plant oil refining is still in the development and research stage.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] (1) Disinfection of seeds: Select green fruits that are in the early stage of maturity and soak them in neutral-concentration washing powder water for 20 minutes, rinse them with tap water for half an hour, take out the seeds, and peel off the outer testa in an ultra-clean workbench. Disinfect with 70% ethanol for 3 seconds, then wash twice with sterile water, then disinfect with 0.1% mercury liter solution for 15 minutes, rinse with sterile water for 3 times and set aside. The statistical pollution rate can be as low as 2% one week after inoculation.

[0017] (2) Embryo callus induction: Cut the seed embryos treated in step (1) into 0.6cm×0.6cm×0.6cm and inoculate them on the induction medium. Illuminated for 8 hours, and the light intensity was 1600x, until embryogenic callus was induced, and the callus induction rate was 65%. The induction medium is ER+5-BA1.0mg / L+NAA0.6mg / L+GA 3 6mg / L + sucrose 12g / L + agar 3.2g / L, pH 5.5.

[0018] (3) Differentiation culture: tra...

Embodiment 2

[0022] (1) Seed disinfection: Select 1.5-year-old immature fruits, soak in washing powder water for 16 minutes, rinse with tap water for 1.5 hours after brushing, take out the immature seeds, peel off the outer testa in an ultra-clean workbench, and first use 75% After disinfecting with ethanol for 9 seconds, wash with sterile water for 4 times, then disinfect with 0.1% mercuric chloride solution for 11 minutes, rinse with sterile water for 5 times and prepare for use. The statistical pollution rate can be as low as 6% one week after inoculation.

[0023] (2) Embryo callus induction: the immature embryos treated in step (1) were cut into 0.4cm×0.4cm×0.4cm and inoculated on the induction medium, and cultured at 26°C in total darkness for 32 days. Then light was applied for 11 hours every day, and the light intensity was 1600x, until embryogenic callus was induced, and the callus induction rate was 65%. The induction medium is ER+8-BA1.0mg / L+NAA0.3mg / L+GA 3 4mg / L + sucrose 15g / ...

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Abstract

The invention discloses a method for establishing a small pine nut tissue culture regeneration system, which is a technical method for obtaining tree seedlings with stable properties through an asexual rapid propagation technology. According to the invention, the small pine nut tissue culture regeneration system is established by taking an immature embryo of small pine nut tree as an explant and through the steps such as seed disinfection, embryo callus induction, differentiation culture, rooting culture, seedling hardening and transplantation; and the problem of shortage in high-quality seedlings of small pine nut tree is solved, and a firm guarantee can be provided for carrying out large-area planting expansion and transformation work of small pine nut trees in future.

Description

technical field [0001] The invention belongs to the field of forestry economic plants, in particular to a method for constructing a tissue culture regeneration system of pine nuts. Background technique [0002] Petroleum is a non-renewable resource. One day the world's underground oil resources will be exhausted. This is also a strategic material, as we all know. Therefore, tidewater power generation, wind power generation, etc. have been developed in the world, while the field of plant oil refining is still in the development and research stage. Small Chinese pine is also called small pine nuts, small pine cones, and its fruits can be used to refine diesel oil with an oil yield of more than 30%. It is a thick, thick and long tree that is easy to manage and plant. Contents of the invention [0003] The object of the present invention is to provide a kind of regeneration system constituted by sterilization, induction, differentiation, cultivation, seedling hardening and tr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/008
Inventor 李军
Owner 李军
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