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Rapid propagation method for cultivating dennstaeditiaceae suspension cells

A suspension cell and bowl fern technology, applied in the field of plants, can solve the problems of unreported research and achieve the effects of high proliferation rate, large yield, and large-scale production

Inactive Publication Date: 2014-12-03
NANJING TONGZE AGRI SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It grows under the forest or by the stream, at an altitude of 1000-2400 meters. At present, the utilization of pod fern adopts wild collection, and there is no report on the study of its suspension cells.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] Take the young leaves of the bowl fern, soak them in bleaching powder for 2 minutes, wash them with running water for 12 minutes, the water flow should not be too large, treat them with 0.1% mercuric chloride on the ultra-clean workbench for 7 minutes, wash them with sterile water 5 times, and inoculate the sterilized leaves into N6 +KT1mg / L+2,4-D2mg / L+0.5g / L activated carbon medium for callus induction, add 6.5g / L agar, 30g / L sucrose, darkroom culture, temperature 20℃, induced bowl Fern callus was inoculated with N6+KT1mg / L+2,4-D1mg / L for subculture of callus, adding 30g / L sucrose, 6.5g / L agar, pH5.8, light 1500lx, temperature 20℃ 0.5g of stable and uniform callus was selected from the callus after proliferation, and inoculated into liquid medium 6,7-V+NAA0.1mg / L+6-BA0.1mg / L+Zn 2+ , Ca 2+ , Mg 2+ Suspension shaking culture was carried out in 0.5mmol / L, using stirring ventilation equipment, temperature 22 ℃, every 60ml cell suspension was put into 250ml Erlenmeyer fla...

Embodiment 2

[0011] Take the young leaves of the bowl fern, soak them in bleaching powder for 2 minutes, wash them with running water for 12 minutes, the water flow should not be too large, treat them with 0.1% mercuric chloride on the ultra-clean workbench for 7 minutes, wash them with sterile water 5 times, and inoculate the sterilized leaves into N6 +KT1mg / L+2,4-D2mg / L+0.5g / L activated carbon medium for callus induction, add 6.5g / L agar, 30g / L sucrose, darkroom culture, temperature 20℃, induced bowl Fern callus was inoculated with N6+KT1mg / L+2,4-D1mg / L for subculture of callus, adding 30g / L sucrose, 6.5g / L agar, pH5.8, light 1500lx, temperature 20℃ 0.5g of stable and uniform callus was selected from the callus after proliferation, and inoculated into liquid medium 6,7-V+NAA0.2mg / L+6-BA0.2mg / L+Zn 2+ , Ca 2+ , Mg 2+ Suspension shaking culture was carried out in 2mmol / L, using stirring ventilation equipment, temperature 22 ℃, every 60ml cell suspension was put into 250ml Erlenmeyer flask...

Embodiment 3

[0013] Take the young leaves of the bowl fern, soak them in bleaching powder for 2 minutes, wash them with running water for 12 minutes, the water flow should not be too large, treat them with 0.1% mercuric chloride on the ultra-clean workbench for 7 minutes, wash them with sterile water 5 times, and inoculate the sterilized leaves into N6 +KT1mg / L+2,4-D2mg / L+0.5g / L activated carbon medium for callus induction, add 6.5g / L agar, 30g / L sucrose, darkroom culture, temperature 20℃, induced bowl Fern callus was inoculated with N6+KT1mg / L+2,4-D1mg / L for subculture of callus, adding 30g / L sucrose, 6.5g / L agar, pH5.8, light 1500lx, temperature 20℃ 0.5g of stable and uniform callus was selected from the callus after proliferation, and inoculated into liquid medium 6,7-V+NAA0.2mg / L+6-BA0.2mg / L+Zn 2+ , Ca 2+ , Mg 2+ Suspension shaking culture in 0.5mmol / L, using stirring ventilation equipment, temperature 22 ℃, each 60ml cell suspension into 250ml Erlenmeyer flask, take out after 4-5 we...

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Abstract

The invention provides a rapid propagation method for cultivating dennstaeditiaceae suspension cells. The method comprises the following steps: obtaining of sterile blades; induction of calluses; proliferation of the calluses; cultivation of suspension cells and the like. The dennstaeditiaceae suspension cells prepared by the method disclosed by the invention are high in proliferation index, short in growth period, and good in genetic stability, and a new path is provided for development and utilization of dennstaeditiaceae medicinal secondary metabolites.

Description

technical field [0001] The invention relates to a rapid multiplication method for cultured suspension cells of fern fern, belonging to the technical field of plants. Background technique [0002] bowl fern, Dennstaedtia scabra (Wall. ex Hook.) T. Moore , Bowl fern family, stems reddish brown horizontally, reddish brown, densely covered with brown transparent nodular hairs, leaves sparse; stalk length 20-35 cm, thick 2-3 mm, reddish brown or light chestnut, slightly shiny, Leaf blades are 20-29-(50) cm long and 15-20 cm wide, triangular-lanceolate or oblong, with 3-4 pinnatifids in the lower part, three pinnatine-parted above the middle, and round sporangia Shaped, located at the top of the small veins of the lobes; the cyst group is tureen-shaped, gray-green, slightly hairy. Distributed in Taiwan, Guangxi, Guizhou, Yunnan, Sichuan, Hunan, Jiangxi and Zhejiang; Nepal, India, Indochina, Japan also. It grows under the forest or by the stream, at an altitude of 1000-2400 mete...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 杨存
Owner NANJING TONGZE AGRI SCI & TECH
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