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Method for castanopsis hystrix immature embryo in-vitro regeneration

A technology for in vitro regeneration and immature embryos, which is applied in the field of in vitro regeneration of red vertebra immature embryos, can solve the problems that there is no red vertebra tissue culture technology and can not maximize the genetic improvement gain, so as to promote the utilization of germplasm resources, Consistent seedling growth and low cost results

Inactive Publication Date: 2020-08-18
YULIN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, sexual reproduction has the problem of differentiation and cannot maximize the genetic improvement gain, while asexual reproduction can overcome the above-mentioned shortcomings of sexual reproduction
At present, there is no report on the tissue culture technology of red vertebrae at home and abroad.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] (1) Collection of explants: collect immature young fruits from good single plants of red vertebrae that grow well in October, soak them in 75% alcohol for 1 min, disinfect with 0.1% mercuric chloride for 15 min, and rinse with sterile water 5 -7 times.

[0038] (2) Callus culture: Peel off the explants of step (1) under aseptic conditions, take out immature embryos, inoculate them in sterilized callus culture medium, and place them at 20℃. It can be induced to form callus after 55 days of dark culture, with an induction rate of 80%.

[0039] The callus culture medium is: modified White medium+1.0mg / L riboflavin+1.0mg / L ascorbic acid+0.2mg / L 6-BA+0.1mg / L NAA+25-30g / L sucrose+ 3.0-4.0g / L agar, pH value is 5.4-5.8.

[0040] (3) Adventitious bud induction culture: The callus cut pieces obtained in step (2) are transferred to adventitious bud culture medium, and adventitious buds are induced after 45 days of full dark culture at 20°C, with an induction rate of 85%.

[0041] The ad...

Embodiment 2

[0049] (1) Collection of explants: collect immature young fruits from good individual plants of red vertebrae that grow well in October, soak them in 75% alcohol for 1 min, disinfect with 0.1% mercuric chloride for 15 min, and rinse with sterile water for 5 -7 times.

[0050] (2) Callus culture: Peel off the explants of step (1) under aseptic conditions, take out immature embryos, inoculate them in sterilized callus culture medium, and place them at 20℃. After 50 days of dark culture, callus formation can be induced, with an induction rate of 83%.

[0051] The callus culture medium is: modified White medium+2.0mg / L riboflavin+3.0mg / L ascorbic acid+0.5mg / L 6-BA+0.3mg / L NAA+25-30g / L sucrose+ 3.0-4.0g / L agar, pH value is 5.4-5.8

[0052] (3) Adventitious bud induction culture: The callus cut pieces obtained in step (2) are transferred to adventitious bud culture medium, and adventitious buds are induced after 40 days of culture at 20°C in dark, with an induction rate of 87%.

[0053] T...

Embodiment 3

[0061] (1) Collection of explants: collect immature young fruits from good individual plants of red vertebrae that grow well in October, soak them in 75% alcohol for 1 min, disinfect with 0.1% mercuric chloride for 15 min, and rinse with sterile water for 5 -7 times.

[0062] (2) Callus culture: Peel off the explants of step (1) under aseptic conditions, take out immature embryos, inoculate them in sterilized callus culture medium, and place them at 20℃. Dark culture can induce callus formation for 40 days, and the induction rate is 85%.

[0063] The callus culture medium is: modified White medium+3.0mg / L riboflavin+5.0mg / L ascorbic acid+0.8mg / L 6-BA+0.5mg / L NAA+25-30g / L sucrose+ 3.0-4.0g / L agar, pH value is 5.4-5.8.

[0064] (3) Adventitious bud induction culture: The callus cut pieces obtained in step (2) are transferred to adventitious bud medium for culture, and adventitious buds are induced after 30 days of culture in the dark at 20°C, with an induction rate of 88%.

[0065] Th...

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Abstract

The invention discloses a method for castanopsis hystrix immature embryo in-vitro regeneration and belongs to the field of plant tissue culture. The method comprises the steps: collecting an immatureyoung fruit on a single castanopsis hystrix plant as an explant; taking out an immature embryo of the explant for culture and formation of a callus; carrying out adventitious bud culture; carrying outstrong seedling culture; carrying out rooting culture; and carrying out transplanting. A plant tissue culture technology is used for establishing the method for castanopsis hystrix immature embryo in-vitro regeneration, a callus induction rate reaches over 80%, an adventitious bud induction rate reaches over 83%, a rooting rate reaches over 85%, and a transplanting survival rate reaches over 90%.The method is characterized by low cost, a high propagation coefficient, relative conformity in seedling growth and the like, the method can be directly used for plant production of castanopsis hystrix seedlings and the method is of great significance for promoting germplasm resource utilization of castanopsis hystrix.

Description

Technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a method for in vitro regeneration of red vertebral embryos. Background technique [0002] Castanopsis hystrix Miq. (Castanopsis hystrix Miq.) is an evergreen broad-leaved tree belonging to the genus Castanopsis. It is also known as Hongchao, Hongke, and evergreen large tree. It has fast growth, straight stem shape, abundant fallen leaves, and good soil improvement effects. Excellent characteristics. The shoots, petioles and inflorescence axis of the year were purple-brown with papery or thin leathery leaves. They are endemic to my country, mainly distributed in southeast Fujian, southwest Hunan, Guangdong, Guangxi, Hainan and other places; they are important in South China Valuable native broad-leaved timber and efficient multi-purpose tree species. It has hard material, strong corrosion resistance, no cracking, no deformation, easy processing, and can be widely used; s...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 莫昭展苏建睦
Owner YULIN NORMAL UNIVERSITY
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