Method for culturing somatic embryos of liquidambar plants into seedlings and breeding method of liquidambar plants

A technology for culturing Liquidambar genus and embryos, which is applied in the field of tree tissue culture in the forestry industry, can solve the problems of high labor consumption, pollution, complicated operation process, etc.

Active Publication Date: 2020-06-05
BEIJING FORESTRY UNIVERSITY +1
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  • Description
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Problems solved by technology

[0008] The object of the present invention is to exist the mature somatic embryo of liquidambar plant (i.e. mature somatic embryo) in the tissue culture rapid propagation method of existing liquidambar plant needs to germinate on the solid germination medium, cultivate adult embryo seedling, and In the process of cultivating adult embryo seedlings on solid germination medium, there is a complicated operation process, which requires a lot of manpower, and at the same time, it is easy to cause pollution, resulting in technical defects such as low germination rate of aseptic seedlings in tissue culture of liquidambar. The invention provides a A method for cultivating mature somatic embryos of liquidambar plants into seedlings. The method of the present invention promotes the transf

Method used

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  • Method for culturing somatic embryos of liquidambar plants into seedlings and breeding method of liquidambar plants
  • Method for culturing somatic embryos of liquidambar plants into seedlings and breeding method of liquidambar plants
  • Method for culturing somatic embryos of liquidambar plants into seedlings and breeding method of liquidambar plants

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Experimental program
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Embodiment 1

[0089] 1. Test method

[0090] The embodiments of the present invention refer to the methods in the following published academic papers: W.A.Vendrame C.P.Holliday S.A.Merkle, etc., Clonal propagation of hybrid sweetgum (Liquidambarstyraciflua × L.formosana) by somatic embryogenesis, Plant Cell Rep (2001) 20:691- 695.

[0091] 2. Test materials

[0092] 1. Immature seeds of hybrid sweetgum

[0093] In the present invention, the immature seeds of the hybrid sweetgum (golden sweetgum as the female parent and the sweetgum eucalyptus as the male parent) are used as explant experimental materials.

[0094] In June and July of each year, immature spherical capsules of hybrid sweetgum (gum xanthus × liquidgum) were collected, and seeds were taken out from them as explant materials.

[0095] 2. Plant growth regulator

[0096] The plant growth regulator used in the present invention adopts 6-benzylaminoadenine (6-BA) and 2,4-dichlorophenoxyacetic acid (2,4-D).

[0097] 3. Medium

...

Embodiment 2

[0117] 1. Sterilization of explants, induction of embryogenic callus, subculture and culture

[0118] Refer to W.A.Vendrame·C.P.Holliday·S.A.Merkle et al., Clonal propagation of hybrid sweetgum (Liquidambar styraciflua×L.formosana) by somaticembryogenesis, Plant Cell Rep (2001) 20:691-695) method for explant sterilization and embryogenesis Callus induction culture.

[0119] After the immature zygotic embryos of the explants were cultured on the induction medium for 3-4 weeks, the growth rate of the embryogenic tissue began to slow down and the explants began to brown. At this time, the embryogenic tissue was selected for subculture Proliferation culture, transferred to the subculture medium for repeated subculture and proliferation culture under dark conditions at (25±2)°C (subculture once every 3 weeks).

[0120] 2. Somatic embryogenesis culture

[0121] Inoculate the embryogenic tissue of the hybrid liquidambaria subculture cultured on the somatic embryogenesis medium, and...

Embodiment 2A1

[0127] Example 2A1, Sterilization of explants, induction of embryogenic callus, subculture and culture

[0128] Same as step 1) of embodiment 2.

[0129] 2. Somatic embryogenesis culture

[0130] Same as step 2) of embodiment 2.

[0131] 3. Somatic embryo liquid maturation culture:

[0132] The embryogenic callus of the initial somatic embryos (i.e. immature somatic embryos, preferably "initial spherical, heart-shaped somatic embryos, torpedo embryos") obtained by culturing on the somatic embryogenesis medium is selected, and inoculated in the somatic embryo liquid In the maturation medium, the somatic embryo liquid maturation medium is placed in a conical flask, and the inoculation amount is 2g (usually 1-5g / 100ml) per 100ml of the somatic embryo maturation liquid medium, and the somatic embryo is grown on a shaker. Suspension liquid maturation culture. The liquid maturation culture of hybrid sweetgum somatic embryos is carried out under dark conditions, the culture tempe...

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Abstract

The invention discloses a method for culturing somatic cell embryos (short for somatic embryos) of liquidambar plants into seedlings and a breeding method of the liquidambar plants, and belongs to thetechnical field of tissue culture of forests. According to the seedling formation method, the mature somatic embryos of the liquidambar plants are subjected to liquid culture (liquid suspension germination culture and diffusion germination culture of the somatic embryos). By adopting the method for conducting liquid culture on the mature somatic embryos of the liquidambar plants, quick germination of the mature somatic embryos of the liquidambar plants in liquid culture media is promoted to form sterile somatic embryo seedlings, and a large number of somatic embryo seedlings can be produced by means of bio-reactors in a short time. The method is high in seedling formation efficiency, the operation method is simple and convenient, the seedlings emerge evenly and uniformly, a transplantingsurvival rate is high, and maintenance and management are convenient. In a whole production process, manpower can be saved, the cost is reduced, and regenerated plants can be continuously obtained without being restricted by seasons, so that the method for culturing the somatic cell embryos (short for somatic embryos) of the liquidambar plants into the seedlings is especially suitable for industrialization production of liquidambar nursery stock.

Description

technical field [0001] The invention relates to a method for plant tissue culture, in particular to a method for culturing mature somatic embryos (somatic embryos for short) of Liquidambar genus plants into seedlings through liquid suspension, and belongs to the technical field of tree tissue culture in the forestry industry. Background technique [0002] Liquidambar spp. species are important forestry resources worldwide, especially sweetgum (L.formosana) and North American sweetgum (L.styraciflua), both of which have high economic value, ornamental value and ecological value, and The hybrid sweetgum obtained by crossing the two has heterosis. [0003] There are many studies on the regenerative reproduction methods of the isolated organs of Liquidambar genus. Among many methods of asexual in vitro reproduction, somatic embryo technology has the characteristics of thorough immaturation and high reproductive efficiency, so it has been widely valued. In recent years, Sommer ...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/008A01H4/001
Inventor 孔立生张金凤赵健李珊珊刘次次皮晨齐帅征张炎范英明
Owner BEIJING FORESTRY UNIVERSITY
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