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Liquid suspension culture method capable of promoting somatic embryo maturation of Liquidambar spp. plants

A technology of suspension culture and Liquidambar genus is applied in the field of plant tissue culture and liquid suspension culture where Liquidambar genus embryos are mature, and can solve the problem of high production cost, low efficiency of Liquidambar genus somatic embryos, and Liquidambar genus somatic embryos. Can not mature normally and other problems, to achieve the effect of reducing production costs and high production efficiency

Active Publication Date: 2020-03-24
BEIJING FORESTRY UNIVERSITY
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to exist in the rapid propagation method of tissue culture of liquidambar tree species that the somatic embryos of liquidambar can not be matured normally in the liquid culture medium, so that the advantages of liquid culture cannot be reflected in the maturation stage of the somatic embryos. There are technical defects in the production of liquidambar somatic embryo seedlings with low efficiency and high production cost. The invention provides a liquid suspension culture method for promoting the maturation of liquidambar somatic embryos (somatic embryos). The inventive method can promote The somatic embryos of Liquidambar are normally matured in the liquid culture medium, and are converted into somatic embryo seedlings in the subsequent culture stage. The method of the present invention can efficiently produce mature somatic embryos, the production volume of mature somatic embryos is large, and the production cost can be reduced. Screen mature somatic embryos; manage somatic embryo development synchronously; improve the consistency of subsequent somatic embryo germination

Method used

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  • Liquid suspension culture method capable of promoting somatic embryo maturation of Liquidambar spp. plants
  • Liquid suspension culture method capable of promoting somatic embryo maturation of Liquidambar spp. plants
  • Liquid suspension culture method capable of promoting somatic embryo maturation of Liquidambar spp. plants

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Effect test

Embodiment 1

[0071] 1. Test method

[0072] The embodiments of the present invention refer to the method 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.

[0073] 2. Test materials

[0074] 1. Immature seeds of hybrid sweetgum

[0075] 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.

[0076] 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.

[0077] 2. Plant growth regulator

[0078] The plant growth regulator used in the present invention adopts 6-benzylaminoadenine (6-BA) and 2,4-dichlorophenoxyacetic acid (2,4-D).

[0079] 3. Medium

[...

Embodiment 2

[0099] 1. Sterilization of explants

[0100] The explants were sterilized according to the method of 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).

[0101] 2. Embryogenic callus induction culture

[0102] Embryogenic callus induction culture was carried out according to the method of 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). figure 1 Embryogenic callus induced from immature hybrid sweetgum seed embryos.

[0103] After the immature zygotic embryos of the explants were cultured on the induction medium for 3-4 weeks, the growth rate of the embryogenic tissues began to slow down and the explants began to brown. At this time, the embryogenic tissues were selected for subculture and transferred to Repeated subcu...

Embodiment 2A

[0131] In addition to step 3, during the embryogenic callus proliferation and culture process, the typical embryogenic callus of the subcultured hybrid sweetgum is inoculated in the embryogenic callus proliferation liquid medium for suspension culture, and the inoculation amount is every 100ml 2-4g in the embryogenic callus proliferation liquid medium; carry out the liquid proliferation culture of the hybrid sweetgum embryogenic callus under dark conditions, wherein, the culture temperature is (25±2)°C, 100-120 revolutions per minute , the 2,4-D used in the embryogenic callus proliferation liquid medium was 0.5mg / L, the 6-BA was 0.25mg / L, the enzymatically hydrolyzed casein was 1g / L, and the sucrose was 40g / L; In the process of sexual callus proliferation and culture, it was subcultured every 3-4 weeks, that is, a new culture was started every 3-4 weeks, that is, 90% fresh medium was added, and the inoculum was kept at 2-4%. The amount of cycle multiplication is 1.5-4 times of...

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Abstract

The invention discloses a liquid suspension culture method capable of promoting somatic embryo maturation of Liquidambar spp. plants, and belongs to the technical field of tree tissue culture. The liquid suspension culture method capable of promoting somatic embryo maturation of the Liquidambar spp. plants comprises the following steps: performing somatic embryogenesis culture on embryogenic calluses of a Liquidambar spp. plant; and then, inoculating a liquid somatic embryo maturation medium with the cultured embryogenic calluses so as to carry out liquid suspension maturation culture of somatic embryos. Being adopted, the liquid suspension culture method capable of promoting somatic embryo maturation of the Liquidambar spp. plants disclosed by the invention is capable of promoting normalmaturation of somatic embryos of Liquidambar spp. plants in liquid media so as to produce a large number of mature somatic embryos of Liquidambar spp. plants in a short period of time; and high seedling efficiency can be achieved after germination of the mature somatic embryos. The liquid suspension culture method capable of promoting somatic embryo maturation of the Liquidambar spp. plants disclosed by the invention is simple, short in regeneration cycle, high in reproductive coefficient, high in survival rate of transplanting, and capable of ensuring neat and consistent seedling emergence soas to facilitate maintenance and management, thereby saving manpower and land resources so as to have low production cost achieved; and moreover, regenerated plants can be continuously obtained without season limitation. Therefore, the liquid suspension culture method capable of promoting somatic embryo maturation of the Liquidambar spp. plants is especially suitable for industrial production ofLiquidambar spp. plants seedlings.

Description

technical field [0001] The invention relates to a method for plant tissue culture, in particular to a liquid suspension culture method for maturing sweetgum body embryos, and belongs to the technical field of cell engineering breeding in the forestry industry. Background technique [0002] Liquidambar spp. species are important forestry resources worldwide, especially Liquidambar formosana and North American sweetgum (L.styraciflua), both of which have high economic value, ornamental value and ecological value, and both The hybrid sweetgum obtained by hybridization of the former has heterosis. [0003] Liquidambar can be reproduced both sexually and asexually. In recent years, with the continuous progress of cutting technology and tissue culture rapid propagation technology, the efficiency of asexual reproduction of liquidambar sweetgum has also been greatly improved. [0004] Tissue culture is an efficient forest tree rapid propagation technology, which is widely used in m...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005A01H4/008
Inventor 孔立生张金凤赵健齐帅征
Owner BEIJING FORESTRY UNIVERSITY
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