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A method for restoring and maintaining embryogenicity of embryogenic callus of larix changbai

A technology of embryogenic callus and long white larch, applied in horticultural methods, botanical equipment and methods, plant regeneration, etc.

Active Publication Date: 2022-03-15
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the embryogenic callus of each type of larix remains embryogenic for a period of time under its suitable conditions, the method of gradually reducing the concentration of NAA combined with high concentration of cytokinins can restore and maintain the embryogenic callus of the embryogenic callus Haven't seen any reports yet

Method used

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  • A method for restoring and maintaining embryogenicity of embryogenic callus of larix changbai
  • A method for restoring and maintaining embryogenicity of embryogenic callus of larix changbai
  • A method for restoring and maintaining embryogenicity of embryogenic callus of larix changbai

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example 1

[0018] Example 1 Restoration and maintenance of embryogenicity during subculture of embryogenic callus

[0019] The subcultured embryogenic callus was inoculated at NAA concentrations ranging from 0.5, 0.4, 0.3, 0.2, 0.1 mg·L -1 Embryogenic recovery culture was carried out in successively lower S medium. Each gradient was subcultured twice and transferred to differentiation medium to induce somatic embryogenesis, and the somatic embryogenesis was counted after 6 weeks.

[0020] Table 2 Effect of embryogenic callus embryogenic recovery treatment on somatic embryogenesis

[0021]

[0022] Note: Duncan's multiple comparisons were used. Different letters indicate significant difference at 0.05 level.

[0023] As shown in Table 2: the somatic embryogenesis rate of the control group was 45.3%, and the number of somatic embryos was 16.5 g -1 . In experimental groups 1, 2, 3, 4, and 5, compared with the control group, substituting NAA for 2,4-D and increasing the concentration...

example 2

[0024] Example 2 Somatic Embryogenesis and Germination

[0025] The embryogenic callus was pre-cultured for 10 days and then inoculated into the differentiation medium, and the somatic embryogenesis rate could reach 100% after 4 weeks. The germination rate of somatic embryos with good growth potential can reach 81.3% after being transferred to the germination medium for 2 weeks. 5 After 2 weeks in the culture medium, the seedling rate can reach 46.2%.

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Abstract

The purpose of the present invention is to restore and maintain the embryogenicity of the embryogenic callus of larch larch by adjusting the hormone concentration in the subculture medium of the embryogenic callus. Embryogenic callus in S+0.1mg·L ‑1 2,4‑D+0.04mg L ‑1 BA+0.02mg·L ‑1 Under the condition of KT hormone, it was found that the somatic embryo differentiation rate of embryogenic callus decreased sharply after 6 months of subculture. Recover embryogenicity by removing 2,4‑D and gradually reducing NAA: inoculate embryogenic callus at NAA concentrations of 0.5, 0.4, 0.3, 0.2, 0.1mg L every 2 weeks ‑1 Carry out embryogenic recovery culture in successively lower S medium, wherein the hormone in S medium also contains 0.5mg·L ‑1 BA, 0.5mg·L ‑1 KT. Finally at S+0.2mg·L ‑1 NAA+0.5mg·L ‑1 BA+0.5mg·L ‑ 1 KT+0.5g·L ‑1 Glutamine+0.5g·L ‑1 Hydrolyzed casein+30g·L ‑1 Sucrose+3g·L ‑1 Under the condition of plant gel culture, the embryogenicity remained good after continuous subculture for 2 years. The somatic embryogenesis rate can reach 100% after the embryogenic callus is differentiated and cultured. The somatic embryos with normal development are transferred to the germination medium, and the seedling rate can reach 46.2% after 4 weeks.

Description

technical field [0001] The invention relates to a method for restoring and maintaining embryogenicity of larch embryogenic callus. The invention belongs to the technical field of forestry biology. Background technique [0002] Changbai larch (Larix olgensis), also known as yellow-flowered larch and Korean larch, belongs to Pinaceae and larch is a tall tree. Its wood is heavy, solid and durable. It is an irreplaceable tree species for short-term natural forest construction in Northeast my country. The embryogenic callus induced by larix immature zygotic embryos has the problem of declining or even disappearing embryogenicity in the continuous subculture process, which seriously restricts the research on the somatic embryogenesis pathway of larix and the genetic transformation of forest trees. [0003] In 1989, Kimaszewska et al. used immature zygotic embryos of Japanese × European hybrid larch (Larix × eurolepis) as materials to induce embryogenic callus. During the continuo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/005A01H4/001
Inventor 由香玲刘建飞
Owner NORTHEAST FORESTRY UNIVERSITY