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Method for recovering and maintaining embryogenic property of larix olgensis embryogenic callus

A technology of embryogenic callus and long white larch, applied in horticultural methods, botany equipment and methods, horticulture, etc.

Active Publication Date: 2021-07-16
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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  • Method for recovering and maintaining embryogenic property of larix olgensis embryogenic callus
  • Method for recovering and maintaining embryogenic property of larix olgensis embryogenic callus
  • Method for recovering and maintaining embryogenic property of larix olgensis embryogenic callus

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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 invention aims to provide a method for recovering and maintaining the embryogenic property of larix olgensis embryogenic callus by adjusting the hormone concentration in an embryogenic callus subculture medium. The somatic embryo differentiation rate of the embryogenic callus is suddenly reduced after the embryogenic callus is subcultured for 6 months under the conditions of S + 0.1 mg.L<-1>2, 4-D + 0.04 mg.L<-1> BA + 0.02 mg.L<-1> KT hormone. The embryogenic property is recovered by a method of removing 2, 4-D and gradually reducing NAA: inoculating the embryogenic callus into an S culture medium of which the NAA concentration is sequentially reduced from 0.5 mg.L<-1>, 0.4 mg.L<-1>, 0.3 mg.L<-1>, 0.2 mg.L<-1> and 0.1 mg.L<-1> every two weeks for embryogenic property recovery culture, wherein hormones in the S culture medium further comprise 0.5 mg.L<-1> BA and 0.5 mg.L<-1>KT. Finally, subculture is carried out continuously for 2 years under the culture conditions of S + 0.2 mg.L<-1> NAA+ 0.5 mg.L<-1>BA +0.5 mg.L<-1> KT+ 0.5 g.L<-1> glutamine, 0.5 g.L<-1> casein hydrolysate+ 30 g.L<-1>sucrose+3 g.L<-1> plant gel, and the embryogenic property is still kept good. After the callus with recovered embryogenicproperty is subjected to differentiation culture, the somatic embryo occurrence rate can reach 100%. The somatic embryos which grow normally are transferred into a germination culture 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] Long white larch (Larix olgensis) alias yellow flower larch, Korea larch , belonging to Pinaceae, larch is a tall tree, its wood is heavy, solid and durable, and it is an irreplaceable tree species for building short-cycle natural forests 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 continuous subcult...

Claims

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Application Information

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