Light green semi-dense callus induction and adventitious bud regeneration method for Fraxinus mandshurica Rupr.

Abstract

The invention provides a method for realizing adventitious bud regeneration and propagation of the hypocotyl and the stem segment of Fraxinus mandshurica Rupr. through a callus approach by a key link,i.e., light green semi-dense callus induction. When a mature embryo is cultured to cotyledon expansion (21 d), the hypocotyl is cut off and is inoculated in an a20 culture medium; the stem segment iscut from an aseptic seedling and is inoculated in a c12 culture medium, through dark / light alternant culture, a light green semi-dense callus is obtained, wherein the inductivity of the hypocotyl andthe inductivity of the stem segment are independently 88.69% and 99.15%; and an adventitious bud is formed on a callus block semi-dense gap, wherein the inductivities are independently 10.52% and 33.33%. According to the method disclosed by the invention, a bottleneck link that the Fraxinus mandshurica Rupr. difficultly realizes adventitious bud differentiation through the callus approach is eliminated, and through efficient callus induction and effective adventitious bud regeneration, a solid foundation is laid for the rapid propagation technology of the Fraxinus mandshurica Rupr..

Description

Technical field: [0001] The invention belongs to the field of genetic breeding of forest trees, the field of plant tissue culture, and high-efficiency propagation technology of Mandshurica mandshurica Background technique: [0002] Mandshurica mandshurica seeds have the characteristics of long growth interval, deep dormancy, and slow reproduction speed. It takes about 20 years for excellent clones and hybrid dominant species to enter the mature stage. Cuttage grafting and vegetative propagation technology is limited by factors such as seasons, and it is impossible to produce excellent seeds. Therefore, it is particularly important to establish an efficient breeding technology. In 2001, Zhang Huijun et al. conducted a preliminary exploration on the induced emergence of explants such as Mandshurica mandshurica buds, stem tips, and seeds; in 2010, Shen Hailong et al. tried to establish a reproductive system based on hypocotyls, but the reproductive coefficient was low and unsta...

AI Technical Summary

Problems solved by technology

[0002] Mandshurica mandshurica seeds have the characteristics of long growth interval, deep dormancy, and slow reproduction speed. It takes about 20 years for excellent clones and hybrid dominant species to enter the mature stage. Cuttage grafting and vegetative propagation technology is limited by factors such as seasons, and it is impossible to produce excellent seeds. Therefore, it is particularly important to establish an efficient breeding technology

In 2001, Zhang Huijun et al. conducted a preliminary exploration on the induced emergence of explants such as Mandshurica mandshurica buds, stem tips, and seeds; in 2010, Shen Hailong et al. tried to establish a reproductive system based on hypocotyls, but the reproductive coefficient was low and unstable; in 2013 Ding Xiaomeng et al. used the petiole of Fraxinus ash as explants to induce the loose callus of Fraxinus ash; in 2018, Liang Xue et al. established the suspension culture system of Fraxinus ash; However, the research on adventitious bud induction differentiation technology of callus is not yet mature

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