Sterile and rapid propagation method for pinus thunbergii

A fast and black pine technology, which is applied in the field of plant tissue culture research to achieve the effect of accelerating the speed of seedling cultivation

Inactive Publication Date: 2017-09-15
SHANDONG UNIV
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AI Technical Summary

Problems solved by technology

At present, the successful induction of regenerated seedlings of black pine all uses zygotic embryos as explants, directly induces the production of adventitious buds from zygotic embryos or induces embryogenic callus to form somatic embryos, and then develops to form sterile seedlings. Technical report on callus proliferation and stem-bud differentiation to form regenerated shoots

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  • Sterile and rapid propagation method for pinus thunbergii

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Embodiment Construction

[0011] A kind of aseptic rapid propagation method of black pine is carried out through the following steps:

[0012] 1. Selection of explants

[0013] Collect young young pine branches from the top of black pine trees less than 7 years old in spring, pull out the pine needles on them, rinse them with running water, and carry out disinfection treatment in an ultra-clean workbench, disinfect with 75% alcohol for 30 seconds, wash with sterile water for 2- 3 times, and then sterilized with 20% hydrogen peroxide for 17-18 minutes, shaking from time to time, washing with sterile water 2-3 times, to obtain sterile pine needles;

[0014] 2. Induction culture of callus

[0015] Cut off 1 cm long fragments from the base of sterile pine needles under sterile conditions, inoculate them in induction medium, and cultivate them at 25°C in the dark. The hormone dosage of induction medium was screened by two-factor design method: MS medium was used as the basic medium, pH=6.4, and 30 g / L suc...

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Abstract

The invention relates to a sterile and rapid propagation method for pinus thunbergii. The method comprises the following steps: (1) selecting a fragment of a needle base part of the pinus thunbergii as an explant; (2) inoculating the fragment onto an MS culture medium + 2 mg/L of 6-BA + 0.05 mg/L of NAA to generate a callus through inducing; (3) inoculating the callus onto 1/2 of the MS culture medium + 0.05 mg/L of NAA + 0.001 mg/L of PSK to realize mass propagation; (4) inoculating the callus onto 1/4 of the MS culture medium + 0.01 mg/L of NAA + 0.002 mg/L of PSK to induce differentiation of adventitious buds; (5) cutting off a single bud, inoculating the single bud onto 1/2 of the MS culture medium + 0.25 mg/L of NAA + 0.25 mg/L of IBA to induce rooting; and (6) hardening a rooted small green seedling, then transplanting the seedling into a matrix prepared from 1/4 of fine sandy soil, 1/4 of vermiculite and 1/2 of humus for further growth. According to the sterile and rapid propagation method for the pinus thunbergii, and the callus can be massively propagated within short time, and the callus is differentiated into regenerated seedlings, so that conservation and mass propagation of high-quality pinus thunbergii individuals can be realized.

Description

technical field [0001] The invention belongs to the field of plant tissue culture research, and the invention relates to a method for rapid aseptic propagation of black pine. Background technique [0002] Black pine (Pinus thunbergii) originated in Japan and was introduced to China in the 1920s. Because of its characteristics of drought resistance, resistance to sea wind and sea monsters, resistance to pine scale and pine caterpillar and other diseases and insect pests, and beautiful tree shape, it has become a coastal defense in the east of my country. It is an irreplaceable dominant species in forest construction, and it is cultivated from Dalian in the north, Lvshun in the south to the coastal areas of Taiwan and some inland areas. When it was first introduced, black pine had the characteristics of fast growth and strong resistance to diseases and insect pests. However, survey data in recent years have shown that black pine is superior in terms of vitality, growth speed, s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/008
Inventor 郑凤英刘雪芹梁中贵
Owner SHANDONG UNIV
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