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Method for Inducing Mature Zygotic Embryos of Pinus sylvestris

A technology of Pinus sylvestris and zygotic embryos, which is applied in the field of induction of somatic embryogenesis of mature embryos of Pinus sylvestris, can solve the problems of seasonal time constraints and failure to achieve success, and achieve the effects of convenient material collection, long storage period, and simple operation

Inactive Publication Date: 2016-08-24
QIQIHAR UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to solve the problem that the use of immature zygotic embryos to induce somatic embryogenesis is limited by season time, and the mature zygotic embryos of Pinus sylvestris are cultivated to obtain somatic embryos in the existing methods. Method for somatic embryogenesis of mature zygotic embryos

Method used

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  • Method for Inducing Mature Zygotic Embryos of Pinus sylvestris
  • Method for Inducing Mature Zygotic Embryos of Pinus sylvestris
  • Method for Inducing Mature Zygotic Embryos of Pinus sylvestris

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specific Embodiment approach 1

[0018] Specific embodiment 1: The method for inducing somatic embryogenesis of mature zygotic embryos of Pinus sylvestris in this embodiment includes the following steps:

[0019] 1. Explant sterilization and inoculation: Take the mature Pinus sylvestris seeds harvested and stored in cold storage in the same year, rinse them with running water, and then soak them in warm water at 40~50℃ for 2h, and then disinfect the surface with alcohol with a mass percentage of 70% for 0.5~1min. , Rinse with sterile water 3 to 5 times, sterilize in a 0.1% by mass mercury solution for 5 minutes, rinse with sterile water 5 to 6 times, peel off the endosperm under ultra-clean conditions, and take out the mature embryo Inoculate into the culture medium;

[0020] 2. Embryogenic callus induction: The zygotic embryos treated in step 1 are induced with DCR medium for embryogenic callus induction. The DCR medium contains 2,4-D at a concentration of 2 mg / L. 1mg / L 6-BA, 500mg / L acid hydrolyzed casein, 3% b...

specific Embodiment approach 2

[0023] Specific embodiment two: This embodiment is different from specific embodiment one in that the mature seeds of Pinus sylvestris var. mongolica in the step one are healthy, full and uniform mature Pinus sylvestris seeds that were harvested in September of that year and stored at 4°C for 1 year. Others are the same as the first embodiment.

specific Embodiment approach 3

[0024] Specific embodiment three: This embodiment is different from specific embodiment one or two in that in the step one, it is rinsed with running water, then soaked in warm water at 45~50℃ for 2h, and the surface is disinfected with 70% alcohol for 0.8~1min. , Rinse with sterile water 3 to 5 times, and sterilize in 0.1% mercury solution for 5 minutes. Others are the same as the first or second embodiment.

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Abstract

The invention discloses a method for inducing mature zygotic embryo somatic cell embryos of Pinus sylvestris sylvestris, which relates to a method for inducing somatic cell embryos of mature zygotic embryos of Pinus sylvestris. The invention aims to solve the problem that the induction of somatic embryogenesis using immature zygotic embryos is limited by season time. The method of the invention includes the following steps: 1. disinfection and inoculation of explants; 2. induction of embryogenic callus; 3. proliferation and maintenance of embryogenic callus; 4. maturation induction of somatic embryos. The explant of the present invention is convenient to obtain materials, the pollution rate is controlled within 5%, the induction rate of the embryogenic callus of Pinus sylvestris is high, and the callus proliferation culture can proliferate 3-4 times, and in the culture medium added with abscisic acid Somatic embryos of Pinus sylvestris were successfully induced. Compared with the mature zygotic embryo explant used in the present invention and the successful immature zygotic embryo of Pinus sylvestris as an explant to induce somatic embryo technology, the somatic embryogenesis technology of mature zygotic embryo has long storage period, abundant materials and no It is limited by factors such as season and plant development period.

Description

Technical field [0001] The invention relates to the field of forest tree seedling propagation, in particular to a method for inducing somatic embryogenesis of mature embryos of Pinus sylvestris. Background technique [0002] Pinus sylvestris Var Mongolica (Pinus sylvestris Var Mongolica) is a tall evergreen tree species of the Pinus genus. It is a geographical variant of European Scots pine. It has the characteristics of developed root system, cold tolerance, drought tolerance, strong soil adaptability, fast growth, and excellent texture. The main fast-growing needles in Northeast China, excellent tree species for protection and greening, and soil and water conservation. [0003] Somatic embryogenesis is an effective method of large-scale asexual reproduction. Somatic embryogenesis can be used to study the mechanism of plant growth and development and organ differentiation, and the law of genetic variation. It can also be used for the large-scale production of artificial seeds thro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 梁艳沈海龙高美玲徐洪国范震宇朱琨陈阳金一锋娄丛艳
Owner QIQIHAR UNIVERSITY
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