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Method for picea balfouriana somatic embryo generation and plant regeneration

A technology of embryogenesis and somatic embryos, which is applied in the field of forestry cell engineering seedling propagation, can solve the problems of low seed maturity rate, slow growth, affecting the genetic improvement process, etc., and achieve the effect of increasing the induction rate

Inactive Publication Date: 2012-11-14
INST OF FORESTRY CHINESE ACAD OF FORESTRY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The growth rate of natural spruce in Western Sichuan is slow, and the seed maturity rate is very low, which causes great difficulties in reproduction and variety selection, and the growth of seedlings and young forests is slow, which seriously affects the process of genetic improvement, especially the process of genetic evaluation.

Method used

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  • Method for picea balfouriana somatic embryo generation and plant regeneration
  • Method for picea balfouriana somatic embryo generation and plant regeneration
  • Method for picea balfouriana somatic embryo generation and plant regeneration

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Isolation of immature zygotic embryos: collecting free-pollinated cones of Picea spruce in western Sichuan at the end of August, which were free-pollinated in spring, and refrigerated at 4°C for 7 days. The cones were treated with 95% ethanol for 15 minutes, and washed 3 times with sterile water. Under sterile conditions, remove the seeds and peel off the seed coat, and immature zygotic embryos (such as figure 1 shown) for the next stage of embryogenic callus induction culture.

Embodiment 2

[0040] Embryogenic callus induction stage: The immature zygotic embryos obtained in Example 1 were cultured for embryogenic callus induction. Using 1 / 2LM basic medium, the additional hormone concentrations were 2.2mg / L for 2,4-D, 1.1mg / L for 6-BA and 0.22mg / L for KT. Also, 1 g / L natural complex enzymatic casein, 1% sucrose, 500 mg / L glutamine (filter sterilized) and 0.2% plant gel (Gelrite, Sigma) were added to the medium. Before sterilization, adjust the pH to 5.7, sterilize at 121°C, pressure 101kp, high temperature, high pressure for 20 minutes, culture temperature 24±1°C, and culture in dark conditions until embryogenic callus is induced. Induction rate: 61.55%.

Embodiment 3

[0042] Embryogenic callus induction stage: The immature zygotic embryos obtained in Example 1 were cultured for embryogenic callus induction. Using 1 / 2LM basic medium, the concentration of additional hormones is 2,4-D 0.55mg / L, 6-BA 0.8mg / L, and adding 1g / L natural complex enzymatic casein to the medium, 1% sucrose, 500mg / L glutamine (filter sterilized) and 0.2% vegetable gel (Gelrite, Sigma). Before sterilization, adjust the pH to 5.7, sterilize at 121°C, pressure 101kp, high temperature, high pressure for 20 minutes, culture temperature 24±1°C, and culture in dark conditions until embryogenic callus is induced. The induction rate was 60.56%.

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Abstract

The invention relates to a method for picea balfouriana somatic embryo generation and plant regeneration, which comprises steps of collecting picea balfouriana open pollination clonal cones, and separating immaturate zygotic embryos out for somatic embryogenetic culture, wherein the somatic embryogenetic culture comprises four stages, i.e. an embryogenetic callus inducing stage, an embryogenetic callus multiplicating stage, a somatic embryo maturity stage and a somatic embryo germination stage. Compared with the prior art, the method has the advantages that the seedling stage is greatly shortened, and natural maturing rate, seed maturation rate, differentiation rate and germination rate are all improved. The seedling culture method having short period, high multiplying rate, closely linked cultivation steps and low cost is provided for popularizing and planting picea balfouriana in a large scale for forestry production, is suitable for a rapid propagation technical system of picea balfouriana cell engineering and satisfies the requirements of markets on picea balfouriana.

Description

technical field [0001] The invention relates to the field of forestry cell engineering seedling propagation, in particular to a method for somatic embryogenesis and plant regeneration of western Sichuan spruce. Background technique [0002] There are about 40 species of Picea dietr plants in the world, mainly distributed in the cold temperate zone or tundra zone of 50°~60° north latitude, and a few are distributed in the temperate or subtropical wet and cold high mountains, that is, most species are circumpolar. Distribution, and in my country and the United States can be distributed to around 30° north latitude, it is an important construction tree species in low-calorie areas. [0003] The spruce tree species has high annual growth per unit area, good wood quality and wide application. It has become an important industrial timber tree species in Western Europe, Northern Europe, Baltic countries, Russia and Canada. There are 17 species and 9 varieties of Picea in my country...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 王军辉
Owner INST OF FORESTRY CHINESE ACAD OF FORESTRY
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