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
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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 s

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
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Example Embodiment

[0037] Example 1

[0038] Isolate immature zygotic embryos: collect free-pollinated cones in spring and free-pollinated clones of western Sichuan spruce at the end of August, and store the cones at 4°C for 7 days at low temperature. Treat the cones with 95% ethanol for 15 minutes and rinse with sterile water 3 times. Under aseptic conditions, remove the seeds and peel off the seed coat, and remove the immature zygotic embryos (such figure 1 Shown) The next stage of embryogenic callus induction culture.

Example Embodiment

[0039] Example 2

[0040] Embryogenic callus induction stage: the immature zygotic embryos obtained in Example 1 were subjected to embryogenic callus induction culture. Using 1 / 2LM minimal medium, the additional hormone concentrations were 2,4-D 2.2mg / L, 6-BA1.1mg / L and KT0.22mg / L. In addition, 1g / L of natural complex enzymatically hydrolyzed casein, 1% of sucrose, 500mg / L of glutamine (filter sterilization) and 0.2% of plant gel (Gelrite, Sigma) were added to the medium. Before sterilization, adjust pH to 5.7, 121°C, pressure 101kp, high temperature, autoclave for 20 minutes, culture temperature 24±1°C, culture in dark condition, until embryogenic callus is induced. Induction rate: 61.55%.

Example Embodiment

[0041] Example 3

[0042] Embryogenic callus induction stage: the immature zygotic embryos obtained in Example 1 were subjected to embryogenic callus induction culture. Use 1 / 2LM minimal medium with additional hormone concentrations of 2,4-D 0.55mg / L and 6-BA 0.8mg / L, and add 1g / L of natural complex enzymolysis casein to the medium. 1% sucrose, 500mg / L glutamine (filter sterilization) and 0.2% vegetable gel (Gelrite, Sigma). Before sterilization, adjust pH to 5.7, 121°C, pressure 101kp, high temperature, autoclave for 20 minutes, culture temperature 24±1°C, culture in dark condition, until embryogenic callus is induced. The induction rate is 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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