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Method for somatic embryogenesis and plant regeneration of Lijiang spruce

A technology of embryogenesis and somatic embryos, applied in plant regeneration, botany equipment and methods, horticultural methods, etc., can solve the problems of low differentiation rate, high concentration of growth regulators, and high cost, so as to improve differentiation rate and reduce cost Effect

Inactive Publication Date: 2012-11-28
INST OF FORESTRY CHINESE ACAD OF FORESTRY
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0004] Regarding the methods of somatic embryogenesis and plant regeneration of Picea spruce Lijiang, there are currently published documents: Somatic embryogenesis of Picea spruce Lijiang (Chen Fang et al., "Forestry Science", Vol. 46, No. 8, pp. 162-167, 2010 In August 2010), the literature selected mature seeds of Picea spruce from Lijiang for embryogenesis. The basic medium used was improved P6, 1 / 2MS and LP medium, and the concentration of growth regulators (hormones) was high, which was not conducive to maintaining embryogenicity. The embryogenic ability of callus, and the risk of losing embryogenic ability is high, the differentiation rate is low, and the cost is also high

Method used

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  • Method for somatic embryogenesis and plant regeneration of Lijiang spruce
  • Method for somatic embryogenesis and plant regeneration of Lijiang spruce
  • Method for somatic embryogenesis and plant regeneration of Lijiang spruce

Examples

Experimental program
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Embodiment 1

[0038] Isolation of mature zygotic embryos: collect the clonal cones obtained by free pollination in spring and at the end of August, and refrigerate the harvested cones 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 mature zygotic embryos (such as figure 1 shown) for the next stage of embryogenic callus induction.

Embodiment 2

[0040] Embryogenic callus induction stage: The mature zygotic embryos obtained in Example 1 were subjected to embryogenic callus induction, using 1 / 2LM basic medium, and the additional hormone concentrations were 2, 4-D2.2mg / L and 6- BA0.55mg / L, add 1g / L natural complex enzymolysis casein, contain 1% sucrose, 500mg / L glutamine (sterilized by filtration) and 0.2% plant gel (Gelrite, Sigma). Before sterilization, adjust the pH to 5.8, 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.

[0041] About 2 to 3 days after the mature zygotic embryo is inserted into the above-mentioned medium, it starts to start, and the grown callus is light yellow and translucent. About 25 days, some of them formed green tumor-like hard callus, which browned after subculture (the subculture method is completely consistent with the conditions of this stage), while some grew...

Embodiment 3

[0043] Embryogenic callus induction stage: The mature zygotic embryos obtained in Example 1 were subjected to embryogenic callus induction, using 1 / 2LM basic medium, and the additional hormone concentrations were 2, 4-D2.2mg / L and 6- BA0.8mg / L, add 1g / L natural complex enzymolysis casein, medium contains 1% sucrose, 500mg / L glutamine (sterilized by filtration) and 0.2% plant gel (Gelrite, Sigma). Before sterilization, adjust the pH to 5.8, 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.

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Abstract

The invention relates to a method for somatic embryogenesis and plant regeneration of Lijiang spruce, comprising the following steps that: open-pollinated clonal cones of Lijiang spruce are collected and mature zygotic embryo is separated out for somatic embryogenesis cultivation; the somatic embryogenesis cultivation is divided into the stages of embryonal callus induction, embryonal callus proliferation, somatic embryo maturity, and somatic embryo germination. Compared with the prior art, the method is beneficial for maintaining the embryonal capacity of embryonal callus and the risk of losing embryonal capacity is reduced and the differentiation rate and the germination rate are both increased. A method for large-scale vegetative propagation seedling raising of Lijiang spruce with short period, high reproductive rate and low cost is provided.

Description

technical field [0001] The invention relates to the field of cell engineering seedling propagation in forestry, in particular to a method for somatic cell embryogenesis and plant regeneration of spruce spruce from Lijiang. Background technique [0002] Lijiang spruce (Picea likiangensis) Pinaceae, Yunshan genus. Evergreen tree, up to 40 meters high. Branchlets yellowish gray, densely hairy. Leaf blades are row, quadrangular or slightly flattened, with stomatal lines on all four sides, usually 0.8-0.5 cm. The cones are cylindrical or ovate-cylindrical, 4-9 meters long, purple-red or purple-black before maturity. Seed scales are thin, orthorhombic, with wavy edentulous margins. Produced in the southwest of Sichuan, China, and the high mountains of northwestern Yunnan, forming a large area of ​​simple forest. The wood is light, soft and durable, and can be used for construction, sleepers, appliances, papermaking and other materials. It is also an ornamental tree. The woo...

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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