Propagation method for pseudotsuga sinensis dode plant through somatic embryogenesis

A technology of embryogenesis and somatic cells, applied in botany equipment and methods, plant regeneration, gardening methods, etc., can solve problems such as cutting difficulties, short cycle reproduction rate, and unsatisfactory seedling propagation technology

Active Publication Date: 2020-12-08
BEIJING FORESTRY UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The main purpose of the present invention is to develop high-efficiency for the current situation that the natural seed setting rate of Douglas fir is low, cutting is difficult, and seedling propagation technology cannot meet the needs of large-scale afforestation. Plant somatic embryo vegetative reproduction technology provides a method with short cycle, high reproduction rate and low cost for the protection and large-scale breeding of Douglas fir and Douglas fir

Method used

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  • Propagation method for pseudotsuga sinensis dode plant through somatic embryogenesis
  • Propagation method for pseudotsuga sinensis dode plant through somatic embryogenesis
  • Propagation method for pseudotsuga sinensis dode plant through somatic embryogenesis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Embodiment 1 test material and culture medium

[0072] 1. Test materials

[0073] 1. Cone collection

[0074] The immature female cones of Douglas fir were harvested in Sanqingshan, Jiangxi Province in mid-to-late July 2019. Before formal sampling, embryo development observation was carried out in mid-June, and a small amount of cones were collected every week after 40 days of flowering and pollination. When it is between the polyembryonic stage and the cotyledon stage, it can be used as an explant. To ensure that the embryos in the seeds were visibly developed but cotyledons had not yet formed at the time of sampling. The cone mother plant selects 3-5 large trees with healthy growth and free pollination. The collected cones were kept refrigerated at 4°C for later use.

[0075] The embryonic age of Douglas chinensis explants had a significant effect on the induction rate of embryogenic tissue. In the invention, the seed embryo whose growth and development period i...

Embodiment 2

[0105] Embodiment 2 (induction of embryogenic tissue)

[0106] 1. Sterilization of explants

[0107] Wash and clean the surface of Douglas fir cones refrigerated at 4°C, then wash them with tap water, cut open the cones, and take out the seeds; use alcohol with a concentration of 75% by volume on the ultra-clean workbench Soak Douglas fir seeds for 30-60s, then scoop out the seeds and rinse them with sterile water for 3-5 times, then put them in 0.1% HgCl 2 Soak in the solution for 5-10min, take out the seeds and rinse them with sterile water for 5-6 times, put the rinsed seeds on the sterilized filter paper to absorb the water, and use tweezers and scissors to open the seeds in the ultra-clean workbench. skin to obtain the female gametophyte. The female gametophytes were further opened to obtain sterile, immature zygotic embryos from Douglas chinensis for future use.

[0108] 2. Embryogenic tissue induction culture

[0109] 1) Inoculate the whole sterile Douglas fir immat...

Embodiment 3

[0116] Embodiment 3 (proliferation culture of embryogenic tissue)

[0117] The cones of the different genotypes (YDF-A, YDF-B) collected from Douglas chinensis were respectively induced and cultured according to the method in Example 2, and induced and cultured for 4-6 weeks to obtain corresponding embryogenic tissues

[0118] 1. Embryogenic tissue proliferation culture

[0119] 1) Solid proliferation culture of embryogenic tissue

[0120] The induced embryogenic tissue was separated from the explant, and divided into small pieces of 0.5 cm × 0.5 cm, and inoculated on the embryogenic tissue solid proliferation medium (such as figure 1 A), the first stage of solid proliferation culture of Douglas fir embryogenic tissue was carried out under dark conditions, wherein the culture temperature was (25±2)°C, and the 2,4-D used in the solid proliferation medium of embryogenic tissue was (2mg / L), 6-BA (1mg / L), hydrolyzed casein (500mg / L), glutamine (500mg / L), sucrose (10g / L), vegeta...

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Abstract

The invention discloses a propagation method for a pseudotsuga sinensis dode plant through somatic embryogenesis. The propagation method comprises the following steps that embryonic tissue induction culture, multiplication culture, embryonic tissue adjustment culture, somatic embryo (somatic embryo) maturation culture, mature somatic embryo germination culture and seedling transfer culture are carried out on a sterile explant of the pseudotsuga sinensis dode plant. Through the method, the stable pseudotsuga gaussenii flous embryogenic cell strain and a large number of mature somatic embryos are obtained through a somatic embryogenesis induction way for the first time, so that a complete somatic embryo plant is obtained, by using the method, a large number of pseudotsuga gaussenii flous somatic embryo seedlings can be produced in a relatively short period, and the technology is suitable for large-scale and industrialized production of the cloned pseudotsuga gaussenii flous seedlings.

Description

technical field [0001] The invention relates to a method for plant tissue culture, in particular to a method for plant regeneration of Douglas chinensis by culturing somatic embryos (somatic embryos), and belongs to the technical field of cell engineering seedling breeding in forestry. Background technique [0002] Pseudotsuga gaussenii Flous is an evergreen tree belonging to the Pinaceae family. There are 7 species in the Douglas genus, 5 of which are endemic to China, and the other 2 are originally distributed in North America. Douglas pine (Pseudotsuga menziesii (Mirb.) Franco), native to North America, is a world-renowned excellent forest tree species. East China fir up to 40 meters high, bark dark gray, leaf pillow top brown, main branches glabrous or sparsely hairy, side branches with brown dense hairs, winter buds ovoid or ovate-conical, brown. Leaf blades are strip-shaped, arranged in two rows or nearly radiating on the main branch, dark green above, with two white...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/008A01H4/001Y02A40/22Y02P60/40
Inventor 孔立生张金凤赵健高英崔莹李珊珊范英明
Owner BEIJING FORESTRY UNIVERSITY
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