Method for performing high frequent induction regeneration of populus euphratica leaves for plants

A technology for high-frequency induction and regeneration of plants, applied in botany equipment and methods, plant regeneration, horticultural methods, etc., can solve the problems of leaf regeneration and transformation system genetic transformation cannot be implemented, leaf regeneration system is difficult to achieve, test-tube seedling yellowing, etc. problems, to achieve the effect of solving the difficulties of tissue culture and regeneration, overcoming yellowing and vitrification, and efficient regeneration

Inactive Publication Date: 2018-08-28
LUDONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Genetic transformation is an important means to study the molecular mechanism of Populus euphratica tolerance to drought and high-salt stress and to improve it, and the biggest bottleneck and obstacle encountered by the most effective Agrobacterium-mediated leaf disk transformation method for plant transgenesis is The establishment of the leaf regeneration system, many plant materials cannot be implement

Method used

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  • Method for performing high frequent induction regeneration of populus euphratica leaves for plants
  • Method for performing high frequent induction regeneration of populus euphratica leaves for plants

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Embodiment 1

[0034] A kind of method of Populus euphratica leaf high-frequency induced regeneration plant of the present embodiment, comprises the following steps:

[0035] 1), the acquisition of explants

[0036] Take the stem segment of Populus euphratica with axillary buds as explants, soak with 0.05% by mass detergent for 10 minutes, and rinse with tap water for 20 to 30 minutes. Transfer to the ultra-clean workbench, rinse with sterile water for 3 times, soak in 75% ethanol for 30 seconds, rinse with sterile water for 3 times, disinfect with 0.1% mercury liter for 5 minutes, wash with sterile water for 3 times, and then use sterile Bacteria absorbent paper to absorb surface moisture, cut into 0.8-1.2cm stem segments with axillary buds;

[0037] 2), acquisition and subculture of sterile test-tube seedlings

[0038] Inoculate the above-mentioned stem segments on the subculture medium to obtain sterile test-tube plantlets, and carry out subculture proliferation of adventitious buds. Th...

Embodiment 2

[0047] A kind of method of Populus euphratica leaf high-frequency induced regeneration plant of the present embodiment, comprises the following steps:

[0048] 1), the acquisition of explants

[0049] Take the stem segment of Populus euphratica with axillary buds as explants, soak with 0.05% by mass detergent for 10 minutes, and rinse with tap water for 20 to 30 minutes. Transfer to the ultra-clean workbench, rinse with sterile water for 3 times, soak in 75% ethanol for 30 seconds, rinse with sterile water for 3 times, disinfect with 0.1% mercury liter for 5 minutes, wash with sterile water for 3 times, and then use sterile Bacteria absorbent paper to absorb surface moisture, cut into 0.8-1.2cm stem segments with axillary buds;

[0050] 2), acquisition and subculture of sterile test-tube seedlings

[0051] Inoculate the above-mentioned stem segments on the subculture medium to obtain sterile test-tube plantlets, and carry out subculture proliferation of adventitious buds. Th...

Embodiment 3

[0069] A kind of method of Populus euphratica leaf high-frequency induced regeneration plant of the present embodiment, comprises the following steps:

[0070] 1), the acquisition of explants

[0071] Take the stem segment of Populus euphratica with axillary buds as explants, soak with 0.05% by mass detergent for 10 minutes, and rinse with tap water for 20 to 30 minutes. Transfer to the ultra-clean workbench, rinse with sterile water for 3 times, soak in 75% ethanol for 30 seconds, rinse with sterile water for 3 times, disinfect with 0.1% mercury liter for 5 minutes, wash with sterile water for 3 times, and then use sterile Bacteria absorbent paper to absorb surface moisture, cut into 0.8-1.2cm stem segments with axillary buds;

[0072] 2), acquisition and subculture of sterile test-tube seedlings

[0073] Inoculate the above-mentioned stem segments on the subculture medium to obtain sterile test-tube plantlets, and carry out subculture proliferation of adventitious buds. Th...

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Abstract

The invention relates to a method for performing high frequent induction regeneration of populus euphratica leaves for plants and belongs to the technical field of populus euphratica leaf regenerationmethods. According to the method disclosed by the invention, by optimizing a basal culture medium and culture conditions, a phenomenon that leaves are yellowed and are difficult to regenerate after three-generation passage of populus euphratica in the past is avoided. The method disclosed by the invention comprises the following steps: on the basis of improving cultivation conditions of tissue culture seedlings, taking leaves as explants, performing adventitious bud induction, subculture, strong seedling culture, rooting culture and other technical links, and finally establishing a method system for performing high-efficiency regeneration induction of the populus euphratica leaves for regeneration plants. The method is capable of realizing high-efficiency regeneration of in-vitro populuseuphratica leaves and has the advantages of excellent reproducibility, high production efficiency and the like, the regeneration rate of adventitious buds can reach 98% or higher, the defect that thepopulus euphratica is difficult in seedling propagation under natural conditions is effectively overcome, rapid propagation in vitro is realized, and a foundation is laid for realizing genetic transformation and planting resource improvement of the populus euphratica.

Description

technical field [0001] The invention relates to a method for high-frequency induction plant regeneration of Populus euphratica leaves, and belongs to the technical field of regeneration methods of Populus euphratica leaves. Background technique [0002] Populus euphratic Oliver (Populus euphratic Oliver) has strong drought resistance and salt-alkali tolerance. It is the only arbor tree species that can form forests in the desert and Gobi regions of northwest my country. It plays an important role in windbreak and sand fixation and maintaining ecological balance. In addition, Populus euphratica has a beautiful shape and can be used as an excellent street tree and garden tree; its wood is hard, and it is a high-quality building and furniture material. More importantly, Populus euphratica is considered to be a model tree species for studying the mechanism of salt tolerance and drought resistance of woody plants, and has become a research hotspot in the aspects of tree salt tole...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 宿红艳王磊张静李晨琦朱世东
Owner LUDONG UNIVERSITY
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