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A kind of ultra-low temperature preservation method of the shoot tip of honeysuckle honeysuckle

A technology of ultra-low temperature preservation and shoot tip, which is applied in the fields of plant preservation, botanical equipment and method, application, etc., can solve the problem that the ultra-low temperature preservation of honeysuckle shoot tips has not been reported yet, and achieves the possibility of reducing plant variation and stability. The effect of high sexuality and low degree of differentiation

Inactive Publication Date: 2020-11-13
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no report on the ultra-low temperature preservation of the shoot tips of Lonicera japonica Lonicerae

Method used

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  • A kind of ultra-low temperature preservation method of the shoot tip of honeysuckle honeysuckle
  • A kind of ultra-low temperature preservation method of the shoot tip of honeysuckle honeysuckle
  • A kind of ultra-low temperature preservation method of the shoot tip of honeysuckle honeysuckle

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A method for ultra-low temperature preservation of the stem tip of Lonicera japonica, the specific operations are as follows:

[0026] 1) Under aseptic conditions, the isolated shoot tip containing 2 leaf primordia with a shoot tip length of 2-3 mm is stripped from the aseptic stem section of Lonicera japonica containing the shoot tip;

[0027] 2) Put the stripped shoot tip into the pre-culture solution for treatment, the optimum concentration of sucrose in the pre-culture solution is MS solution of 0.3mol / L, and pre-culture at 4°C for 3 days;

[0028] 3) Put the pre-cultured isolated shoot apex in the LS loading solution, and load it at room temperature 25°C for 10-50 minutes; the composition of the loading solution is: 2M glycerol+0.4mol / L sucrose+MS solution;

[0029] 4) Put the loaded stem tip into PVS2 protection solution and dehydrate for 20-100 minutes at 0°C; the composition of the PVS2 protection solution is: 30% glycerol+15% ethylene glycol+15% dimethyl sulfox...

Embodiment 2

[0034] To test the effect of sucrose concentration and pre-cultivation time in the pre-medium on the cryopreservation of Lonicera Lonicerae shoot tips: only changing the sucrose concentration and pre-cultivation time in the pre-medium, first put the stripped shoot tips into containers with different concentrations (0.1-0.6mol / L) sucrose MS solution for pre-cultivation; the pre-cultivation temperature condition is 4°C, and the culture time is 0-5d; then the shoot tip survival rate is counted. The results of the study showed that (such as figure 1 figure 2 Shown) After putting the stripped shoot tip into the pre-culture solution with different sucrose concentrations for different time, the survival rate of the shoot tip was different. When the sucrose concentration was 0.3mol / L, the When cultured for 3 days, the shoot tip survival rate can reach up to 60%. It can be seen that pre-cultivation is the key factor affecting the survival rate of Lonicera Lonicera shoot tips cryopre...

Embodiment 3

[0036] Test the effect of LS loading solution treatment time on the survival rate of shoot tips after cryopreservation: In a sterile ultra-clean workbench, put the pre-cultured shoot tips into the LS loading solution, and load different shoot tips at room temperature at 25°C. Time (0, 10, 20, 30, 40, 50 min). It can be seen from the results of the study (such as image 3 shown), there was a significant difference between the control group treated with loading solution and the control group without treatment, and the shoot tip survival rate without loading solution treatment was very low, and the shoot tip survival rate was only 3.3%; , the survival rate of the shoot tip showed a trend of first increasing and then decreasing, reaching the highest value of 63.67% when the LS was loaded for 40 minutes, and there was a significant difference with the survival rate of the shoot tip under other loading times. When the LS loading time was 50 minutes , the shoot tip survival rate dro...

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Abstract

The invention relates to a super-low-temperature storage method for honeysuckle stem tips. The method successively comprises the following steps: peeling and carrying out pre-treatment on the honeysuckle stem tips; loading; dehydrating; liquid nitrogen storage; defreezing and recovery culture. The method can keep excellent characteristics and is convenient to operate and high in feasibility, and excellent germplasm resources are stored favorably. The genetic materials of the stem tips stored at the super-low temperature are relatively stable, and most stem tips can form intact small plants, sothat the method has an important using value in honeysuckle germplasm resource storage and other related research aspects.

Description

technical field [0001] The invention relates to a method for long-term preservation of stem tips, in particular to a method for ultra-low temperature preservation of the stem tips of honeysuckle honeysuckle. Background technique [0002] Honeysuckle, also known as honeysuckle, belongs to the family Lonicera and is one of the good ornamental shrubs in my country. The bark often has irregular columns and is off-white to gray-brown in color. Honeysuckle flowers are axillary, and every 2 small flowers are born on a main flower stalk. The single flower is a lip-shaped corolla. The flower is white during the opening period, and gradually turns light yellow after opening, blooming in layers, gold and silver complement each other. The fruit is a red berry, often clustered on long branches, and it is ornamental in autumn, with numerous red fruits. Honeysuckle flowers are elegant and fragrant, and it is an excellent honey source tree species; honeysuckle has strong adaptability to t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N3/00
CPCA01N3/00
Inventor 宋红田晓璇李萍李享
Owner NORTHEAST FORESTRY UNIVERSITY
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