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Upland grass tissue-culture rapid propagation method for ecological restoration use

A tissue culture and ecological restoration technology, applied in horticultural methods, botanical equipment and methods, applications, etc., can solve the problems of low embryogenic callus acquisition rate, lack of tissue culture system for highland grass, and few creeping bentgrass.

Inactive Publication Date: 2020-03-27
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In June 2006, Gansu Agricultural University conducted a preliminary study on the tissue culture process of creeping bentgrass; however, the regeneration rate and the acquisition rate of embryogenic callus were relatively low, and the callus browning problem that occurred resulted in usable callus Less amount of injured tissue and less high quality creeping bentgrass available for further acquisition
At present, the highland grass lacks a set of efficient tissue culture system. The present invention aims at this and lays the foundation for ecological restoration and other issues.

Method used

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  • Upland grass tissue-culture rapid propagation method for ecological restoration use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1) The sterilized and sterilized seeds of creeping bentgrass are used as explants after vernalization, and first inoculated on the callus induction medium to induce callus

[0031] Explant selection and sterilization, vernalization

[0032] A. select the creeping bentgrass highland seed full of particles to be the tissue culture explant;

[0033] B. Soak for 2 minutes with 70% ethanol solution for the disinfection of the seed epidermis, and wash with distilled water for 3 times after disinfection to remove residual ethanol;

[0034] C. Soak in 70% sodium hypochlorite for 10 minutes, add 0.1ml Tween20, place on a magnetic stirrer for stirring, after sterilization, clean it with 500ml sterilized water on the ultra-clean workbench to remove residual sodium hypochlorite, then add 50ml sterile water Put it in a refrigerator at 4°C for low-temperature vernalization for 24 hours;

[0035] D. Put the vernalized seeds on the filter paper of the ultra-clean bench and blow dry. ...

Embodiment 2

[0052] 1) The sterilized and sterilized seeds of creeping bentgrass are used as explants after vernalization, and first inoculated on the callus induction medium to induce callus

[0053] Explant selection and sterilization, vernalization

[0054] A. Select creeping bentgrass highland seeds with full particles as tissue culture explants

[0055] B. Soak for 2 minutes with 70% ethanol solution for the disinfection of the seed epidermis, and wash with distilled water for 3 times after disinfection to remove residual ethanol;

[0056] C. Soak in 70% sodium hypochlorite for 10 minutes, add 0.1ml Tween20, place on a magnetic stirrer for stirring, after sterilization, clean it with 500ml sterilized water on the ultra-clean workbench to remove residual sodium hypochlorite, then add 50ml sterile water Put it in a refrigerator at 4°C for low-temperature vernalization for 24 hours;

[0057] D. put the vernalized seeds on the filter paper of the ultra-clean workbench and blow dry;

[...

Embodiment 3

[0075] 1) The sterilized and sterilized seeds of creeping bentgrass are used as explants after vernalization, and first inoculated on the callus induction medium to induce callus

[0076] Explant selection and sterilization, vernalization

[0077] A. select the creeping bentgrass highland seed full of particles to be the tissue culture explant;

[0078] B. Soak for 2 minutes with 70% ethanol solution for the disinfection of the seed epidermis, and wash with distilled water for 3 times after disinfection to remove residual ethanol;

[0079] C. Soak in 60% sodium hypochlorite for 10 minutes, add 0.1ml Tween 20, place on a magnetic stirrer for stirring, after sterilization, clean it with 500ml sterilized water on the ultra-clean workbench to remove residual sodium hypochlorite, then add 50ml Sterile water was placed in a 4°C refrigerator for low-temperature vernalization for 24 hours;

[0080] D. put the vernalized seeds on the filter paper of the ultra-clean workbench and blow...

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Abstract

The invention provides an upland grass tissue-culture rapid propagation method for ecological restoration use. The upland grass tissue-culture rapid propagation method for the ecological restoration use comprises the following steps: 1) taking vernalized creeping bentgrass seeds subjected to sterilization and disinfection as explants, inoculating a callus induction medium with the explants so as to induce callus; 2) inoculating a subculture medium with the callus so as to carry out several rounds of proliferation culture; 3) inoculating a bud induction and differentiation medium with the callus subjected to the proliferation culture so as to carry out differentiation culture; 4) inoculating a rooting medium with adventitious buds obtained after the differentiation culture so as to carry out rooting; and 5) carrying out acclimatizing and transplanting. According to the upland grass tissue-culture rapid propagation method for the ecological restoration use provided by the invention, an efficient upland grass tissue culture system is established so as to achieve maximum callus induction rate at 66.2%, embryogenic callus rate at 65.4%, bud differentiation rate at 80.3%, rooting rate at85.7%, and survival rate of tissue culture seedlings transferred into soil at 98.5%.

Description

technical field [0001] The invention belongs to the field of plant tissue culture, and particularly relates to a rapid propagation method of highland grass tissue culture for ecological restoration. Background technique [0002] Creeping bentgrass (Agrostis stolonifera), whose English name is Creeping bentgrass, is a perennial cool-season turfgrass belonging to the genus Bentgrass in the Poaceae family. Creeping bentgrass is native to Eurasia, and it is distributed in Northeast my country, North China, Northwest China and other regions. Highland grass belongs to a species of creeping bentgrass. The leaves of highland grass are soft and tender, with strong ornamental value, strong cold tolerance, drought tolerance, shade tolerance, trampling tolerance, and low pruning tolerance. Commonly used in golf course greens, tees and other high-quality lawns. In recent years, highland grass has also been widely used in environmental restoration and other aspects. Highland grass has ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 晁跃辉李殷睿智贾丝俏刘卓成
Owner BEIJING FORESTRY UNIVERSITY
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