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Exosomatic breeding method of switchgrass

A technology for switch branches and stem nodes is applied in the field of in vitro propagation of switchgrass, which can solve the problems of biotechnology limitation, long culture period, genotype differences, etc., and achieve the effects of improved reproduction, reduced experimental cost, and reduced cost.

Inactive Publication Date: 2007-12-05
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, similar to the tissue culture of most gramineous plants, switchgrass also has genotype differences in artificially induced callus, long culture period, low regeneration rate, high cost and other shortcomings, which make the application of biotechnology in switchgrass limited. certain limit

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Four switchgrass varieties, Alamo, Blackwell, GA-001A and N.Switchgrass, were propagated in vitro. The specific production process is:

[0054] (1) Cut out the stem node tissue

[0055] When the four varieties of switchgrass grow to have 5-6 nodes respectively, remove the 2 stem nodes at the top and the base, and manually cut the 3-4 stem nodes in the middle, and each stem node includes the upper and lower regions of the stem node The length of each segment is 1-1.5 cm.

[0056] For each variety, 100 stem node tissues from different plants were excised.

[0057] (2) Induce regenerated shoots

[0058] a. Place the stem node tissue in a petri dish, and use 70% alcohol surface disinfection for 1 minute;

[0059] b. discard the alcohol, adopt 20% mass fraction of commercial sodium hypochlorite (adding 0.1% Tween 80) to sterilize for 15 minutes;

[0060] c. Rinse 3 times with sterile water, artificially induce primary regenerated shoots on MBP medium, place 2 stem node ...

Embodiment 2

[0079] Introduced leaf senescence suppressor gene P SAG12 -IPT T 1 The Alamo transgenic plants were propagated in vitro. The specific production process is:

[0080] (1) Cut out the stem node tissue

[0081] After PCR and Southern hybridization detection, it was identified to contain P SAG12 -T for the IPT target gene 1 Generation of Alamo transgenic plants. When the transgenic plants grow and develop to have 6 nodes, remove the 2 stem nodes at the top and the base, and manually cut out the middle 4 stem nodes. A total of 100 stem node tissues of transgenic plants from different sources were excised.

[0082] (2) Induce regenerated shoots

[0083] a. Place the stem node tissue in a petri dish, and use 70% alcohol surface disinfection for 1 minute;

[0084] b. discard the alcohol, adopt 20% mass fraction of commercial sodium hypochlorite (adding 0.1% Tween 80) to sterilize for 15 minutes;

[0085] c. Rinse 3 times with sterile water, artificially induce primary regener...

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PUM

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Abstract

The present invention discloses the in vitro propagation process of switchgrass. The in vitro propagation process includes the following steps: cutting 3-4 middle stem nodes after switchgrass grows to possess 5-6 stem nodes, and artificially inducing regeneration bud in MBP culture medium; excising the growing point of cespitose bud repeatedly for 4 or 5 times until obtaining cespitose bud tuber with diameter of about 0.5 cm, and inducing in new MBP culture medium to obtain large cespitose bud; culturing the regenerated plant in rooting culture medium SMO to obtain seedling; and transplanting to greenhouse or field. The in vitro propagation process of switchgrass has expanded tissue culture and genetic transformation range of switchgrass, shorted culture period, raised regeneration frequency and lowered experiment cost. The present invention provides effective measure for the fast propagation of transgenic switchgrass.

Description

technical field [0001] The invention relates to the biotechnology field of bioenergy plant switchgrass, and further relates to a method for in vitro propagation of switchgrass. Background technique [0002] Switchgrass (English name Switchgrass, Latin name Panicum virgatum Linn) belongs to Poaceae, Panicum genus, is a perennial high-quality forage grass, and has a long history of cultivation in my country. In recent years, because of its simple cultivation techniques, wide adaptability, strong ability of windbreak and sand fixation, resistance to barrenness, high biological yield, and the ability to use low-quality or desertified soils, it has been identified internationally as the first choice for the production of fuel ethanol by replacing food resources. energy plant. At present, many countries represented by the United States have invested a lot of manpower and material resources in the research and development of switchgrass. Among them, the use of biotechnology to en...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00C12N5/04
Inventor 奚亚军刘曙东
Owner NORTHWEST A & F UNIV
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