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A method for establishing a high-efficiency regeneration system of Pseudomonas chinensis by using young panicles as explants

A high-efficiency regeneration and explant technology, applied in the field of plant biotechnology research, can solve the problem of high-efficiency genetic transformation, poor callus differentiation ability, and callus that are not suitable for large-scale screening of somatic mutants in micropropagation. Long induction period and other problems, to achieve the effect of improving callus induction rate, induction efficiency and easy implementation

Active Publication Date: 2020-08-14
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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Problems solved by technology

[0006] The technical tasks proposed by the present invention and the technical problems to be solved are to overcome the long induction period and low induction efficiency of the callus in the existing Pseudomonas in vitro culture technology, and the induced callus differentiation ability is poor, which is not suitable for use. Defects in large-scale micropropagation, large-scale screening of somatic mutants, and high-efficiency genetic transformation mediated by Agrobacterium

Method used

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  • A method for establishing a high-efficiency regeneration system of Pseudomonas chinensis by using young panicles as explants
  • A method for establishing a high-efficiency regeneration system of Pseudomonas chinensis by using young panicles as explants
  • A method for establishing a high-efficiency regeneration system of Pseudomonas chinensis by using young panicles as explants

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Experimental program
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Embodiment Construction

[0038] (1) Experimental materials

[0039] Take the fresh and tender young spikes (inflorescences) that have not drawn out the leaf sheaths at the beginning of flowering stage of E092-1 as the explant material (such as figure 1 shown);

[0040] (2) Medium

[0041] The basic medium was MS medium composed of macroelements, trace elements and vitamins, supplemented with 30.0 g / L sucrose (Sigma) and 8.0 g / L agar (Japan);

[0042] Callus induction medium MS1: add 2.0 mg / L of 2,4-D (Sigma D-7299) or 4.0 mg / L of 2,4-D (Sigma D-7299) or 6.0 mg / L of 2,4-D (Sigma D-7299), 0.1 mg / L of BAP (Sigma B3408);

[0043] Subculture medium MS2: Add 1.5 mg / L 2,4-D (Sigma D-7299) and 0.1 mg / L BAP (Sigma B3408) to the basic medium;

[0044]Green shoot differentiation medium MS3: Add 2.0 mg / L of KT (Sigma K-3378), 0.1 mg / L of NAA (Sigma N0640), or 2.0 mg / L of BAP (Sigma B3408), 0.1 mg / L of NAA (SigmaN0640), or 2.0 mg / L of CPPU (Sigma C2791), 0.1 mg / L of NAA (Sigma N0640);

[0045] Test-tube see...

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Abstract

The invention relates to a method for establishing an eremochloa ophiuroides highly efficient regeneration system with a young spike as an explant. The method includes the steps of explant selection and sterilization, callus induction and proliferation, callus differentiation, and rooting induction. The method provided by the invention is simple in explant sterilization, high in callus induction rate, easy to implement, and short in cycle of callus induction, obtained callus adventitious buds have high differentiation rate, test-tube plantlets are easy to root, and the method is suitable for eremochloa ophiuroides germplasm innovation, genetic inheritance transformation and other modern biological technology operations.

Description

technical field [0001] The invention relates to a method for establishing a high-efficiency regeneration system of Pseudomonas officinale in vitro, and belongs to the field of plant biotechnology research. Background technique [0002] False Thrift Grass [ Eremochloa ophiuroides (Munro.) Hack.] is a perennial C4 herbaceous plant of the genus Centipede in the Poaceae Panicaceae, and it is also the only excellent warm-season turfgrass plant in this genus that can be used as turfgrass. Except for a small amount of distribution in Southeast Asia, False Thriftgrass is mainly distributed in the Yangtze River Basin and the south of China. The central and southern part of my country is its natural distribution center. It is recognized as the best lawn grass originating in China at home and abroad, so it is called Known as "Chinese lawn grass" (Hanna et al. 1995). Compared with other lawn grasses, this grass is known for its tolerance to barrenness, few diseases and insect pests, a...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/008
Inventor 李建建马晶晶刘建秀张兵汪毅郭海林宗俊勤陈静波李丹丹李玲
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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