Method for increasing efficiency of tissue culture of Medicago sativa L.

A technique of tissue culture and alfalfa, applied in the field of plant biology, can solve problems such as insufficient callus formation and differentiation, long induction time of alfalfa tissue culture, explant contamination, etc., to retain differentiation ability and increase callus induction rate , The effect of low seed pollution rate

Inactive Publication Date: 2012-06-20
GRASSLAND RES INST OF CHINESE ACAD OF AGRI SCI
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Problems solved by technology

[0013] At present, in the tissue culture of alfalfa, there are mainly two technical problems: serious contamination of explants, insufficient callus formation and differentiation, which lead to long induction time, low efficiency, poor quality and serious browning of alfalfa tissue culture.

Method used

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  • Method for increasing efficiency of tissue culture of Medicago sativa L.
  • Method for increasing efficiency of tissue culture of Medicago sativa L.
  • Method for increasing efficiency of tissue culture of Medicago sativa L.

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Embodiment

[0036] 1. Seed sterilization and acquisition of sterile vaccines

[0037] Select seeds with full grains, soak in 75% alcohol for 1 minute, soak in 1% mercuric acid for 5 minutes, then soak in 30% hydrogen peroxide for 30 minutes, then rinse with distilled water for 3-4 times, put the rinsed seeds on filter paper to absorb the water , inoculated in solid medium (pH=6.0). Medium formula: MS+agar (7.5g / L)+sucrose (30g / L). Culture conditions, 25°C, light intensity 2500lk, divided into light and dark cycle 16 / 8h, cultured for 5-7 days.

[0038] By adopting the compound disinfection method of mercuric chloride and hydrogen peroxide, the disinfection effect is thorough, the pollution rate is low, and the damage to the seeds is minimized, finally solving the current serious problem of explant contamination.

[0039] 2. Induction of Callus

[0040] Select robust seedlings, and intercept the hypocotyls (the part above the root and below the stem). Cut it into 1cm segments and place ...

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Abstract

The invention belongs to the technical field of plant biology, in particular relates to a method of tissue culture of Medicago sativa L. By way of developing a novel tissue culture technical method, the invention focuses on improvement and innovation of two technical nodes (seed disinfection and callus formation and differentiation) in culture steps. With adoption of a mercury bichloride and hydrogen peroxide compound disinfection method, the invention realizes the purposes of thorough disinfection effect and low pollution rate and minimum damage to seeds. Finally, the invention solves the problem of severe pollution to current explants. Meanwhile, a method of TDZ and 6-BA is adopted, thereby, the callus induction rate and the formation rate are remarkably increased, the callus quality is improved, the differentiation capacity of the callus is reserved to a maximum extent, the formation of embryoid sprouts is promoted so that the novel technical method of the tissue culture of the Medicago sativa L. which is efficient, saves time, and has low toxicity and low browning, is obtained.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, in particular to a method for tissue culture of alfalfa (Medicago sativa L.). The present invention develops a new tissue culture technology method, focusing on improving and innovating the two major technical nodes of seed disinfection and callus formation and differentiation in the culture steps, so as to obtain a new type of alfalfa tissue culture with high efficiency, low toxicity and low browning. Technical method. Background technique [0002] Alfalfa (Medicago sativa L.) is an important leguminous forage, known as the "king of forage" because of its high nutritional value and good palatability. Tissue culture of alfalfa mainly refers to the technical method of using the isolated organs of alfalfa (cotyledon, radicle, etc.) to carry out plant regeneration through artificial cultivation. The method is of great significance for the reproduction and preservation of the precious genetic mat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 任卫波郭慧琴李晶王茅雁刘雅学张文静赵海霞解继红
Owner GRASSLAND RES INST OF CHINESE ACAD OF AGRI SCI
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