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Method of using spermine to induce tobacco embryoid

A technology of embryoid body and tobacco, which is applied in the field of plant bioengineering tissue culture, can solve the problems of long induction period and low efficiency of tobacco embryoid body, and achieve the effect of reducing the cost of cultivation, simplifying the preparation procedure, and good induction effect

Active Publication Date: 2017-05-10
广西壮族自治区烟草公司百色市公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can be seen that the induction medium of tobacco embryoid body is all compounded by multiple inducer components, and there is no report on the successful induction of tobacco embryoid body with a single inducer
2. The induction period of the existing culture medium for tobacco embryoid bodies is long and the efficiency is low

Method used

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  • Method of using spermine to induce tobacco embryoid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] A method for inducing tobacco embryoid bodies with spermine, comprising the following steps:

[0020] Step 1, explant preparation: take the sterile tobacco plants obtained from tobacco seeds, under aseptic conditions, cut the tobacco leaves into square slices as explants, and set aside, the square slices can be cut into about 5mm×5mm in size ;

[0021] Step 2. Preparation of induction medium: use MS as the basic medium, add spermine 3.2 mg / L, adjust the pH of the medium to 5.8-6.0, and sterilize for later use;

[0022] Step 3, induction culture: insert the explants of tobacco outer leaves obtained in step 1 into the induction medium of step 2, and culture for 20 days.

Embodiment 2

[0024] Step 1, explant preparation: take the sterile tobacco plant obtained from tobacco seeds, when it grows to 5 or 6 leaves, take the third leaf from top to bottom, and cut it into square pieces under aseptic conditions As an explant, spare, the square piece can be cut into a size of about 5mm×5mm;

[0025] Step 2. Preparation of induction medium: use MS as the basic medium, add spermine 3.5 mg / L, adjust the pH of the medium to 5.8-6.0, and sterilize for later use;

[0026] Step 3, induction culture: insert the tobacco leaf explants obtained in step 1 into the induction medium of step 2, and culture for 20 days.

[0027] When the sterile tobacco plant grows to 5 or 6 leaves, the third leaf from top to bottom is a fully mature and robust functional leaf, with vigorous cell metabolism and strong vitality. Differentiate and redifferentiate to form embryoid bodies.

Embodiment 3

[0029] Step 1, explant preparation: take the sterile tobacco plant obtained from tobacco seeds, when it grows to 5 or 6 leaves, take the third leaf from top to bottom, and cut off 2mm above the petiole under aseptic conditions and the blade part 2mm below the tip, and leave the middle part of the blade, and then cut it into 5mm×5mm square slices along the two sides of the main vein as explants for future use. It should be noted that the aforementioned section along the main vein refers to The part of the leaf on both sides of the leaf near the edge is discarded, as long as the part of the leaf on both sides of the main vein;

[0030] Step 2. Preparation of induction medium: use MS as the basic medium, add spermine 4.0 mg / L, adjust the pH of the medium to 5.8-6.0, and sterilize it for later use;

[0031] Step 3, induction culture: insert the tobacco leaf explants obtained in step 1 into the induction medium of step 2, and culture for 20 days.

[0032] When the sterile tobacco ...

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Abstract

The invention discloses a method of using spermine to induce tobacco embryoid. The method includes: using spermine as an inducer to prepare an inducing culture medium for embryoid induction of tobacco leaf explant. Compared with conventional culture media, the inducing culture medium prepared by taking spermine as the inducer is high in tobacco embryoid forming speed, yield and quality, and the objective of producing tobacco embryoid can be achieved directly without transferring original explant into other bottles for subculture.

Description

technical field [0001] The invention relates to the technical field of plant bioengineering tissue culture. More specifically, the present invention relates to a method for inducing tobacco embryoid bodies with spermine. Background technique [0002] "Embryoid" (embryoid) is a somatic tissue or germ cell tissue material under special culture conditions, which has not undergone sexual fertilization process, but has undergone embryonic development process, and has the special physiological function of zygotic embryo. The structure is actually the prototype of the future plant. Embryoid bodies are also called somatic (cell) embryos or asexual embryos, which can be directly used in production to make artificial seeds and produce regenerated plants (test-tube plantlets); in theoretical research, it is to explore cell differentiation, organ building, individual development, and Important material for its related anatomical and physiological biochemical processes. Therefore, the...

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 周凤珏龚银花周文亮许鸿源赖洪敏王五权韦忠陈念平许皓翔林北森罗刚覃迎姿刘春萍
Owner 广西壮族自治区烟草公司百色市公司
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