Method for inducing tobacco callus by spermine

A callus and tobacco technology, applied in the field of plant bioengineering tissue culture, can solve the problems of cumbersome and labor-intensive medium preparation procedures, affecting the redifferentiation of proliferative organs, and low callus yield, so as to reduce the cost of culture and produce Faster and save material and manpower

Active Publication Date: 2017-01-04
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

These traditional techniques for obtaining tobacco callus have three disadvantages: 1. The preparation procedure of the medium is cumbersome and labor-intensive; 2. The yield of callus is not high; 3. 2,4-D, NAA and IBA all have obvious toxicity and residues, especially 2,4-D is originally a herbicide, which can easily cause browning and hardening of the newly obtained callus, resulting in a decline in quality and directly affecting its subsequent proliferation and organ redifferentiation capabilities
However, in order to show the success of respective technologies, relevant reports often only emphasize its "healing rate %", that is, how high is the number of explants / inoculated explants × 100% (according to different reports, culture 25d- 30d can reach 75%-100%), but few people clearly inform readers of the actual yield and quality of their callus
The fact is that, according to the reports that have been investigated, there are almost no exceptions. After obtaining a small amount of callus from explants, they must be transferred to another special proliferation medium for expansion in time. The operation is cumbersome and the cost is correspondingly increased.

Method used

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  • Method for inducing tobacco callus by spermine

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Experimental program
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Effect test

Embodiment 1

[0018] A method for inducing tobacco callus with spermine, comprising the following steps:

[0019] 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 ;

[0020] Step 2, preparation of induction medium: use MS as the basic medium, add spermine 0.5 mg / L, adjust the pH of the medium to 5.8-6.0, and sterilize for later use;

[0021] Step 3, induction culture: the tobacco leaf explants obtained in step 1 are inserted into the induction medium of step 2, and cultured for 10-20 days.

Embodiment 2

[0023] 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;

[0024] Step 2, preparation of induction medium: use MS as the basic medium, add spermine 2 mg / L, adjust the pH of the medium to 5.8-6.0, and sterilize for later use;

[0025] Step 3, induction culture: the tobacco leaf explants obtained in step 1 are inserted into the induction medium of step 2, and cultured for 10-20 days.

[0026] When the leaves of sterile tobacco plants grow to 5 or 6 leaves, the third leaf from top to bottom is a fully mature and robust functional leaf with strong cell metabolism and strong vitality. Compared with other tobacco leaves, it is easier to cultivate as explants Dedifferentiation occurs to form callus.

Embodiment 3

[0028] 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 edge leaf parts on the left and right sides of the leaves are discarded, only the leaf parts on both sides of the main vein;

[0029] Step 2. Preparation of induction medium: use MS as the basic medium, add spermine 2.5 mg / L, adjust the pH of the medium to 5.8-6.0, and sterilize for later use;

[0030] Step 3, induction culture: the tobacco leaf explants obtained in step 1 are inserted into the induction medium of step 2, and cultured for 10-20 days.

[0031] When the leaves of the steri...

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Abstract

The invention discloses a method for inducing tobacco callus by spermine. The method applies spermine as the inductive agent and prepares the inducing medium to carry out the callus induction of tobacco leaf explant. Compared with the traditional medium, the inducing medium prepared by using spermine as the inductive agent is quick in callus rate, high in output, and excellent in quality; moreover, the medium can be directly applied to the proliferation and propagation expansion of the callus, and reach multiple applications of one medium; thus the overall working efficiency can be significantly improved.

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 of inducing tobacco callus with spermine. Background technique [0002] Tobacco callus is not only an important material for obtaining regenerated plants, but also for obtaining genetic variation and improving agronomic traits. At the same time, tobacco is a model plant for plant biotechnology research, and its callus is also an important experimental material for basic research in cell biology, molecular biology and genetics. Therefore, there are many technical research reports on how to obtain tobacco callus at home and abroad. But to sum up, the vast majority of its basic medium is MS (Murashige and Skoog, 1962), and occasionally H (Bourgin and Nitsch, 1967). Inducers were almost entirely restricted to two classes of 5 plant growth substances. That is, three of the auxins: 2,4-D (2,4-dichlorophenoxyace...

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 周文亮周凤珏许鸿源赖洪敏龚银花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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