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Potato isolated culture one-step seedling culture medium and optimization method and seedling method thereof

A technology of in vitro culture and culture medium, applied in the biological field, can solve the problems of short regeneration cycle and achieve the effect of short regeneration cycle, simple culture procedure and easy operation

Inactive Publication Date: 2010-08-04
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The one-step seedling-forming method has the advantages of short regeneration period, high reproduction coefficient and regeneration frequency, easy operation, and simple cultivation procedure, and there is no report on one-step seedling-forming of potato in vitro.

Method used

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  • Potato isolated culture one-step seedling culture medium and optimization method and seedling method thereof
  • Potato isolated culture one-step seedling culture medium and optimization method and seedling method thereof
  • Potato isolated culture one-step seedling culture medium and optimization method and seedling method thereof

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Experimental program
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Effect test

Embodiment 1

[0035] A method for optimizing the one-step seedling growth medium for potato in vitro culture, the medium enables the one-step seedling growth of potato test-tube plantlet leaves in vitro culture, comprising the following steps:

[0036] Taking the MS medium without any hormone as the control, three factors and three levels of L 9 (3 4 ) Orthogonal experiment design, adding different concentrations and combinations of 6-benzylaminopurine, naphthalene acetic acid and 2,4-dichlorophenoxyacetic acid to MS medium to prepare 10 different potato one-step seedling growth medium. In each group, the concentrations and combinations of 6-benzylaminopurine, naphthaleneacetic acid and 2,4-dichlorophenoxyacetic acid are shown in Table 1

[0037] Table 1 Orthogonal experiment design L of potato one-step seedling growth medium with different hormone compositions 9 (3 4 )

[0038]

[0039] Sterilize the above-mentioned 10 kinds of potato one-step seedling growth medium, and take an app...

Embodiment 2

[0054] In order to avoid the error of the experimental results caused by human factors, the applicant asked different researchers to use the potato one-step seedling growth medium identical to the embodiment to carry out the same experimental operation as in Example 1 under the same conditions. The experimental results are shown in Table 4 and Table 5.

[0055] Table 4 Effects of different hormone combinations on leaf callus induction

[0056]

[0057] Table 5 Effects of different hormone combinations on the differentiation of adventitious buds in leaves

[0058]

[0059]

[0060] Analysis of the experimental data obtained in Table 4 shows that, except for the control group, each group of potato one-step seedling growth medium can better induce potato leaves, and its induction rate all reaches more than 85%. group, the induction rate reached 100%; while the 3rd, 4th, 5th, 6th, 8th, and 9th groups all had good callus volume, and among them, the 6th, 9th group had the ...

Embodiment 3

[0068] Utilize described potato one-step seedling-forming medium, make the cultivation method of one-step seedling-forming of potato test-tube plantlet leaf culture in vitro, comprise the steps:

[0069] (1), prepare potato one-step seedling growth medium: in 1LMS medium, add 2.0mg 6-benzylaminopurine and 0.4mg naphthaleneacetic acid, carry out sterilization treatment, stand-by;

[0070] (2), select the aseptic test-tube plantlet of potato with the same source, under aseptic conditions, cut out the blade about 0.5cm * 0.5cm that the top fully expands, stick on the culture medium described in step 1 with the blade back side;

[0071] (3), placing the inoculated culture medium obtained in step (2) at a temperature of 22±2° C. in an incubator, cultivating in total darkness, and cultivating under 16 h / d light after 7 to 10 days, with a light intensity of 2000 lx;

[0072] (4) After culturing for 60 days, regenerated potato plants were obtained.

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Abstract

The invention discloses a method for optimizing a potato isolated culture one-step seedling culture medium. The culture medium makes leaves of a potato test tube plantlet undergo isolated culture to form a seedling in one step. The one-step seedling culture medium takes an MS culture medium as a basal medium, and 6-benzyladenine, naphthyl acetic acid and 2,4-dichlorphenoxyacetic acid with different concentrations and combinations are supplied in 1L of the MS culture medium; adventitious buds and adventitious roots are directly differentiated through callus induction; and the potato regenerated seedling is obtained in one step. The callus obtained by induction in a primary culture medium is unnecessarily inoculated to a differential medium for regenerating the seedling. Compared with an isolated culture multi-step regenerated seedling method, the method of the invention has the advantages of simplified steps, short culture period, high repeatability, and high seedling survival rate.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a medium for one-step seedling growth in vitro culture of potatoes, and a method for making seedlings of potato test-tube seedlings in one step by using the medium. Correspondingly, the present invention also provides a method for optimizing the Medium method. Background technique [0002] Potato (Solanun tuberosum L.) is an important grain, vegetable, feed and industrial raw material and crop, and occupies a very important position in my country's agricultural production. In order to innovate germplasm resources, tissue culture technology has been widely used in potato breeding research, and has achieved remarkable results. [0003] Plant tissue culture generally adopts several cultivation procedures such as dedifferentiation, redifferentiation buds and rooting strong seedlings, which are called multi-step seedling formation. In order to simplify the cultivation proced...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01G1/00A01H4/00C12N5/04
Inventor 王西瑶倪苏黄雪丽刘帆杨先泉王俊胡小弦
Owner SICHUAN AGRI UNIV
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