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Preparation method and application of PEG culture medium with solid-liquid two phases

A liquid culture medium and culture medium technology, applied in the field of plant drought resistance research, can solve the problems of limited use of PEG, inconvenient use of vertical infiltration method, etc., and achieve the effect of simple production method, long storage time, and continuous and effective stress effect

Active Publication Date: 2015-12-16
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] In order to solve the technical problems of limited use of high-concentration common solid PEG and inconvenient use of the vertical osmosis method, the present invention specially designs a solid-liquid two-phase PEG medium for horizontal osmosis

Method used

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  • Preparation method and application of PEG culture medium with solid-liquid two phases
  • Preparation method and application of PEG culture medium with solid-liquid two phases
  • Preparation method and application of PEG culture medium with solid-liquid two phases

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preparation example Construction

[0039] The preparation method of solid-liquid two-phase PEG culture medium of the present invention, at first preparation contains 3% sucrose, 0.8% agar powder, pH is the solid MS culture medium of 5.8, in each 230mL glass culture bottle (diameter 6.5cm, height 9.5cm ) into 50mL, autoclaved and cooled for 30-40min to form a solid gel. Half of the gel was cut vertically along the diameter of the culture bottle, and an equal volume of PEG-containing MS culture solution was added to the gap, the concentration of PEG in the PEG-containing MS liquid medium was 0-25% (W / V), Obtain solid-liquid two-phase PEG culture medium of the present invention (as figure 1 shown). The medium is composed of solid-phase MS gel and liquid-phase PEG solution, each accounting for half of the total volume of the medium, and is a solid-liquid two equal-volume PEG medium.

[0040]The solid-liquid two-phase PEG medium of the present invention is a solid PEG medium of the horizontal osmosis method, compr...

Embodiment 1

[0045] In the following, it is verified by experiments that the solid-liquid two-phase PEG medium can be applied to the determination of the drought resistance difference of test-tube plantlets of different potato varieties.

[0046] Get four different kinds of potatoes and cultivate them on the solid-phase MS medium in the solid-liquid two-phase PEG medium of the present invention, configure several groups of PEG-containing MS liquid medium, wherein the concentration of PEG is respectively 0%, 5% %, 10%, 15%, 20%, 25%, to observe the growth state of the test-tube plantlets. The four varieties of potatoes are Shuangfeng 5, Xia Bodi, Atlantic and Kexin 18, respectively. The specific culture conditions are as follows: use potato plantlets that grow robustly and have the same height and growth as materials, cut stems with a length of about 2 cm and contain terminal buds as explants, and insert 3 stems into each bottle of solid medium , arranged in a row, the position is 0.5cm aw...

Embodiment 2

[0054] The solid-liquid two-phase PEG medium of the invention can also be applied to the determination of the drought resistance of transgenic test-tube plantlets.

[0055] In the PEG simulated drought test, the test-tube plantlets obtained by transforming the Porphyra variegata trehalose-6-phosphate trehalose synthase gene (TPS) into the potato variety Desiree were used as materials, and the non-transgenic test-tube plantlets were used as controls. The invented solid-liquid two-phase PEG medium was subjected to stress culture, cultured for 4 weeks, the growth of the plants was observed and parameters such as plant height, root length, root number, fresh weight and dry weight were measured. In the determination of water shortage and drought stress, the test-tube seedlings of transgenic and non-transgenic Desiree potatoes were acclimated for one week, and then transplanted into paper cups containing nutrient soil. When the plant height reaches 20cm, carry out the drought stress...

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Abstract

The invention provides a preparation method and application of PEG culture medium with solid-liquid two phases. The preparation method comprises: preparing a solid MS culture medium at the bottom of a culture bottle; carrying out autoclaved sterilization and cooling to form a solid gel; removing part of the solid gel in a vertical direction, preferably half of the solid gel in volume along the diameter of the culture bottle, to form a cavity at the bottom of the culture bottle; and adding an MS liquid culture medium containing PEG into the cavity, wherein the volume of the added MS liquid culture medium containing PEG is as same as the volume of the removed solid gel. The culture medium provided by the invention is composed of a solid phase MS culture medium and a liquid phase PEG culture solution, wherein liquid PEG seeps into the solid gel along the horizontal direction and a potato test tube seedling stem section is inoculated to the solid phase culture medium for drought resistance test. The culture medium has the characteristics of being simple to manufacture, short in required time, convenient to inoculate stem sections and the like.

Description

technical field [0001] The invention belongs to the field of plant drought resistance research, and in particular relates to a preparation method and application of a solid-liquid two-phase PEG medium. Background technique [0002] Potato is the fourth largest food crop in the world, and it is also a potential staple food in my country, and it has the characteristics of both vegetables and food. The growth of potato requires a lot of water, and water shortage and salt stress have a great impact on it. Adversity stress such as drought is an important limiting factor for potato production, and its drought resistance and other adversity research is of great value. [0003] Bansal et al. (1991) found that the resistance of potato leaf discs to PEG6000 (polyethylene glycol, molecular weight 6000) solution was consistent with the drought resistance of donor varieties, and large molecular weight PEG can be used as a screening agent and determination of potato drought resistance ag...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00A01G9/10A01G7/00
Inventor 郭宝太宋志成牛倩雅孟庆芹赵春梅王晶珊李广存
Owner QINGDAO AGRI UNIV
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