A kind of preparation method and application of solid-liquid two-phase PEG medium
A technology of liquid culture medium and culture medium, applied in the field of plant drought resistance research, can solve the problems of inconvenient use of vertical osmosis method and limited use of PEG, and achieve the advantages of simple production method, long existence time, and meeting the needs of identification and screening. Effect
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[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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