Fluorescence tracking method for transportation direction and route of assimilate of in-vitro lily plant

A technique of fluorescent tracing and assimilation, which is applied in the field of plant cell physiology research, can solve the problems of unreported fluorescent staining of isolated lily bulbs, easy damage to the internal structure of the plant, and inability to achieve the labeling effect, and is not easy to damage. Plant body, high quantum yield, not easy to evaporate

Inactive Publication Date: 2019-03-26
ZHEJIANG UNIV
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Problems solved by technology

5(6)-CF has been widely used as a fluorescent indicator for the study of phloem symplast unloading and phloem translocation in plants. At present, assimilates during fruit development have been studied in walnuts, grapes, apples, pears, and other fruits. From the source-leaf, the path and unloading of transport to the sink-fruit, and also used CF to study the phloem symplast unloading path in the barley and tobacco sink leaves, micropropagation tissue culture as a variety of plants, especially lily, production and commonly used system, but there is no report on CFDA fluorescent staining of isolated lily bulbs
[0005] However, when the applicant used isolated lily bulblets as research materials, he found that because isolated lily bulblets have higher requirements for a sterile environment, and during the CFDA labeling process, if the current injection method adopted on other crops, The pressure injection method and the microinjection method require special equipment and high technical requirements, while the Eppendorf tube cotton thread infiltration method and leaf smearing method do not require special equipment, but the operation process is cumbersome and easily damages the internal structure of the plant. The solution evaporates, and the ideal marking effect cannot be achieved.

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  • Fluorescence tracking method for transportation direction and route of assimilate of in-vitro lily plant

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Embodiment Construction

[0032] The following describes the implementation process of the present invention through specific implementation examples:

[0033] 1. Prepare the plant and dye

[0034] 1. Select isolated lily bulbs grown in the medium as the plants to be treated, and healthy and sterile isolated lily plants at any growth stage can be used in the operation of the present invention;

[0035] 2. Use acetone or a mixture of DMSO and pure water with a volume ratio of 1:1 to dissolve the fluorescent dye CFDA, and prepare a CFDA solution of 1 to 2 mg / mL, protected from light. The fluorescent dye CFDA is insoluble in water. It can be stored in a refrigerator at -20°C and protected from light after being divided into aliquots according to the amount used.

[0036] 2. Introducing dyes into plants

[0037] 1. Select the scale of the isolated lily bulb as the introduction site, and use a double-sided knife to cut a 0.3mm wide wedge-shaped incision in the middle of the scale, the depth shall be subject to the p...

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Abstract

The invention relates to the research technique of plant stoichiology and aims at providing a fluorescence tracking method for the transportation direction and route of assimilate of an in-vitro lilyplant. The fluorescence tracking method comprises the following steps: applying medical absorbent cotton wetted by a CFDA solution to outer-layer scale notches of bulbs or cross sections of vascular bundles of leaves of an in-vitro lily, carrying out packaging by virtue of a paraffin sealing film, and covering by virtue of tin foil paper; continuing to carry out culturing for 72 hours under the original culture condition; carrying out freehand section sampling at an observed part, observing distribution images of CF in the notches by virtue of a fluorescence microscope, and taking a picture. Afluorescent dye CFDA adopted in the invention has no toxin or chemical inertness to cells, the quantum yield is relatively high, the transportation direction and route of the assimilate in the lily bulbs in different developmental stages can be properly tracked in a cell level, and compared with a traditional microradioactive autographing technique, the method is relatively safe and simple. The special sample treatment is omitted, the cost is low, and an ideal marking result can be rapidly and effectively achieved.

Description

Technical field [0001] The invention relates to the research technology of plant cell physiology, in particular to a fluorescent tracing method for the transport direction and path of assimilation during the development of lily bulblets based on an in vitro model system. Background technique [0002] Lily is known as the "king of bulbous flowers" and is the collective name of all species, subspecies, varieties, variants and varieties of Lilium in the Liliaceae family. Lily is mainly for viewing flowers and is one of the five largest cut flowers in the world. Lily can not only view flowers, but its bulbs are rich in starch. Some varieties can be used for edible or medicinal purposes, and can also be used for extracting spices. It has a wide range of applications. Flower bulbs have a low natural reproduction rate and a long childhood period, so the breeding cycle of new varieties is longer, for example, lilies are about 12-15 years, and tulips are about 20-30 years. The robust gr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428
Inventor 闵芮涵夏宜平张琳吴昀李世琦
Owner ZHEJIANG UNIV
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