Convenient observation method of orchid mycorrhiza microstructure

A microstructure and plant root technology, applied in the agricultural field, can solve the problems of complex and cumbersome, poor observation effect, long time-consuming, etc., and achieve the effect of convenient and fast operation, good observation effect and reliable results

Inactive Publication Date: 2014-11-05
SHANGHAI ACADEMY OF LANDSCAPE ARCHITECTURE SCI & PLANNING
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a method for conveniently observing the

Method used

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  • Convenient observation method of orchid mycorrhiza microstructure
  • Convenient observation method of orchid mycorrhiza microstructure
  • Convenient observation method of orchid mycorrhiza microstructure

Examples

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Embodiment 1

[0025] Example 1 Isolation and identification of orchid mycorrhizal fungi:

[0026] 1. Separation of root materials: Take the potted Chunlan from Shanghai Academy of Agricultural Sciences and move it to a shaded greenhouse for later use.

[0027] 2. Separation and identification medium:

[0028] Use malt extract medium (MEA), the formula is: malt extract 20g, tryptone 1g, glucose 20g, agar 20g, distilled water to 1000ml, then autoclave at 121°C for 20 minutes, cool to 60 When the temperature is below 0.03 g streptomycin is added.

[0029] 3. Separation and morphological identification methods:

[0030] Isolation of mycorrhizal fungal strains: take several sections of roots of fresh and healthy orchid plants, rinse them under tap water until there is no sludge on the surface of the roots. Cut the roots into several small sections about 10cm long, wash them three times with sterile water, soak the filter paper in 75% alcohol for 60s, then immerse in 2% sodium hypochlorite for...

Embodiment 2

[0044] Example 2 Screening of suitable dyeing methods

[0045] Take out the orchid tissue culture seedlings, cut off the 3-6 cm long roots with a blade, rinse them with deionized water, blot the water and fix them in FAA fixative solution for more than 24 hours. Prepare a petri dish containing deionized water, make freehand slices of the fixed roots, the thickness of the slices is less than 1cm, pick thinner slices with uniform thickness and put them into the collection tube, immerse in 10% KOH solution, choose 10% KOH Different time gradients for dissociation of the dissociation solution, specifically 5min, 10min, 30min and 60min. Then place them in a 90°C water bath for 10 minutes, pour off the hydrolyzate, add 2% trypan blue staining solution, place them at room temperature and immerse them in different time gradients, specifically 6h, 12h and 24h, and put the experimental materials Put it into a glass slide, drip a drop of glycerin, cover with a cover glass at an angle of...

Embodiment 3

[0050] 1. Orchid tissue culture seedling materials: collect tissue culture seedlings of orchid "Red Beauty", the selected tissue culture seedlings are basically the same, with a fresh weight of about 0.6g and a plant height of about 8cm.

[0051] 2. Cultivation substrate: choose peat that is fully sterilized and dried: yellow sand mixed in a certain proportion as the cultivation substrate, the specific formula is peat: yellow sand 1:2, pour MMN liquid medium, 500mL / 1500g cultivation substrate, mix After uniformity, it is divided into 640mL cultivation flats, autoclaved and cooled to room temperature.

[0052] 3. Inoculation method: transplant 3 orchid seedlings per bottle under aseptic conditions, inoculate 6 bottles of orchid seedlings for each treatment as a repetition, and place them in the tissue culture room for aseptic culture. After one week of growth, check for contamination, and inoculate the strains to be tested in non-polluted tissue culture bottles, that is, place ...

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Abstract

A convenient observation method of an orchid mycorrhiza microstructure comprises the following steps: Step 1, preparing a root segment material, a stationary liquid, a dissociation buffer and a staining solution; Step 2, fixing the root segment, and completely immersing the orchid root segment in the stationary liquid; Step 3, slicing the root segment, and shifting the sliced root segment into a clear water-containing container for later use; Step 4, a step of dissociation of the root material: immersing the root material into a KOH solution for dissociation; Step 5, a staining step of the root material: the dissociated root material is placed into a trypan blue solution for staining; and Step 6, carrying out preforming and mounting on the root material, and carrying out microscopic examination with an optical microscope. According to the invention, whether cells in sight are infected by mycorrhiza fungi can be observed clearly, and whether orchid root is infected and the infection degree are detected by statistics of the proportion of the infected cells in sight in total cells in sight. Thus, essential data is provided for the research on coupled growth of orchid.

Description

Technical field: [0001] The invention relates to the field of agriculture, in particular to a mycorrhizal plant of the Orchidaceae plant, in particular to a method for conveniently observing the microscopic structure of the mycorrhizal plant of the Orchidaceae plant. Background technique: [0002] Orchidaceae are elegant and fragrant, and they are precious flowers and medicinal plants with high ornamental value and economic value. With the rapid development of flower industry and natural medicine industry, the demand for orchidaceae resources is increasing, but Many orchid plants have weak reproductive ability and slow growth (Chen Ruirui, Lin Xiangui, Shi Yaqin, "Research Progress of Orchidaceae Mycorrhiza", "Journal of Applied Environmental Biology", 2003, 9(1): 97-101). [0003] At present, the propagation of orchids mostly adopts the method of tissue culture, but the growth of transplanted aseptic seedlings is slow, and the growth is obviously inferior to the plants grow...

Claims

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Application Information

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IPC IPC(8): G01N21/84G01N1/28G01N1/30
Inventor 黄芳张春英王强尹丽娟高建红张杰
Owner SHANGHAI ACADEMY OF LANDSCAPE ARCHITECTURE SCI & PLANNING
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