Single-spore propagation method for Gigaspora rosea mycorrhiza fungi

A technology of giant cyst mold and fungus, which is applied in the field of single spore propagation of giant cyst mold rhizogenes, can solve problems such as increased labor costs, inability to know the infection status of inoculated plants mycorrhiza, and inability to monitor the cultivation process in real time.

Inactive Publication Date: 2020-07-14
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional potted propagation method involves the process of seedling raising, inoculation and later plant growth management, and the operation is relatively complicated
Importantly, the entire cultivation process after inoculation cannot be monitored in real time, and the mycorrhizal infection status of the inoculated plants cannot be known until the final sample is collected
When the single-spore inoculation method is adopted and there are many inoculated plants, but the proportion of plants actually infected is not high, the potted method will undoubtedly increase a lot of unnecessary labor costs
In addition, the traditional potting method mostly adopts an open or semi-open greenhouse environment, and the proliferating bacteria are easily polluted by other miscellaneous bacteria.

Method used

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  • Single-spore propagation method for Gigaspora rosea mycorrhiza fungi
  • Single-spore propagation method for Gigaspora rosea mycorrhiza fungi
  • Single-spore propagation method for Gigaspora rosea mycorrhiza fungi

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Preparation: Mycorrhizal fungus agent of Gigaspora rosea, which is obtained from polyspore propagation by conventional pot planting method. It is a mixture of spores, hyphae, infected root segments and soil and sand substrates. It is approved by the French National Scientific Research Institute. Provided by Professor Christophe ROUX of the center; carrot hairy roots, provided by Mr. Li Airong, Kunming Institute of Botany, Chinese Academy of Sciences, and preserved in our research group with M medium.

[0035] (1) Separation, screening and surface disinfection of the mycorrhizal fungus spores of Megacystis rosea rubrum: Weigh 5 g of the fungal agent, and use the wet sieve decantation method to collect the spores between the 80 mesh and 180 mesh sampling sieves; , use a pipette to absorb spores with relatively uniform color and texture for surface disinfection; the disinfection method is: soaking in 0.02g / ml ammonium chloride T solution on ice twice, each time for 10min, a...

Embodiment 2

[0047] According to the method of embodiment 1, carry out the mycorrhizal fungus propagation of Megacystis rosea rubrum, the difference is: step (4) has chosen another high-yield spore single-spore propagation line, and numbering is GRspore-3 to carry out enlarged cultivation (expanded cultivation effect is as follows figure 1 shown on the right).

[0048]The detection of the number of spores in the multiplication spore suspension: in the ultra-clean bench, use a sterile dropper to respectively draw 1ml of the multiplication spore suspension obtained in the step (5) of embodiment 1 and embodiment 2 and place it in two petri dishes, add an appropriate amount of After dilution with distilled water, the number of spores was counted under a stereomicroscope, and the results are shown in Table 1. Wherein, the formula for calculating the total number of spores for propagation is: number of spores in 1 ml of suspension × volume of total spore suspension (ml).

[0049] Detection of t...

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Abstract

The invention provides a single-spore propagation method for Gigaspora rosea mycorrhiza fungi. The method includes the following steps: inoculating the Gigaspora rosea mycorrhiza fungi single spores into a sucrose-free M culture medium for culture to obtain germinated single spores; inoculating the germinated single spores into a carrot hairy root-M culture medium system, culturing and screening to obtain a high-spore-yield Gigaspora rosea mycorrhiza fungi single-spore propagation system; re-inoculating the single-spore propagation system into a carrot hairy root-M culture medium system, and expanding culture to obtain a solid culture; and mixing and incubating the solid culture with a gel dissolving solution, leaching and sieving to obtain a propagated Gigaspora rosea mycorrhiza fungi spore suspension. The method can quickly obtain a large number of progeny spores from a single spore, has good expanded spore germination and good host infection activity, and can provide ideal inoculantmaterials for basic research and application research such as arbuscular mycorrhizal fungi genetics and molecular biology.

Description

technical field [0001] The invention belongs to the technical field of microorganism culture, and in particular relates to a single-spore propagation method of the mycorrhizal fungus Megacystis roseum. Background technique [0002] Arbuscular mycorrhizal fungi (AMF) can significantly promote the uptake of soil phosphorus by host plants, so they have the potential to be used as biofertilizers. In addition, research on the interaction between arbuscular mycorrhizal fungi and host plants is still one of the current biological research hotspots. The current basic and applied research on arbuscular mycorrhizal fungi mainly focuses on a few species of Glomerales, which is mainly related to the ease of cultivation and propagation of species in this order. In fact, there are many species of arbuscular mycorrhizal fungi, and there may be significant physiological and ecological differences between each branch. Therefore, its basic and related applied research needs to include more s...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N3/00C12R1/645
CPCC12N1/14C12N3/00
Inventor 唐年武许雯珺于富强
Owner KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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