Long-term preservation method of mycorrhiza fungus piriformospora indica

A technology of Pyromorpha spp. and mycorrhizal fungi, applied in the field of microbial applications, can solve the problems of loss of plant root symbiosis, easy loss of vitality, and bacterial source pollution, so as to reduce the growth of miscellaneous bacteria, reduce the variation of bacterial sources, and maintain symbiotic ability Effect

Active Publication Date: 2018-04-06
三明市农业科学研究院 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Pyriformis indica can be amplified and preserved in artificial medium without using host plants. The traditional method is often preserved in PDA and other medium, which is easy to lose vitality. It needs to be subcultured once every 2-3 weeks at room temperature, It also needs to be subcultured every 2-3 months for low-temperature storage in the refrigerator at 6°C.
Frequent subculture not only brings labor costs, but also easily causes contamination and variation of bacterial sources
At the same time, due to the long-term preservation of Pyriformis indica in artificial medium, it may lose the ability to symbiosis with plant roots

Method used

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  • Long-term preservation method of mycorrhiza fungus piriformospora indica
  • Long-term preservation method of mycorrhiza fungus piriformospora indica
  • Long-term preservation method of mycorrhiza fungus piriformospora indica

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Preservation medium preparation of Pyrocystis indica

[0030] For each liter of storage medium, add 1 / 2 MS basic medium, sucrose 12g / L, mannose 17g / L, agar powder 6g / L, distilled water to volume, boil, dilute hydrochloric acid to adjust the pH value to 5.8, and pack in 625 mL blue For vases, pack 10 bottles per liter of culture medium, seal with a ventilated cap, sterilize by autoclaving at 121°C for 20 minutes, and cool for later use.

[0031] 2. Preparation of sterile orchids

[0032] In a sterile environment, inoculate sterile 3 cm seedlings of Oncidium with roots on the medium described in 1, place them at 25°C, 2000 lx, and 14 h / d light conditions, and cultivate them for 8 days for later use.

[0033] 3. Inoculation and storage

[0034] Open the oncidium culture bottle under aseptic conditions, and use a puncher to inoculate the vigorous bacterial mass around the ring of Pyromorpha indica cultured in PDA medium for a week to the root of the oncidium plant, and...

Embodiment 2

[0042] 1. Preservation medium preparation of Pyrocystis indica

[0043] For each liter of preservation medium, add 1 / 2 MS basic medium, sucrose 10g / L, mannose 15g / L, agar powder 5g / L, distilled water to volume, boil, dilute hydrochloric acid to adjust the pH value to 5.8, and pack in 625 mL blue For vases, pack 9 bottles per liter of culture medium, seal with a ventilated cap, sterilize by autoclaving at 121°C for 20 minutes, and cool for later use.

[0044] 2. Preparation of sterile orchids

[0045] In a sterile environment, inoculate the sterile clematis stems with roots in the preservation medium, place them at 20°C, 2000 lx, 14 h / d light conditions, and cultivate for 7 days for later use.

[0046] 3. Inoculation and storage

[0047] Under aseptic conditions, open 2 described clematis culture bottles, and use a puncher to inoculate the vigorous bacterium block around the Pyromorpha indica bacterium ring that is newly cultivated in the PDA medium for one week to the root o...

Embodiment 3

[0055] 1. Preservation medium preparation of Pyrocystis indica

[0056] For each liter of storage medium, add 1 / 2 MS basic medium, sucrose 15g / L, mannose 20g / L, agar powder 6g / L, distilled water to volume, boil, dilute hydrochloric acid to adjust the pH value to 5.8, and pack in 625 mL blue For vases, pack 11 bottles per liter of medium, seal them with a 0.5cm air-permeable film, sterilize them by high-pressure steam sterilization at 121°C for 20 minutes, and cool them down for later use.

[0057] 2. Preparation of sterile orchids

[0058] In a sterile environment, inoculate the sterile seedlings of Dendrobium officinale with roots in the preservation medium, place them in 28°C, 2000 lx, 14 h / d light conditions, and cultivate them for 10 days for later use.

[0059] 3. Inoculation and storage

[0060] Under aseptic conditions, open the Dendrobium officinale culture bottle described in 2, and use a puncher to inoculate the vigorous bacterium block around the Pyrosporium indic...

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PUM

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Abstract

The invention provides a long-term preservation method of mycorrhiza fungus piriformospora indica. According to the long-term preservation method, mycorrhiza fungus piriformospora indica is inoculatedwith the root of sterile orchidaceae, a culture medium is optimized and preserved, the purpose of long-term (longer than 12 months) preservation at a normal temperature is achieved, the method has advantage of long-term preservation at normal temperatures when being compared with conventional plate culture medium preservation, the risks of contamination and bacterium source variations caused by frequent subculture are reduced, and meanwhile by recovering culture of a culture medium, bacterial strains with symbiosis properties can be rapidly separated.

Description

technical field [0001] The invention belongs to the field of microorganism application, and in particular relates to a long-term preservation method of mycorrhizal fungus Pyromorpha indica. Background technique [0002] Indian piriformis ( Piriformospora indica ) is an endophytic fungus isolated from the Thar Desert in India, belonging to Basidiomycota, Agaricomycetes, Sebacinales, Sebacinaceae, Pyromorpha ( Piriformospora ), is an endophytic fungus that can colonize the roots of a variety of host plants. Its function is very similar to that of mycorrhizal fungi. It can promote plant growth, accelerate plant absorption of nitrogen, phosphorus and other minerals, and improve crop Tolerance of adversity stress, induction of plant systemic resistance, etc. [0003] Pyriformis indica can be amplified and preserved in artificial medium without using host plants. The traditional method is often preserved in PDA and other medium, which is easy to lose vitality. It needs to be s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04C12N1/14C12R1/645
CPCC12N1/04C12N1/14
Inventor 叶炜邱如永江金兰曹奕鸯李永清张燕芳杨学雷伏贵邱天飚张瑶周建金颜沛沛莫智龙严胜泽
Owner 三明市农业科学研究院
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