A kind of salt stress resistant fungal strain and its breeding method and its application

A salt stress and strain technology, applied in the application, fungi, horticultural methods and other directions, can solve the problems such as limitations, the production and application methods of salt stress resistant fungi have not been reported, and achieve high yield, good stress resistance, and promote plant growth. effect of growth

Active Publication Date: 2017-11-10
北京瑞斯达生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research on the symbiotic relationship between endophytic fungi and plants has become a hot spot, but most of the research is limited to biological phenomena. More valuable salt-resistant endophytic fungi are yet to be discovered, and the production and application of salt stress-resistant fungi for production applications The method has not been reported

Method used

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  • A kind of salt stress resistant fungal strain and its breeding method and its application
  • A kind of salt stress resistant fungal strain and its breeding method and its application
  • A kind of salt stress resistant fungal strain and its breeding method and its application

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Experimental program
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Effect test

Embodiment approach

[0043] The medium used in the present invention is:

[0044] The composition of the liquid seed medium is: NaNO 3 2 g, K 2 HPO 4 1 g, KCl 0.5 g, MgSO 4 0.5 g, FeSO 4 0.01 g, 30 g of sucrose, 1 g of yeast extract, dissolved in water to 1000 ml, aliquoted, sterilized at 121°C for 20 min, and then used.

[0045] The composition of the primary screening medium is as follows: peeled potatoes (200 g), chopped into cubes, boiled with water for 30 min, filtered off the residue with gauze, and added water to the filtrate to make up to 1000 ml; g of glucose was dissolved, dispensed, and sterilized at 121°C for 20 minutes before use. The one without agar is the corresponding liquid medium (PDB).

[0046] Solid medium: bran, 15 g; KH 2 PO 4 , 2 g; MgSO 4 , 0.5 g; distilled water 10 mL, pH 6.0-6.5.

Embodiment 1

[0048] Arthrinium sp. The screening process of NX002:

[0049] Take the root of the higher halophyte Salicornia pysicolor, and use 1% Tween80, 5%H 2 o 2 Treat for 5 minutes respectively, wash with sterile water, grind aseptically and add to liquid seed medium (plus 80ppm streptomycin) for enrichment culture for 1 day, then dilute the enriched culture solution to 10 -4 , take the diluted solution, and spread it on the primary screening medium for plate culture; after 4-7 days of culture, use an inoculation needle to pick out different forms of fungal colonies on the same plate, and then transfer them to the plate for single colony isolation, and finally get multiple strains Bacteria; each strain was inoculated in liquid seed medium and cultured in a constant temperature incubator at 28°C for 7 days. Collect the silk fermentation broth, centrifuge at 5000g for 15min at 4°C. The mycelia and 50ml of pure fermentation broth were immediately stored at -70°C. The mycelium was ...

Embodiment 2

[0051] Arthrinium sp. NX002 is produced and applied to the method for plant resistance to salt stress:

[0052] (1) Fungal culture

[0053] Will Arthrinium sp. NX002 was inoculated on the slant primary screening medium, cultured at 27°C for 2 days, then transferred to liquid seed medium, and cultivated at 27°C for 5 days;

[0054] (2) Treatment of salt stress-resistant fungi and determination of metabolites

[0055] Freeze and centrifuge the fermentation broth at a speed of 5000g for 15min, collect mycelium, inoculate it on a solid medium (which is a fermentation medium), incubate at 27°C for 40 days, freeze-dry the fermentation product, crush it, and dissolve it in 1mL sterile water 10ug to obtain the salt stress-resistant fungal product NX002, which is used to inoculate plants in reverse; the composition of the solid medium is: bran, 15 g; KH 2 PO 4 , 2 g; MgSO 4 , 0.5 g; distilled water 10 mL, pH 6.0-6.5.

[0056] The solid fermentation product is extracted by et...

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Abstract

The invention relates to the fields of microbiology, botany, fermentation engineering, etc., and discloses a fungal strain resistant to salt stress, a breeding method and its application. The strain preservation number is CCTCC No.M2014462, and the classification is named Arthrinium sp NX002; The root of the halophyte Salicornia pilaris is isolated and cultured, and the secondary metabolites of the strain include gibberellin, helmectinol and helmectin. The present invention provides the rDNA sequences of the identified strain cultures. The invention also provides the production medium and culture conditions of the bacterial strain, as well as the application method of the bacterial strain. The stress-resistant endophytic fungus produced by the invention has low production cost, is easy to ferment on a large scale, and can be widely used in the application of plant resistance to salt stress.

Description

technical field [0001] The invention relates to the fields of microbiology, botany, fermentation engineering and the like, and in particular relates to a fungal strain resistant to salt stress and a method for breeding and application thereof. The fungus bred by the invention and its products are mainly used in the fields of plant stress resistance, ecological restoration, biocatalysis and the like. Background technique [0002] Halophytes are stress plants that live in soils that are salinized. It has been widely reported that endophytic fungi improve plant salt tolerance. Frank et al found a fungus isolated from the Indian desert Piriformospora indica After infecting plant roots, it can significantly improve the salt tolerance and yield of monocot plants (PNAS, 2005, 13386–13391); Helmut et al. further found that plant infection Piriformospora indica The improvement of post-salt tolerance is related to the enhancement of antioxidant enzyme activities (DHAR, CAT, MDHAR,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14A01N63/04A01P21/00A01G7/06C12R1/645
CPCA01G7/06A01N63/30C12N1/14C12N1/145C12R2001/645
Inventor 周峰霍光明顾祝军华春
Owner 北京瑞斯达生物科技有限公司
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