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Walnut endophyte and use thereof

An endophyte and walnut technology, applied in the field of applied microorganisms, can solve the problems of increased prevention and control costs, pesticide residues, deterioration of the ecological environment, etc., and achieve the effects of easy colonization and stable prevention and control effects.

Inactive Publication Date: 2015-05-06
SHANXI AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the growth of social economy, people gradually realized a series of problems caused by the long-term and large-scale use of pesticides: firstly, the natural enemies of pests were killed, and the population of microorganisms in the soil decreased, which destroyed the ecological balance; secondly, due to perennial use, pathogenic bacteria and pests produced In addition, the control cost increases and the control effect decreases; there is also the problem of pesticide residues. On the one hand, it leads to the deterioration of the ecological environment, and on the other hand, the pesticide residues in food eventually threaten human health.
Biocontrol strains are isolated from the soil and the rhizosphere of plants. Most of the strains with antagonistic effects can not colonize the surface or body of the host when applied outdoors. At the same time, they are affected by changes in environmental conditions and plant rhizosphere microorganisms under different ecological conditions. Affected by factors such as population changes, its biological control effect is often unstable, and sometimes even ineffective

Method used

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  • Walnut endophyte and use thereof
  • Walnut endophyte and use thereof
  • Walnut endophyte and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Isolation, screening and identification of embodiment 1 walnut endophytic bacteria

[0051] Basal medium: NA medium: peptone 0.5g, beef extract 0.3g, agar 2g, distilled water 100mL, pH7. NB culture medium: liquid medium of NA medium. LB medium: tryptone 10g, NaCl 10g, yeast powder 5g, water 1000mL. PDA medium: fresh potato 20g, glucose 2g, agar 2g, distilled water 100mL. PBA culture medium: liquid medium of PDA medium.

[0052] 1. Isolation of walnut endophytic strains

[0053] Using the tissue separation method, the non-lignified branches, leaves and green fruits collected from the Walnut Resource Garden of the Institute of Fruit Trees, Shanxi Academy of Agricultural Sciences and the Botanical Garden of Shanxi Agricultural University were washed with tap water. Weigh 3g according to different collection sites and different collection sites, first disinfect the surface with 75% alcohol solution for 1-2min, then disinfect with sodium hypochlorite solution for 0.5-1mi...

Embodiment 2

[0096] The bacteriostasis of embodiment 2 walnut endophytic strain Ht-q6 to walnut anthracnose

[0097] Walnut endophytic strain: Bacillus amyloliquefaciens Ht-q6, pathogenic fungus: walnut anthracnose (G.fructigenum).

[0098] Culture medium: NA, NB, PDA, PBA refer to the basal medium in Example 1; Chapi culture medium: NaNO 3 2g, K 2 HPO 4 1g, KCl0.5g, MgSO 4 ·7H 2 O, FeSO 4 0.01g, 30g sucrose, 1000mL distilled water.

[0099] 1. Preparation of sterile filtrate of walnut endophytic strain Ht-q6

[0100] Transfer 1mL of the Ht-q6 strain stored at -20°C to 10mL / 150mL NB culture medium, 28°C, 170r min -1 Shake culture overnight to obtain seed solution. Transfer the seed liquid to the new NB culture liquid according to the inoculum amount of 1 mL, shake and culture at 28 °C and 170 r·min-1 for 48 h to obtain the fermentation liquid. Place the fermentation broth in a refrigerated high-speed centrifuge at 4°C, 12000r min -1 Centrifuge for 10 minutes to remove the preci...

Embodiment 3

[0128] Example 3 The optimization of the fermentation conditions for the production of metabolites by the antagonistic strain Ht-q6

[0129] Seed culture solution: 1% peptone, 1% glucose, 0.2% NaH 2 PO 4 2H 2 O, 0.4% Na 2 HPO 4 2H 2 O, 0.05% MgSO 4 ·7H 2 O, 100mL distilled water, pH7.0-7.2.

[0130] Basic fermentation medium: 1% peptone, 1% glucose, 0.2% NaH 2 PO 4 2H 2 O, 0.4% Na 2 HPO 4 2H 2 O, 0.05% MgSO 4 ·7H 2 O, 0.02% CaCl 2 , 100mL distilled water, pH7.0-7.2.

[0131] NB culture medium was used for the activation of antagonistic strains, and PDA medium and PBA culture medium were used for the cultivation of walnut anthracnose bacteria. Reagents: glucose, D-fructose, yeast extract powder, mung bean powder, KCl, NaCl, etc.

[0132] 1. Preparation of seed solution

[0133] Draw 1mL of the activated Ht-q6 strain and inoculate it into 50mL / 250mL seed culture solution, 28°C, 180r·min -1 Constant temperature shaking culture 16-24h.

[0134] Second, the det...

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Abstract

The invention discloses walnut endophyte and a use thereof. The walnut endophyte is Bacillus amyloliquefaciens Ht-q6, is preserved in China general microbiological culture collection center in No.2, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Z. C. 100101 and has a preservation number of 9401. The walnut endophyte is separated and selected from walnut green peel, is Bacillus amyloliquefaciens and is named as Ht-q6. The walnut endophyte has antagonistic action on plant pathogenic fungi. The invention determines the optimal formula and fermentation conditions of the fermentation nutrient solution and provides a bacterial strain resource and a theoretical basis for development of an antibiological inoculants. The walnut endophyte is separated from plant tissue so that the endophyte easily colonize in plants and especially has stable effects of prevention and control of walnut diseases.

Description

technical field [0001] The invention belongs to the technical field of applied microorganisms, and in particular relates to a walnut endophyte and its application. Background technique [0002] For a long time, plant diseases and insect pests have been one of the main culprits threatening agricultural production. Chemical agents have been widely used since the 1970s due to their quick effect, low cost, and convenient use, although they have a significant effect on increasing the yield of crops. However, with the growth of social economy, people gradually realized a series of problems caused by the long-term and large-scale use of pesticides: firstly, the natural enemies of pests were killed, and the population of microorganisms in the soil decreased, which destroyed the ecological balance; secondly, due to perennial use, pathogenic bacteria and pests produced In addition, there is also the problem of pesticide residues. On the one hand, it leads to the deterioration of the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P3/00C12R1/07
CPCA01N63/00A01N63/10C12N1/205C12R2001/07
Inventor 王美琴胡春艳赵立娟张红娟吴红玉郝晓娟刘慧平韩巨才
Owner SHANXI AGRI UNIV
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