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A rhizosphere probiotic strain Klebsiella zh07 and its application

A Klebsiella, ZH07 technology, applied in the field of microorganisms, can solve the problems affecting seed germination, plant growth, peanut production reduction, etc.

Active Publication Date: 2022-04-08
SHANDONG AGRICULTURAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the existing studies, there are six main autotoxic substances secreted by peanut roots, namely: benzoic acid, p-hydroxybenzoic acid, salicylic acid, cinnamic acid, coumaric acid and vanillic acid. Toxic effect, affecting seed germination, plant growth, causing a large reduction in peanut yield

Method used

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  • A rhizosphere probiotic strain Klebsiella zh07 and its application
  • A rhizosphere probiotic strain Klebsiella zh07 and its application
  • A rhizosphere probiotic strain Klebsiella zh07 and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Example 1: Isolation and identification of microorganisms degrading peanut autotoxic substances

[0026] The isolation of microorganisms degrading peanut autotoxic substances mainly includes four parts: enrichment, selective culture, purification and domestication.

[0027] (1) Enrichment

[0028] Obtain the soil of continuous cropping peanuts in saline-alkali land for ten years, directly inoculate 1g of peanut soil into 100ml of inorganic salt medium containing 1g / L benzoic acid, shake at 30°C and 160r / min. In this way, the strains capable of degrading benzoic acid can be enriched, which is convenient for subsequent selective cultivation and purification. Similarly, microorganisms that use p-hydroxybenzoic acid, salicylic acid, cinnamic acid, vanillic acid and coumaric acid as the only carbon source can be enriched. The experiment uses inorganic salt medium, the composition includes (NH 4 ) 2 SO 4 2g / L, KH 2 PO 4 2g / L, Na 2 HPO 4 1.3g / L, carbon source 1g / L, ...

Embodiment 2

[0048] Embodiment 2: Determination of the degradation rate of peanut autotoxic substances

[0049] The degradation rate was determined by taking six kinds of autotoxic substances as the sole carbon source, absorbing 10 μL of the strains in the -80 °C refrigerator and adding them to 5 mL of LB liquid medium for activation, and culturing them on a shaker at 37 °C for 12 hours to complete the activation. Adjust OD with LB medium 600 to 0.5, take 1mL OD 600 The bacterial solution with a concentration of 0.5 was added to 49 mL of inorganic salt culture solution (1 mL of sterile water was added for the control), and the corresponding autotoxic substance was used as the only carbon source in the culture solution at a concentration of 5 g / L, and cultured on a shaking table at 37°C. Taking LB liquid medium as a control, take 1mL culture medium every 12h to measure OD 600 The value is taken as the relative number of degrading bacteria proliferation. Take another small amount of bacte...

Embodiment 3

[0055] Embodiment 3: Determination of promoting growth of rhizosphere probiotics to peanut growth

[0056] Greenhouse pot experiments were used to verify the ability of Klebsiella ZH07 to promote peanut growth.

[0057] The ratio of nutrient solution is as follows (1L): ferric citrate 0.075g, Ca(NO3) 2 0.03g, KCL 0.075g, MgSO 4 ·7H 2 O 0.06g, K 2 HPO 4 0.136g, CaSO 4 0.46g, trace elements 1mL.

[0058] The ratio of trace elements is as follows (1L): H 3 BO 3 2.86g, MnSO 4 1.81g, CuSO 4 ·5H 2 O 0.8g, ZnSO 4 0.22g, H 2 MoO 4 0.02g.

[0059] Disinfect peanut seeds with 0.1% sodium hypochlorite solution for 10 min, and then rinse the seeds with sterile water three times. The washed seeds were soaked in sterile water for 10 hours to fully absorb water. Mix the nutrient solution into the sterilized vermiculite to reach a wet state, then divide it into plastic cups, about 100g per cup, put a peanut seed in the middle of the vermiculite, and uniformly adjust the ...

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Abstract

The invention discloses a rhizosphere probiotic Klebsiella ZH07, which is a microorganism capable of degrading peanut autotoxic substances isolated from peanut soil that has been continuously cropped for ten years. The Klebsiella ZH07 provided by the present invention has degradative effect on peanut autotoxic phenolic acids including benzoic acid, p-hydroxybenzoic acid, salicylic acid, cinnamic acid, coumaric acid and vanillic acid, and has degradative effect on six autotoxic phenolic acids. The degradation rates of toxic substances are all over 50%; after being cultured with the Klebsiella ZH07 bacteria solution, the dry weight of peanuts increases by about 32%, the seedling height of the aboveground part increases by about 32%, and the root length of the underground part increases by about 6%. %, it can be seen that Klebsiella ZH07 has an obvious growth-promoting effect on peanuts and can promote the growth of peanut plants.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a rhizosphere probiotic Klebsiella ZH07 and an application thereof. Background technique [0002] As one of the four major oil crops in my country, peanut is an important economic crop. China has always been the world's largest peanut producer, consumer and exporter. In recent years, with the adjustment of agricultural planting structure, peanut planting has become large-scale and intensive. The continuous cropping area is getting larger and larger, and the continuous cropping obstacle has become one of the important factors for the low and unstable peanut yield. During peanut continuous cropping, the plants are thin, short, poor in growth, pods become smaller, quality decline, total biomass and pod yield are significantly reduced. At present, the problem of peanut continuous cropping obstacles seriously threatens the sustainable development of peanut planting and related...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/20A01P21/00C12R1/22
CPCC12N1/20A01N63/20
Inventor 解志红王丹丹殷志秋孙丽孔波王洪凤王庆彬
Owner SHANDONG AGRICULTURAL UNIVERSITY
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