Pseudomonas chloroaphis with effects of effective zea mays sheath blight control and zea mays growth promotion

A technology for Pseudomonas chloropinus and corn sheath blight, applied in the field of microorganisms, can solve problems such as adverse effects on human and animal health, destruction of ecological balance, and unreasonable large-scale use of chemical pesticides.

Inactive Publication Date: 2014-03-05
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the comprehensive control of agricultural pests and diseases, chemical control plays an important role, but the long-term and irrational u...

Method used

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  • Pseudomonas chloroaphis with effects of effective zea mays sheath blight control and zea mays growth promotion
  • Pseudomonas chloroaphis with effects of effective zea mays sheath blight control and zea mays growth promotion
  • Pseudomonas chloroaphis with effects of effective zea mays sheath blight control and zea mays growth promotion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Isolation and preliminary screening of rhizosphere growth-promoting bacteria

[0026] The method of Fang Zhongda (1979) was used to isolate the growth-promoting bacteria in the rhizosphere. Take 10 g of the collected soil sample, add it to a conical flask filled with 90 mL of sterile water, place the conical flask on a shaker, and vibrate at 180 r / min for 20 min to make a soil suspension. Let the prepared soil suspension stand for 5 minutes, take the supernatant, and use the 10-fold serial dilution method to successively dilute to 10 -2 、10 -3 、10 -4 and 10 -5 , pipette 200 μL of each concentration on the NA plate, spread evenly, and repeat each concentration three times. Then the coated NA plates were incubated in a constant temperature incubator at 28°C for 24h.

[0027] When bacterial colonies appear on the NA plate, Geotrichum candidum (Geotrichum candidum) spore suspension (10 5 spores / mL) evenly sprayed on the NA plates that have grown colonies. ...

Embodiment 2

[0029] Embodiment 2: the mensuration of antibacterial spectrum of rhizosphere growth-promoting bacteria

[0030] Use the confrontation culture method (Sijam et al., 2005). In the center of a PDA plate with a diameter of 90mm, connect bacteria cakes with a diameter of 9mm of corn sheath blight Rhizoctonia solani, wheat head blight Gibberella zeae and corn small spot fungus Bipolaris maydis, then seal the plate with a sealing film, and place it at 28°C Cultured in the incubator for 2 days. Dip the sterilized filter paper with a diameter of 6mm into the prepared bacterial solution (10 8 cfu / mL), moved to a distance of 15mm from the edge of the pathogenic bacteria, placed symmetrically, and the control was only connected to the bacterial cake of the pathogenic bacteria, sealed, and finally placed in a 28°C incubator for cultivation. Each treatment was repeated 3 times. After 3 days, measure the colony diameter of the pathogenic bacteria at the place where the bacteria are conne...

Embodiment 3

[0032] Embodiment 3: the acquisition of bacterial strain 16S rRNA sequence

[0033] Using the genome of Pseudomonas choloaphis as a template, its 16S rDNA was amplified by PCR. The upstream and downstream primers used were 16S-F: 5′-AGAGTTTGATYMTGGCTCAG-3′, 16S-R: 5′-AGGGYTACCTTGTTACGACTT-3′, respectively. The PCR reaction program was 94°C for 3min; 30 cycles of 94°C for 30s, 54°C for 45s, and 72°C for 2min; extension at 72°C for 10min; storage at 4°C. After the reaction, the PCR products were placed on a 1.0% agarose gel for electrophoresis analysis. The PCR product was recovered with reference to the PCR Purification and Recovery Kit of AXYGEN, USA, and ligated with pMD18-T overnight in a water bath at 16°C. The ligation reaction system is: 10×T4DNA Ligase buffer 1μL, pMD18-T (50ng / μL) 0.3μL, target DNA 7.7μL, T4DNA Ligase (350U / μL) 1μL.

[0034] The ligation product was transformed into competent cells by heat shock method. The specific method is: put 100 μL of E. coli ...

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Abstract

The invention relates to a strain of Pseudomonas chloroaphis with effects of effective zea mays sheath blight control and zea mays growth promotion, and belongs to the technical field of microorganisms. The strain is Pseudomonas chloroaphis PSJ1, is isolated from wheat rhizosphere, and has the preservation number of CGMCC NO.7932. The Pseudomonas chloroaphis PSJ1 has effects of zea mays growth promotion, Rhizoctonia solani growth inhibition, and sheath blight occurrence reduction.

Description

technical field [0001] The invention belongs to the field of microorganisms, and relates to a kind of Pseudomonas chlororaphis and its application, in particular to a kind of Pseudomonas chlororaphis which can effectively inhibit Rhizoctonia solani (Rhizoctonia solani) and can promote the growth of corn . Background technique [0002] Corn (Zea mays) is the main grain, feed crop and important industrial raw material in my country. It occupies an important position in my country's national economy and agricultural production. Corn production is directly related to my country's food strategic security. In recent years, diseases such as corn sheath blight, small spot and Curvularia leaf spot have become more and more common, and the damage is increasing, which seriously restricts the development of corn production. [0003] In the comprehensive control of agricultural pests and diseases, chemical control plays an important role, but the long-term and irrational use of chemical...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01P3/00C12R1/38
Inventor 李向东王晓强毕涛
Owner SHANDONG AGRICULTURAL UNIVERSITY
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