B. vallismortis HSB-2 and application thereof

A technology of HSB-2 and grain sprouts, which is applied to Bacillus spp. HSB-2 and its application fields, can solve the problems of few bacillus, and achieve the effect of reducing continuous cropping obstacles and promoting growth.

Active Publication Date: 2019-02-22
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are few studies on the prevention and ...

Method used

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  • B. vallismortis HSB-2 and application thereof
  • B. vallismortis HSB-2 and application thereof
  • B. vallismortis HSB-2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Isolation, purification and screening of Bacillus death valley HSB-2

[0033] 1. Isolation and purification of antagonistic strains

[0034] Rhizosphere bacteria were separated by the following modified dilution coating plate method: Fresh soil samples were passed through a sieve with a diameter of 3-4 mm to remove impurities in the soil. Weigh 5 g of rhizosphere soil, add it to a triangular flask (with glass beads) filled with 45 mL of sterile water, vibrate at 180 rpm for 30 min, let it stand for 5 min, and serially dilute to 1×10 with sterile water. -4100 μL of the supernatant of the gradient dilution was spread on the LB plate, repeated three times, and cultured upside down in a constant temperature incubator at 28°C. After a single colony grows in the plate, pick a single colony of different shapes and streak on a new LB plate for purification. At the same time, all purified single colonies were inoculated in liquid LB medium (tryptone 10.0g, yeast extract 5.0g, ...

Embodiment 2

[0044] Taxonomic identification of HSB-2 strain

[0045] 1. Morphological and physiological and biochemical identification

[0046] The isolated HSB-2 strain was cultured on LB medium at 28°C for 24 hours, and when a single colony appeared, the morphological characteristics of the colony were observed. Crystal violet was used for Gram staining, and the shape and size of bacteria were observed with Nikon fluorescence microscope BX-51 and scanning electron microscope SU-8010. For the scanning electron microscope sample preparation process, refer to the method of Lin Ying et al. Physiological and biochemical characteristics were analyzed according to the methods of "Bergey's Bacterial Identification Manual (Second Edition)" and "Common Bacterial System Identification Manual". Each index was detected 3 times, and the test was repeated 2 times.

[0047] like figure 1 As shown, after the HSB-2 bacterial strain was cultured on the LB medium for 24 hours, the single colony was milky...

Embodiment 3

[0061] Double antibody labeling and colonization of antagonistic bacteria

[0062] 1. Double antibody labeling of antagonistic bacteria

[0063] With reference to the method of Chen Changqing et al., the method was modified, and the strain HSB-2 was first applied to the -1 After growing a single colony, select a resistant strain with good growth and the same shape as the original colony and transfer it to the next double concentration LB plate, and gradually screen out the strains containing 350 μg·mL -1 Stable growth of the resistant strain HSB-2 on the rifampicin LB plate, on this basis, use the same method to induce the resistance of the resistant strain HSB-2 to streptomycin sulfate, the initial streptomycin sulfate mass The concentration is 0.5μg·mL -1 , gradually increase the concentration of streptomycin sulfate to 320 μg·mL -1 , and finally obtain the double antibacterial strain of HSB-2.

[0064] 2. Detection of genetic stability of HSB-2 double antibacterial stra...

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Abstract

The invention relates to B. vallismortis HSB-2 and application thereof. The preservation number of the B. vallismortis HSB-2 is CGMCC No.15305, and the sequence of 16S rDNA is shown as SEQ.ID.NO.1. The invention further relates to the application of the B. vallismortis HSB-2 to preparation of bacterial fertilizer for relieving the successive cropping obstacle of apples. The B. vallismortis HSB-2 has an obvious antagonistic effect on four types of Fusarium fusarium pathogenic bacteria causing the successive cropping obstacle of the apples; the bacterial fertilizer manufactured through the B. vallismortis HSB-2 can promote growing of the overground parts and the underground parts of Malus malus hupehensis seedlings, and at the rhizosphere, colonization can be stabilized, and growing of the four types of Fusarium fusarium pathogenic bacteria of solani, proliferatum, verticillioides and oxysporum, causing the successive cropping obstacle of the apples, in soil can be remarkably restrained;and activity of soil urease, phosphatase, sucrase and root system protective enzymes, namely SOD, POD and CAT can be remarkably improved, and the successive cropping obstacle can be relieved.

Description

technical field [0001] The invention relates to a bacillus dead valley HSB-2 and an application thereof, belonging to the field of fruit tree cultivation. Background technique [0002] Limited by land resources, the phenomenon of apple continuous cropping is common, which leads to the occurrence of apple continuous cropping obstacles. The study found that the main pathogenic bacteria causing apple continuous cropping obstacles in Italy were Fusarium solani (Fusarium solani) and F. The main pathogens of apple continuous cropping obstacles in the Bohai Bay area; Xu Wenfeng and Liu Zhi respectively isolated a large number of F. oxysporum (F.oxysporum) and Verticillium verticillium (F.verticillioides), Fusarium solani (F.solani) and Fusarium proliferatum (F.proliferatum), and verified that they are highly pathogenic to Pingyi sweet tea seedlings. Verticillium, rot and Fusarium exfoliates are harmful pathogenic fungi that hinder apple continuous cropping. [0003] At present, ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F11/08C12R1/07
CPCC05F11/08C12N1/205C12R2001/07
Inventor 尹承苗段亚楠毛志泉陈学森沈向胡艳丽
Owner SHANDONG AGRICULTURAL UNIVERSITY
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