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Bacterial chemotactic substance screening glass slide and application thereof

A technology of glass slides and chemoattractants, which is applied in the field of plant-microbe interaction research, can solve the problems of high cost of microfluidic chip devices, large errors in capillary-like methods, and long experimental cycles of swimming plate methods. To achieve the effect of shortening the experimental cycle, reducing steps, and reducing errors

Pending Publication Date: 2022-03-11
QIQIHAR UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the experimental cycle of the swimming plate method is longer, the error of the capillary-like method is larger, and the cost of the microfluidic chip device is larger (Ling, N. et al. Identification and role of organic acids in watermelon rootexudates for recruiting Paenibacillus polymyxa SQR -21in therhizosphere.European Journal of Soil Biology.2011,47(6):374-379)

Method used

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  • Bacterial chemotactic substance screening glass slide and application thereof
  • Bacterial chemotactic substance screening glass slide and application thereof
  • Bacterial chemotactic substance screening glass slide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Preparation of Bacterial Chemotactic Substance Screening Slides

[0030] The bacterial chemoattractant material screening glass slide 4 provided by the present embodiment is made of glass, with a length of 8 cm, a width of 5.5 cm, and a thickness of 1 cm. There are seven cylindrical grooves on the surface, including a central groove 2 and surrounding all The 6 peripheral grooves 1 of the central groove 2 described above have a groove diameter of 0.5 cm and a height of 0.5 cm, and a connecting groove 3 with a width of 0.1 cm is passed between the central groove 2 and each peripheral groove 1 Connection, the depth of the connection groove 3 is also 0.5cm. Among them, the central groove 2 is used to drop the bacterial solution in the experiment, the four peripheral grooves 1 are used to drop the chemoattractant solution, and the remaining two peripheral grooves 1 are used to drop sterile water and 0.9% w / v NaCl respectively as negative and positive controls. Th...

Embodiment 2

[0031] The collection of embodiment 2 rice rhizosphere exudates

[0032] 1. Test materials

[0033]The strain used is Bacillus altitudinis LZP02 isolated from rice rhizosphere soil in Northeast China. It is preserved in the China Center for Type Culture Collection at Wuhan University. . The bacterial strain has been recorded in a patent application whose publication number is CN109536401A, and the title of the invention is a compound microbial fertilizer and its preparation method and its application in promoting rice growth. B.altitudinisLZP02 in LB medium (peptone, 10g L -1 ; Sodium chloride, 8g L -1 and yeast extract 5g L -1 ) for 10 h, centrifuged at 10000 rpm and 4° C. for 2 min, and stored at -80° C. with 15% glycerol for future use. Rice seeds (Longgeng 46 rice) were provided by the Rice Research Institute of Heilongjiang Academy of Agricultural Sciences.

[0034] 2. Test method

[0035] 2.1 Cultivate rice seedlings

[0036] Rice seeds were sterilized with 30% h...

Embodiment 3

[0046] Embodiment 3 utilizes swimming plate method to carry out chemotaxis test to 16 kinds of chemoattractants

[0047] 1. Test materials: The bacterial strains used are the same as in Example 2, and the reagents used are all standard products with a content of more than 98%, all purchased from Nanjing Jiancheng Bioengineering Research Institute.

[0048] 2. Test method

[0049] Prepare swimming medium (peptone, 10g / L; sodium chloride, 8g / L; agar 4%). The culture medium was sterilized at 121°C for 20min, and then cooled to 40°C-50°C. In the experimental group, different dilutions of chemoattractant aqueous solutions were added to the culture medium 1:1 until the final concentrations were 25 μM, 50 μM, and 100 μM. The control group was prepared with an equal amount of sterile water. Dispense the medium into Petri dishes after mixing well, keep inverted and allow to stand at room temperature. Next, take cultures to OD 600 =0.5 B.altitudinis LZP02 bacterial solution 5μL on ...

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Abstract

The invention discloses a bacterial chemotactic substance screening glass slide and application thereof. A cylindrical central groove is formed in the surface of the glass slide, one or more cylindrical peripheral grooves are further formed around the central groove, and the central groove is connected with each peripheral groove through a connecting groove. When the glass slide is used as a chemotactic experiment device, a large number of complex steps can be reduced. And through multiple comparisons of experimental results, the result of the chemotactic experiment carried out by using the device is consistent with that of a traditional swimming plate method, and meanwhile, multiple defects of the traditional swimming plate method can be overcome. For example, the error of plate counting is reduced, the experimental period is greatly shortened, and the cost of experimental materials is reduced. The invention provides a new technical means for screening bacterial chemotactic substances.

Description

technical field [0001] The invention relates to a screening glass slide for bacterial chemotactic substances, which can quickly identify chemotactic substances that can recruit bacteria in plant rhizosphere exudates, and also relates to its application in the study of plant-microbe interaction. Background technique [0002] The chemotaxis of plant growth-promoting rhizobacteria (PGPR) to plant rhizosphere exudates is of great significance in providing beneficial functions for plant-microbe interactions (Shen Yifei, E Yaoyao, Yang Fang. Watermelon root exudates Effects of Amino Acid Components on Chemotaxis and Rhizosphere Colonization of Paenibacillus polymyxa SQR-21[J]. Journal of Nanjing Agricultural University, 2017, 40(01): 101-108; Belas, R. Biofilms, flagella , and mechanosensing of surfaces by bacteria. Trends Microbiol. 2014, 22(9):517-527). [0003] A class of low-molecular-weight compounds in plant root exudates induce chemotactic movement of PGPR to the root surf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12Q1/06
CPCC12M23/04C12M23/38C12Q1/06
Inventor 王志刚焦慧文徐伟慧胡云龙陈文晶
Owner QIQIHAR UNIVERSITY
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