Field monitoring of plant-growth-promoting rhizobacteria by colony immunoblotting

Author:

Krishnen Ganisan12,Kecskés Mihály L.1,Rose Michael T.1,Geelan-Small Peter1,Amprayn Khanok-on1,Pereg Lily3,Kennedy Ivan R.1

Affiliation:

1. SUNFix Centre for Nitrogen Fixation, Faculty of Agriculture, Food and Natural Resources, Biomedical Building, 1 Central Avenue, Eveleigh, The University of Sydney, New South Wales 2006, Australia.

2. Strategic Resources Research Centre, MARDI Head Quarters, 43400 Serdang, Selangor, Malaysia.

3. School of Science and Technology, University of New England, Armidale New South Wales, Australia.

Abstract

Inoculant plant-growth-promoting bacteria are emerging as an important component of sustainable agriculture. There is a need to develop inexpensive methods for enumerating these organisms after their application in the field, to better understand their survival and impacts on yields. Immunoblotting is one potential method to measure viable cells, but the high cost of the conventionally used nylon membranes makes this method prohibitive. In this study, less expensive alternative materials such as filter papers, glossy photo papers, and transparencies for the purpose of colony immunoblotting were evaluated and the best substance was chosen for further studies. Whatman filter paper No. 541 combined with a 0.01 mol·L–1 H2SO4 rinsing step gave similar results to nylon membranes but <20% of the overall cost of the original colony immunoblotting assay. The application of the modified immunoblot method was tested on nonsterile clay soil samples that were spiked with high numbers (>107 CFU·g–1) of the plant-growth-promoting bacteria Pseudomonas fluorescens , Azospirillum brasilense , or Rhizobium leguminosarum . The modified protocol allowed the identification and recovery of over 50% of the inoculated cells of all three strains, amidst a background of the native soil microflora. Subsequently, the survival of P. fluorescens was successfully monitored for several months after application to field-grown rice at Jerilderie, New South Wales, Australia, thus validating the procedure.

Publisher

Canadian Science Publishing

Subject

Genetics,Molecular Biology,Applied Microbiology and Biotechnology,General Medicine,Immunology,Microbiology

Cited by 6 articles. 订阅此论文施引文献 订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献

1. References;New Insights, Trends, and Challenges in the Development and Applications of Microbial Inoculants in Agriculture;2024

2. UnravelingAzospirillum’s colonization ability through microbiological and molecular evidence;Journal of Applied Microbiology;2023-04

3. Different strategies for colonization and prevalence after inoculation with plant growth-promoting rhizobacteria revealed by a monitoring method;Soil Science and Plant Nutrition;2022-06-21

4. Current opinion and perspectives on the methods for tracking and monitoring plant growth‒promoting bacteria;Soil Biology and Biochemistry;2019-03

5. Bioprospecting PGPR Microflora by Novel Immunobased Techniques;Crop Improvement Through Microbial Biotechnology;2018

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