Isolation and identification of maize phyllosphere bacteria and determination of their plant growth promoting traits

Document Type : Complete scientific research article

Authors

1 Dept. of Soil Sci., College of Agric., Shiraz Univ.

2 Soil Science Department, College of Agriculture, Shiraz University

3 Soil and Water Research Institute, Agriculture Research, Education and Extension Organization

4 Department of Soil Science, Shiraz University

5 Shiraz university

6 Department of Plant pathology, Shiraz University

Abstract

Background and Objectives: The plant growth promoting bacteria (PGPB) inhabiting the phyllosphere of different plants provides a tool for more efficient use of bacteria as a source of biofertilizers. The use of molecular technologies leads to in-depth research of the phyllosphere microorganisms and may provide new avenues for the development and validation of modern agricultural operations and significantly increase productivity. The aim of this research was to investigate the plant growth promoting (PGP) traits of bacteria isolated from maize (Zea mays L.) phyllosphere cultivated in different geographical zones of country (Iran) and their molecular identification by 16Sr RNA sequencing.

Materials and Methods: In September (2016), 116 samples of healthy mature leaves were collected from the maize fields located in 6 provinces of Iran (Fars, Alborz, Qazvin, Khuzestan, Tehran and Khorasan) and then, phyllosphere bacteria were isolated from them. Growth promoting traits of bacteria including Indole-3 Acetic acid (IAA) production, organic phosphate dissolution, siderophore production, exopolysaccharides (EPS) production and also nitrogenase enzyme activity were measured using standard methods. Superior strains were identified based on 16S rRNA sequencing method.

Results: The 10 from 242 bacterial strains isolated from maize phyllosphere were selected based on their growth promoting traits. All strains selected demonstrated nitrogenase activity, organic phosphate dissolution and IAA and EPS production capability. The highest nitrogenase activity (70.31 nM ml-1 h-1) and organic phosphate dissolution (75.05 g l-1) were observed in THE15 strain, while the lowest amount of these properties belonged to KHO57 (5.55 nM ml-1 h-1) and KHO51 (15.90 g l-1) strains, respectively. The QAZ26 and TEH15 strains had the highest amount of IAA production (31.56 and 30.00 µg ml-1, respectively) and the ALB32 strain produced the lowest amount of IAA (4.50 µg ml-1). The EPS production by strains ranged from 0.120 to 1.805 g l-1. Among the strains investigated, only KUU32, KHU53, ALB32 and QAZ26 were able to produce siderophore. 16S rRNA sequencing revealed that the 10 strains selected belong to Bacillus, Microbacterium, Pseudarthrobacter, Pseudomonas, Stenotrophomonas, Enterobacter and Kocuria genera. The Bacillus with 4 different species (including Bacillus subtilis, Bacillus megaterium, Bacillus paralicheniformis and Bacillus pumilus) was the most dominant genus.

Conclusion: Based on the results of this study, Bacillus was the most dominant genus of plant growth promoting bacteria in the maize phyllosphere. Among the isolated bacteria, Enterobacter hormaechei TEH15 was highly capable of nitrogenase enzyme activity, IAA and organic phosphate solubility. Therefore, this strain deserves further research in greenhouse and field studies of maize cultivation and maize family plants.

Keywords

References

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Journal of Soil Management and Sustainable Production
Volume 10, Issue 2
September 2020
Pages 47-67

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