Optimization of gamma polyglutamic acid (γ-PGA) production by Bacillus velezensis and its effect on increasing wheat growth and biocontrol of Bipolaris sorokiniana causal agent of common root rot of wheat

Document Type : Complete paper

Authors

1 Department of Plant Protection, University of Tehran, Karaj, Iran.

2 Department of Biology, Soil and Water Institute, Karaj, Iran.

3 Department of Plant Protection, University of Tehran, Karaj, Iran

Abstract

Modern agriculture in the 21st century must provide food security for this growing population, which means producing more food and fiber with less labor. Therefore, agriculture in this century requires smart investment based on more efficient and environmentally friendly production methods. Common wheat root rot caused by B. sorokiniana is one of the most important pathogens of wheat, which causes economic losses every year. In this research, from 180 Bacillus isolates from the samples collected from the wheat fields of Hamadan province, wastes of edible mushrooms, palm wastes and wood wastes, 9 isolates were able to produce γ-PGA with different production rates. 2 isolates were able to produce γ-PGA independently of glutamic acid. Bacillus velezensis UTB97 with a production rate of 40 g/L γ-PGA was selected as the superior producer isolate. UTB97 could prevent 65% growth of pathogenic fungi in laboratory conditions by dual culture method. Greenhouse result showed that the treatment of UTB97 and UTB97 + γ-PGA was able to control Common wheat root rot disease by 59 and 73%, respectively, compared to the control treatment, and also the treatment of UTB97 + γ-PGA was able to increase the growth index of wheat. (stem length and root dry weight), in the presence and absence of the pathogenic fungus.

Keywords

Main Subjects

Extended Abstract

Introduction

Agriculture in the 21st century must provide food security for this growing population, which means producing more food and fiber with less labor. Therefore, agriculture in this century requires smart investment based on more efficient and environmentally friendly production methods. In modern agriculture, by using biological fertilizers and adopting appropriate strategies, it is possible to prevent nitrogen losses in the soil and reduce the efficiency of chemical fertilizers. In recent years, the development of the use of biological control agents as an alternative to chemical fungicides for the biological control of pathogens has been considered. Among them, Bacillus has an advantage over other antagonistic microorganisms due to its broad antibacterial properties, significant growth promoting effect by producing a wide range of secondary metabolites, Poly gamma glutamic acid (γ-PGA) is an anionic, biodegradable, non-toxic and hydrophilic polypeptide produced by various Bacillus species. Glutamic acid D and L are the basic units of this polymer which are created by amide bonds, numerous studies have proven that γ-PGA significantly increases plant performance and absorption of nutrients N, P, K by strengthening the absorption capacity of roots and regulating the availability of food through changing the microbial and enzymatic characteristics can also increase the ability of the biocontrol agent to control plant pathogens.

 

Materials and methods:

Isolation of γ-PGA producing bacteria was done from samples collected from wheat fields of Hamadan province, edible mushroom waste, palm waste and wood waste; The media culture was used for the production of γ-PGA includes: 20 grams of glucose, 20 grams of glutamic acid, 0.5 grams of KH2PO4, 0.1 grams of MgSo4, 0.15 grams of CaCl2, 2 grams of yeast extract, 0.5 grams of ZnSo4 and 0.1 grams of MnSo4 per litter. The γ-PGA production rate was evaluated by using cetyltrimethylammonium bromide (CTAB). Identification of bacteria using a DNA extraction kit and amplify the 16S rDNA region (Blank et al., 2018). In order to investigate the effect of bacteria and γ-PGA on the control of crown and root rot disease of wheat, cross-culture method was used in laboratory conditions and Peng et al.'s (2020) method was used in greenhouse conditions.

 

Results

In this research, from 180 Bacillus isolates from the samples collected from the wheat fields of Hamadan province, wastes of edible mushrooms, palm wastes and wood wastes, 9 isolates were able to produce γ-PGA with different production rates. 2 isolates were able to produce γ-PGA independently of glutamic acid. Bacillus velezensis UTB97 with a production rate of 40 g/L γ-PGA was selected as the superior producer isolate. UTB97 could prevent 65% growth of pathogenic fungi in laboratory conditions by dual culture method. Greenhouse result showed that the treatment of UTB97 and UTB97 + γ-PGA was able to control Common wheat root rot disease by 59 and 73%, respectively, compared to the control treatment, and also the treatment of UTB97 + γ-PGA was able to increase the growth index of wheat. (stem length and root dry weight), in the presence and absence of the pathogenic fungus.

 

Conclusion

The results of this research showed that B.Velezensis UTB97 is a powerful strain for gamma polyglutamic acid production. This strain can produce γ-PGA at the rate of 40 g/liter in optimal media culture. also use of UTB97 and γ-PGA caused the control of common crown and root rot disease of wheat and increased the growth index of wheat. Therefore, in order to increase the yield and production of wheat, simultaneous application of bacteria and γ-PGA can be promising.

References

Ahmadzadeh, M and Sharifi Tehrani, A.  (2021) Plant probiotic bacteria. University of Tehran Press,
Allali, K., Goudjal, Y., Zamoum, M., Bouznada, K., Sabaou, N., & Zitouni, A. (2019). Nocardiopsis dassonvillei strain MB22 from the Algerian Sahara promotes wheat seedlings growth and potentially controls the common root rot pathogen Bipolaris sorokiniana. Journal of Plant Pathology, 101, 1115-1125.
Al-Sadi, A. M. (2021). Bipolaris sorokiniana-induced black point, common root rot, and spot blotch diseases of wheat: A review. Frontiers in cellular and infection microbiology, 11, 584899.      
Ashiuchi, M. and H. Misono (2002). "Biochemistry and molecular genetics of poly-γ-glutamate synthesis." Applied microbiology and biotechnology 59: 9-14.
Cai, D., He, P., Lu, X., Zhu, C., Zhu, J., Zhan, Y., ... & Chen, S. (2017). A novel approach to improve poly-γ-glutamic acid production by NADPH regeneration in Bacillus licheniformis WX-02. Scientific Reports, 7(1), 43404.
Chatterjee, A., Huma, B., Shaw, R., & Kundu, S. (2017). Reconstruction of Oryza sativa indica genome scale metabolic model and its responses to varying RuBisCO activity, light intensity, and enzymatic cost conditions. Frontiers in plant science, 8, 2060.
Chen, L., Fei, L., Mohamed, K. S., Liu, L., Wang, Z., Zhong, Y., & Dai, Z. (2018). The effects of ploy (γ-glutamic acid) on spinach productivity and nitrogen use efficiency in North-West China. Plant, Soil and Environment, 64(11), 517-522.
Cho, S. M., Kang, B. R., Han, S. H., Anderson, A. J., Park, J. Y., Lee, Y. H., ... & Kim, Y. C. (2008). 2R, 3R-butanediol, a bacterial volatile produced by Pseudomonas chlororaphis O6, is involved in induction of systemic tolerance to drought in Arabidopsis thaliana. Molecular plant-microbe interactions, 21(8), 1067-1075.   
Clapp, J., Moseley, W. G., Burlingame, B., & Termine, P. (2022). The case for a six-dimensional food security framework. Food Policy, 106, 102164.
Fu, Y., Wang, S., Gao, S., Wang, S., Gao, Z., & He, Z. (2022). Effect of a Superabsorbent Polymer (Poly-Gamma-Glutamic Acid) on Water and Salt Transport in Saline Soils under the Influence of Multiple Factors. Polymers, 14(19), 4056.
Ghosh, K., Ray, M., Adak, A., Halder, S. K., Das, A., Jana, A., ... & Mondal, K. C. (2015). Role of probiotic Lactobacillus fermentum KKL1 in the preparation of a rice based fermented beverage. Bioresource technology, 188, 161-168.
Gondal, M. A., Dastageer, M. A., & Khalil, A. (2009). Synthesis of nano-WO3 and its catalytic activity for enhanced antimicrobial process for water purification using laser induced photo-catalysis. Catalysis Communications, 11(3), 214-219.
Guo, J., Zhang, J., Zhang, K., Li, S., & Zhang, Y. (2023). Effect of γ-PGA and γ-PGA SAP on soil microenvironment and the yield of winter wheat. Plos one, 18(7), e0288299.
Guo, Y., Gao, P., Li, F., & Duan, T. (2019). Effects of AM fungi and grass endophytes on perennial ryegrass Bipolaris sorokiniana leaf spot disease under limited soil nutrients. European Journal of Plant Pathology, 154, 659-671.    
Haas, D. and C. Keel (2003). "Regulation of antibiotic production in root-colonizing Pseudomonas spp. and relevance for biological control of plant disease." Annual review of phytopathology 41(1): 117-153.        
Halmschlag, B., Putri, S. P., Fukusaki, E., & Blank, L. M. (2020). Poly-γ-glutamic acid production by Bacillus subtilis 168 using glucose as the sole carbon source: a metabolomic analysis. Journal of bioscience and bioengineering, 130(3), 272-282.  
Harishchandra, H. and P. Perumpuli (2023). "IDENTIFICATION OF ACETIC ACID BACTERIA FROM NATURALLY FERMENTED BANANA VINEGAR." Self-Sustaining Agriculture: Way Forward for Food Security and Safety: 105.  
Hu, Y., Shao, Y., Wu, C., Yuan, C., Ishimura, G., Liu, W., & Chen, S. (2018). γ-PGA and MTGase improve the formation of ε-(γ-glutamyl) lysine cross-links within hairtail (Trichiurus haumela) surimi protein. Food Chemistry, 242, 330-337.         
Izano, E. A., Sadovskaya, I., Wang, H., Vinogradov, E., Ragunath, C., Ramasubbu, N., ... & Kaplan, J. B. (2008). Poly-N-acetylglucosamine mediates biofilm formation and detergent resistance in Aggregatibacter actinomycetemcomitans. Microbial pathogenesis, 44(1), 52-60.
Kimura, K., Tran, L. S. P., Uchida, I., & Itoh, Y. (2004). Characterization of Bacillus subtilis γ-glutamyltransferase and its involvement in the degradation of capsule poly-γ-glutamate. Microbiology, 150(12), 4115-4123.
Kocianova, S., Vuong, C., Yao, Y., Voyich, J. M., Fischer, E. R., DeLeo, F. R., & Otto, M. (2005). Key role of poly-γ-DL-glutamic acid in immune evasion and virulence of Staphylococcus epidermidis. The Journal of clinical investigation, 115(3), 688-694.
Layek, J., Das, A., Idapuganti, R. G., Sarkar, D., Ghosh, A., Zodape, S. T., ... & Meena, R. S. (2018). Seaweed extract as organic bio-stimulant improves productivity and quality of rice in eastern Himalayas. Journal of Applied Phycology, 30, 547-558.   
Liang, J., Shi, W., He, Z., Pang, L., & Zhang, Y. (2019). Effects of poly-γ-glutamic acid on water use efficiency, cotton yield, and fiber quality in the sandy soil of southern Xinjiang, China. Agricultural Water Management, 218, 48-59.
Liu, W., Xie, Y., Ma, J., Luo, X., Nie, P., Zuo, Z., ... & Ren, J. (2015). IBS: an illustrator for the presentation and visualization of biological sequences. Bioinformatics, 31(20), 3359-3361.
Miljaković, D., Marinković, J., & Balešević-Tubić, S. (2020). The significance of Bacillus spp. in disease suppression and growth promotion of field and vegetable crops. Microorganisms, 8(7), 1037.
O’Sullivan, C. A., Belt, K., & Thatcher, L. F. (2021). Tackling control of a cosmopolitan phytopathogen: Sclerotinia. Frontiers in Plant Science, 12, 707509.
Paul, S. K., Mahmud, N. U., Gupta, D. R., Surovy, M. Z., Rahman, M., & Islam, M. T. (2021). Characterization of Sclerotium rolfsii causing root rot of sugar beet in Bangladesh. Sugar Tech, 23, 1199-1205. 
Peng, Y. P., Chang, Y. C., Chen, K. F., & Wang, C. H. (2020). A field pilot-scale study on heavy metal-contaminated soil washing by using an environmentally friendly agent—poly-γ-glutamic acid (γ-PGA). Environmental Science and Pollution Research, 27, 34760-34769.
Pötter, M., Oppermann-Sanio, F. B., & Steinbüchel, A. (2001). Cultivation of bacteria producing polyamino acids with liquid manure as carbon and nitrogen source. Applied and environmental microbiology, 67(2), 617-622.
Rehm, B. H. (2010). "Bacterial polymers: biosynthesis, modifications and applications." Nature Reviews Microbiology 8(8): 578-592.      
Shih, I. L., Van, Y. T., & Shen, M. H. (2004). Biomedical applications of chemically and microbiologically synthesized poly (glutamic acid) and poly (lysine). Mini reviews in medicinal chemistry, 4(2), 179-188. 
Thammasittirong, S. N. R. (2017). The potential of Bacillus subtilis BAS114 for in vitro biocontrol of Fusarium oxysporum. Advances in Environmental Biology, 11(1), 46-51.
Villa-Rodriguez, E., Parra-Cota, F., Castro-Longoria, E., López-Cervantes, J., & de los Santos-Villalobos, S. (2019). Bacillus subtilis TE3: a promising biological control agent against Bipolaris sorokiniana, the causal agent of spot blotch in wheat (Triticum turgidum L. subsp. durum). Biological control, 132, 135-143.
Villa-Rodriguez, E., Lugo-Enríquez, C., Ferguson, S., Parra-Cota, F. I., Cira-Chávez, L. A., & de los Santos-Villalobos, S. (2022). Trichoderma harzianum sensu lato TSM39: A wheat microbiome fungus that mitigates spot blotch disease of wheat (Triticum turgidum L. subsp. durum) caused by Bipolaris sorokiniana. Biological Control, 175, 105055.
Wang, L., Mao, J., Zhao, H., Li, M., Wei, Q., Zhou, Y., & Shao, H. (2016). Comparison of characterization and microbial communities in rice straw-and wheat straw-based compost for Agaricus bisporus production. Journal of Industrial Microbiology and Biotechnology, 43(9), 1249-1260.          
Yi, Y., Shan, Y., Liu, S., Yang, Y., Liu, Y., Yin, Y., ... & Li, R. (2021). Antagonistic strain Bacillus amyloliquefaciens XZ34-1 for controlling Bipolaris sorokiniana and promoting growth in wheat. Pathogens, 10(11), 1526.      
Yin, A., Jia, Y., Qiu, T., Gao, M., Cheng, S., Wang, X., & Sun, Y. (2018). Poly-γ-glutamic acid improves the drought resistance of maize seedlings by adjusting the soil moisture and microbial community structure. Applied Soil Ecology, 129, 128-135.
Zabed, H. M., Akter, S., Yun, J., Zhang, G., Awad, F. N., Qi, X., & Sahu, J. N. (2019). Recent advances in biological pretreatment of microalgae and lignocellulosic biomass for biofuel production. Renewable and Sustainable Energy Reviews, 105, 105-128.     
Zhang, L., Yang, X., Gao, D., Wang, L., Li, J., Wei, Z., & Shi, Y. (2017). Effects of poly-γ-glutamic acid (γ-PGA) on plant growth and its distribution in a controlled plant-soil system. Scientific reports, 7(1), 6090.       
Zhang, P., Guo, G., Wu, Q., Chen, Y., Xie, J., Lu, P., ... & Liu, Z. (2020). Identification and fine mapping of spot blotch (Bipolaris sorokiniana) resistance gene Sb4 in wheat. Theoretical and Applied Genetics, 133, 2451-2459.    
Zhang, X., Wu, Y., & Gu, B. (2015). Urban rivers as hotspots of regional nitrogen pollution. Environmental Pollution, 205, 139-144.
BIOLOGICAL CONTROL OF PESTS AND PLANT DISEASES
Volume 11, Issue 2
January 2024
Pages 69-82

Files

Share

How to cite

Statistics