Isolation, Molecular Characterization and Application of Aspergillus niger and Penicillium chrysogenum with Biofertilizer Potentials to Enhance Rice Growth

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Oluwole M. David
Ayobami C. Olawusi
Olusola A. Oluwole
Pius O. Adeola
Adebowale T. Odeyemi


The use of chemical fertilizers has been associated with a persistent decline in soil fertility, which is also detrimental to soil health. Biofertilizers have been reported to be better alternatives to chemical fertilizers. In this study, the mycofertilizer potentials of the species Aspergillus and Penicillium were investigated. The fungi were isolated from the rice (Oryza sativa Linn) rhizosphere and identified using cultural and molecular methods. The fungal isolates were examined for protease synthesis, nitrogen fixation, cellulose breakdown, and phosphate solubilization using standard methods. The mycofertilizer potentials of the isolates were screened for in an in-situ experiment that was carried out in the greenhouse using a pot experimental method. Isolates that solubilized phosphate and also produced cellulase and protease were selected for the greenhouse experiment. Aspergillus niger and Penicillium chrysogenum proved to be the best candidates among the isolates. The results of the greenhouse pot experiment showed that after 30 days of planting, rice (O. sativa) in the control group had the best performance, but after 63 days of planting, the rice in the pot inoculated with both A. niger and P. chrysogenum had the best performance, followed by the plant inoculated with A. niger, while the plant in the control group had the least average growth. Plants in the test groups had significant growth compared to the plants in the control group. These isolates could be used in the production of mycofertilizer for the growth of grain crops that are known not to fix nitrogen.

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David, O. M., Olawusi, A. C., Oluwole, O. A., Adeola, P. O., & Odeyemi, A. T. (2023). Isolation, Molecular Characterization and Application of Aspergillus niger and Penicillium chrysogenum with Biofertilizer Potentials to Enhance Rice Growth: Tropical Journal of Natural Product Research (TJNPR), 7(4), 2790–2795. Retrieved from


Franc M, Hannula SE, Bełka M, Jedryczka M. Fungal biodiversity and their role in soil health. Front. Microbiol. 2018; 9.

Bhattacharyya PN, Jha DK. Plant growth-promoting rhizobacteria (PGPR) emergence in agriculture. World J. Microbiol. Biotechnol, 2012; 28:1327-1350.

Gururani MA, Venkatesh J, Upadhyaya CP, Nookaraju A, Pandey SK, Park SW. Plant disease resistance genes: Current status and future directions. Physiol. Mol. Plant Pathol., 2012; 78:51-65.

Kumar S, Diksha A, Sindhu SS, Kumar R. Biofertilizers: An ecofriendly technology for nutrient recycling and environmental sustainability. Curr. Res. Microb. Sci., 2022; 3:100094.

Sylvia DM, Fuhrmann JJ, Hartel PG, Zuberer DA. Principles and Applications of Soil Microbiology. Pearson Prentice Hall, Upper Saddle River, New Jersey, 2005.

Yuvaraj M, Ramasamy M. Role of Fungi in Agriculture. In: S. M. Mirmajlessi, R. Radhakrishnan (Eds.), Biostimulants in Plant Science. IntechOpen, 2020.

Fasusi OA, Cruz C, Babalola O.O. Agricultural sustainability: microbial biofertilizers in rhizosphere management. Agriculture, 2021; 11:163. https://

Gnanachitra M. Studies on the development of acid tolerant Azospirillum for tea. Ph.D. Thesis, Department of Agricultural Microbiology, Tamilnadu Agricultural University, Coimbatore. 2000; pp. 164.

Kour D, Rana KL, Yadav N, Yadav AN, Singh J, Rastegari AA, Saxena AK. Agriculturally and industrially important fungi: Current developments and potential biotechnological applications. In: Yadav A N, Singh S, Mishra S, Gupta A (eds.) Recent Advancement in White Biotechnology

Through Fungi. Vol. 2. Perspective for Value-Added Products and Environments. Springer, Cham. 2019; 1–64.

Kour D, Rana KL, Yadav AN, Yadav N, Kumar M, Kumar V, Vyas P, Dhaliwal HS, Saxena AK. Microbial biofertilizers: Bioresources and eco-friendly technologies for agricultural and environmental sustainability. Biocatal. Agric. Biotechnol., 2020; 23: 101487.

Hayat R, Ali S, Amara U. Soil beneficial bacteria and their role in plant growth promotion: a review. Ann. Microbiol., 2010; 60: 579–598.

Mendes R, Garbeva P, Raaijmakers JM. The rhizosphere microbiome: significance of plant beneficial, plant pathogenic, and human pathogenic microorganisms. FEMS Microbiol. Rev., 2013; 37(5):634–663.

Kaewchai S, Soytong K, Hyde KD. Mycofungicides and fungal biofertilizers. Fungal Diversity, 2009; 38:25-50.

Itelima JU, Bang WJ, Onyimba IA. A review: Biofertilizer; a key player in enhancing soil fertility and crop productivity. J. Microbiol. Biotechnol. Rep., 2018; 2(1):22-28.

Singh R, Srivastava P, Verma P, Singh P, Bhadouria R, Singh V, Singh H, Raghubanshi A. Exploring soil responses to various organic amendments under dry tropical agroecosystems. Climate Change and Soil Interactions. 2020; 583-611.

Khan N, Bano A. Role of plant growth promoting rhizobacteria and Ag-Nano particle in the bioremediation of heavy metals and maize growth under municipal wastewater irrigation. Inter. J. Phytoremed., 2016;18: 211-221.

1Zhu L, Li T, Wang C. The effects of dark septate endophyte (DSE) inoculation on tomato seedlings under Zn and Cd stress. Environ. Sci. Pollut. Res., 2018; 25:35232–35241.

Elias F, Woyessa D, Muleta D. Phosphate solubilization potential of rhizosphere fungi isolated from plants in Jimma Zone, Southwest Ethiopia. Inter. J. Microbiol, 2016; 54:72601.

Teather RN, Wood PJ. Use of congo red-polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from the bovine rumen. Appl. Environ. Microbiol. 1982; 43:777-780.

Odeyemi AT, Oluwole OA, Oso MO. Characterization of bacteria isolated from rumen substrates of goat (Capra aegagrushircus). Afr. J. Biol. Sci. 2020; 2(2):100-111.

Kumar C.M.S, Jacob TK, Devasahayam S, Thomas S, Geethu C. Multifarious plant growth promotion by an entomopathogenic fungus Lecanicillium psalliotae. Microbiol. Res., 2018; 207:153–160.

Park M, Kim C, Yang J, Lee H, Shin W, Kim S, Sa T. Isolation and characterization of diazotrophic growth promoting bacteria from rhizosphere of agricultural crops of Korea. Microbiol. Res, 2005;160(2):127–133.

Choudhary DK. Plant growth-promotion (PGP) activities and molecular characterization of rhizobacterial strains isolated from soybean (Glycine max L. Merril) plants against charcoal rot pathogen, Macrophomina phaseolina. Biotechnol. Lett. 2011; 33:2287.

Nenwani V, Doshi P, Saha T, Rajkumar S. Isolation and characterization of a fungal isolate for phosphate solubilization and plant growth promoting activity. J. Yeast Fungal Res., 2010; 1:9-14.

Bhattacharyya C, Banerjee S, Acharya U. Evaluation of plant growth promotion properties and induction of antioxidative defense mechanism by tea rhizobacteria of Darjeeling, India. Sci. Rep., 2020; 10:15536.

Frac M, Jezierska-tys S, Takashi Y. Occurrence, detection, and molecular and metabolic characterization of heatresistant fungi in soils and plants and their risk to human health. Adv. Agron., 2015; 132:161–204.

Bagyaraj DJ, Ashwin R. Soil biodiversity: role in sustainable horticulture. Biodivers. Hortic. Crops. 2017; 5:1-18. doi: 10.1016/j.jenvman.2017.08.001

Kumar A, Maurya BM, Raghuwanshi R. The microbial consortium of indigenous rhizobacteria improving plant health, yield and nutrient content in wheat (Triticum aestivum). J. Plant Nutr., 2021; 44:1942–1956. doi: 10.1080/01904167.2021.1884706.

Mittal V, Singh O, Nayyar H, Kaur J, Tewari R. Stimulatory effect of phosphate-solubilizing fungal strains (Aspergillus awamori and Penicillium citrinum) on the yield of chickpea(Cicer arietinum L. cv. GPF2). Soil Biol. Biochem., 2008; 40:718-727.

Yadav J, Verma JP, Yadav SK, Tiwari KN. Effect of salt concentration and pH on soil inhabiting fungus Penicillium citrinum Thom. for solubilization of tricalcium phosphate.Microbiol. J., 2011; 1:25-32.

Onyia CE, Anyawu CU, Ikegbunam MN. Ability of fungi, isolated from Nsukka peppers and garden-eggplant rhizospheres, to solubilize phosphate and tolerate cadmium. Adv. Microbiol. 2015; 5(7):500-506.

Yadav BK, Verma A. Phosphate solubilization and mobilization in soil through microorganisms under arid ecosystems. The Function Ecosyst. 2012; 6:94-108.

Iman M. Effect of phosphate solubilizing fungi on growth and nutrient uptake of soyabean (Glycine max L.) plants. J. Appl. Sci. Res. 2008; 4:592-598.

Alam S, Khalil S, Ayub N, Rashid M. In vitro solubilization of inorganic phosphate by phosphate solubilizing microorganism (PSM) from maize rhizosphere. Inter. J. Agricult. Biol., 2002; 4:454–458.

Mundim GDSM, Maciel GM, Mendes GDO. Aspergillus niger as a biological input for improving vegetable seedling production. Microorganisms. 2022; 10(4): 674.

Araujo CV, Rossati KF, Xavier LV, De Oliveira VA, Carmo GJ, De Assis GA, Mendes GM. Enhanced growth in nursery of coffee seedlings inoculated with the rhizosphere fungus Aspergillus niger for field transplantation. Rhizosphere. 2020; 15: 100236. doi: 10.1016/j.rhisph.2020.100236.

Chezang D, Ngawang, C. Effects of different planting methods on rice (Oryza sativa L.) crop performance and cost of production. Bhutanese J. Agricult., 2018; 1(1):13-22.

Wang X, Wang C, Sui J, Liu Z, Li Q, Ji C, Song X, Hu Y, Wang C, Sa R, Zhang J, Du J, Liu X. Isolation and characterization of phosphofungi, and screening of their plant growth-promoting activities. AMB Express, 2018; 8(1):63.

Subowo YB. The effect of biofertilizer fungi on ciherang rice growth at some level of soil salinity. J. Biological Res. 2009;18:106 - 110.

Pradhan N, Sukla LB. Solubilization of inorganic phosphates by fungi isolated from agriculture soil. Afr. J. Biotechnol., 2015; 5:850-854.