SavedAdd your journal

Journals / Türk Tarım ve Doğa Bilimleri Dergisi / 2026 / Volume: 13 Issue: 1

Molecular Characterization of Phosphate Solubilizing Fungi Aspergillus niger and Penicillium expansum using the Species-Specific PCR Based Molecular Markers

Muhammed Tatar(SİVAS BİLİM VE TEKNOLOJİ ÜNİVERSİTESİ, TARIM BİLİMLERİ VE TEKNOLOJİLERİ FAKÜLTESİ, BİTKİSEL ÜRETİM VE TEKNOLOJİLERİ BÖLÜMÜ, BİTKİ SAĞLIĞI ANABİLİM DALI)0000-0002-8312-8434
Amjad Ali(SIVAS SCIENCE AND TECHNOLOGY UNIVERSITY)0000-0003-0816-1872
Zemran Mustafa(SIVAS SCIENCE AND TECHNOLOGY UNIVERSITY)0000-0002-1754-6320
Fatih Ölmez(SİVAS BİLİM VE TEKNOLOJİ ÜNİVERSİTESİ, TARIM BİLİMLERİ VE TEKNOLOJİLERİ FAKÜLTESİ, BİTKİ KORUMA BÖLÜMÜ)0000-0001-7016-2708
Tolga Karaköy(SIVAS SCIENCE AND TECHNOLOGY UNIVERSITY)0000-0002-5428-1907
Pages
—
Publication Date
2026-01-22
🏛️Bibliographic metadata and full text access for this article are provided via DergiPark.
DergiPark Record ↗
✨

AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

Future work will couple PCR-based genotyping with phosphate-solubilization assays and pathogenicity tests to identify elite biofertilizer candidates while monitoring and managing high-risk postharvest lineages.

📌 3 Key Takeaways

🎯
Objective & Hypothesis:Phosphorus (P) is a vital macronutrient, yet most soil P occurs in insoluble forms, driving intensive use of chemical fertilizers.
📊
Key Findings & Data:A total of 126 fungal isolates (98 putative P. expansum, 28 putative A. niger) were cultured on PDA for macro- and micromorphological assessment and subjected to CTAB-based DNA extraction and PCR with species-specific primers.
💡
Conclusion & Impact:Future work will couple PCR-based genotyping with phosphate-solubilization assays and pathogenicity tests to identify elite biofertilizer candidates while monitoring and managing high-risk postharvest lineages.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:phosphate-solubilizing fungi · Aspergillus niger · Penicillium expansum

Abstract

Phosphorus (P) is a vital macronutrient, yet most soil P occurs in insoluble forms, driving intensive use of chemical fertilizers. Phosphate-solubilizing fungi (PSF), including Aspergillus niger and Penicillium expansum, can enhance P availability but also comprise important postharvest pathogens. This study aimed to characterize apple-associated isolates of both species using morphology and species-specific PCR based markers to provide a basis for future functional screening. A total of 126 fungal isolates (98 putative P. expansum, 28 putative A. niger) were cultured on PDA for macro- and micromorphological assessment and subjected to CTAB-based DNA extraction and PCR with species-specific primers. A. niger formed rapidly growing, granular powdery colonies that darkened to brown–black, with spherical vesicles densely covered by biseriate phialides and rough-walled, dark globose conidia in chains. P. expansum produced moderately fast, velvety colonies that shifted from white to blue–green with concentric rings and showed typical brush-like penicilli with terverticillate branching and smooth, spherical conidia. PCR amplification yielded the expected 404 bp product in 95/98 P. expansum isolates (96.93%) and the 197 bp product in 24/28 A. niger isolates (85.71%), indicating strong concordance between morphological and molecular identification. The integrated morphological-molecular approach reliably resolved apple-associated A. niger and P. expansum populations and establishes a validated genetic resource. Future work will couple PCR-based genotyping with phosphate-solubilization assays and pathogenicity tests to identify elite biofertilizer candidates while monitoring and managing high-risk postharvest lineages.

🔖

Cite This Article

Tatar, M., Ali, A., Mustafa, Z., Ölmez, F., Karaköy, T. (2026). Molecular Characterization of Phosphate Solubilizing Fungi Aspergillus niger and Penicillium expansum using the Species-Specific PCR Based Molecular Markers. Türk Tarım ve Doğa Bilimleri Dergisi, Volume: 13 Issue: 1. https://doi.org/10.30910/turkjans.1841672

📄 Full Text PDF Viewer

📄

PDF görüntüleyici hazırlanıyor...

References (50)

  1. Chakdar, H., Singha, A., & Satya, P. (2017). New generation markers for fingerprinting and structural analysis of fungal community. In Molecular Markers in Mycology: Diagnostics and Marker Developments (pp. 123-150). Cham: Springer International Publishing. https://doi.org/10.1007/978-3-319-34106-4_6
  2. Desire, M. H. (2025). Diversity of the Phosphate Solubilizing Fungi (PSFs) Population from the Soybeans Rhizosphere in the Noun Valley, Western Highlands of Cameroon. Journal of Plant Sciences, 13(3), 132-144. https://doi.org/10.11648/j.jps.20251303.15
  3. Doyle JJ, Doyle JL. Isolation of plant DNA from fresh tissue. Focus. 1990;12:13–5.
  4. do Nascimento, J. M., Netto, J. A. F. V., Valadares, R. V., de Oliveira Mendes, G., da Silva, I. R., Vergütz, L., & Costa, M. D. (2021). Aspergillus niger as a key to unlock fixed phosphorus in highly weathered soils. Soil Biology and Biochemistry, 156, 108190. https://doi.org/10.1016/j.soilbio.2021.108190
  5. Doilom, M., Guo, J. W., Phookamsak, R., Mortimer, P. E., Karunarathna, S. C., Dong, W.,. & Xu, J. C. (2020). Screening of phosphate-solubilizing fungi from air and soil in Yunnan, China: four novel species in Aspergillus, Gongronella, Penicillium, and Talaromyces. Frontiers in microbiology, 11, 585215. https://doi.org/10.3389/fmicb.2020.585215
  6. El-Banna, A. A., Pitt, J. I., & Leistner, L. (1987). Production of mycotoxins by Penicillium species. Systematic and Applied Microbiology, 10(1), 42-46. https://doi.org/10.1016/S0723-2020(87)80008-5
  7. Frisvad, J. C., & Samson, R. A. (2004). Polyphasic taxonomy of Penicillium subgenus Penicillium — A guide to identification of food and air-borne terverticillate penicillia and their mycotoxins. Studies in Mycology, 49, 1-173.
  8. Gal, T. E., Alexa, E. C., Șumălan, R. M., Dascălu, I., & Iordănescu, O. A. (2025). Factors Affecting Patulin Production by Penicillium expansum in Apples. Foods, 14(13), 2310. https://doi.org/10.3390/foods14132310
  9. Geiser, D. M., Klich, M. A., Frisvad, J. C., Peterson, S. W., Varga, J., & Samson, R. A. (2007). The current status of species recognition and identification in Aspergillus. Studies in mycology, 59, 1-10. https://doi.org/10.3114/sim.2007.59.01
  10. Kalayu, G. (2019). Phosphate solubilizing microorganisms: promising approach as biofertilizers. International Journal of Agronomy, 2019(1), 4917256. https://doi.org/10.1155/2019/4917256
  11. Kansu, B. and Tunalı, B. (2016). The Determination of Prevalence and Definition of Some Important Toxigenic Fungi in Maize Production Area of Turkey. 6th International Plant Protection Congress, 5-8 September, Konya-Türkiye, p777
  12. Khan, I. H., & Javaid, A. (2021). Molecular characterization of Penicillium expansum associated with blue mold disease of apple in Pakistan. Pak. J. Bot, 53(6), 2299-2303. http://dx.doi.org/10.30848/PJB2021-6(34
  13. Khan, K., Ali, S. Z., Nisa, T., Rasool, G., & Qadir, A. (2024). Identification and In vitro management of apple blue mold disease in Balochistan. Pak-Euro Journal of Medical and Life Sciences, 7(1), 25-30. http://dx.doi.org/10.31580/pjmls.v7i1.2923
  14. Kichu, A., Ajungla, T., Walling, M., Temjen, W., & Yeptho, L. (2022). Seasonal variation of soil fungi in tea garden, mokokchung district, Nagaland. The Journal of Plant Science Research, 38(1), 151-158. http://dx.doi.org/10.32381/JPSR.2022.38.01.15
  15. Klich, M. A. (2002). Identification of common Aspergillus species. Utrecht, Netherlands: Centraalbureau voor Schimmelcultures.
  16. Luciano‐Rosario, D., Keller, N. P., & Jurick, W. M. (2020). Penicillium expansum: biology, omics, and management tools for a global postharvest pathogen causing blue mould of pome fruit. Molecular Plant Pathology, 21(11), 1391-1404. https://doi.org/10.1111/mpp.12990
  17. Ma, Y., Chen, S., Liu, S., Guo, L., Zhang, C., Ye, X., & Tian, D. (2025). Phosphate solubilizing fungi enhance insoluble phosphate dissolution via organic acid production: mechanisms and applications. Frontiers in Microbiology, 16, 1600231. https://doi.org/10.3389/fmicb.2025.1600231
  18. Marek, P., Annamalai, T., & Venkitanarayanan, K. (2003). Detection of Penicillium expansum by polymerase chain reaction. International Journal of Food Microbiology, 89(2-3), 139-144. https://doi.org/10.1016/S0168-1605(03)00115-6
  19. Nassef, K., Sahli, A., Bouhdid, S., Mezzoug, N., Abrini, J., & Khay, E. O. (2025). Phosphate solubilizing microorganisms and their use in sustainable agriculture: A review. Geomicrobiology Journal, 42(3), 224-243. https://doi.org/10.1080/01490451.2025.2457660
  20. Oliveira, M., & Azevedo, L. (2022). Molecular markers: An overview of data published for fungi over the last ten years. Journal of Fungi, 8(8), 803. https://doi.org/10.3390/jof8080803
  21. Pan, T., Brookes, J., Cui, C., Qin, B., Ding, K., Zhang, Y., & Zhu, G. (2025). Decoupling phytoplankton biomass-sediment phosphorus interaction induced by lower incident radiation mainly drives the attenuation of harmful algal blooms in lake Taihu, China. Water Resources Research, 61(6), e2024WR039014. https://doi.org/10.1029/2024WR039014
  22. Pitt, J. I. (1980). The genus Penicillium and its teleomorphic states Eupenicillium and Talaromyces. Academic Press (pp. 634-pp).
  23. Pitt, J. I., & Hocking, A. D. (1985). Interfaces among genera related to Aspergillus and Penicillium. Mycologia, 77(5), 810-824. https://doi.org/10.1080/00275514.1985.12025167Pitt, J. I., & Hocking, A. D. (2009). Fungi and food spoilage (3rd ed.). (Vol. 519, p. 388). New York: Springer. https://doi.org/10.1007/978-0-387-92207-2
  24. Sanzani, S. M., Montemurro, C., Di Rienzo, V., Solfrizzo, M., & Ippolito, A. (2013). Genetic structure and natural variation associated with host of origin in Penicillium expansum strains causing blue mould. International Journal of Food Microbiology, 165(2), 111-120. https://doi.org/10.1016/j.ijfoodmicro.2013.04.024
  25. Samson, R. A., Houbraken, J., Thrane, U., Frisvad, J. C., & Andersen, B. (2010). Food and indoor fungi. CBS laboratory manual series 2. Utrecht: CBS-Fungal Biodiversity Centre, 2(1), 276-390.
  26. Samson, R. A., & Pitt, J. I. (2003). Integration of modern taxonomic methods for Penicillium and Aspergillus classification. CRC Press. https://doi.org/10.1201/9781482284188
  27. Samson, R. A., Visagie, C. M., Houbraken, J., Hong, S. B., Hubka, V., Klaassen, C. H.,. & Frisvad, J. (2014). Phylogeny, identification and nomenclature of the genus Aspergillus. Studies in mycology, 78(1), 141-173. https://doi.org/10.1016/j.simyco.2014.07.004
  28. Šimonovičová, A., Vojtková, H., Nosalj, S., Piecková, E., Švehláková, H., Kraková, L.,. & Pangallo, D. (2021). Aspergillus niger environmental isolates and their specific diversity through metabolite profiling. Frontiers in Microbiology, 12, 658010. https://doi.org/10.3389/fmicb.2021.658010
  29. Toma, M. A., Nazir, K. N. H., Mahmud, M. M., Mishra, P., Ali, M. K., Kabir, A.,. & Alim, M. A. (2021). Isolation and identification of natural colorant producing soil-borne Aspergillus niger from Bangladesh and extraction of the pigment. Foods, 10(6), 1280. https://doi.org/10.3390/foods10061280
  30. Vassileva, M., Mendes, G. D. O., Deriu, M. A., Benedetto, G. D., Flor-Peregrin, E., Mocali, S.,. & Vassilev, N. (2022). Fungi, P-solubilization, and plant nutrition. Microorganisms, 10(9), 1716. https://doi.org/10.3390/microorganisms10091716
  31. Vico, I., Duduk, N., Vasić, M., & Nikolić, M. (2014). Identification of Penicillium expansum causing postharvest blue mold decay of apple fruit. Pesticidi i fitomedicina, 29(4), 257-266. http://dx.doi.org/10.2298/PIF1404257V
  32. Visagie, C. M., Houbraken, J., Frisvad, J. C., Hong, S. B., Klaassen, C. H. W., Perrone, G.,. & Samson, R. A. (2014). Identification and nomenclature of the genus Penicillium. Studies in mycology, 78(1), 343-371. https://doi.org/10.1016/j.simyco.2014.09.001
  33. von Hertwig, A. M., Sant'Ana, A. S., Sartori, D., da Silva, J. J., Nascimento, M. S., Iamanaka, B. T.,. & Taniwaki, M. H. (2018). Real-time PCR-based method for rapid detection of Aspergillus niger and Aspergillus welwitschiae isolated from coffee. Journal of microbiological methods, 148, 87-92. https://doi.org/10.1016/j.mimet.2018.03.010
  34. Walter, R., & Kortekamp, A. (2023). Molecular identification of Penicillium species associated with blue mold on grapes in German vineyards. Vitis, 62(1). https://doi.org/10.5073/vitis.2023.62.1-9
  35. Wang, K., Ngea, G. L. N., Godana, E. A., Shi, Y., Lanhuang, B., Zhang, X.,. & Zhang, H. (2023). Recent advances in Penicillium expansum infection mechanisms and current methods in controlling P. expansum in postharvest apples. Critical Reviews in Food Science and Nutrition, 63(15), 2598-2611. https://doi.org/10.1080/10408398.2021.1978384
  36. Wang, Y., Yuan, L., Wang, Y., Lang, J., Ye, M., Liu, Q.,. & Zhou, N. (2025). Phosphorus-solubilizing fungi promote the growth of Fritillaria taipaiensis PY Li by regulating physiological and biochemical reactions and protecting enzyme system-related gene expression. Frontiers in Genetics, 15, 1459191. https://doi.org/10.3389/fgene.2024.1459191
  37. Xiao, C., Chi, R., He, H., Qiu, G., Wang, D., & Zhang, W. (2009). Isolation of phosphate-solubilizing fungi from phosphate mines and their effect on wheat seedling growth. Applied biochemistry and biotechnology, 159(2), 330-342. https://doi.org/10.1007/s12010-009-8590-3
  38. Žebeljan, A., Duduk, N., Vučković, N., Jurick, W. M., & Vico, I. (2021). Incidence, speciation, and morpho-genetic diversity of Penicillium spp. causing blue mold of stored pome fruits in Serbia. Journal of Fungi, 7(12), 1019. https://doi.org/10.3390/jof7121019
  39. Zhang, Y., Chen, F. S., Wu, X. Q., Luan, F. G., Zhang, L. P., Fang, X. M.,. & Ye, J. R. (2018). Isolation and characterization of two phosphate-solubilizing fungi from rhizosphere soil of moso bamboo and their functional capacities when exposed to different phosphorus sources and pH environments. PloS one, 13(7), e0199625. https://doi.org/10.1371/journal.pone.0199625
  40. Zhu, Y., Xing, Y., Li, Y., Jia, J., Ying, Y., & Shi, W. (2024). The role of phosphate-solubilizing microbial interactions in phosphorus activation and utilization in plant-soil systems: A review. Plants, 13(19), 2686. https://doi.org/10.3390/plants13192686
  41. Doilom, M., Guo, J. W., Phookamsak, R., Mortimer, P. E., Karunarathna, S. C., Dong, W.,. & Xu, J. C. (2020). Screening of phosphate-solubilizing fungi from air and soil in Yunnan, China: four novel species in Aspergillus, Gongronella, Penicillium, and Talaromyces. Frontiers in microbiology, 11, 585215. https://doi.org/10.3389/fmicb.2020.585215
  42. Samson, R. A., Visagie, C. M., Houbraken, J., Hong, S. B., Hubka, V., Klaassen, C. H.,. & Frisvad, J. (2014). Phylogeny, identification and nomenclature of the genus Aspergillus. Studies in mycology, 78(1), 141-173. https://doi.org/10.1016/j.simyco.2014.07.004
  43. Šimonovičová, A., Vojtková, H., Nosalj, S., Piecková, E., Švehláková, H., Kraková, L.,. & Pangallo, D. (2021). Aspergillus niger environmental isolates and their specific diversity through metabolite profiling. Frontiers in Microbiology, 12, 658010. https://doi.org/10.3389/fmicb.2021.658010
  44. Toma, M. A., Nazir, K. N. H., Mahmud, M. M., Mishra, P., Ali, M. K., Kabir, A.,. & Alim, M. A. (2021). Isolation and identification of natural colorant producing soil-borne Aspergillus niger from Bangladesh and extraction of the pigment. Foods, 10(6), 1280. https://doi.org/10.3390/foods10061280
  45. Vassileva, M., Mendes, G. D. O., Deriu, M. A., Benedetto, G. D., Flor-Peregrin, E., Mocali, S.,. & Vassilev, N. (2022). Fungi, P-solubilization, and plant nutrition. Microorganisms, 10(9), 1716. https://doi.org/10.3390/microorganisms10091716
  46. Visagie, C. M., Houbraken, J., Frisvad, J. C., Hong, S. B., Klaassen, C. H. W., Perrone, G.,. & Samson, R. A. (2014). Identification and nomenclature of the genus Penicillium. Studies in mycology, 78(1), 343-371. https://doi.org/10.1016/j.simyco.2014.09.001
  47. von Hertwig, A. M., Sant'Ana, A. S., Sartori, D., da Silva, J. J., Nascimento, M. S., Iamanaka, B. T.,. & Taniwaki, M. H. (2018). Real-time PCR-based method for rapid detection of Aspergillus niger and Aspergillus welwitschiae isolated from coffee. Journal of microbiological methods, 148, 87-92. https://doi.org/10.1016/j.mimet.2018.03.010
  48. Wang, K., Ngea, G. L. N., Godana, E. A., Shi, Y., Lanhuang, B., Zhang, X.,. & Zhang, H. (2023). Recent advances in Penicillium expansum infection mechanisms and current methods in controlling P. expansum in postharvest apples. Critical Reviews in Food Science and Nutrition, 63(15), 2598-2611. https://doi.org/10.1080/10408398.2021.1978384
  49. Wang, Y., Yuan, L., Wang, Y., Lang, J., Ye, M., Liu, Q.,. & Zhou, N. (2025). Phosphorus-solubilizing fungi promote the growth of Fritillaria taipaiensis PY Li by regulating physiological and biochemical reactions and protecting enzyme system-related gene expression. Frontiers in Genetics, 15, 1459191. https://doi.org/10.3389/fgene.2024.1459191
  50. Zhang, Y., Chen, F. S., Wu, X. Q., Luan, F. G., Zhang, L. P., Fang, X. M.,. & Ye, J. R. (2018). Isolation and characterization of two phosphate-solubilizing fungi from rhizosphere soil of moso bamboo and their functional capacities when exposed to different phosphorus sources and pH environments. PloS one, 13(7), e0199625. https://doi.org/10.1371/journal.pone.0199625