Diversity of inulinase-producing fungi associated with two Asteraceous plants, Pulicaria crispa (Forssk.) and Pluchea dioscoridis (L.) growing in an extreme arid environment

  • Doaa M. A. Khalil Botany Department, Faculty of Sciences, Aswan University, Aswan 81528, Egypt
  • Mohamed S. Massoud Botany Department, Faculty of Sciences, Aswan University, Aswan 81528, Egypt
  • Mostafa Abdelrahman Botany Department, Faculty of Sciences, Aswan University, Aswan 81528, Egypt; Graduate School of Life Sciences, Tohoku University, Sendai 9808577, Japan
  • Soad A. El-Zayat Botany Department, Faculty of Sciences, Aswan University, Aswan 81528, Egypt
  • Magdi A. El-Sayed Botany Department, Faculty of Sciences, Aswan University, Aswan 81528, Egypt; Unit of Environmental Studies and Development, Aswan University, Aswan 81528, Egypt
Keywords: Microbial inulinase, Arid land, Pluchea dioscoridis, Pulicaria crispa


Inulinases are potentially valuable enzymes catalyze the hydrolysis of plant’s inulin into high fructose syrups as sweetening ingredients for food industry and ethanol production. The high demands for inulinase enzymes have promoted interest in microbial inulinases as the most suitable approach for biosynthesis of fructose syrups from inulin. Arid land ecosystem represents a valuable bioresource for soil microbial diversity with unique biochemical and physiological properties. In the present study, we explored the fungi diversity associated with the rhizosphere and rhizoplane of two desert medicinal plants namely Pluchea dioscoridis and Pulicaria crispa growing in the South-Eastern desert of Aswan, Egypt. A total of 180 fungal isolates were screened based on their ability to grow on potato dextrose agar medium supplemented with 1% inulin. The isolated fungal colonies were morphologically identified according to cultural characteristics and spore-bearing structure. In addition, the inulinase activity of the isolated fungi was examined spectrophotometrically. Among these, Aspergillus terreus var. terreus 233, Botrytis cinerea, Aspergillus aegyptiacus, Cochliobolus australiensis 447 and Cochliobolus australiensis exhibited high inulinase activity ranging from 5.05 to 7.26 U/ml. This study provides a promising source of microbial inulinase, which can be scaled up for industrial applications.

DOI: http://dx.doi.org/10.5281/zenodo.1205649


1. Simões B, Conceição N, Matias AC, Bragança J, Kelsh RN, Cancela ML. Molecular characterization of cbfβ gene and identification of new transcription variants: implications for function. Arch Biochem Biophys. 2015; 567: 1-12.

2. Liu L, Zhang K, Sandoval H, Yamamoto S, Jaiswal M, Sanz E, et al. Glial lipid droplets and ROS induced by mitochondrial defects promote neurodegeneration. Cell. 2015; 160(1-2): 177-190.

3. Abdelrahman M, Abdel-Motaal F, El-Sayed M, Jogaiah S, Shigyo M, Ito S. Dissection of Trichoderma longibrachiatum-induced defense in onion (Allium cepa L.) against Fusarium oxysporum f. sp. cepa by target metabolite profiling. Plant Sci. 2016; 246: 128-138.

4. Ali AA, Abdelrahman M, Usama R, Soad E, El-Sayed M. Effect of Thermomyces fungal endophyte isolated from extreme hot desert adapted plant on heat stress tolerance of cucumber. Appl Soil Ecol. 2017: 1-8.

5. Jogaiah S, Abdelrahman M, El-Sayed M, Burritt DJ, Tran LP. The STAYGREEN trait and phytohormone signaling networks in plants under heat stress. Plant Cell Rep. 2017; 36: 1009-1025.

6. Cavaglieri LR, Keller KM, Pereyra CM, González-Pereyra ML, Alonso VA, Rojo FG, et al. Mycotoxicity of barley rootlets and malt used as pig feedstuff ingredients. J Stored Prod Res. 2009; 4: 147-150.

7. Wang L, Li H, Zhao C, Li S, Kong L, Wu W, et al. The inhibition of protein translation mediated by AtGCN1 is essential for cold tolerance in Arabidopsis thaliana. Plant Cell Environ. 2017; 40(1): 56-68.

8. Shivanna MB, Vasanthakumari MM. Temporal and spatial variability of rhizosphere and rhizoplane fungal communities in grasses of the subfamily Chloridoideae in the Lakkavalli region of the Western Ghats in India. Mycosphere. 2011; 2(3): 255-271.

9. Cardoso EJBN, Nogueira MA. A rizosfera e seus efeitos na comunida de microbianaena nutrição de plantas. In: Silveira APD, Freitas SS, eds. Microbiota do soloe qualidade ambiental. Instituto Agronômico, Campinas. 2007: 79-96.

10. Mwajita MR, Murage H, Tani A, Kahangi EM. Evaluation of rhizosphere, rhizoplane and phyllosphere bacteria and fungi isolated from rice in Kenya for plant growth promoters. Springer Plus. 2013; 2: 606.

11. Srivastava V, Kumar A. Biodiversity of mycoflora in rhizosphere and rhizoplane of some Indian herbs. Biol Forum Int J. 2013; 5(2): 123-125.

12. Kumar RS, Sivakumar T, Sunderam RS, Gupta M, Mazumdar UK, Gomathi P, et al. Antioxidant and antimicrobial activities of Bauhinia racemosa L. stem bark. Braz J Med Biol Res. 2005; 38: 10015-1024.

13. Flores-Gallegos AC, Contreras-Esquivel JC, Morlett-Chávez JA, Aguilar CN, Rodríguez-Herrera R. Comparative study of fungal strains for thermostable inulinase production. J Biosci Bioeng. 2015; 2(2): 1-6.

14. Abd Al-Aziz SAA, El-Metwally MM, Saber WEIA. Molecular identification of a novel inulinolytic fungus isolated from and grown on tubers of Helianthus tuberosus and statistical screening of medium components. World J Microbiol Biotechnol. 2012; 28: 3245-3254.

15. Sheng J, Chi ZM, Li J, Gao LM, Gong F. Inulinase production by the marine yeast Cryptococcus aureus G7a and inulin hydrolysis by the crude inulinase. Process Biochem. 2007; 42: 805-811.

16. Gill PK, Manhas RK, Singh P. Purification and properties of a heat-stable exoinulinase isoform from Aspergillus fumigatus. Biores Technol. 2006; 97: 894-902.

17. Zhao J, Lin W, Ma X, Lu Q, Ma X, Bian G, Jiang L. The protein kinase Hal5p is the high-copy suppressor of lithium-sensitive mutations of genes involved in the sporulation and meiosis as well as the ergosterol biosynthesis in Saccharomyces cerevisiae. Genomics. 2010; 95(5): 290-298.

18. El-Naggar SA, Alm-Eldeen AA, Germoush MO, El-Boray KF, Elgebaly HA. Ameliorative effect of propolis against cyclophosphamide-induced toxicity in mice. Pharm Biol. 2014; 53: 235-241.

19. Timon MI. The interaction of higher plants and soil microorganism. I - Microbial population of rhiosphere of seedling of certain cultivated plants. Can J Res. 1940; 18: 307-317.

20. Moubasher AH, Abdel-Hafez SII. Effect of soil amendements with three organic substances on soil, rhizosphere and rhizoplane fungi and on the incidence of damping off-disease of cotton seedling in Egypt. Naturalia Monspeliensia Ser Bot. 1986; 50: 91-108.

21. Abdel-Hafez AII, Moharram AM, Abdel-Gawad KM. Survey of keratinophilic and saprobic fungi in the cloven-hooves and horns of goats and sheep from Egypt. J Basic Microbiol. 1990; 30(1): 13-20.

22. Raper KB, Thom C. A manual of the Penicillia. Williams and Wilkins, Baltimore, USA, 1949.

23. Raper KB, Fennell PI. The genus Aspergillus. Williams and Wilkins, Baltomore, USA, 1965.

24. Ellis MM. Dematiaceae Hyphomycetes. Common wealth Mycological Institute, Kew, Surrey, England, 1976.

25. Booth C. Fusarium Laboratory Guide to the identification of major species. CMI, Kew, Surrey, England, 1977.

26. Christensen M, Raper KB. Synoptic key to Aspergillus nidulans group species and related Emericella species. Transact Brit Mycol Soc. 1978; 71(2): 177-191.

27. Pitt JI. A laboratory guide to common Penicillium species. Common wealth Scientific and Industerial Research Organization, Division of Food Research, North Ryde, N.S.W. Australia, 1985.

28. Moubasher AH. Soil fungi in Qatar and other Arab Countries. The Scientific and Applied Research Center, University of Qatar, Doha, Qatar, 1993.

29. Singh RS, Sooch BS, Puri M. Optimization of medium and process parameters, for the production of inulinase from a newly isolated Kluyveromyces marxianus YS-1. Biores Technol. 2007; 98: 2518-2525.

30. Rozpądek P, Wężowicz K, Nosek M, Ważny R, Tokarz K, Lembicz M, et al. The fungal endophyte Epichloe typhina improves photosynthesis efficiency of its host orchard grass (Dactylis glomerata). Planta. 2015; 242: 1025-1035.

31. Alphei J, Bonkowski M, Scheu S. Protozoa, nematoda and lumbricidae in the rhizosphere of Hordelymus europaeus (Poaceae): Faunal interactions, response of microorganisms and effects on plant growth. Oecologia. 1996; 106: 111-126.

32. Abdel-Motaal FF. The role of secondary metabolites of the medicinal Solanacies plant (Hyoscyamus muticus L.) and its associated fungi in plant - fungal interaction. Ph. D thesis, Tottori University, Japan, 2010: 1-161.

33. Lima TEF, Bezerra GL, Queiroz Cavalcanti MA. Fungi from the rhizosphere and rhizoplane from the grapevine Vitis labrusca in Pernambuco, Brazil. Nova Hedwigia. 2014; 99: 531-540.

34. Klich MA. Biogeography of Aspergillus species in soil and litter. Mycologia. 2002; 94(1): 21-27.

35. Qureshi SA, Sutlana V, Haque SE, Athar M. Isolation and identification of fungi associated with the rhizosphere and rhizoplane of wild and cultivated plants of Pakistan. Sida. 2004; 21(2): 1019-1053.

36. Deyab AS. Ecological studies on mycoflora in Wadi Allaqi Biosphere Reserve, Egypt. M.Sc. Thesis, South Valley University. Aswan, 2006: 119 & 145.

37. Gherbawy Y, Maghraby T, Yassmin S. Seasonal variation of Fusarium species in wheat fields in Upper Egypt. Phytopathol Plant Protect. 2006; 39(5): 365-377.

38. Seddek NH. Fungi associated with some wild plants. M. Sc. Thesis, Department of Botany, Faculty of Science, Assiut, University, Egypt, 2007.

39. Jain SC, Jain PC, Kango N. Production of inulinase from Kluyveromyces marxianus using Dahlia tuber extract. Braz J Microbiol. 2012; 43(1): 62-69.

40. Miller GL. Use of dinitrosalysalic acid reagent for determination of reducing sugar. Anal Chem. 1959; 31: 426-428.

41. Souza-Motta CM, Queiroz-Cavalcanti MA, Santos-Fernands MJ, Massa-Lima DM, Nascimento JP, Laranjeira D. Identification and characterization of filamentous fungi isolated from the sunflower (Hellanthu annus L.) Rhizosphere according to their capacity to hydrolyse inulin. Braz J Microbiol. 2003; 34: 273-280.

42. Abdel-Hafez SII, Ismail MA, Hussein NA, Nafady NA. The diversity of Fusarium species in Egyptian soils, with three new record species. The first International Conference of Biological Sciences, March 4-5th 2009, Faculty of Science, Assiut University, Assiut, Egypt. Assiut Univ J Bot. 2009; (Special 1): 129-147.

43. Ismail MA, Abdel-Hafez SII, Hussein NA, Nafady NA. Monthly fluctuations of Fusarium species in cultivated soil, with a new record species. The first International Conference of Biological Sciences, March 4-5th 2009, Faculty of Science, Assiut University, Assiut, Egypt. Assiut Univ J Bot. 2009; (Special 1): 117-128.

44. Bhat PR, Kaveriappa KM. Rhizoplane mycoflora of some species of Myristicaceae of the Western Ghats, India. Trop Ecol. 2011; 52(2): 163-175.

45. Gomathi S, Ambikapathy V, Panneerselvam A. Studies on soil mycoflora in Chilli Field of Thiruvarur District. Asian J Res Pharm Sci. 2011; 1(4): 117-122.

46. Kumar VV, Premkumar MP, Sathyaselvabala VK, Dineshkirupha S, Nandagopal J, Sivanesan S. Aspergillus niger exo-inulinase purification by three phase partitioning. Eng Life Sci. 2011; 11(6): 607-614.

47. Silva MF, Rigo D, Mossi V, Dallago RM, Henrick P, Kuhn GO, et al. Evaluation of enzymatic activity of commercial inulinase from Aspergillus niger immobilized in polyurethane foam. Food Bioprod Process. 2013; 91: 54-59.

48. Zhang L, Zhao C, Zhu D, Ohta Y, Wang Y. Purification and characterization of inulinase from Aspergillus niger AF10 expressed in Pichia pastoris. Protein Exp Purif. 2004; 35: 272-275.

49. Coitnho JB, Guimaraes V M, De Almeida MN, Falkoskl DL, De Queiroz JH, De Rezende ST. Characterization of an exoinulinase produced by Aspergillus terreus CCT 4083 grown on sugar cane bagasse. J Agri Food Chem. 2010; 58(14): 8386-8391.

50. Noda J, Brito N, Gonzalez C. The Botrytis cinerea xylanase Xyn11A contributes to virulence with its necrotizing activity, not with its catalytic activity. BMC Plant Biol. 2010; 10: 38.
How to Cite
Khalil, D., Massoud, M., Abdelrahman, M., El-Zayat, S., & El-Sayed, M. (2018). Diversity of inulinase-producing fungi associated with two Asteraceous plants, Pulicaria crispa (Forssk.) and Pluchea dioscoridis (L.) growing in an extreme arid environment. European Journal of Biological Research, 8(2), 42-55. Retrieved from http://journals.tmkarpinski.com/index.php/ejbr/article/view/13
Research Articles