Combination on endophytic fungal as the Plant Growth-Promoting Fungi (PGPF) on cucumber (Cucumis sativus)




Abstract. Syamsia S, Idhan A, Firmansyah AP, Noerfitryani N, Rahim I, Kesaulya H, Armus R. 2021. Combination on endophytic fungal as the Plant Growth-Promoting Fungi (PGPF) on Cucumber (Cucumis sativus). Biodiversitas 22: 1194-1202. Endophytic fungi are known to stimulate plant growth by producing secondary metabolites, including phytohormones (IAA and Gibberellins), siderophore, phosphate-solubilizing metabolites. In this study, a total of six endophytic fungi were successfully isolated from local rice plants and showed different abilities in producing secondary metabolites, during single isolates testing. These six isolates were then combined to obtain 15 combinations for analysis, to determine the best combination for application as a plant growth promoter. Subsequently, each combination was tested for phytohormones (IAA, gibberellins) and siderophore (quantitatively)-producing activity, phosphate-solubilizing ability, and the effect on cucumber (Cucumis sativus L) plant growth. F13 showed activity in producing IAA and produced the highest gibberellin levels, while F1 exhibited the highest phosphate-solubilizing activity. In addition, F11 (Na-salicylate) and F1 (catechol) showed the highest siderophore activity, while a combination of F6, F8, F9, and F12 successfully increased plant height growth. Also, F4 increased the root growth, while the fresh weight of cucumber was increased by F8 treatment, under controlled conditions. Molecular analysis showed the tested isolates have close similarity to Daldinia eschscholtzii, Sarocladium oryzae, Rhizoctonia oryzae, Penicillium allahabadense, and Aspergillus foetidus. The combination of endophyte fungal isolates showed potential as plant growth promoters, however, further testing on several plant types is required before the combination is to be widely applied.


Abdel-Motaal, Fatma, Noha Kamel, Soad El-Zayat, and Mohamed Abou-Ellail. 2020. “Early Blight Suppression and Plant Growth Promotion Potential of the Endophyte Aspergillus Flavus in Tomato Plant.” Annals of Agricultural Sciences 65 (2): 117–23.
Ahmed, E., and S. J.M. Holmström. 2014. “Siderophores in Environmental Research: Roles and Applications.” Microbial Biotechnology 7 (3): 196–208.
Ahmed, A., Hasnain, S., 2010. Auxin-producing Bacillus sp.: Auxin quantification and effect on the growth ofSolanum tuberosum. Pure and Applied Chemistry 82, 313–319
Alori, Elizabeth T., Bernard R. Glick, and Olubukola O. Babalola. 2017. “Microbial Phosphorus Solubilization and Its Potential for Use in Sustainable Agriculture.” Frontiers in Microbiology 8 (JUN): 1–8.
Aramsirirujiwet, Yaovapa, and Chenchira Gumlangmak and Vichien Kitpreechavanich. 2016. “Studies on Antagonistic Effect Against Plant Pathogenic Fungi from Endophytic Fungi Isolated from Hottuynia Cordata Thunb. and Screening for Siderophore And.” KKU Res. J. 21 (1): 55–66. kkurj.2016.5
Barka AE, Gognies S, Nowak J, Audran JC, Belarbi A. 2002. Inhibitory effect of endophyte bacteria on Botrytis cinerea and its influence to promote the grapevine growth. Biol Control. 24 (2):135–142. doi: 10.1016/S1049-9644(02)00034-8
Casimiro, I., A. Marchant, R.P. Bhalerao, T. Beeckman, S. Dhooge, R. Swarup, N. Graham, D. Inze, G. Sandberg, P.J. Casero, M. Bennett., 2001. Auxin transport promotes Arabidopsis lateral root initiation. Plant Cell, 13: 843–852. DOI:
Duhan, Priti, Poonam Bansal, and Sulekha Rani. 2020. “Isolation , Identi Fi Cation and Characterization of Endophytic Bacteria from Medicinal Plant Tinospora Cordifolia.” South African Journal of Botany 000: 1–7.
El-Maraghy, Saad Shehata, Tohamy Anwar Tohamy, and Khalid Abdallah Hussein. 2020. “Role of Plant-Growth Promoting Fungi (PGPF) in Defensive Genes Expression of Triticum Aestivum against Wilt Disease.” Rhizosphere 15 (May): 100223.
Elias, Firew, Delelegn Woyessa, and Diriba Muleta. 2016. “Phosphate Solubilization Potential of Rhizosphere Fungi Isolated from Plants in Jimma Zone, Southwest Ethiopia.” International Journal of Microbiology 2016 (1).
Elsharkawy, Mohsen Mohamed, and Nagwa Mohamed Mohamed El-Khateeb. 2019. “Antifungal Activity and Resistance Induction against Sclerotium Cepivorum by Plant Growth-Promoting Fungi in Onion Plants.” Egyptian Journal of Biological Pest Control 29 (1).
Fadiji, Ayomide Emmanuel, and Olubukola Oluranti Babalola. 2020. “Exploring the Potentialities of Beneficial Endophytes for Improved Plant Growth.” Saudi Journal of Biological Sciences, no. xxxx.
Ghosh, Swapan Kr, Subhankar Banerjee, and Chandan Sengupta. 2017. “Siderophore Production by Antagonistic Fungi (Coleoptera: Chrysomelidae: Bruchinae) © 527 Bioassay, Characterization and Estimation of Siderophores from Some Important Antagonistic Fungi.” JBiopest 10 (2): 105–12.
Glickman E, Dessaux Y. A critical examination of the specificaty of the Salkowski reagent for indolic compoumds produced by phytopatogenic bacteria. Appl Environ Microbial 1995; 61:793-796.
Hamayun, Muhammad, Sumera Afzal Khan, Abdul Latif Khan, Nadeem Ahmad, Yasmin Nawaz, Hasan Sher, and In Jung Lee. 2011. “Gibberellin Producing Neosartorya Sp. CC8 Reprograms Chinese Cabbage to Higher Growth.” Scientia Horticulturae 129 (3): 347–52.
Hilarino, M. P. A., Silveira, F. A. de O. e, Oki, Y., Rodrigues, L., Santos, J. C., Corrêa Junior, A., Rosa, C. A. (2011). Distribution of the endophytic fungi community in leaves of Bauhinia brevipes (Fabaceae). Acta Botanica Brasilica, 25(4), 815– 821. 400008
Hiraishi,A., Kamagata, Y., & Nakamura, N. 1995. Polymerase chain reaction amplification and restriction fragment length polymorphism analysis of 16S rRNA genes from methanogens. Journals of Fermentation Bioengineering. 79: 523--529.
Jogaiah, S., Abdelrahman, M., Tran, L.-S.P., Ito, S.-I., 2018. Different mechanisms of Trichoderma virens-mediated resistance in tomato against Fusarium wilt involve the jasmonic and salicylic acid pathways. Mol. Plant Pathol. 19, 870–882.
Kawaide, H., 2006. Biochemical and molecular analysis of gibberellin biosynthesis in fungi. Biosci. Biotechnol. Biochem. 70, 583–590
Kesaulya, H., J. V. Hasinu, and G. N.C. Tuhumury. 2018. “Potential of Bacillus Spp Produces Siderophores Insuppressing Thewilt Disease of Banana Plants.” IOP Conference Series: Earth and Environmental Science 102 (1).
Khan, A.L., Hussain, J., Al-Harrasi, A., et al., 2013. Endo- phytic fungi: a source of gibberellins and crop resistance to abiotic stress. Crit. Rev. Biotech., 35(1):1-13. 07388551. 2013.800018
Leitão, Ana Lúcia, and Francisco J. Enguita. 2016. “Gibberellins in Penicillium Strains: Challenges for Endophyte-Plant Host Interactions under Salinity Stress.” Microbiological Research 183: 8–18.
Lu H ,Zou WX, Meng JC, Hu J, Tan RX. 2000. New bioactive metabolites produced by Collelotrichum sp., an endophytic fungi in Artemisia annua. Plant Sci. 151 (1): 67-73. doi: 10.1016/S0168-9452(99)00199-5.
MacMillan, J., 2002. Occurence of gibberellins in vascular plants, fungi and bacteria. J. Plant Growth Reg. 20, 21, 242–243.
Manganyi, M.C., Regnier, T., Tchatchouang, C.D.K., Bezuidenhout, C.C., Ateba, C.N., 2019. Antibacterial activity of endophytic fungi isolated from Sceletium tortuosum L. (Kougoed). Ann. Microbiol. 69 (6), 659–663.
Malinowski, D.P., Belesky, D.P., 1999. Neotyphodium coenophialum-endophyte infection affects the ability of tall fescue to use sparingly available phosphorus. J. Plant Nutr. 22, 835–853. 01904169909365675
Mishra, B.S., Singh, M., Aggrawal, P., Laxmi, A., 2009. Glucose and auxin signaling inter- action in controlling Arabidopsis thaliana seedlings root growth and development. PLoS One 4, e4502.
Navale, A. M., D. B. Shinde, B. R. Vaidya, and S. B. Jadhav. 1995. Effects of Azotobacter and Azospirillum inoculation under graded levels of nitrogen on growth and yield of sugarcane (Saccharum officinarim). Ind. J. Agron. 40:665–669.
Numponsak, Tosapon, Jaturong Kumla, Nakarin Suwannarach, Kenji Matsui, and Saisamorn Lumyong. 2018. “Biosynthetic Pathway and Optimal Conditions for the Production of Indole-3-Acetic Acid by an Endophytic Fungus, Colletotrichum Fructicola CMU-A109.” Edited by Sabrina Sarrocco. PLOS ONE 13 (10): e0205070.
Neilands, J.B. 1952. A Crystalline Organo- iron pigment from a rust fungus (Ustilago sphaerogena). Journal of The American Chemical Society, 74: 4846.
O`Donnell, K. 1993. Fusarium and its near relatives. In: Reynolds, D.R. & Taylor, J.W. (eds). The fungal holomorph: Mitotic, meiotic, and pleomorphic specification in fungal systematics. CAB International, Wallingford, pp. 225--233.
Pieterse, C.M., Van der Does, D., Zamioudis, C., Leon-Reyes, A., and Van Wees, S.C. (2012). Hormonal modulation of plant immunity. Annu. Rev. Cell Dev. Biol. 28:489–521.
Renshaw, Joanna C., Verity Halliday, Geoffrey D. Robson, Anthony P.J. Trinci, Marilyn G. Wiebe, Francis R. Livens, David Collison, and Robin J. Taylor. 2003. “Development and Application of an Assay for Uranyl Complexation by Fungal Metabolites, Including Siderophores.” Applied and Environmental Microbiology 69 (6): 3600–3606.
Sapareng, Sukriming, Ambo Ala, Tutik Kuswinanti, and Burhanuddin Rasyid. 2017. “Capability of Rot Fungus Isolates from Oil Palm Empty Bunches in the Production of Indole Acetic Acid (IAA).” International Journal of Current Microbiology and Applied Sciences 6 (11): 2174–80. ijcmas.2017.611.256.
Saxena, Jyoti, Amita Saini, Indu Ravi, Shalini Chandra, and Veena Garg. 2015. “Consortium of Phosphate-Solubilizing Bacteria and Fungi for Promotion of Growth and Yield of Chickpea (Cicer Arietinum).” Journal of Crop Improvement 29 (3): 353–69.
Syed, Shameer, N. V.K.V.Prasad Tollamadugu, and Bin Lian. 2020. “Aspergillus and Fusarium Control in the Early Stages of Arachis Hypogaea (Groundnut Crop) by Plant Growth-Promoting Rhizobacteria (PGPR) Consortium.” Microbiological Research 240 (1): 126562.
Sharma, S. B., Sayyed, R. Z., Trivedi, M. H., and Gobi, T. A. (2013). Phosphate solubilizing microbes: sustainable approach for managing phosphorus deficiency in agricultural soils. Springerplus 2, 587–600. doi: 10.1186/2193- 1801-2-587
Singh, S., and Mathur, N. (2010). Biological control of root-knot nematode, Meloidogyne incognita infesting tomato. Biocontrol Sci. Technol. 20, 865–874. doi: 10.1080/09583157.2010.487935
Spellberg, B.R., Guidos, D., Gilbert, J., Bradly, H.W., Boucher, W.M., Scheld, J.G., Bartlett Jr., J.E., 2008. The epidemic of antibiotic-resistant infections: a call to action for the medical community from the infectious diseases society of America. Clin. Infect. Dis. 46, 155–164.
Srinivasan, R., Yandigeri, M. S., Kashyap, S., and Alagawadi, A. R. (2012). Effect of salt on survival and P-solubilization potential of phosphate solubilizing microorganisms from salt affected soils. Saudi J. Biol. Sci. 19, 427–434. doi: 10.1016/j.sjbs.2012.05.004
Techaoei, Surachai, Chariya Jirayuthcharoenkul, Khemjira Jarmkom, Thisakorn Dumrongphuttidecha, and Warachate Khobjai. 2020. “Chemical Evaluation and Antibacterial Activity of Novel Bioactive Compounds from Endophytic Fungi in Nelumbo Nucifera.” Saudi Journal of Biological Sciences 27 (11): 2883–89.
Vandenbussche, F., Fierro, A.C., Wiedemann, G., Reski, R., Van Der Straeten, D., 2007. Evolutionary conservation of plant gibberellin signaling pathway components. BMC Plant Biol. 7, 65.
Vasundhara, M., Kumar, A., Reddy, M.S., 2016. Molecular approaches to screen bioactive compounds from endophytic fungi. Front. Microbiol. 7, 1–12.
Verma, V.C., Singh, S.K., and Prakash, S. (2011) Bio-control and plant growth promotion potential of siderophore pro- ducing endophytic Streptomyces from Azadirachta indica A. Juss. J Basic Microbiol 51: 550–556.
Waqas, Muhammad, Abdul Latif Khan, Muhammad Kamran, Muhammad Hamayun, Sang Mo Kang, Yoon Ha Kim, and In Jung Lee. 2012. “Endophytic Fungi Produce Gibberellins and Indoleacetic Acid and Promotes Host-Plant Growth during Stress.” Molecules 17 (9): 10754–73. molecules 170910754.
White, T.J., Bruns, T.D., Lee, S.B. & Taylor, J.W. 1990. Amplification and direct sequencing of fungal RNA genes for phylogenetics. In: Innis, M.A., Gelfand, D.H., Sninsky, J.J. & White, T.J. (eds). PCR protocols. Academic, San Diego, pp. 315--322.
Widowati, Tiwit, Nuryanah Nuriyanah, Indah Budi Asih, and Harsatini Sukiman. 2016. “THE POTENCY OF Endophytic Fungi Isolated From Taro ( Colocasia Esculenta (L.) Schott) As Plant Growth Promoting AgenT.” In The 6th International Symposium for Sustainable Humanosphere, 15–16.
Wulandari, Reine Suci, and Rosa Suryantini. 2019. “Endophytic Fungi in Paraserianthes Falcataria: Production of Indole Acetic Acid.” Microbiology Indonesia 13 (1): 16–24.
Yuan, Z. L., Zhang, C. L., & Lin, F. C. (2010). Role of diverse non-systemic fungal endophytes in plant performance and response to stress: progress and approaches. Journal of Plant Growth Regulation, 29,116-126.
Zhang, Chunlei, Youming Zhang, Zhaojun Ding, and Yang Bai. 2019. “Contribution of Microbial Inter-Kingdom Balance to Plant Health.” Molecular Plant 12 (2): 148–49.
Zhou, Li Si, Kun Tang, and Shun Xing Guo. 2018. “The Plant Growth-Promoting Fungus (PGPF) Alternaria Sp. A13 Markedly Enhances Salvia Miltiorrhiza Root Growth and Active Ingredient Accumulation under Greenhouse and Field Conditions.” International Journal of Molecular Sciences 19 (1): 1–15.