Biochar and microbial bio-fertilizers, such as endophytic fungi and plant growth-promoting endophytes (PGPEs), offer sustainable alternatives to chemical fertilizers by enhancing soil fertility and plant performance. However, their synergistic effects particularly those involving auxin (IAA) biosynthesis and nutrient uptake, remain underexplored in calcareous soils. This study investigates how the integration of biochar with auxin-producing microbial inoculants influences soil-plant interactions and canola productivity. A pot experiment was conducted using two Brassica napus (canola) cultivars (DGL and Faisal canola) grown under alkaline calcareous soil. Treatments included individual and combined applications of A. baumannii MN24 and P. indica, with and without biochar. Physiological traits, auxin concentrations (IAA), root and shoot biomass, nutrient uptake (N, P, K), seed quality, and soil microbial and enzymatic activities were measured. Results revealed that the combined application of biochar, MN24, and P. indica significantly enhanced plant biomass, stem diameter, and grain yield by up to 203%, 127%, and 212%, respectively, in Faisal canola compared to the control. Biochar also amplified the microbial colonization and enzymatic activity in soil, leading to higher microbial biomass, enhanced nutrient uptake, and reduced electrolyte leakage and osmotic potential, indicating improved plant performance. Notably, these improvements translated to higher canola seed quality, with elevated fat, ash, and fiber contents. Enhanced auxin synthesis, particularly in the presence of l-tryptophan, was linked to improved root architecture, microbial colonization, and reduced plant stress indicators. These findings suggest that biochar-microbe-l-tryptophan synergy offers a powerful tool for sustainable crop intensification in challenging soils by optimizing hormone-mediated soil-plant processes.

Centre for Water and Environmental Studies King Faisal University Al Ahsa 31982 Saudi Arabia

Date Palm Research Center of Excellence King Faisal University Al Ahsa 31982 Saudi Arabia

Department of Agrochemistry Soil Science Microbiology and Plant Nutrition Faculty of AgriSciences Mendel University in Brno Zemedelska 1 Brno 61300 Czech Republic

Guangdong Provincial Key Laboratory of Applied Botany South China Botanical Garden Chinese Academy of Sciences Guangzhou 510650 China

Institute of Soil and Environmental Sciences University of Agriculture Faisalabad 38040 Pakistan

Key Laboratory of Arable Land Conservation Ministry of Agriculture and Rural Affairs Huazhong Agricultural University Wuhan 430070 China

Key Laboratory of Grassland Ecology School of Ecology and Environment Inner Mongolia University Hohhot 010020 China

Zobrazit více v PubMed

Mohamed I. A. Shalby N. El-Badri A. M. Awad-Allah E. F. Batool M. Saleem M. H. Wang Z. Wen J. Ge X. Xu Z. Wang J. J. Agric. Food Res. 2025:101794.

Borges C. E. Von dos Santos Veloso R. Da Conceição C. A. Mendes D. S. Ramirez-Cabral N. Y. Shabani F. Shafapourtehrany M. Nery M. C. Da Silva R. S. Sci. Rep. 2023;13:12656. PubMed PMC

Secchi M. A. Fernandez J. A. Stamm M. J. Durrett T. Prasad P. V. V. Messina C. D. Ciampitti I. A. Field Crops Res. 2023;293:108848.

Naveed M. Abid I. Mustafa F. Ahmad H. R. Alamri S. Siddiqui M. H. Alfagham A. T. Mustafa A. Chem. Biol. Technol. Agric. 2024;11:168.

Hudhaifa A. H. Aljumaily M. M. Al-Obaidi M. A. Rev. Agric. Neotrop. 2024;11:e8543.

Younas N. Naveed M. Yaseen M. Younas M. Mumtaz M. Z. Babar M. H. Nadeem M. Soysal S. Al-Ashkar I. Mustafa A. Agronomy. 2024;14:1289.

Muzaffar M. U. Naveed M. Naseem Z. Abid I. Amir K. Z. Alamri S. Siddique M. Brtnicky M. Mustafa A. Chemosphere. 2024;368:143797. PubMed

Xing Y. Wang X. Mustafa A. Ecotoxicol. Environ. Saf. 2025;289:117703. PubMed

Zhang F. Hu B. Meng L. Pang M. Chen Y. Song W. Feng Y. Ma Y. Soil Use Manag. 2025;41:e70044.

Holatko J. Kucerik J. Mustafa A. Lonova K. Siddiqui M. H. Naveed M. Hammerschmiedt T. Kintl A. Malicek O. Chorazy T. Baltazar T. BMC Plant Biol. 2025;25:323. PubMed PMC

Zou Q. Zhao L. Guan L. Chen P. Zhao J. Zhao Y. Du Y. Xie Y. Front. Plant Sci. 2024;15:1501400. PubMed PMC

Iqbal M. Naveed M. Sanaullah M. Brtnicky M. Hussain M. I. Kucerik J. Holatko J. Mustafa A. J. Soils Sediments. 2023;23:1233–1249.

Virk Z. A. Al Farraj D. A. Iqbal M. Lewińska K. Hussain S. Sustainability. 2022;14:6975.

Saeed Q. Xiukang W. Haider F. U. Kučerik J. Mumtaz M. Z. Holatko J. Naseem M. Kintl A. Ejaz M. Naveed M. Brtnicky M. Int. J. Mol. Sci. 2021;22:10529. PubMed PMC

Shahzad R. Bilal S. Imran M. Khan A. L. Alosaimi A. A. Al-Shwyeh H. A. Almahasheer H. Rehman S. Lee I. J. Biochem. J. 2019;476:3385–3400. PubMed

Nazli F. Mustafa A. Ahmad M. Hussain A. Jamil M. Wang X. Shakeel Q. Imtiaz M. El-Esawi M. A. Sustainability. 2020;12:9056.

Kour A. Shawl A. S. Rehman S. Sultan P. Qazi P. H. Suden P. Khajuria R. K. Verma V. World J. Microbiol. Biotechnol. 2008;24:1115–1121.

Mourouzidou S. Ntinas G. K. Tsaballa A. Monokrousos N. Sustainability. 2023;15:5959.

Akhtar J., Upadhyay P. and Kumar H., in Hybrid Seed Production for Boosting Crop Yields: Applications, Challenges and Opportunities, Springer Nature Singapore, Singapore, 2025, pp. 177–224

Javed T. I I. Singhal R. K. Shabbir R. Shah A. N. Kumar P. Jinger D. Dharmappa P. M. Shad M. A. Saha D. Anuragi H. Front. Plant Sci. 2022;13:877544. PubMed PMC

Mushtaq T. Bano A. Ullah A. J. Plant Growth Regul. 2024:1–22.

Varma A., Singh A., Sahay N. S., Sharma J., Roy A., Kumari M., Rana D., Thakran S., Deka D., Bharti K., Piriformospora indica: an axenically culturable mycorrhiza-like endosymbiotic fungus, in Fungal Associations, Springer Berlin Heidelberg, Berlin, Heidelberg, 2001, pp. 125–150

Sarwar M. Arshad M. Martens D. A. Frankenberger W. Plant Soil. 1992;147:207–215.

Sánchez M. E. Lindao E. Margaleff D. Martínez O. Morán A. J. Anal. Appl. Pyrolysis. 2009;85:142–144.

Vance E. D. Brookes P. C. Jenkinson D. S. Soil Biol. Biochem. 1987;19:703–707.

Sanaullah M. Blagodatskaya E. Chabbi A. Rumpel C. Kuzyakov Y. Appl. Soil Ecol. 2011;48:38–44.

Reiter B. Pfeifer U. Schwab H. Sessitsch A. Appl. Environ. Microbiol. 2002;68:2261–2268. PubMed PMC

Steel R. G., Torrie J. H., Dicky D. A., Principles and Procedures of Statistics, 2nd edn, 1997

Li C. Huang Q. Zhang H. Wang Q. Xue R. Guo G. Hu J. Li T. Wang J. Hu S. Int. J. Environ. Res. Public Health. 2021;18:11413. PubMed PMC

Mona S. Saini N. Kumar S. Sharma A. Yadav A. Yadav N. Deepak B. Bioresour. Technol. Rep. 2024;27:101893.

Hou J. Pugazhendhi A. Sindhu R. Vinayak V. Thanh N. C. Brindhadevi K. Chi N. T. Yuan D. Environ. Res. 2022;14:113909. PubMed

Nurhidayati N. Ansari A. S. Sholihah A. Chiangmai P. N. J. Hortic. Res. 2022;30:55–66.

Yeboah E. Asamoah G. Ofori P. Amoah B. Agyeman K. O. Open Agric. 2020;5:352–360.

Ajeng A. A. Abdullah R. Ling T. C. Ismail S. Lau B. F. Ong H. C. Chew K. W. Show P. L. Chang J. S. Environ. Technol. Innov. 2020;20:101168.

Głodowska M. Schwinghamer T. Husk B. Smith D. Agric. Sci. 2017;8:1048–1064.

Bolan S. Hou D. Wang L. Hale L. Egamberdieva D. Tammeorg P. Li R. Wang B. Xu J. Wang T. Sun H. Sci. Total Environ. 2023;886:163968. PubMed

Ma H. Shurigin V. Jabborova D. dela Cruz J. A. dela Cruz T. E. Wirth S. Bellingrath-Kimura S. D. Egamberdieva D. Plants. 2022;11:423. PubMed PMC

Lu J. Zhang W. Liu X. Khan A. Wang W. Ge J. Yan S. Xiong Y. C. Environ. Technol. Innov. 2024;35:103683.

Rafique M. Ortas I. Ahmed I. A. Rizwan M. Afridi M. S. Sultan T. Chaudhary H. J. J. Environ. Manage. 2019;247:672–680. PubMed

Liu J. Chen Z. Wu S. Sun H. Xing J. Zhang Z. Plants. 2024;13:614. PubMed PMC

Sharifi M. Hajiaghaei-Kamrani M. Can. J. Soil Sci. 2022;103:200–212.

Winarso S. Hermiyanto B. Romadhona S. Pandutama M. H. Setiawati T. C. Indasah I. J. Degraded Min. Lands Manage. 2020;7:2263.

Sarapat S. Longtonglang A. Umnajkitikorn K. Girdthai T. Boonkerd N. Tittabutr P. Teaumroong N. J. Plant Interact. 2020;15:322–334. PubMed PMC

Singh M. Qureshi K. A. Jaremko M. Rajput M. Singh S. K. Pandey K. D. Ferreira L. F. Kumar A. Plants. 2022;11:1787. PubMed PMC

Yang H. G Q. H B. Chen H. P X. F R. Du J. J. Agric. Resour. Environ. 2023;40:15.

Aggangan N. S. Aguilar E. A. Philipp. J. Crop Sci. 2019;44:89–97.

Azeem M. Hassan T. U. Tahir M. I. Ali A. Jeyasundar P. G. Hussain Q. Bashir S. Mehmood S. Zhang Z. Appl. Soil Ecol. 2021;157:103732.

Kocsis T. Kotroczó Z. Kardos L. Biró B. Environ. Technol. Innov. 2020;20:101191.

Ullah Z. Ali S. Muhammad N. Khan N. Rizwan M. Khan M. D. Khan N. Khattak B. Alhaithloul H. A. Soliman M. H. Ullah H. Arab. J. Geosci. 2020;13:757.

Schmidt H. P. Kammann C. Niggli C. Evangelou M. W. H. Mackie K. A. Abiven S. Agric. Ecosyst. Environ. 2014;191:117–123.

Schulz H. Glaser B. J. Plant Nutr. Soil Sci. 2012;175:410–422.

Yu H. Zou W. Chen J. Chen H. Yu Z. Huang J. Tang H. Wei X. Gao B. J. Environ. Manage. 2019;232:8–21. PubMed

Chen H. Ma J. Wei J. Gong X. Yu X. Guo H. Zhao Y. Sci. Total Environ. 2018;635:333–342. PubMed

Liu D. Ding Z. Ali E. F. Kheir A. M. Eissa M. A. Ibrahim O. H. Sci. Rep. 2021;11:8739. PubMed PMC

Alori E. T. Dare M. O. Babalola O. O. Sustain. Agric. Rev. 2017:281–307.

Alori E. T. Babalola O. O. Front. Microbiol. 2018;9:2213. PubMed PMC

Salomon M. J. Demarmels R. Watts-Williams S. J. McLaughlin M. J. Kafle A. Ketelsen C. Soupir A. Bücking H. Cavagnaro T. R. van der Heijden M. G. Appl. Soil Ecol. 2022;169:104225.

Abid M. Ali S. Qi L. K. Zahoor R. Tian Z. Jiang D. Snider J. L. Dai T. Sci. Rep. 2018;8:4615. PubMed PMC

Igbokwe N. A. Sokoto J. Vet. Sci. 2018;16:1–23.

Valentovic P. S. Luxova M. S. Kolarovic L. S. Gasparikova O. S. Plant Soil Environ. 2006;52:184.

Çığ F. Sönmez F. Nadeem M. A. Sabagh A. E. Agronomy. 2021;11:2418.

Ijaz M. Tahir M. Shahid M. Ul-Allah S. Sattar A. Sher A. Mahmood K. Hussain M. Braz. J. Microbiol. 2019;50:449–458. PubMed PMC

Saeed M. Ilyas N. Jayachandran K. Shabir S. Akhtar N. Shahzad A. Sayyed R. Z. Bano A. Environ. Pollut. 2022;305:119282. PubMed

Lalay G. Ullah A. Iqbal N. Raza A. Asghar M. A. Ullah S. Environ. Dev. Sustain. 2024;26:3457–3480.

Zhang M. Liu Y. Wei Q. Gou J. Liu L. Gu X. Wang M. Horticulturae. 2023;9:1104.

Desoky E. S. Mahmood M. Feng D. Ahmed A. F. Al Masoudi L. M. Alahmari A. S. Eid R. S. Rady M. M. Ayyoub A. Taha N. A. BMC Plant Biol. 2025;25:236. PubMed PMC

Javeed H. M. Naeem R. Ali M. Qamar R. Sarwar M. A. Nawaz F. Shehzad M. Farooq A. Khalid S. Mubeen K. Masood N. Acta Physiol. Plant. 2022;44:110.

Pallavi D. C., Sharma A. K., in Plant-Microbe Interactions, Springer, 2017, pp. 607–626

Xie Y. Wang H. Guo Y. Wang C. Cui H. Xue J. Waste Manage. 2023;171:512–521. PubMed

Ren H. Wang Z. Lv C. Huang B. Sun X. Qin F. Forests. 2024;15:1091.

Wang K. Wang S. Zhang X. Wang W. Wang X. Kong F. Xi M. Environ. Res. 2024;240:117426. PubMed

Egamberdieva D. Hua M. Reckling M. Wirth S. Bellingrath-Kimura S. D. Environ. Sustain. 2018;1:19–24.

Mahmood M. Wang Y. Ahmed W. Mehmood S. Ayyoub A. Elnahal A. S. Li W. Zhan X. Front. Plant Sci. 2023;14:1224583. PubMed PMC

Qi W. Y. Chen H. Wang Z. Xing S. F. Song C. Yan Z. Wang S. G. J. Hazard. Mater. 2023;458:131921. PubMed

Lehmann J. Rillig M. C. Thies J. Masiello C. A. Hockaday W. C. Crowley D. Soil Biol. Biochem. 2011;43:1812–1836.

Kannan P. Paramasivan M. Marimuthu S. Swaminathan C. Agric. Ecosyst. Environ. 2021;308:107258.

Schoebitz M., López Belchí M. D., in Bioformulations, Springer, 2016, 251–265

Hale L. Luth M. Crowley D. Soil Biol. Biochem. 2015;81:228–235.

Jaiswal A. K. Alkan N. Elad Y. Sela N. Graber E. R. Frenkel O. Sci. Rep. 2020;10:13934. PubMed PMC

Yang E. Meng J. Hu H. Cheng D. Zhu C. Chen W. Ecotoxicol. Environ. Saf. 2019;170:338–345. PubMed

Hossain M. Z. Bahar M. M. Sarkar B. Donne S. W. Ok Y. S. Palansooriya K. N. Kirkham M. B. Chowdhury S. Bolan N. Biochar. 2020;2:379–420.

Yang W. Zhao Y. Yang Y. Zhang M. Mao X. Guo Y. Li X. Tao B. Qi Y. Ma L. Liu W. J. Soils Sediments. 2023;23:947–957.

Andrenelli M. C. Maienza A. Genesio L. Miglietta F. Pellegrini S. Vaccari F. P. Vignozzi N. Agric. Water Manage. 2016;163:190–196.

Asif M. Ahmad F. Ansari T. Khan A. J. Agric. Sci. 2017:2017.

Hossain M. K. Strezov V. Nelson P. F. Pedosphere. 2015;25:680–685.

Cong M. Hu Y. Sun X. Yan H. Yu G. Tang G. Chen S. Xu W. Jia H. Front. Plant Sci. 2023;14:1172425. PubMed PMC

Wu C. Zhi D. Yao B. Zhou Y. Yang Y. Zhou Y. Environ. Res. 2022;212:113226. PubMed

Grichar W. J. Janak T. W. McGinty J. A. Brewer M. J. Agronomy. 2023;13:1429.

Means N. E. Starbuck C. J. Kremer R. J. Jett L. W. Compost Sci. Util. 2005;13:116–121.

Alixandre R. D. Lima P. A. Almeida T. F. Oliveira J. S. Pereira M. B. Alixandre F. T. Jacomino G. R. Dias R. S. Alexandre R. S. Ferreira A. Passos R. R. Braz. J. Biol. 2023;83:e277437. PubMed

Caldara M. Gullì M. Graziano S. Riboni N. Maestri E. Mattarozzi M. Bianchi F. Careri M. Marmiroli N. Sci. Total Environ. 2024;917:170168. PubMed

Primaz C. T. Ribes-Greus A. Jacques R. A. Energy. 2021;235:121363.

Hu X. Song J. Wang H. Zhang W. Wang B. Lyu W. Wang Q. Liu P. Chen L. Xing J. Water Sci. Technol. 2019;80:2260–2272. PubMed

Hamzah Z. Shuhaimi S. N. IOP Conf. Ser. Earth Environ. Sci. 2018;215:012011.

Olszyk D. Shiroyama T. Novak J. Cantrell K. Sigua G. Watts D. Johnson M. G. Agrosyst. Geosci. Environ. 2020;3:e20067. PubMed PMC

Ali J. Mohammed A. S. Mekonnen A. B. Adv. Agric. 2023;2023:1483976.

Khan Z. Yang X. J. Fu Y. Joseph S. Khan M. N. Khan M. A. Alam I. Shen H. Biochar. 2023;5:52.

Zhang Y. Ding J. Wang H. Su L. Zhao C. BMC Plant Biol. 2020;20:288. PubMed PMC

Vázquez Vázquez M. D. César S. Barea Navarro J. M. Microb. Ecol. 2000;39:200–208.

Querejeta G. A. Int. J. Methodol. 2023;2:34–40.

Trivedi P. Singh K. Pankaj U. Verma S. K. Verma R. K. Patra D. D. Ecol. Eng. 2017;102:127–136.

Ayyar S. Appavoo S. Basker M. Pandiyarajan P. Kavimani R. Int. J. Curr. Microbiol. Appl. Sci. 2019;8:1804–1814.

Cherni A. E. Trabelsi D. Chebil S. Barhoumi F. Rodríguez-Llorente I. D. Zribi K. Water, Air, Soil Pollut. 2015;226:145.

Frankenberger Jr W. T. Johanson J. B. Soil Biol. Biochem. 1982;14:433–437.

Liu X. Zhang S. Plant Soil. 2019;440:11–24.

Haider F. U. Coulter J. A. Cheema S. A. Farooq M. Wu J. Zhang R. Shuaijie S. G. Liqun C. Ecotoxicol. Environ. Saf. 2021;214:112112. PubMed

Li M. Cai L. Sustainability. 2021;13:3244.

Rafique M. Ortas I. Rizwan M. Chaudhary H. J. Gurmani A. R. Munis M. F. Chemosphere. 2020;238:124710. PubMed

Abdulrahman D. K. Othman R. Saud H. M. Malays. J. Soil Sci. 2016;20:177–194.

Saranya K. Kumutha K. Krishnan P. S. Madras Agric. J. 2011;98:158–164.