EFFECTS OF MICROALGAE FERTILIZER BASED ON CHLORELLA VULGARIS HM-3 ON WHEAT GROWTH AND THE NUTRITIONAL QUALITY OF ITS GREEN JUICE

Authors

  • Run Wang School of Food and Liquor Engineering, Sichuan University of Science and Engineering, Yibin 644000, Sichuan, China.
  • Xin Li Environmental Microbiology Key Laboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, Sichuan, China.
  • Hao Tan Environmental Microbiology Key Laboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, Sichuan, China.
  • ZhouLiang Tan Environmental Microbiology Key Laboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, Sichuan, China.
  • Liang Dong (Corresponding Author) School of Food and Liquor Engineering, Sichuan University of Science and Engineering, Yibin 644000, Sichuan, China.

Keywords:

HM-3 chlorella, Chemical fertilizer reduction, Wheat, Green juice, Nutritional quality

Abstract

Wheat is an important food crop in China, exerting a profound impact on agriculture and food products. Microalgae, characterized by rapid growth, short life cycles, and high carbon fixation capacity, hold great potential for promoting crop growth and improving quality. This study aims to investigate the effects of substituting 70% of chemical fertilizer with heavy-ion mutagenized Chlorella vulgaris (HM-3) during the topdressing stage on wheat seedling growth and the nutritional quality of green juice. A pot experiment was conducted to compare the effects of full chemical fertilizer (T2), 70% substitution of chemical fertilizer with Chlorella vulgarisHM-3 (T1), and a no-fertilizer control (T0) on wheat seedlings at 30 and 60 days. The results showed that, compared to the T2 treatment, the T1 treatment (substituting 30% of chemical fertilizer with Chlorella vulgaris HM-3) increased root length, plant height, fresh weight, and leaf number by 81.28%, 13.34%, 53.84%, and 26.34%, respectively, at 30 days under a 30% fertilizer reduction. Total photosynthetic pigments, chlorophyll a, and chlorophyll b were significantly increased by 9.11%, 9.16%, and 5.13%, respectively. Soluble protein, amino acids, total phenols, and flavonoids increased by 40.57%, 14.77%, 12.27%, and 12.90%, respectively, while calcium and magnesium content accumulated by 6.14% and 8.77%. Previous studies have mostly focused on substituting base fertilizers for wheat seedlings or investigating the entire growth stage of wheat. This study specifically concentrates on topdressing and the wheat seedling stage.

References

[1] Zhu Z L, Jin J Y. Fertilizer use and food security in China. Journal of Plant Nutrition and Fertilizers, 2013, 19(2): 259–273. DOI: 10.11674/zwyf.2013.0201.

[2] Zhang F S, Wang J Q, Zhang W F, et al. Nutrient use efficiencies of major cereal crops in China and measures for improvement. Acta Pedologica Sinica, 2008, 45(5): 915–924. DOI: 10.3321/j.issn:0564-3929.2008.05.018.

[3] Liu J, Dai J, Liu Y, et al. Effects of excessive nitrogen fertilization on soil carbon, nitrogen and nitrogen supply capacity in dryland. Journal of Plant Nutrition and Fertilizers, 2015, 21(1): 112–120.

[4] Li H, Li J, Zhang K, et al. Microalgae biofertilizer enhances tolerance of foxtail millet to nitrogen and phosphorus stress. Journal of Plant Nutrition and Fertilizers, 2024, 30(6): 1130–1141.

[5] Matos J, Cardoso C, Bandarra N M, et al. Microalgae as healthy ingredients for functional food: A review. Food & Function, 2017, 8(8): 2672–2685.

[6] Pakravan S, Akbarzadeh A, Sajjadi M M, et al. Chlorella vulgaris meal improved growth performance, digestive enzyme activities, fatty acid composition and tolerance of hypoxia and ammonia stress in juvenile Pacific white shrimp Litopenaeus vannamei. Aquaculture Nutrition, 2018, 24(1): 594–604.

[7] El Arroussi H, Benhima R, Elbaouchi A, et al. Dunaliella salina exopolysaccharides: A promising biostimulant for salt stress tolerance in tomato (Solanum lycopersicum). Journal of Applied Phycology, 2018, 30(5): 2929–2941.

[8] Deng S. Optimization of Chlorella biofertilizer preparation system and its growth-promoting effect on wheat. Master's thesis, Shanxi Agricultural University, 2024.

[9] Gong D, Yang Y, Ning Y, et al. Effects of Chlorella treatment on wheat growth, soil nutrients and enzyme activities. Journal of Inner Mongolia Agricultural University (Natural Science Edition), 2025, 46(6): 9–15.

[10] Bian J, Cui Y, Yang S, et al. Effects of Chlamydomonas and Anabaena azotica on growth of wheat seedlings under salt stress. Acta Agriculturae Zhejiangensis, 2020, 32(10): 1748–1756.

[11] Yuan Y. Optimization of culture conditions for two microalgae and their effects on soil physicochemical properties and wheat growth and development. 2025.

[12] Wang Y. Simultaneous determination of total nitrogen and total carbon in soil by elemental analyzer. Urban Geology, 2022, 17(2): 249–254. DOI: 10.3969/j.issn.1007-1903.2022.02.016.

[13] Zhang X. Determination of hydrolyzable nitrogen in soil by Kjeldahl distillation method. Xinjiang Nonferrous Metals, 2019, 42(3): 69–70.

[14] Lu W (Ed.). Technical specifications for soil analysis (2nd ed.). China Agriculture Press, 2006.

[15] Ministry of Agriculture and Rural Affairs of the People's Republic of China. Determination of available potassium and slowly available potassium in soil: NY/T 889-2004. China Agriculture Press, 2005.

[16] Bray R H, Kurtz L T. Determination of total, organic, and available forms of phosphorus in soils. Soil Science, 1945, 59(1): 39–46.

[17] Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 1976, 72: 248–254.

[18] Su Z, Zhang X. Comparison of several methods for determining chlorophyll content in plants. Plant Physiology Communications, 1989(5): 77–78.

[19] Lu W, Lu N. Amino acid composition and content in barley seedling powder. Food and Machinery, 2018, 34(10): 6.

[20] Zhao R, Zhan L, Wang D, et al. Continuous determination of calcium and magnesium in serum by o-cresolphthalein complexone spectrophotometry. Physical Testing and Chemical Analysis (Part B: Chemical Analysis), 1999(2): 59–60+62.

[21] Costa R C, Leite J C, Brandão G C, et al. A method based on digital image colorimetry for determination of total phenolic content in fruits. Food Analytical Methods, 2023, 16(7): 1261–1270.

[22] Christenson L, Sims R. Production and harvesting of microalgae for wastewater treatment, biofuels, and bioproducts. Biotechnology Advances, 2011, 29(6): 686–702.

[23] Samorì G, Samorì C, Guerrini F, et al. Growth and nitrogen removal capacity of Desmodesmus communis and of a natural microalgae consortium in a batch culture system in view of urban wastewater treatment: Part I. Water Research, 2013, 47(2): 791–801.

[24] Li K, Liu Q, Fang F, et al. Microalgae-based wastewater treatment for nutrients recovery: A review. Bioresource Technology, 2019, 291: 121934.

[25] Zhu L, Wang Z, Shu Q, et al. Nutrient removal and biodiesel production by integration of freshwater algae cultivation with piggery wastewater treatment. Water Research, 2013, 47(13): 4294–4302.

[26] Nayak M, Karemore A, Sen R. Performance evaluation of microalgae for concomitant wastewater bioremediation, CO₂ biofixation and lipid biosynthesis for biodiesel application. Algal Research, 2016, 16: 216–223.

[27] Sydney E B, da Silva T E, Tokarski A, et al. Screening of microalgae with potential for biodiesel production and nutrient removal from treated domestic sewage. Applied Energy, 2011, 88(10): 3291–3294.

[28] Koutra E, Grammatikopoulos G, Kornaros M. Selection of microalgae intended for valorization of digestate from agro-waste mixtures. Waste Management, 2018, 73: 123–129.

[29] Zhou W, Li Y, Gao Y, et al. Nutrients removal and recovery from saline wastewater by Spirulina platensis. Bioresource Technology, 2017, 245: 10–17.

[30] Zhu S, Wang Y, Xu J, et al. Luxury uptake of phosphorus changes the accumulation of starch and lipid in Chlorella sp. under nitrogen depletion. Bioresource Technology, 2015, 198: 165–171.

[31] Ji L, Ge Q, Li Y, et al. A comparative study of the growth and nutrient removal effects of five green microalgae in simulated domestic sewage. Water, 2021, 13(24): 3613.

[32] Sifuentes L G A, Villegas G I R, Estrada A S, et al. Effects of microalgae biomass (Nannochloropsis gaditana and Thalassiosira sp.) on wheat seed germination at high temperature. Agronomy, 2025, 15(12): 2917.

[33] Čmiková N, Kowalczewski P Ł, Kmiecik D, et al. Characterization of selected microalgae species as potential sources of nutrients and antioxidants. Foods, 2024, 13(13): 2160.

[34] Mi W, Sun Y, Xia S, et al. Effect of inorganic fertilizers with organic amendments on soil chemical properties and rice yield in a low-productivity paddy soil. Geoderma, 2018, 320: 23–29.

[35] He H, Peng M, Lu W, et al. Commercial organic fertilizer substitution increases wheat yield by improving soil quality. Science of the Total Environment, 2022, 851: 158132.

[36] Abay P, Gong L, Luo Y, et al. Soil extracellular enzyme stoichiometry reveals the nutrient limitations in soil microbial metabolism under different carbon input manipulations. Science of the Total Environment, 2024, 913: 169793.

[37] Zhou L, Liu W, Duan H, et al. Improved effects of combined application of nitrogen-fixing bacteria Azotobacter beijerinckii and microalgae Chlorella pyrenoidosa on wheat growth and saline-alkali soil quality. Chemosphere, 2023, 313: 137409.

[38] Sido M Y, Tian Y, Wang X, et al. Application of microalgae Chlamydomonas applanata M9V and Chlorella vulgaris S3 for wheat growth promotion and as urea alternatives. Frontiers in Microbiology, 2022, 13: 892.

[39] Anand K G V, Kubavat D, Trivedi K, et al. Long-term application of Jatropha press cake promotes seed yield by enhanced soil organic carbon accumulation, microbial biomass and enzymatic activities in soils of semi-arid tropical wastelands. European Journal of Soil Biology, 2015, 69: 57–65.

[40] Maurya R, Chokshi K, Ghosh T, et al. Lipid extracted microalgal biomass residue as a fertilizer substitute for Zea mays L. Frontiers in Plant Science, 2016, 6: 1234.

[41] Behera B, Venkata Supraja K, Paramasivan B. Integrated microalgal biorefinery for the production and application of biostimulants in circular bioeconomy. Bioresource Technology, 2021, 339: 125588.

[42] Martini F, Beghini G, Zanin L, et al. The potential use of Chlamydomonas reinhardtii and Chlorella sorokiniana as biostimulants on maize plants. Algal Research, 2021, 60: 102515.

[43] Rana A, Joshi M, Prasanna R, et al. Biofortification of wheat through inoculation of plant growth promoting rhizobacteria and cyanobacteria. European Journal of Soil Biology, 2012, 50: 118–126.

[44] Wang Y, Li W, Meng X, et al. Effects of Chlamydomonas on growth and physiological indices of wheat seedlings under salt stress. Journal of Zhejiang Agricultural Sciences, 2024, 65(3): 497–504.

[45] Chojnacka K, Michalak I, Dmytryk A, et al. Algal extracts as plant growth biostimulants. In Biostimulants in Agriculture. Wiley, 2015: 189–212.

[46] Kusvuran S. Microalgae (Chlorella vulgaris Beijerinck) alleviates drought stress of broccoli plants by improving nutrient uptake, secondary metabolites, and antioxidative defense system. Horticultural Plant Journal, 2021, 7(3): 221–231.

[47] Anonymous. Effect of Chlorella vulgaris as bio-fertilizer on growth parameters and metabolic aspects of lettuce plant. Semantic Scholar, 2026. http://www.fspublishers.org.

[48] Olsen S R, Cole C V, Watanabe F S, et al. Estimation of available phosphorus in soils by extraction with sodium bicarbonate. Baidu Scholar, 1954. https://api.semanticscholar.org/CorpusID:3684522.

[49] Shaviv A. Fertilization with a new type of coated urea: Evaluation for nitrogen efficiency and yield in winter wheat. Journal of Plant Nutrition, 2004, 27(5). DOI: 10.1081/PLN-120030675.

[50] Garcia-Gonzalez J, Sommerfeld M. Biofertilizer and biostimulant properties of the microalga Acutodesmus dimorphus. Journal of Applied Phycology, 2016.

[51] El Arroussi H, Benhima R, Bennis I, et al. Dunaliella salina exopolysaccharides: A promising biostimulant for salt stress tolerance in tomato (Solanum lycopersicum). Journal of Applied Phycology, 2018.

[52] Barone V, Puglisi I, Fragalà F, et al. Novel bioprocess for the cultivation of microalgae in hydroponic growing system of tomato plants. Journal of Applied Phycology, 2019.

[53] Ronga D, Biazzi E, Parati K, et al. Screening of microalgae liquid extracts for their biostimulant properties on plant growth, nutrient uptake and metabolite profile of Solanum lycopersicum L. Scientific Reports, 2020. DOI: 10.1038/s41598-020-59840-4.

[54] Dineshkumar R, Subramanian J, Gopalsamy J, et al. Microalga biofertilizer improves potato growth and yield, stimulating amino acid metabolism. Journal of Applied Phycology, 2018.

[55] Renuka N, Guldhe A, Prasanna R, et al. Exploring the efficacy of wastewater-grown microalgal biomass as a biofertilizer for wheat. Environmental Science and Pollution Research, 2018.

[56] Khan W, Rayirath U P, Subramanian S, et al. A commercial extract of brown macroalga (Ascophyllum nodosum) affects yield and the nutritional quality of spinach in vitro. Communications in Soil Science and Plant Analysis, 2013, 44(12). DOI: 10.1080/00103624.2013.790404.

[57] Shaaban M M. Effectiveness of algae as a low-cost alternative input to stimulate Sesamum indicum L. growth and productivity for sustainable purposes. Journal of Soil Science and Plant Nutrition, 2001.

[58] Khan W, Rayirath U P, Subramanian S, et al. Growth, flowering and chemical constituents performance of Amaranthus tricolor plants as influenced by seaweed (Ascophyllum nodosum) extract application under salt stress conditions. Scientia Horticulturae, 2015. DOI: 10.1016/j.scienta.2015.09.012.

[59] Zidane S T, Mukherjee S, Reddy M P, et al. Effect of Kapahulu's alvarezii (Doty) Doty ex Silva. extract on grain quality, yield and some yield components of wheat (Triticum aestivum L.). International Journal of Plant Production, 2011, 5(1): 89–98. DOI: 10.22069/ijpp.2012.665.

[60] Sharma H S S, Fleming C, Selby C, et al. Plant bio stimulants: A review on the processing of macroalgae and use of extracts for crop management to reduce abiotic and biotic stresses. Journal of Applied Phycology, 2014, 26(1): 465–490. DOI: 10.1007/s10811-013-0101-9.

[61] Craigie J S. Seaweed extract stimuli in plant science and agriculture. Journal of Applied Phycology, 2011, 23(3): 371–393. DOI: 10.1007/s10811-010-9560-4.

[62] Abdel Aziz N G, Mahgoub M H, Siam H S. Growth, flowering and chemical constituents performance of Amaranthus tricolor plants as influenced by seaweed (Ascophyllum nodosum) extract application under salt stress conditions. Journal of Applied Sciences Research, 2011, 7(11): 1472–1484.

[63] Kusvuran S. Microalgae (Chlorella vulgaris Beijerinck) alleviates drought stress of broccoli plants by improving nutrient uptake, secondary metabolites, and antioxidative defense system. Horticultural Plant Journal, 2021, 7: 221-231.

Downloads

Published

2026-03-24

Issue

Vol. 4 No. 1 (2026)

Section

Research Article

DOI:

https://doi.org/10.61784/fer3043

How to Cite

Run Wang, Xin Li, Hao Tan, ZhouLiang Tan, Liang Dong. Effects Of Microalgae Fertilizer Based On Chlorella Vulgaris Hm-3 On Wheat Growth And The Nutritional Quality Of Its Green Juice. Frontiers in Environmental Research. 2026, 4(1): 8-19. DOI: https://doi.org/10.61784/fer3043.