The Effect of Using Mealworm Frass and Spraying of Zinc Lignosulfonate on the Yield and Some Properties of Green Mung Bean (Vigna radiata) Seeds

Document Type : Research Article

Authors

N. Mirzababaei

A. Hassani

M. Babaakbari Sari

A. Golchin

Department of Soil Science, Faculty of Agriculture, University of Zanjan, Zanjan, Iran

https://doi.org/10.22067/jsw.2026.93966.1486

Abstract

Introduction
Increasing the quality of food grains is one of the goals of sustainable agriculture. Mung bean (Vigna radiata L.) is one of the most important legume crops cultivated on more than 6 million hectares worldwide. There are few studies on the effect of mealworm frass application and its effectiveness on plant growth and yield. On the other hand, given the soil and climatic conditions of Iran, its high potential in legume production, and its economic justification, it seems necessary to pay attention to feeding mung beans with organic fertilizers. The cultivation of mung bean is increasingly developing in Iran, and its use for food and medicinal purposes is also increasing. Therefore, considering the possible positive effect of mealworm frass and zinc fertilizer on the growth of this plant, the present study was conducted to investigate the effect of these fertilizers on growth, yield indices, and the molar ratio of phytic acid to zinc in the seeds.
 
Materials and Methods
In this study, an experiment was conducted as a factorial experiment in a randomized complete block design with three replications. The aim was to investigate the effect of different levels of mealworm frass and zinc lignosulfonate foliar application on growth characteristics, yield, zinc concentration, the molar ratio of phytic acid to zinc, and some biochemical factors of mung bean seeds. The treatments included four levels of mealworm frass (0, 500, 1000 and 1500 kg ha^-1) and three levels of zinc foliar application during the flowering period (foliar application with irrigation water as a control and foliar application with concentrations of 2.5 and 5 g L^-1 of zinc lignosulfonate). Growth and yield indices including plant height, number of secondary stems, number of pods per plant, number of seeds per pod, pod length, and seed yield were measured. Also, some biochemical properties such as molar ratio of phytic acid to zinc, flavones, total flavonoids, total phenols, tannin concentration and percentage of antioxidant activity were measured. Nitrogen, phosphorus and potassium of mung bean seeds were also measured.
 
Results and Discussion
The results showed that the simple effect of mealworm frass was significant for all measured traits except the number of secondary stems, pod length, and number of seeds per pod. The simple effect of zinc lignosulfonate was significant for all measured traits except the number of secondary stems, pod length, number of seeds per pod and total phenol concentration, and seed tannin. The interaction effect of mealworm frass and zinc lignosulfonate for all measured traits except pod length and number of seeds per pod was significant. The highest grain yield (115.2 g m^-2) was related to the treatment of 1500 kg of mealworm frass and 5 ppt zinc lignosulfonate foliar spray, which was in the same group as the treatment of 1500 kg mealworm frass with 0 and 2.5 g L^-1 zinc foliar spray, and the treatment of 1000 kg mealworm frass and 5 g L^-1 zinc foliar spray. While the lowest amount (80.1 g m^-2) was observed in the control treatment. The highest plant height (37.05 cm) was measured in the treatment of 500 kg mealworm frass with zero zinc application, and the lowest stem length (24.68 cm) was observed in the treatment of 500 kg of mealworm frass with the application of 5 g L^-1 zinc. The highest molar ratio of phytic acid to zinc (30.8) was observed in the treatment of 500 kg of mealworm frass and no foliar application of zinc lignosulfonate, which was not significantly different from the treatments of 0 kg of mealworm frass and 0 and 2.5 g L^-1 of zinc lignosulfonate application. While the lowest ratio (24.1) was observed in the treatment of 0 kg of mealworm frass and 5 g L^-1 zinc lignosulfonate application, which was not significantly different compared to the treatment of 1500 kg of mealworm frass and 2.5 and 5 g L^-1 of zinc lignosulfonate application. Foliar application of zinc reduced the molar ratio of phytic acid to zinc in the grain (maximum 21%), and application of mealworm frass increased this ratio up to 500 kg and then decreased it. Based on the results of this study, the application of 1000 kg of mealworm frass without zinc application was more effective in increasing the concentration of flavonoids and antioxidant activity. However, zinc foliar application was effective in increasing the concentration of phenolic compounds and tannin. The highest antioxidant activity (72.44%) was observed in the treatment of 1000 kg of mealworm frass without foliar application of zinc, and the lowest (58.18%) was measured in the treatment of 1500 kg of mealworm frass and application of 2.5 g L^-1 zinc lignosulfonate, followed by treatments of 500 kg of mealworm frass without foliar application of zinc lignosulfonate and foliar application of 2.5 g L^-1 zinc lignosulfonate. Zinc foliar spraying was effective in increasing the concentration of phenolic compounds and tannin content. Foliar application of zinc lignosulfonate increased the zinc and nitrogen content of the grain but had no effect on the phosphorus and potassium concentrations. Application of mealworm frass increased the nitrogen, phosphorus, and potassium content of the mung bean grain but simultaneously reduced the zinc concentration.
 
 Conclusion
In general, the results of this study showed that the use of mealworm frass increased mung bean grain yield, and foliar spraying of zinc lignosulfonate increased the grain zinc concentration and reduced the molar ratio of phytic acid to zinc. Overall, the use of mealworm frass increased the yield, and the use of zinc lignosulfonate improved the grain quality.

Keywords

Antioxidant activity

Flavonoids

Molar ratio of phytic acid to zinc

Tannin

Total phenol

Subjects

• Soil science

---

Authors retain the copyright. This is an open access article distributed under Creative Commons Attribution 4.0 International License (CC BY 4.0).

References

1. Ali, N.M., Mohd Yusof, H., Yeap, S.K., Ho, W.Y., Beh, B.K., Long, K., & Alitheen, N.B. (2014). Anti‐inflammatory and antinociceptive activities of untreated, germinated, and fermented mung bean aqueous extract. Evidence‐Based Complementary and Alternative Medicine, (1), 350507.
2. Alloway, B.J. (2008). Zinc in soils and crop nutrition (2^th). Brussels: international zinc association (IZA), 136p.
3. Álvarez-Fernández, A., Orera, I., Abadía, J., & Abadía, A. (2007). Determination of synthetic ferric chelates used as fertilizers by liquid chromatography-electrospray/mass spectrometry in agricultural matrices. Journal of American Society of Mass Spectrom, 18, 37–47. https://doi.org/10.1016/j.jasms.2006.08.018
4. Azizian-Shermeh, O., Einali A., & Valizadeh, J. (2018). Physiological and biochemical responses of basil (Ocimum basilicum) seedlings to different concentrations of zinc. Iranian Journal of Plant Biology, 10(2), 35-56. (In Persian with English abstract). https://doi.org/10.22108/IJPB.2018.105679.1043
5. Barakoti, L., & Bains, K. (2007). Effect of household processing on the in vitro bioavailability of iron in mungbean (Vigna radiata). Food and Nutrition Bulletin, 28(1), 18-22. https://doi.org/10.1177/156482650702800102
6. Boye, J., Zare, F., & Pletch, A. (2010). Pulse proteins: processing, characterization, functional properties and applications in food and feed. Food Research International, 43(2), 414-431. https://doi.org/10.1016/j.foodres. 2009.09.003
7. Benedicto, A., Hernández-Apaolaza, L., Rivas, I., & Lucena, J.J. (2011). Determination of 67Zn distribution in navy bean (Phaseolus vulgaris) after foliar application of 67Zn–lignosulfonates using isotope pattern deconvolution. Journal of Agricultural and Food Chemistry, 59(16), 8829-8838.
8. Blakstad, J.I., Strimbeck, R., Poveda, J., Bones, A.M., & Kissen, R. (2023). Frass from yellow mealworm (Tenebrio molitor) as plant fertilizer and defense priming agent. Biocatalysis and Agricultural Biotechnology, 53, 102862.
9. Bonnet, M., Camares, O., & Veisseire, P. (2000). Effect of zinc and influence of Acremonium lolli on growth parameters, chlorophyll a fluorescence and antioxidant enzyme activity of ryegrass. Experimental Botany, 51(346), 945-953. https://doi.org/10.1093/jxb/51.346.945
10. Borowiak, K., Gasecka, M., Mleczek, M., Dabrowski, J., Chadzinikolau, T., Magdziak, Z., Golinski, P., Rutkowski, P., & Kozubik, T. (2015). Photosynthetic activity in relation to chlorophylls, carbohydrates, phenolics and growth of a hybrid Salix purpurea × triandra × viminalis 2 at various Zn concentrations. Acta Physiologiae Plantarum, 37(8), 155. https://doi.org/10.1007/s11738-015-1904-x
11. Broadhurst, R.B., & Jones, W.T. (1978). Analysis of condensed tannins using acidified vanillin. Journal of the Science of Food and Agriculture, 29(9), 788–794. https://doi.org/10.1002/jsfa.2740290908
12. Broadley, M.R., White, P.J., Hammond, J.P., Zelko, I., & Lux, A. (2007). Zinc in plants. New Phytologist, 173(4), 677-702. https://doi.org/abs/10.1111/j.1469-8137.2007.01996.x
13. Dahiya, P.K., Linnemann, A.R., Van Boekel, M.A.J.S., Khetarpaul, N., Grewal, R.B., & Nout, M.J.R. (2015). Mung bean: Technological and nutritional potential. Critical Reviews in Food Science and Nutrition, 55(5), 670-688. https://doi.org/10.1080/10408398.2012.671202
14. Dangles, O. (2012). Antioxidant activity of plant phenols: chemical mechanisms and biological significance. Current Organic Chemistry, 16(6), 692-714. https://doi.org/10.2174/138527212799957995
15. Di Paola, A., Rulli, M.C., & Santini, M. (2017). Human food vs. animal feed debate. A thorough analysis of environmental footprints. Land Use Policy, 67, 652-659. https://doi.org/10.1016/j.landusepol.2017.06.017
16. EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP). (2015). Scientific Opinion on the safety and efficacy of lignosulphonate as a feed additive for all animal species. EFSA Journal, 13(7), 4160. https://doi.org/10.2903/j.efsa.2015.4160
17. Feiziasl, V., & Valizadeh, Gh. (2004). Effects of phosphorus and zinc fertilizer applications on nutrient concentrations in plant and grain yield in cv. Sardari "Triticum aestivum" under dryland conditions. Iranian Journal of Crop Sciences, 6(3), 223- 235. (In Persian with English abstract). https://doi.org/20.1001.1.15625540. 1383.6.3.5.4
18. Fujita, M., & Hasanuzzaman, M. (2022). Approaches to enhancing antioxidant defense in plants. Antioxidants, 11(5), 925. https://doi.org/10.3390/antiox11050925
19. Gan, R.Y., Lui, W.Y., Wu, K., Chan, C.L., Dai, S.H., Sui, Z.Q., & Corke, H. (2017). Bioactive compounds and bioactivities of germinated edible seeds and sprouts: An updated review. Trends in Food Science & Technology, 59, 1-14. https://doi.org/10.1016/j.tifs.2016.11.010
20. Gargari, B.P., Mahboob, S., & Razavieh, S.V. (2007). Content of phytic acid and its mole ratio to zinc in flour and breads consumed in Tabriz, Iran. Food Chemistry, 100(3), 1115-1119. https://doi.org/10.1016/j.foodchem. 2005.11.018
21. Gonzalez, D., Obrador, A., López Valdivia, L.M., & Álvarez, J.M. (2008). Effect of zinc source applied to soils on its availability to navy bean. Soil Science Society of America, 72, 641–649. https://doi.org/10.2136/sssaj 2007.0099
22. Haug, W., & Lantzsch, H.J. (1983). Sensitive method for the rapid determination of phytate in cereals and cereal products. Journal of the Science of Food and Agriculture, 34(12), 1423-1426. https://doi.org/10.1002/jsfa.274034 1217
23. Houben, D., Daoulas, G., Faucon, M.P., & Dulaurent, A.M. (2020). Potential use of mealworm frass as a fertilizer: Impact on crop growth and soil properties. Scientific Reports, 10(1), 4659. https://doi.org/10.1038/s41598-020-61765-x
24. Houben, D., Daoulas, G., & Dulaurent, A.M.(2021). Assessment of the short-term fertilizer potential of mealworm frass using a pot experiment Front. Sustain Food Syst., 5 Article 714596, https://doi.org/10.3389/fsufs.2021.714596
25. Hussain, M., Shahid, M.Z., Mehboob, N., Minhas, W.A., & Akram, M. (2021). Zinc application improves growth, yield and grain zinc concentration of mung bean (Vigna radiata). Semina: Ciências Agrárias, 42(2), 487-500.
26. Hussain, T., Tontisirin, K., & Chaowanakarnkit, L. (1983). Protein digestibility of weaning foods prepared from rice-minced meat and rice-mungbean [Vigna radiata] combination in infants using a short term nitrogen balance method. Journal of Nutritional Science and Vitaminology (Japan), 29(4). https://doi.org/10.3177/jnsv.29.497
27. Hummel, M., Talsma, E.F., Taleon, V., Londoño, L., Brychkova, G., Gallego, S., & Spillane, C. (2020). Iron, zinc and phytic acid retention of biofortified, low phytic acid, and conventional bean varieties when preparing common household recipes. Nutrients, 12(3), 658. https://doi.org/10.3390/nu12030658
28. Jalilian, J., Khade, A., & Pirzad, A. (2014). Effect of Fe and Zn spraying on some characteristics of mungbean using chemical and organic fertilization. Journal of Crops Improvement, 16(3), 725-732. https://doi.org/10.22059/jci.2014.53272
29. Kalra, Y. (Ed.). (1997). Handbook of reference methods for plant analysis. CRC press.
30. Kaya, M., Küçükyumuk, Z., & Erdal, I. (2009). Phytase activity, phytic acid, zinc, phosphorus and protein contents in different chickpea genotypes in relation to nitrogen and zinc fertilization. African Journal of Biotechnology, 8, 4508-4513.
31. Kumar Dahiya, P., Nout, M.J.R., A. van Boekel, M., Khetarpaul, N., Bala Grewal, R., & Linnemann, A. (2014). Nutritional characteristics of mung bean foods. British Food Journal, 116(6), 1031-1046. https://doi.org/10.1108/ BFJ-11-2012-0280
32. Li, L., Zhao, Z., & Liu, H. (2013). Feasibility of feeding yellow mealworm (Tenebrio molitor) in bioregenerative life support systems as a source of animal protein for humans. Acta Astronautica, 92(1), 103-109. https://doi.org/ 10.1016/j.actaastro.2012.03.012
33. Liyanage, R., Kiramage, C., Visvanathan, R., Jayathilake, C., Weththasinghe, P., Bangamuwage, R., & Vidanarachchi, J. (2018). Hypolipidemic and hypoglycemic potential of raw, boiled, and sprouted mung beans (Vigna radiata Wilczek) in rats. Journal of Food Biochemistry, 42(1), e12457.
34. Malakouti, M.J. (2011). Towards improving the quality of consumed breads in Iran, a review. Journal of Food Science and Technology, 8(32), 11-21. (In Persian with English abstract). https://fsct.modares.ac.ir/article-7-1738-en.html
35. Mangal, M., Agarwal, M., & Bhargava, D. (2013). Effect of cadmium and zinc on growth and biochemical parameters of selected vegetables. Journal of Pharmacognosy and Phytochemistry, 2, 106-114. https://doi.org/ 10.1371/journal.pone.0087582
36. Martin-Ortiz, D., Hernandez-Apaolaza, L., & Garate, A. (2009). Efficiency of a NPK fertilizer with adhered zinc lignosulfonate as a zinc source for maize (Zea mays). Journal of Agricultural and Food Chemistry, 57(19), 9071-9078.
37. Mehdiniya Afra, J., & Manavi Amri, SS. (2015). The effects of interaction between the elements phosphorous and zinc are some traits of soybean cultivars of Sari. Iranian Journal of Dynamic Agriculture, 11(4), 309-315. (In Persian with English abstract)
38. Menichini, F., Tundis, R., Bonesi, M., Loizzo, M.R., Conforti, F., Statti G., Di Cindi, B., Houghton, P.J., & Menichini, F. (2009). The influence of fruit ripening on the phytochemical content and biological activity of Capsicum chinense Habanero. Food Chemistry, 114, 553-560. https://doi.org/10.1016/ j.foodchem.2008.09.086
39. Motalebifard, R. (2017). Effects of zinc and phosphorus levels on yield, nutrients uptake and zinc recovery and agronomic efficiency in potato. Journal of Water and Soil, 31(3), 886-899. (In Persian with English abstract). https://doi.org/10.22067/JSW.V31I3.54513
40. Motesharezadeh, B., &Savaghebi, R. (2012).The effect of balanced fertilization on nutrients’ concentration and phytic acid to zinc molar ratio in Iranian red been (Phaseolus calcaratus L.) cultivars at different stages of seed development. Journal of Science and Technology of Greenhouse Culture, 3(1), 73-84. (In Persian with English abstract). https://dorl.net/dor/20.1001.1.20089082.1391.3.1.6.4   
41. Mubarak, A.E. (2005). Nutritional composition and antinutritional factors of mung bean seeds (Phaseolus aureus) as affected by some home traditional processes. Food Chemistry, 89(4), 489-495. https://doi.org/10.1016/ j.foodchem.2004.01.007
42. Mukhopadhyay, M.S., Das, A., Subba, P., Bantawa, P., Sarkar, B., Ghosh, P. & Mondal, TK. (2013). Structural, physiological, and biochemical profiling of tea plants under zinc stress. Biologia Plantarum, 57, 474-480. https://doi.org/10.1007/s10535-012-0300-2
43. Nair, R.M., Yang, R.Y., Easdown, W.J., Thavarajah, D., Thavarajah, P., Hughes, J.D.A., & Keatinge, J.D.H. (2013). Biofortification of mungbean (Vigna radiata) as a whole food to enhance human health. Journal of the Science of Food and Agriculture, 93(8), 1805-1813. https://doi.org/10.1002/jsfa.6110
44. Oke, F., Aslim, B., Ozturk, S., & Altundag, S. (2009). Essential oil composition, antimicrobial and antioxidant activities of Satureja cuneifolia Food Chemistry, 112, 874-879. https://doi.org/10.1016/ j.foodchem.2008.06.061
45. Ojo, M.A. (2022). Tannins in foods: nutritional implications and processing effects of hydrothermal techniques on underutilized hard-to-cook legume seeds–a review. Preventive Nutrition and Food Science, 27(1), 14. https://doi.org/10.3746/pnf.2022.27.1.14
46. Pandey, N., Pathak, G.C., & Sharma, C.P. (2006). Zinc is critically required for pollen function and fertilization in lentil. Journal of Trace Elements in Medicine and Biology, 20, 89-96. https://doi.org/10.1016/j.jtemb.2005.09.006
47. Popova, M., Bankova, V., Butovska, D., Petkov, V., Nikolova-Damyanova, B., Sabatini, A.G., Marcazzan, G.L., & Bogdanov, S. (2004). Validated methods for the quantification of biologically active constituents of poplar-type propolis. Phytochemistry Analysis, 15, 235-240. https://doi.org/10.1002/pca.777
48. Poveda, J., Jiménez-Gómez, A., Saati-Santamaría, Z., Usategui-Martín, R., Rivas, R., & García-Fraile, P. (2019). Mealworm frass as a potential biofertilizer and abiotic stress tolerance-inductor in plants. Applied Soil Ecology, 142, 110-122. https://doi.org/10.1016/j.apsoil.2019.04.016
49. Ryan, M.H., McInerney, J.K., Record, I.R., & Angus, J.F. (2008). Zinc bioavailability in wheat grain in relation to phosphorus fertiliser, crop sequence and mycorrhizal fungi. Journal of the Science of Food and Agriculture, 88(7), 1208-1216. https://doi.org/10.1002/jsfa.3200
50. Sánchez Jiménez, S., & Lucena, J.J. (2015). Characterization of zinc fertilizers. adjustment to the European and Spanish regulations (in Spanish). Phytoma,272, 47–52.
51. Sandberg, A.S. (2002). Bioavailability of minerals in legumes. British Journal of Nutrition, 88(S3), 281-285.
52. Tajoddin, M.D., Shinde, M., & Lalitha, J. (2011). In vivo reduction the phytic acid content of mung bean (Phaseolus aureus) cultivars during germination. Am.-Eurasian Journal of Agriculture Environment Science, 10, 127–132. http://www.idosi.org/aejaes/jaes10(1)/20.pdf
53. Vassilev, A., Nikolova, A., Koleva, L., & Lidon, F. (2011). Effect of excess zinc on growth and photosynthetic performance of young bean plants. Journal of Phytology, 3, 58-62.
54. Verardi, A., Sangiorgio, P., Della Mura, B., Moliterni, S., Spagnoletta, A., Dimatteo, S., & Errico, S. (2025). Tenebrio molitor Frass: A cutting-edge biofertilizer for sustainable agriculture and advanced adsorbent precursor for environmental remediation. Agronomy, 15(3), 758. https://doi.org/10.3390/agronomy15030758
55. Vitosh, M.L., Warncke, D.D., & Lucas, R.E. (1994). Secondary and micronutrients for vegetable and field crops. extension bulletin E-486, Michigan State University Extension Service, 18 p.
56. Wang, X., Yang, R., Jin, X., Chen, Z., Zhou, Y., & Gu, Z. (2015). Effect of germination and incubation on Zn, Fe, and Ca bioavailability values of soybeans (Glycine max) and mung beans (Vigna radiate L.). Food Science and Biotechnology, 24, 1829-1835. https://doi.org/10.1007/s10068-015-0239-0
57. Weaver, C.M., & Kannan, S. (2001). Phytate and mineral bioavailability. In Food phytates(pp. 227-240). CRC Press.
58. World Health Organization. (1996). Trace elements in human nutrition and health. WHO Library Cataloguing in Publication Data, 105-122.
59. Wojdylo, A., Oszmianski, J., & Czemerys, R. (2007). Antioxidant activity and phenolic compound in 32 selected herbs. Food Chemistry, 1005: 940-949. https://doi.org/10.1016/j.foodchem.2007.04.038
60. Zare Dehabadi, S., & Asrar, Z. (2009). Effect of excess zinc on the concentration of some mineral element and antioxidant responses of spearmint (Mentha spicata L.). Iranian Journal of Medicinal and Aromatic Plants, 24(4), 530-540. (In Persian with English abstract)
61. Zim, J., Aitikkou, A., EL Omari, M.H., EL Malahi, S., Azim, K., Hirich, A., & Oumouloud, A. (2022). A new organic amendment based on insect frass for zucchini (Cucurbita pepo) cultivation. Environmental Sciences Proceedings, 16(1), 28.
62. Zunzunegui, I., Martín-García, J., Santamaría, Ó., & Poveda, J. (2024). Analysis of yellow mealworm (Tenebrio molitor) frass as a resource for a sustainable agriculture in the current context of insect farming industry growth. Journal of Cleaner Production, 460, 142608. https://doi.org/10.1016/j.jclepro.2024.142608