Effect of Pseudomonas fluorescence strains and humic acid on some morphological ‎and physiological characteristics of marshmallow (Altheae officinalis L.) ‎

Document Type : Full Paper


1 M. Sc. Graduate, Faculty of Agriculture, University of Birjand, Birjand, Iran ‎

2 Associate Professor, Faculty of Agriculture, University of Birjand, Birjand, Iran ‎

3 Professor, College of Agriculutre & Natural Resources, University of Tehran, Karaj, Iran

4 Associate Professor, Faculty of Agriculture, University of Birjand, Birjand, Iran‎


In order to investigate the effect of plant growth promoting rhizo-bacteria (PGPRs) and humic acid on some morphological and physiological characteristics of marshmallow (Altheae officinalis L.) a pot experiment was carried out under greenhouse condition at Faculty of Agriculture, University of Birjand, in 2017. The experiment was conducted using a complete randomized blocks design based on factorial arrangement with three replications. First factor was seed inoculation with strains of Pseudomonas fluorescence bacteria at four levels (non- inoculation, inoculation with strains 51, 79 and 163) and second factor was humic acid at four levels (0, 150, 300 and 450 mg/kg soil). Results showed that Pseudomonas fluorescence strains and humic acid had significant effects on morphological and physiological measured traits. The most effects of inoculation created by strain 163 of Pseudomonas fluorescenss bacteria and it increased leaf dry weight, leaf area, leaf chlorophyll index, carotenoids and protoin leaf compare to control. The result also showed that increasing of humic acid concentration up to 150 mg/kg of soil increased, leaf area, leaf chlorophylls index, chlorophyll fluorescence ­index and protein leaf compare to control. Combination use of Pseudomonas fluorescenss bacteria (strain 163) and humic acid (150 mg/kg soil) had the most effect on increasing of leaf fresh weight, leaf area, chlorophyll a, chlorophyll b, total chlorophyll, carotenoids and phosphor leaf compare to control. Due to these results, application of biological fertilizers containing Pseudomonas fluorescence and humic acid can improve morphological and physiological characteristics of marshmallow.


  1. Abdol-Jaleel, C., Manivannan, P., Sankar, B., Kishorekumar, A., Gopi, R., Somasundaram, R. & Panneerselvam, R. (2002). Pseudomonas fluorescens enhances biomas yield and ajmalicine production in Catharanthus roseus under water deficit stress. Colloids and Surfaces B: Biointerfaces, 60, 7-11
  2. Ahmad, A. G., Orabi, S. A. & Gaballah, M. (2010). Effect of bio-N-P fertilizer on the growth, yield and some biochemical components of two sunflower cultivars. International Journal Academic Research, 2, 271-277.
  3. Ahmad, I., Usman, R. S., Muhammad, S., Ahmad, S. K. & Muhammad, Y. (2013). Humic acid cultivar effects on growth, yield vase life, and corn characteristics of gladiolus. Chilean Journal of Agricultural Research, 74(3), 339-344.
  4. Ali, A., Rehman, S. U., Reza, S. & Butt, S. J. (2014). Combined effect of humic acid and NPK on growth and flower development of Tulipa gesneriana in Faisalabad. Pakistan Journal of Ornamental Plants, 4(4), 227-236.
  5. Ali, S. K. Z., Sandhya, V., Grover, M., Kishore, N., Rao, L. V. & Venkateswarlu, B. (2009(. Pseudomonas Strain AKM-P6 enhances tolerance of sorghum seedlings to elevated temperatures. Biology and Fertility of Soils, 46, 45-55.
  6. Allavi zadeh, N. & Nazari Deljou, M. (2014). Effect of humic acid on morph-physiological traits, nutrients uptake and postharvest vase life on pot marigold cut flower (Calendula officinalis cv. Crysantha) in hydroponic system. Journal of Science and Technology of Greenhouse Culture, 5(18), 133-143. (in Farsi).
  7. Amal, G. A., Orabi, S. & Gomaa, A. M. (2010). Bio-organic farming of grain sorghum and its effect on growth, physiological and yield parameters and antioxidant enzymes activity. Research Journal of Agriculture and Biological Sciences, 6(3), 270-279.
  8. Arnon, D. I. (1949). Copper enzymes in isolated chloroplasts, polyphenol oxidase in (Beta vulgaris). American Society of Plant Physiologists, 24(1), 1.
  9. Asgari, M., Habibi, D., & Brojerdi, G. (2011). Study on the application of vermin compost, growth stimulating bacteria and humic acid on growth index of peppermint in central province. Journal of Agronomy and Plant Breeding, 7(4), 41-54.
  10. Ashrafuzzaman, M. F. A., Hossen, I. M., Razi, H. M., Anamul, I. M., Zahurul, S. M., Shahidullah, M. & Sariah, M. (2009). Efficiency of plant growth- promoting rhizobacteria (PGPR) for the enhancement of rice growth. African Journal of Biotechnology, 8, 1247-1252.
  11. Azarpour, E., Khosravi Danesh, R., Mohammadi, S., Bozorgi, H. & Moraditochaee, M. (2011). Effects of nitrogen fertilizer under foliar spraying of humic acid on yield and yield components of cowpea (Vigna unguiculata). World Applied Sciences Journal. 13(6), 1445-1449.
  12. Bahrami, S., Soleimani, A. & Habibi, F. (2015). The effect of humic acid on the mineral composition level, yield and fruit quality apple variety Granny Smith (Malus domestica cv. Granny Smith). Journal of Crops, 17(2), 517-529. (In Farsi).
  13. Barghmadi, K. & Najafi, S. H. (2015). Nitroxin and different levels of humic acid on some essential characteristics of small and ajowan (Trachyspermum copticum (L.) Link). Journal of Horticultural Science, 29, 332-341. (in Farsi).
  14. Banchio, E., Bagino, P., Zygadlo, J. & Giordano, W. (2008). Plant growth promoting rhizobacteria improve growth and essential oil yield in Origanum majorana Biochemistry Systemic Ecology. 36: 766-771.
  15. Boroumand, A., Sajedi, N. & Changizi, M. (2012). Combined effects of chemical fertilizers and plant growth promoting bacteriaon yield and yield components of maize in Arak weather conditions. New Findings in Agriculture, 4, 296-307.
  16. Bromand Sivieri, M., Heidary, M., Gholami, A. & Ghorbani, H. (2021). Effects of foliar application of nano iron oxide and biofertilizers on the activity of antioxidant enzymes and some physiological characteristics of the root and aerial parts in black cumin (Nigella sativa ). Iranian Journal of Horticultural Science. 51(4), 939-953.(in Farsi).
  17. Celik, H. A. V., Katkat, B. B., Asik M. A. & Turan, M. A. (2010). Effect for foliar-applied humic acid to dry weight and mineral nutrient uptake of maize under calcareous soil conditions. Soil Sciences, Plant Annual, 42(2), 29-38.
  18. Chamani, A., Bonyadi, M. & Ghanbari, A. (2015). Effect of salicylic acid and humic acid on vegetative indices of periwinkle (Catharanthus roseus). Journal of Horticultural Science, 29(40), 631-641.
  19. Darzi, M. T., Ghalavand, A., Rejali, F. & Sefidkon, F. (2006). Effects of biofertilizers application on yield and yield ­components in fennel (Foeniculum vulgar ). Iranian Journal of Medicinal and Aromatic Plants, 22(4), 276-292. (in Farsi).
  20. Davis, K. F., Gephart, J. A. & Gunda, T. (2016). Sustaining food self-sufficiency of a nation: The case of Sri lanken rice production and related water and fertilizer demands, Ambio, 45(3), 302-312.
  21. Delfin, S., Tognetti, R., Desideri, E., & Alrin, A. (2005). Effect of foliar application of N and humic acids on growth and yield of durum wheat, Agronomy for Sustainable Development, 25, 183-191
  22. (2016). FAOSTAT. Food and Agriculture Organization of the United Nation. Available in: http:// faostat.fao.org/country profiles/.
  23. Farahani, A. & Madani, H. (2014). Evaluate the usefulness of humic acid organic matter in comparison to chemical fertilizer and manure and their combination in summer savory (Satureja hortensis). New Finding in Agriculture, 8(4), 324-337. (in Farsi).
  24. Fernandez, L., Zalba, P., Gomez, M. & Sagardoy, M. (2007). Phosphate-solubilization activity of bacterial strains in soil and their effect on soybean growth under greenhouse conditions. Biology and Fertility of Soil, 43, 805-809.
  25. Ganji Arjenaki, F., Jabbari, R. & Morshedi, A. (2012). Evaluation of drought stress on relative water content, chlorophyll content and mineral elements of wheat (Triticum aestivum ) varieties. International Journal of Agriculture and Crop Sciences, 4(11), 726-729.
  26. Gehan, G. M. & Abo-Baker A. (2010). Effect bio chemical fertilization on growth of sunflower (Helianthus annuus) at south valley area. Asian Journal of Crop Science 2, 137-146.
  27. Gorbani, S., Khazaei, H. R., Kafi, M. & Bannayan Aval, M. (2010). Effects of humic acid application in irrigation water on yield components of maize. Journal of Agroecology, (2), 123-131.
  28. Golami, A., Shahsavani, S. & Nezarat, M. (2009). The effect of growth promoting rhizobacteria on germination, seedling growth on yield of maize. International Journal of Biological Life Sciences, 1(1), 35-40.
  29. Golmohammadzade, S., Ghanbari, S., Hosseini, R. & Hasannia, H. (2015). Impact of vermicompost and chemical fertilizer on yield, growth and essential oil of garlic (Allium sativum L.). International Journal of Life Science, 9(4), 44-48.
  30. Goswami, D., Thakker, J. N. & Dhandhukia, P. S. (2016). Portraying mechanics of plant growth promoting rhizobacteria (PGPR): A review. Cogent Food & Agriculture, 2(1), 127-500.
  31. Haghighi, M., Kafi, M. & Fang, P. (2012). Photosynthetic activity and N metabolism of lettuce as affected by humic acid. International Vegetable Science, 18, 182-189. (in Farsi)
  32. Hatami, M., Khanizadeh, P., Alsadat Abtahi, F. & Abaszadeh Dehgi, P. (2021). Influence of plant growth promoting rhizobacteria and hydro-priming on some physiological indices of lemonbalm (Melissa officinalis). Iranian Journal of Horticultural Science. 52(1), 11-21.(in Farsi).
  33. Harper, S. M., Kerven, G. L., Edwards, L. & Zostakek Boczyski, Z. (2000). Characterizyion on folic and humic acids from leaves of eucalyptus comaldulensis and from decomposed hey. Soil Biochemistry, 32, 1331-1336.
  34. Jahan, M., Nassiri Mahallati, M., Amiri, M. B. & Ehyayi, H. R. (2013). Radiation absorption and use efficiency of sesame as affected by bio fertilizers inoculation in a low input cropping system. Industrial Crops and Products, 43, 606-611.
  35. Jalili, F., Khavazy, K. & Asadi Rahmani, E. (2011). The effect of adenosine on fluorescent enzyme activity ACC-canola growth under saline conditions. Journal of Soil Science. 2: 175-188.
  36. Khaled, H. & Fawy, H. (2011). Effect of different levels of humic acids on the nutrient content, plant growth and soil properties under conditions of salinity. Soil and Water Research, 6(1), 21-2.
  37. Mahfouz, S. A. & Sharaf-Eldin, M. A. (2007). Effect of mineral vs. biofertilizer on growth, yield, and essential oil content of fennal (Foeniculum vulgare Mill.). Indian Agrophysics. 21, 361-366.
  38. Merchan, F., Breda, C., Hormaeche, J. P. Sousa, C., Kondorosi, A., Aguilar, O. M. & Cnespi, M. (2003). A Kruppel-like transcription factor gene is involved in salt stress responses in Medicaga spp. Plant and Soil, 257(1), 1-9.
  39. Mia, M. A. B., Shamusuddin, Z. H., Wahab, Z., & Marziah, M. (2010). Rhizobacteria as bioenhancer and biofertilizer for growth and yield of banana (Musa spp. Cv. “Berangan”). Scientia Horticulturae, 126, 80-87.
  40. Mirzaie, H. & Ebrahemi Monfared, K. (2011). Medicine herbs: Packaging and marketing. Iranian Agricultural Press, Tehran. P.192. (in Farsi).
  41. Montanez, A., Radriguze- Blanco, A., Barlocco, C., Beracochea, M. & Sicardi, M. (2012). Characterization of cultivable putative endophytic plant growth promoting bacteria associated with maize cultivars (Zea mays) and effects in vitro. Applied Soil Ecology, 58, 21-28.
  42. Nadeem, S. M., Ahmad, M., Zahir, Z. A., Javaid, A. & Ashraf, M. (2014). The role of mycorrhizae plant growth promoting rhizobacteria (PGPR) in improving crop productivity under stressful environments. Biotechnology Advances, 32(2), 429-448.
  43. Naghdibadi, H., Omidi, H., Rezazadeh, S. H. & Zeinali Mobarakeh, Z. (2012). Morphological, agronomical and phytochemical changes in borage (Borago officinalis) under biological and chemical fertilizers. Journal of Medicinal Plants, 2(42), 145-156. (in Farsi).
  44. Narolia, G. P., Shivran, A. C. & Reager, M. I. (2013). Growth and quality of isabgol (Plantago ovate forsk) influenced by phosphorus, PSB and zinc. International Journal of Plant Science, 8(1), 160-162.
  45. Omidbeigi, R. (2009). Approaches to Production and Processing of Medicinal Plants. Astane-Ghodse-Razavi Publications. Mashhad. 423 P. (in Farsi).
  46. Perez-Montano, F., Alias-Villegas, C., Bellogin, R. A., Del Cerro, P., Espuny, M., Jimenez-Guerrero, I. & Cubo, T. (2014). Plant growth promotion in cereal and leguminous agricultural important plant: from microorganism capacities to crop production. Microbiological Research, 169(5), 325-336
  47. Rahi, A., Davoodifar, M., Azizi, F. & Habibi, D. (2013). Effects of different amounts of humic acid and response curves in the Dactylis glomerata. Agronomy and Plant Breeding Journal, 8(3), 15-28.
  48. Rahimzadeh, S. T., Sohrabi, Y., Heydari, G. R. & Pirzad, A. R. (2012). The effect of application of biofertilizers on some morphological characteristic and yield of Dracocephalum moldavica Iranian Journal of Horticultural Science, 25(3), 335-343. (in Farsi).
  49. Rehman, S. U., Reza, S. & Butt, S. J. (2014). Combined effect of humic acid and NPK on growth and flower development of Tulipa gesneriana in Faisalabad. Pakistan. Journal of Ornamental Plants, 4(4), 227-236.
  50. Rathi, M. & Gaur, N. (2016). Phosphate solubilizating bacteria as biofertilitizer and its applications. Journal of Pharmancy Research, 10(3), 146-148.
  51. Rezvani Moghadam, P., Lashkari, A., & Amin Ghafouri, A. (2014). Estimation of cardinal temperatures of Echium amienum Fisch. with application of regression models. Iranian Journal of Field Crops Research, 12(2), 164-169. (in Farsi).
  52. Sadat, A. (2007). Effects of vesicular arbuscular mycorhiza and plant growth promoting bacteria on nutrient uptake and yield of wheat under salinity condition. M.Sc. Thesis, University of Tehran, Iran. (in Farsi).
  53. Salehi Surmaghi, M. H. (2008). Medicinal plants and Phytotheraphy (Vol. 1). Donyaye-Taghziye Press, Tehran, 406p. (in Farsi)
  54. Salwa, A. (2011). Effect of amendments humic and amino acids on increases soils fertility yields and seeds quality of peanut and sesame on sandy soils. Agriculture and Biological Science, 7(1), 115-125
  55. Shirmardi, M., Savaghebi, G. R., Khavazi, K., Akbarzadeh, A., Farahbakhsh, M., Rejali, F. & Sadat, A. (2010). Effect of microbial inoculants on uptake of nutrient elements in two cultivars of sunflower (Helianthus annuus ). Saline Soils, 2, 57-66. (in Farsi).
  56. Shaharoona, B., Arshad, M., Zahir, Z. A. & Khalid A. (2006). Performance of Pseudomonas containing ACC-deaminase for improving growth and yield of maize (Zea mays L.) in the presence of nitrogenous fertilizer. Soil Biology and Biochemistry, 38, 2971-2975.
  57. Stancheva, I. & Dinev, N. (2003). Effect of inoculation of maize and species of Tribe triticeae with Azospirillum brasilense. Journal of Plant Physiology. 4, 550-552.
  58. Tahamy Zarandi, M., Rezvani Moghadam, P., & Jahan, M. (2010). Comparing the effect of organic and chemical fertilizers on yield and essential oil percentage of basil (Ocimum basilicum). Journal of Agricultural Ecology, 2(1), 63-74. (in Farsi).
  59. Tejada, M. & Gonzalez, J. L. (2003). Influence of foliar fertilization with amino acids and humics acid on productivity and quality of asparagus. Biological Agriculture and Horticulture, 21(3), 277-291.
  60. Wahyudi, J., Astuti, I. & Giyanto, R. (2011). Screening of Pseudomonas isolated from rhizosphere of soybean plant as plant growth promoter and bio-control Agent. American Journal of Agricultural and Biological Sciences 6(1), 134-144.
  61. Wu, F., Wan, J. H. C., Wu, S. & Wong, M. (2012). Effect of earthworms and plant growth-promoting rhizobacteria (PGPR) on availability of nitrogen, phosphorus, and potassium in soil. Journal of Plant Notrition and Soil Science, 175(3), 423-433.
  62. Zhao, J., Ni, T., Li, J., Lu, Q., Fang, Z., Huang, Q. & Shen, Q. (2016). Effects of organic inorganic compound fertilizer with reduced chemical fertilizer application on crop yields, soil biological activity and bacterial community structure in a rice-wheat cropping system. Applied Soil Ecology, 99, 1-12.