Journal of Soils and Sediments

, Volume 20, Issue 1, pp 452–460 | Cite as

Concomitant biocontrol of pepper Phytophthora blight by soil indigenous arbuscular mycorrhizal fungi via upfront film-mulching with reductive fertilizer and tobacco waste

  • Shaowei Hou
  • Yu Zhang
  • Minghui Li
  • Homgmin Liu
  • Fuyong WuEmail author
  • Junli HuEmail author
  • Xiangui Lin
Soils, Sec 5 • Soil and Landscape Ecology • Research Article



Both reductive disinfestation and germicide can suppress Phytophthora blight, while soil arbuscular mycorrhizal (AM) fungi also have biocontrol effects on soilborne diseases. However, the combined effects of reductive disinfestation and botanical germicide [e.g., tobacco (Nicotiana tabacum L.) waste] on pepper (Capsicum annuum L.) Phytophthora blight and soil AM fungi are at present unclear. The purposes of this work were to develop application strategy for dealing with pepper Phytophthora blight, and to explore the concomitant contribution from soil indigenous AM fungi.

Materials and methods

A field experiment with four treatments was carried out in a pepper continuous planting field, including normal film-mulching with common fertilizer (control), normal film-mulching with reductive fertilizer (RF), upfront film-mulching with reductive fertilizer (UM+RF), and upfront film-mulching with reductive fertilizer and tobacco waste (UM+RF+TW). Phytophthora blight severity index, root mycorrhizal colonization rate, and the biomass and nutrient (N, P, and K) concentrations of shoots, roots, and fruits of pepper were measured. Soil pH, organic C, mineral N, available P, available K, acid phosphatase activity, and AM fungal abundance were also tested. The Pearson correlation analysis was carried out among plant and soil parameters.

Results and discussion

RF tended to increase pepper fruit yield compared with control, and UM+RF tended to decrease Phytophthora blight severity in relative to RF, while UM+RF+TW tended to decrease blight severity and increase fruit yield compared with UM+RF, and had a significantly (P < 0.05) lower blight severity and a significantly higher fruit yield in comparison with control. UM+RF+TW also significantly decreased soil pH, and significantly increased AM fungal population and colonization, as well as soil acid phosphatase activity and available P concentration. In addition, UM+RF+TW had a significantly higher fruit K accumulation ratio, which negatively correlated with blight severity and positively correlated with fruit yield. However, fruit K accumulation ratio positively correlated with fruit P accumulation ratio, which was greatly elevated by the enhanced mycorrhizal colonization.


The coalition of reductive disinfestation (upfront film-mulching with reductive fertilizer) and tobacco waste had the greatest suppression of pepper Phytophthora blight, and the highest fruit yield and AM fungal population. It suggests that combined application of reductive disinfestation and botanical germicide has superposition in inhibiting Phytophthora blight and increasing fruit yield, and there seems to be a concomitant biocontrol by soil indigenous AM fungi which could enhance P and K transfer from plant to fruit.


Botanical germicide Fruit K accumulation ratio Fruit P accumulation ratio Reductive disinfestation Soil acid phosphatase 



We are grateful to Mr. Wendong Ge, Yangguo Tan, and Jiu’an Tan for their support on the field experiment, and to two anonymous reviewers for their useful suggestions on manuscript revision.

Funding information

This work was financially supported by the National Key R & D Program (2017YFD0200603) and National Natural Science Foundation (No.41671265) of China, the Knowledge Innovation Program (ISSASIP1634) of Chinese Academy of Sciences (CAS), and the Development Center of Characteristic Agricultural Industries, Shizhu county, Chongqing city, China. Junli Hu is supported by the Youth Innovation Promotion Association (No. 2016285), CAS.


  1. Azcón-Aguilar C, Barea J (1998) Arbuscular mycorrhizas and biological control of soil-borne plant pathogens-an overview of the mechanisms involved. Mycorrhiza 6:457–464Google Scholar
  2. Bosmans L, De BI, De MR (2016) Agar composition affects in vitro screening of biocontrol activity of antagonistic microorganisms. J Microbiol Methods 127:7–9Google Scholar
  3. Botella MA, Arevalo L, Mestre TC, Rubio F, Garcia-Sanchez F, Rivero RM, Martinez V (2017) Potassium fertilization enhances pepper fruit quality. J Plant Nutr 40:145–155Google Scholar
  4. Buurman P (1997) Soil organic matter: analysis and interpretation. Sci Hortic 70:261–262Google Scholar
  5. Candemır F, KutlukYılmaz ND, Gülser C (2012) The effect of tobacco waste application on tobacco mosaic virus (TMV) concentration in the soil. Zemdirbyste 99:99–104Google Scholar
  6. Chuah AM, Lee YC, Yamaguchi T, Takamura H, Yin LJ, Matoba T (2008) Effect of cooking on the antioxidant properties of colored peppers. Food Chem 111:20–28Google Scholar
  7. Costa TR, Fernandes OFL, Santos SC, Olibeira CMA, Hferri P (2000) Antifungal activity of volatile constituents of Eugenia dysenterica leaf oil. J Ethnopharmacol 72:111–117Google Scholar
  8. Cuenca G, Azcón R (1994) Effects of ammonium and nitrate on the growth of vesicular-arbuscular mycorrhizal Erythrina poeppigiana O.I. cook seedlings. Biol Fertil Soils 18:249–254Google Scholar
  9. Dalal RC (1973) Estimation of available phosphorus in soils by extraction with sodium hydroxide-sodium carbonate solution. J Austr Inst Agric Sci 39:142–143Google Scholar
  10. Davey JF, Gregory NF, Mulrooney RP (2008) First report of Mefenoxam-resistant isolates of Phytophthora capsici from Lima bean pods in the Mid-Atlantic Region. Plant Dis 92:656–656Google Scholar
  11. Depret G, Houot S, Allard MR, Breuil MC (2010) Long-term effects of crop management on Rhizobium leguminosarum biovar viciae populations. FEMS Microbiol Ecol 51:87–97Google Scholar
  12. Duan SZ, Du YM, Hou XD, Li DD, Ren X (2015) Inhibitory effects of tobacco extracts on eleven phytopathogenic fungi. Nat Prod Res Dev 3:470–474Google Scholar
  13. García de León D, Davison J, Moora M (2018) Anthropogenic disturbance equalizes diversity levels in arbuscular mycorrhizal fungal communities. Glob Chang Biol 24:2649–2659Google Scholar
  14. Giovannetti M, Mosse B (1980) An evaluation of techniques for measuring vesicular-arbuscular mycorrhizal infection in roots. New Phytol 84:489–500Google Scholar
  15. Gobena D, Roig J, Galmarini C, Hulvey J, Lamour K (2012) Genetic diversity of Phytophthora capsici isolates from pepper and pumpkin in Argentina. Mycologia 104:102–107Google Scholar
  16. Gomes SEC, Donnici CL, Camargos ERDS (2013) Effects of Copaifera duckei Dwyer oleoresin on the cell wall and cell division of Bacillus cereus. J Med Microbiol 62:1032–1037Google Scholar
  17. Goud JKC, Termorshuizen AJ, Blok WJ (2004) Long-term effect of biological soil disinfestation on Verticillium wilt. Plant Dis 88:688–694Google Scholar
  18. Hageahmed K, Krammer J, Steinkellner S (2013) The intercropping partner affects arbuscular mycorrhizal fungi and Fusarium oxysporum f. sp. lycopersici, interactions in tomato. Mycorrhiza 23:543–549Google Scholar
  19. Hodge A, Campbell CD, Fitter AH (2001) An arbuscular mycorrhizal fungus accelerates decomposition and acquires nitrogen directly from organic material. Nature 413:297–299Google Scholar
  20. Holmes GJ, Ojiambo PS, Hausbeck MK, Quesada-Ocampo L, Keinath AP (2015) Resurgence of cucurbit downy mildew in the United States: a watershed event for research and extension. Plant Dis 99:428–441Google Scholar
  21. Hu SQ (2009) Research of the nicotine to disinfect pathogenic bacteria affects. Biotechnology 19:73–75Google Scholar
  22. Hu J, Li M, Liu H, Zhao Q, Lin X (2019) Intercropping with sweet corn (Zea mays L. var. rugosa Bonaf.) expands P acquisition channels of chili pepper (Capsicum annuum L.) via arbuscular mycorrhizal hyphal networks. J Soils Sediments 19:1632–1639Google Scholar
  23. Huang X, Liu L, Wen T, Zhu R, Zhang J, Cai Z (2015) Illumina MiSeq investigations on the changes of microbial community in the Fusarium oxysporum f.sp. cubense infected soil during and after reductive soil disinfestation.Microbiol Res 181:33–42Google Scholar
  24. Katan J (2000) Physical and cultural methods for the management of soilborne pathogens. Crop Prot 19:725–731Google Scholar
  25. Kayikçioglu HH, Okur N (2011) Evolution of enzyme activities during composting of tobacco waste. Waste Manag Res 29:1124–1133Google Scholar
  26. Keinath AP (2007) Sensitivity of populations of Phytophthora capsici from South Carolina to Mefenoxam, Dimethomorph, Zoxamide, and Cymoxanil. Plant Dis 91:743–748Google Scholar
  27. Keitaro T, Ryouta H, Tadao W (2012) Inoculation of arbuscular mycorrhizal fungi can substantially reduce phosphate fertilizer application to Allium fistulosum L. and achieve marketable yield under field condition. Biol Fertil Soils 48:839–843Google Scholar
  28. Kennedy BS, Severson RF, Nielsen MT (1995) Biorationals from Nicotiana protect cucumbers against Colletotrichum lagenarium & halst disease development. J Chem Ecol 21:221–231Google Scholar
  29. Khan MA, Cheng Z, Khan AR, Rana SJ, Ghazanfar B (2015) Pepper-garlic intercropping system improves soil biology and nutrient status in plastic tunnel. Int J Agric Biol 17:869–880Google Scholar
  30. Kousik CS, Donahoo RS, Hassell R (2012) Resistance in watermelon rootstocks to crown rot caused by Phytophthora capsici. Crop Prot 39:18–25Google Scholar
  31. Larkin RP, Griffin TS (2007) Control of soilborne potato diseases using Brassica green manures. Crop Prot 26:1067–1077Google Scholar
  32. Lim JH, Kim SD (2010) Biocontrol of phytophthora blight of red pepper caused by Phytophthora capsici, using Bacillus subtilis, AH18 and B. Licheniformis, K11 formulations. J Korean Soc Appl Biol Chem 53:766–773Google Scholar
  33. Liu Y, He J, Shi G, An L, Öpik M, Feng H (2011) Diverse communities of arbuscular mycorrhizal fungi inhabit sites with very high altitude in Tibet Plateau. FEMS Microbiol Ecol 78:355–365Google Scholar
  34. Lu RK (2000) Analytical methods for soil and agro-chemistry. China Agricultural Science and Technology Press, BeijingGoogle Scholar
  35. Mardanluo S, Souri MK, Ahmadi M (2018) Plant growth and fruit quality of two pepper cultivars under different potassium levels of nutrient solutions. J Plant Nut 41:1–11Google Scholar
  36. Mccarty DG, Inwood SEE, Ownley BH, Sams CE, Wszelaki AL, Butler DM (2014) Field evaluation of carbon sources for anaerobic soil disinfestation in tomato and bell pepper production in Tennessee. Hortscience 49:272–280Google Scholar
  37. Meisinger JJ, Bandel VA, Angle JS, O'Keefe BE, Reynolds CM (1992) Presidedress soil nitrate test evaluation in Maryland. Soil Sci Soc Am J 56:1527–1532Google Scholar
  38. Messiha NAS, Diepeningen ADV, Wenneker M (2007) Biological Soil Disinfestation (BSD), a new control method for potato brown rot, caused by Ralstonia solanacearum race 3 biovar 2. Eur J Plant Pathol 117:403–415Google Scholar
  39. Momma N (2008) Biological soil disinfestation of soilborne pathogens and its possible mechanisms. Japan Agric Res Quart 42:7–1Google Scholar
  40. Momma N, Yamamoto K, Simandi P, Shishido M (2006) Role of organic acids in the mechanisms of biological soil disinfestation (BSD). J Gen Plant Pathol 72:247–252Google Scholar
  41. Momma N, Momma M, Kobara Y (2010) Biological soil disinfestation using ethanol: effect on Fusarium oxysporum f. sp lycopersici and soil microorganisms. J Gen Plant Pathol 5:336–344Google Scholar
  42. Momma N, Kobara Y, Momma M (2011) Fe2+, and Mn2+, potential agents to induce suppression of Fusarium oxysporum for biological soil disinfestation. J Gen Plant Pathol 77:331–335Google Scholar
  43. Momma N, Kobara Y, Uematsu S (2013) Development of biological soil disinfestations in Japan. Appl Microbiol Biotechnol 97:3801–3809Google Scholar
  44. Nadeem SM, Ahmad M, Zahir ZA, Javaid A, Ashraf M (2014) The role of mycorrhizae and plant growth promoting rhizobacteria in improving crop productivity under stressful environments. Biotechnol Adv 32:429–448Google Scholar
  45. Nannipieri P, Giagnoni L, Landi L, Renella G (2011) Role of phosphatase enzymes in soil. In: Bünemann E, Oberson A, Frossard E (eds) Phosphorus in Action. Soil Biology. Springer, Berlin, Heidelberg, pp. 215–243Google Scholar
  46. Ozgonen H, Erkilic A (2007) Growth enhancement and Phytophthora blight (Phytophthora capsici Leonian) control by arbuscular mycorrhizal fungal inoculation in pepper. Crop Prot 26:1682–1688Google Scholar
  47. Phillips JM, Hayman DS (1970) Improved procedures for clearing roots and staining parasitic and vesicular arbuscular mycorrhizal fungi for rapid assessment of infection. Trans Br Mycol Soc 55:158–161Google Scholar
  48. Pinkas Y, Kariv A, Katan J (1984) Soil solarization for the control of Phytophthora cinnamomi: thermal and biological effects. Phytopathology 74:796–798Google Scholar
  49. Polo-López MI, Oller I, Fernández-Ibáñez P (2013) Benefits of photo-Fenton at low concentrations for solar disinfection of distilled water. A case study: Phytophthora capsici. Catal Today 209:181–187Google Scholar
  50. Qiu L, Bi YL, Jiang B, Wang ZG (2017) Effects of plastic film mulching and inoculation with AM fungi on soil physicochemical properties of maize rhizosphere in semiarid areas. Mycorrhiza 36:904–913Google Scholar
  51. Reifschneider FJB, Boiteuxl LS, Vecchia PTD, Poulos JM, Kuroda N (1992) Inheritance of adult-plant resistance to Phytophthora capsici in pepper. Euphytica 62:45–49Google Scholar
  52. Ren L, Zhang N, Wu P, Huo H, Xu G, Wu G (2015) Arbuscular mycorrhizal colonization alleviates Fusarium wilt in watermelon and modulates the composition of root exudates. Plant Growth Regul 77:77–85Google Scholar
  53. Reyes-Tena A, Rincón-Enríquez G, López-Pérez L, Quiñones-Aguilar EE (2017) Effect of mycorrhizae and actinomycetes on growth and biocontrol of Capsicum annuum L. against Phytophthora capsici. Pak J Agric Sci 54:513–522Google Scholar
  54. Rosskopf EN, Burelle N, Hong J, Butler DM, Noling JW, He Z, Booker B, Sances F (2014) Comparison of anaerobic soil disinfestation and drip-applied organic acids for raised-bed specialty crop production in Florida. Acta Hortic 1044:221–228Google Scholar
  55. Rosskopf EN, Serrano-Pérez P, Hong J, Shrestha U, Martin K, Kokalis-Burelle N, Shennan C, Muramoto J, Butler D (2015) Anaerobic soil disinfestation and soilborne pest management. In: Meghvansi M, Varma A (eds) Organic amendments and soil suppressiveness in plant disease management. Springer International Publishing, Switzerland, pp 277–305Google Scholar
  56. Shinmura A (2004) Principle and effect of soil sterilization method by reducing redox potential of soil. Phytopathol Soc Jpn, Soilborne Disease Workshop Report 22:2–12Google Scholar
  57. Sivakumar N (2013) Effect of edaphic factors and seasonal variation on spore density and root colonization of arbuscular mycorrhizal fungi in sugarcane fields. Ann Microbiol 63:151–160Google Scholar
  58. Smith SE, Read DJ (2008) Mycorrhizal symbiosis, 3rd edn. Academic Press, LondonGoogle Scholar
  59. Strauss SL, Kluepfel DA (2015) Anaerobic soil disinfestation: a chemical-independent approach to pre-plant control of plant pathogens. J Integr Agric 14:2309–2318Google Scholar
  60. Sunwoo JY, Lee YK, Hwang BK (1996) Induced resistance against Phytophthora capsici in pepper plants in response to DL-X-aminon-butyric acid. Eur J Plant Pathol 102:663–670Google Scholar
  61. Tabatabai MA (1982) Soil enzymes. In: Page AL, Miller RH, Keeney DR (eds) Methods of soil analyses, Part 2, Chemical and microbiological properties, 2nd edn. American Society of Agronomy, Madison, pp 903–947Google Scholar
  62. Tabin T, Arunachalam A, Shrivastava K, Arunachalam K (2009) Effect of arbuscular mycorrhizal fungi on damping-off disease in Aquilaria agallocha Roxb. seedlings. Trop Ecol 50:243–248Google Scholar
  63. Verbruggen E, Van DHMGA, Kowalchuk GA (2012) Community assembly, species richness and nestedness of arbuscular mycorrhizal fungi in agricultural soils. Mol Ecol 21:2341–2353Google Scholar
  64. Verma RK, Arya ID (1998) Effect of arbuscular mycorrhizal fungal isolates and organic manure on growth and mycorrhization of micropropagated Dendrocalamus asper plantlets and on spore production in their rhizosphere. Mycorrhiza 8:113–116Google Scholar
  65. Vierheilig H, Steinkellner S, Khaosaad T, García-Garrido JM (2008) The biocontrol effect of mycorrhization on soil borne fungal pathogens and the autoregulation of the AM symbiosis: one mechanism, two effects. In: Varma A (ed) Mycorrhiza: genetics and molecular biology, eco-function, biotechnology, ecophysiology, structure and systematics. Springer-Verlag, Heidelberg, pp 307–320Google Scholar
  66. Vigo C, Norman JR, Hooker JE (2000) Biocontrol of the pathogen Phytophthora parasitica by arbuscular mycorrhizal fungi is a consequence of effects on infection loci. Plant Pathol 49:509–514Google Scholar
  67. Wang FY, Lin XG, Yin R, Wu LH (2006) Effects of arbuscular mycorrhizal inoculation on the growth of Elsholtzia splendens and Zea mays and the activities of phosphatase and urease in a multi-metal contaminated soil under unsterilized conditions. Appl Soil Ecol 31:110–119Google Scholar
  68. Wang F, Adarms CA, Shi Z, Sun Y (2018) Combined effects of ZnO NPs and Cd on sweet sorghum as influceds by an arbuscular mycorrhizal fungus. Chemosphere 209:421–429Google Scholar
  69. Willis A, Rodrigues BF, Harris PJC (2013) The ecology of arbuscular mycorrhizal fungi. Crit Rev Plant Sci 32:1–20Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Key Laboratory of Plant Nutrition and the Agri-environment in Northwest China, Ministry of Agriculture and Rural Affairs; College of Natural Resources and EnvironmentNorthwest A&F UniversityYanglingPeople’s Republic of China
  2. 2.State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil ScienceChinese Academy of SciencesNanjingPeople’s Republic of China
  3. 3.University of Chinese Academy of SciencesBeijingPeople’s Republic of China

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