PLANT BIOLOGY
ANIMAL BIOLOGY
SUBSCRIPTION
E-SUBSCRIPTION
 
MAP
MAIN PAGE

 

 

 

 

doi: 10.15389/agrobiology.2019.5.1041eng

UDC: 633.11:632.4:632.9:579.64

Acknowledgements:
Supported financially by the Kazakhstan Ministry of Education and Science (grant project No. АР05131526)

 

CELLULOLYTIC BACTERIA AND ASSOCIATION OF EFFECTIVE MICROORGANISMS FOR BIOCONTROL OF ROOT ROT INFECTIONS IN SUGAR BEET (Beta vulgaris L.)

I.E Smirnova, A.K. Sadanov

LP Sсientific Production Center of Microbiology and Virology, 105, Bogenbai Batyr str., Almaty, 050010 Kazakhstan, e-mail: iesmirnova@mail.ru (✉ corresponding author), a.sadanov@inbox.ru

ORCID:
Smirnova I.E orcid.org/0000-0001-5854-1529
Sadanov A.K. orcid.org/0000-0002-2593-6302

Received June 20, 2019

 

At the present time, many farmers growing sugar beet (Beta vulgaris L.) reduce application of fertilizers and crop rotations, which leads to accumulation of phytopathogens. The main pathogens of sugar beet causing root rot are fungi from genera Fusarium Lk.:Fr. and Alternaria (Fr.) Keissi. Chemicalfungicidesare used worldwide to protect crops but plant pathogenic fungi acquire resistance against conventional chemicals. Therefore, the biological methods of plant protection are relevant. In our previous works, we have designed the association of effective microorganisms (EM Association) which includes nitrogen-fixing Azotobacter chroococcum and phosphate-mobilizing Bacillus megaterium bacteria. This association can increase the productivity of sugar beet plants but does not possess antifungal activity against sugar beet root rot. Cellulolytic bacteria are an important component of microbiocenoses. They play a significant role in soil processes; their number is an indicator of soil fertility and ecological quality. Colonizing rhizosphere of plants, they synthesize bioactive substances, including antifungal metabolites. Among cellulolytic bacteria there are active antagonists of fungal root rot causative agents. In this paper we present our research findings on the antifungal properties of a new cellulolytic strain Bacillus sp. C-82/3 and the first effective association of this strain with nitrogen-fixing and phosphate-mobilizing bacteria which promotes sugar beet growth and yield. The goal of the research was to evaluate antifungal activity of the novel strain of cellulolytic bacteria Bacillus sp. C-82/3 isolated from soil rhizosphere of healthy sugar beet plants in the South-East of Kazakhstan (Zhambyl region),to enrich the EM association developed with this strain, and to assess the plant growth promoting activity of the improved EM Association and its ability to biocontrol rot root infections under field condition. Antifungal activity was determined in agar block diffusion testsagainst Alternaria alternata (Fr.) Keissl, Fusarium solani (Mart.) Sacc.and F. oxysporum Schlecht.The strain was grown on the Hutchinson’s medium (1.0 g/l К2НРО4, 0.1 g/l CaCl2, 0.3 g/l MgS04, 2.5 g/l NaNO3, 1.0 g/l NaCl, 0.01 g/l FеСl3, 20 g/l wheat straw, 5,0 g/l yeast extract; pH 7.0). Blocks with growing culture were cut out, and put on Petri dishes with potato-dextrose agar earlier inoculated with fungi, and cultured at 28 °С for 3 days. Antifungal activity was assessed by the diameter of growth inhibition zone. To study the plant growth promoting activity of the EM association with Bacillus sp. C-82/3, the cv. Aisultan seeds were treated with the bacterial suspension (107 cells/ml) at 23 °С for 2 hours. The stem and root length measured in the inoculated seedling after 30-day growing in a climatic chamber (Constant Сlimate Сhambe rHPP750, Memmert GmbH + Co. KG, Germany) were compared to the control. Field tests were conducted in the South-East of Kazakhstan (Zhambyl region, Kaiyndy farm) in 2017-2018. The results of lab screening showed high antifungal activityof the novel strain Bacillus sp. C-82/3 with the mean inhibition halos of 28.9±0.2 mm for F. oxysporum, 38.2±0.3 mm for F. solani, and 46.6±0.9 mmfor A. alternate. The improved EM Association which includes three strains (Bacillus megateriumAzotobacter chroococcum and Bacillus sр. С-82/3) was characterized by high growth-promoting activity. Germination of the inoculated seeds was 7-16 % higher, and stem and root length increased 1.2-1.5-fold and 1.1-2.0-fold, respectively, as compared to control (р ≤ 0.05). We also revealed the high ability of the EM Association containing Bacillus sр. С-82/3 strain to suppress sugar beet root rot pathogens in soil biocenosis. Seed inoculation with the microbial association decreased the damage to seedlings 2.3 times, to roots 3.0 times. The yield of sugar beet was 34.2±2.3 c/ha higher compared to control (р ≤ 0.05). Thus, our data are the first evidence that the EM Association with Bacillus sp. C-82/3, a new cellulolytic strain with high antifungal activity that we have detected, is effective against root rot infection and promotes an increase in sugar beet yield under field condition.

Keywords: Beta vulgaris L., sugar beet, biological control, cellulolytic bacteria, effective microorganisms association, antifungal activity, growth-promoting activity, phytopathogenic fungi, root rot.

 

REFERENCES

  1. Solomon S., Quirk R.G., Shukla S.K. Green management for sustainable sugar industry. Sugar Tech., 2019, 21(2): 183-185 CrossRef
  2. Tereshchenkova I.A. Vestnik Belorusskoi gosudarstvennoi sel'skokhozyaistvennoi akademii, 2015, 4: 11-14 (in Russ.).
  3. Kirillov N.A., Volkov A.I., Prokhorova L.H. Sakharnaya svekla, 2013, 1: 23-27 (in Russ.).
  4. Paramasivan M., Chandrasekaran A., Mohan S., Muthukrishnan N. Ecological management of tropical sugar beet (TSB) root rot by rhizosphere Trichoderma species. Archives of Phytopathology and Plant Protection, 2014, 47(13): 1629-1644 CrossRef
  5. Mahmoud A.F. Suppression of sugar beet damping-off caused by Rhizoctonia solani using bacterial and fungal antagonists. Archives of Phytopathology and Plant Protection, 2016, 49(19-20): 575-585 CrossRef
  6. Maui A.A. Novosti nauki Kazakhstana, 2014, 2(120): 63-70 (in Russ.).
  7. Merzaliev K., Kul'keev E.E., Al'dekov N.A., Amanova K.S. Vestnik sel'skokhozyaistvennykh nauk Kazakhstana, 2016, 1-2: 21-25 (in Russ.).
  8. Abd-El-Khair H., Abd-El-Fattah A.I., El-Nagdi W.M.A. Evaluation of five sugar beet varieties for root-knot nematode and root-rot fungal infection. Arch. Phytopath. Plant Prot.,2013, 46(18): 2163-2173 CrossRef
  9. Mahmoud A.F. Occurrence of Fusarium wilt on summer squash caused by Fusarium oxysporum in Assiut, Egypt. Journal of Phytopathology and Pest Management, 2016, 3(1): 34-45.
  10. Mahmoud A.F. Evaluation of certain antagonistic fungal species for biological control of faba bean wilt disease incited by Fusarium oxysporum. Journal of Phytopathology and Pest Management, 2016, 3(2): 1-14.
  11. Maui A.A., Ismukhambetov Zh.D. Kompleksnaya sistema zashchity posevov sakharnoi svekly ot vreditelei, boleznei i sornyakov dlya uslovii yuga i yugo-vostoka Kazakhstana [A comprehensive system for the protection of sugar beet crops from pests, diseases and weeds in the south and southeast Kazakhstan]. Almaty, 2012 (in Russ.).
  12. Selivanova G.A. Zemledelie, 2013, 4: 31-37 (in Russ.).
  13. Stognienko O.I., Shamin A.A. Zashchita i karantin rastenii, 2014, 8: 12-15 (in Russ.).
  14. Shamin A.A., Stognienko O.I., Borotov O.K. Zemledelie, 2013, 4: 35-38 (in Russ.).
  15. Strausbaugh C.A., Gillen A.M. Sugar beet root rot at harvest in the US Intermountain West. Canadian Journal of Plant Pathology, 2009, 31(2): 232-240 CrossRef
  16. Karimi E., Sadeghi A., Dahaji P.A., Dalvand Y., Omidvari M., Nezhad M.K. Biocontrol activity of salt tolerant Streptomyces isolates against phytopathogens causing root rot of sugar beet. Biocontrol Science and Technology, 2012, 22(3): 333-349 CrossRef
  17. Webb K.M., Brenner T., Jacobsen B.J. Temperature effects on the interactions of sugar beet with Fusarium yellows caused by Fusarium oxysporum f. sp. betae. Canadian Journal of Plant Pathology, 2015, 37(3): 353-362 CrossRef
  18. Gossen B.D., Carisse O., Kawchuk L.M., van der Heyden H., McDonald M.R. Recent changes in fungicide use and the fungicide insensitivity of plant pathogens in Canada. Canadian Journal of Plant Pathology, 2014, 36(3): 327-340 CrossRef
  19. Prior R., Mittelbach M., Begerow D. Impact of three different fungicides on fungal epi- and endophytic communities of common bean (Phaseolus vulgaris) and broad bean (Vicia faba). Journal of Environmental Science and Health, Part B, 2017, 52(6): 376-386 CrossRef
  20. Wedge D.E., Surry J.K., Kreiser B., Curry A., Abril M., Smith B.J. Fungicide resistance profiles for 13 Botrytis cinerea isolates from strawberry in Southeastern Louisiana. International Journal of Fruit Science, 2013, 13(4): 413-429 CrossRef
  21. Shabaev V.P. Sel'skokhozyaistvennaya biologiya [Agricultural Biology], 2005, 3: 55-61 (in Russ.).
  22. Trenozhnikova L.P., Balgimbaeva A.S., Ultanbekova G.D., Galimbaeva R.Sh. Antifungal activity against pathogens of cereals and characterization of antibiotics of Streptomyces sp. strain K-541 isolated from extreme ecosystems in Kazakhstan. Agricultural Biology [Sel'skokhozyaistvennaya biologiya], 2018, 53(1): 96-102 CrossRef
  23. Abeer H., Asma A.H., Allah A., Qarawi A., Shalawi A., Dilfuza E. Impact of plant growth-promoting Bacillus subtilis on growth and physiological parameters of Bassia indica (Indian Bassia) grown under salt stress. Pakistan Journal of Botany, 2015, 47(5): 1735-1741.
  24. Bjelić D., Marinković J., Tintor B., Mrkovački N. Antifungal and plant growth promoting activities of indigenous rhizobacteria isolated from maize (Zea mays L.) rhizosphere. Communications in Soil Science and Plant Analysis,2018, 49(1): 88-98 CrossRef
  25. Talaat N.B. Effective microorganisms improve growth performance and modulate the ROS-scavenging system in common bean (Phaseolus vulgaris L.) plants exposed to salinity stress. J. Plant Growth Regul., 2015, 34(1): 35-46 CrossRef
  26. Bzdyk R.M., Olchowik J., Studnicki M., Oszako T., Sikora K., Szmidla H., Hilszczańska D. The impact of effective microorganisms (EM) and organic and mineral fertilizers on the growth and mycorrhizal colonization of Fagus sylvatica and Quercus robur seedlings in a bare-root nursery experiment. Forests, 2018, 9: 597-610 CrossRef
  27. Hu C., Qi Y. Long-term effective microorganisms application promote growth and increase yields and nutrition of wheat in China. European Journal of Agronomy, 2013, 46: 63-67 CrossRef
  28. Ndona R.K., Friede J.K., Spornberger A., Jezik K. Effective microorganisms (EM): an effective plant strengthening agent for tomatoes in protected cultivation. Biological Agriculture & Horticulture, 2011, 27(2): 189-204 CrossRef
  29. Mayer J., Scheid S., Widmer F., Fliebach A., Oberholzer H.-R. How effective are effective microorganisms (EM). Results from a field study in temperate climate. Applied Soil Ecology, 2010, 46(2): 230-239 CrossRef
  30. Kleiber T., Starzyk J., Bosiacki M. Effect of nutrient solution, effective microorganisms (EM-A), and assimilation illumination of plants on the induction of the growth of lettuce (Lactuca sativa L.) in hydroponic cultivation. Acta Agrobot., 2013, 66(1): 27-38 CrossRef
  31. Chudasama K.S., Thaker V.S. Screening of potential antimicrobial compounds against Xanthomonas campestris from 100 essential oils of aromatic plants used in India: an ecofriendly approach. Archives of Phytopathology and Plant Protection,2012, 45(7): 783-795 CrossRef
  32. Zameer M., Zahid H., Tabassum B., Ali Q., Nasir I.A., Saleem M., Butt S.J. PGPR potentially improve growth of tomato plants in salt-stressed environment. Turkish Journal of Agriculture - Food Science and Technology, 2016, 4(6): 455-463 CrossRef
  33. Saini J.K., Saini R., Tewari L. Simultaneous isolation and screening of cellulolytic bacteria: selection of efficient medium. Journal of Pure and Applied Microbiology,2012, 6(3): 1339-1344.
  34. Zhao Y.-N., Zhang Y.-Q., Du H.-X., Wang Y.-H., Zhang L.-M., Shi X.-J. Carbon sequestration and soil microbes in purple paddy soil as affected by long-term fertilization. Toxicological & Environmental Chemistry, 2015, 97(3-4): 464-476 CrossRef
  35. Shmidt K.N., Khudaigulov G.G. Vestnik YuUrGU. Seriya Pishchevye i biotekhnologii, 2016, 4(4): 54-63 (in Russ.).
  36. Naplekova N.N. Metabolity aerobnykh tsellyulozoliticheskikh mikroorganizmov i ikh rol' v pochvakh [Metabolites of aerobic cellulolytic microorganisms and their role in soils]. Novosibirsk, 2010 (in Russ.).
  37. Ang S.K., Yahya A., Aziz S.A., Salleh M.M. Isolation, screening, and identification of potential cellulolytic and xylanolytic producers for biodegradation of untreated oil palm trunk and its application in saccharification of lemongrass leaves. Preparative Biochemistry and Biotechnology,2015, 45(3): 279-305 CrossRef
  38. Smirnova I.E. Mikologiya i fitopatologiya, 2004, 38(2): 89-93 (in Russ.).
  39. Smirnova I.E., Sadanov A.K., Galimbaeva R.Sh. Biological method for improving germinating and productivity of melilot. In: Recent trends in PGPR research for sustainable crop productivity. M.S. Reddy, R.I. Ilao, P.S. Faylon (eds). Jodhpur-Delhi-Germany, 2016: 21-28.
  40. Egorov N.S. Osnovy ucheniya ob antibiotikakh [Basics of the antibiotics doctrine]. Moscow, 2004 (in Russ.).
  41. Semenov A.V., Sgibnev A.V., Cherkasov S.V., Bukharin O.V. Byulleten' eksperimental'noi biologii i meditsiny, 2007, 144: 702-705 (in Russ.).
  42. Vessey J.K. Plant growth promoting rhizobacteria as biofertilizers. Plant Soil, 2003, 255(2): 571-586 CrossRef
  43. Borovikov V.P. Populyarnoe vvedenie v sovremennyi analiz dannykh v sisteme STATISTICA [A popular introduction to modern data analysis in the STATISTICA system]. Moscow, 2013 (in Russ.).
  44. Felske A.D.M. Ecology of Bacillus species in soil. In: Bacterial spore formers: probiotics and emerging applications. E. Ricca, A.O. Henriques, S.M. Cutting (eds.). Horizon Bioscience, Norfolk, 2004: 35-44.
  45. Fira D., Dimkić I., Berić T., Lozo J., Stanković S. Biological control of plant pathogens by Bacillus species. Journal of Biotechnology, 2018, 285: 44-55 CrossRef
  46. Cao Y., Xu Z., Ling N., Yuan Y., Yang X., Chen L., Shen B., Shen Q. Isolation and identification of lipopeptides produced by B. subtilis SQR 9 for suppressing Fusarium wilt of cucumber. Scientia Horticulturae, 2012, 135: 32-39 CrossRef
  47. Guo Q., Dong W., Li S., Lu X., Wang P., Zhang X., Wang Y., Ma P. Fengycin produced by Bacillus subtilis NCD-2 plays a major role in biocontrol of cotton seedling damping-off disease. Microbiological Research, 2014, 169(7-8): 533-540 CrossRef
  48. Pérez-García A., Romero D., de Vicente A. Plant protection and growth stimulation by microorganisms: biotechnological applications of Bacilli in agriculture. Current Opinion in Biotechnology, 2011, 22(2): 187-193 CrossRef
  49. Hinarejos E., Castellano M., Rodrigo I., Bellés J.M., Conejero V., López-Gresa M.P., Lisón P. Bacillus subtilis IAB/BS03 as a potential biological control agent. Eur. J. Plant Pathol., 2016, 146(3): 597-608 CrossRef
  50. Smirnova I.E., Koishibaev M.K., Talip Zh.Sh. Novosti nauki Kazakhstana, 2008, 2: 124-126 (in Russ.).
  51. Dardanelli M.S., De Córdoba F.J.F., Espuny M.R., Carvajal M.A.R., Díaz M.E.S., Serrano A.M.G., Okon Y., Megías M. Effect of Azospirillum brasilense coinoculated with Rhizobium on Phaseolus vulgaris flavonoids and Nod factor production under salt stress. Soil Biology and Biochemistry, 2008, 40(11): 2713-2721 CrossRef
  52. Askary M., Mostajeran A., Amooaghaei R., Mostajeran M. Influence of the coinoculation Azospirillum brasilense and Rhizobium meliloti plus 2,4-D on grain yield and N, P, K content of Triticum aestivum (cv. Baccros and Mahdavi). American-Eurasian J. Agric. Environ. Sci., 2009, 5(3): 296-307.
  53. Packialakshmi N., Yasotha C. Role of effective microorganism in unfertile soil. Int. J. Phytopharm., 2014, 4(1): 25-27.
  54. Gobbetti M., Cagno R.D., De Angelis M. Functional microorganisms for functional food quality. Critical Reviews in Food Science and Nutrition, 2010, 50(8): 716-727 CrossRef
  55. Kopteva T.S., Erina N.V. Zaikina I.A. Nauchnyi zhurnal KubGAU, 2015, 114(10): 1-10 (in Russ.).
  56. Nautiyal C.S., Srivastava S., Chauhan P.S., Seem K., Mishra A., Sopory S.K. Plant growth-promoting bacteria Bacillus amyloliquefaciens NBRISN13 modulates gene expression profile of leaf and rhizosphere community in rice during salt stress. Plant Physiology and Biochemistry, 2013, 66: 1-9 CrossRef
  57. Iriti M., Scarafoni A., Pierce S., Castorina G., Vitalini S. Soil application of Effective Microorganisms (EM) maintains leaf photosynthetic efficiency, increases seed yield and quality traits of bean (Phaseolus vulgaris L.) plants grown on different substrates. International Journal of Molecular Sciences, 2019, 20(9): 2327-2335 CrossRef
  58. Ncube L., Minkeni P.N.S., Brutsch O. Agronomic suitability of effective microorganisms for tomato production. African Journal of Agricultural Research, 2011, 6(3): 650-654.
  59. Souza R., Ambrosini A., Passaglia L.M.P. Plant growth-promoting bacteria as inoculants in agricultural soils. Genetics and Molecular Biology, 2015, 38(4): 401-419 CrossRef
  60. Patkowska E., Konopiński M. Antagonistic activity of selected bacteria occurring in the soil after root chicory cultivation. Plant, Soil and Environment, 2018, 60(7): 320-324 CrossRef
  61. Singh R., Kumar M, Mittal A., Mehta R.K. Microbial metabolites in nutrition, healthcare and agriculture. 3 Biotech, 2017, 7(1): 4-14 CrossRef

 

back

 


CONTENTS

 

 

Full article PDF (Rus)

Full article PDF (Eng)