Cytophysiological manifestations of wheat’s defense reactions against stem rust induced by the biofungicide Novochizol

Biologization is a priority direction of agricultural production. One of the promising approaches to solve the biologization problem is the use of chitosan-based biopreparations to stimulate plant growth and protect plants from a wide range of pathogens. Currently, active work is underway to create and test new chitosan preparations. Novochizol was obtained as a result of intramolecular crosslinking of linear chitosan molecules and has a globular shape. Previously, a Novochizol-stimulating effect on the growth and development of common wheat was demonstrated. However, the induced resistance mechanisms against rust diseases have not been studied before. The reported studies have revealed the dose effect of the preparation on the development of wheat stem rust. The best results of visual estimation of plant reactions were obtained with 0.125 and 0.75 % Novochizol pretreatment four days before rust infection. After pretreatment of susceptible cv. Novosibirsk 29 seedlings, a resistant reaction appeared and the urediniopustule density was decreased. Cytophysiological studies have shown that 0.75 % Novochizol stimulated an intensive accumulation of hydrogen peroxide H₂O₂ in the leaves of the infected and healthy plants within 48 hours post inoculation (h p/in). During the period of 48–144 h p/in, H₂O₂ gradually disappeared from tissues, but its content increased significantly at the sporulation stage around pustules. However, Novochizol did not induce the hypersensitivity reaction in infected plants. The preparation induced an earlier and more intensive (compared with untreated plants) accumulation of phenolic substances with different autofluorescence in the zones around pathogen colonies. Novochizol induced a change in the ratio of phenols with different spectral characteristics towards compounds with an increased content of syringin derivatives. This work is the first stage in the study of Novochizol effects on wheat defense mechanisms against stem rust. The research will be continued using molecular genetics, biochemical and cytophysiological methods.

1. Abd El-Kareem F., Haggag W. Chitosan and citral alone or in combination for controlling early blight disease of potato plants under field conditions. Res J Pharm Biol Chem Sci. 2014;5(6):941-949. https://rjpbcs.com/pdf/2014_5(6)/%5B141%5D.pdf

2. Badanova E.G., Davletbaev I.M., Sirotkin A.S. Preparations based on chitosan for agriculture. Vestnik Tekhnologicheskogo Universiteta = Herald of Technological University. 2016;19(16):89-95 (in Russian)

3. Bellameche F., Jasim M., Mauch-Mani B., Mascher F. Histopathological aspects of resistance in wheat to Puccinia triticina, induced by Pseudomonas protegens CHA0 and β-aminobutyric acid. Phytopathol Mediterr. 2021;60(3):441-453. doi 10.36253/phyto13123

4. Boller T., Keen N.T. Perception and transduction of elisitor signals in host-pathogen interactions. In: Slusarenko A.J., Fraser R.S.S., van Loon L.C. (Eds) Mechanisms of Resistance to Plant Diseases. Dordrecht: Springer, 2000;189-230. doi 10.1007/978-94-011-3937-3_7

5. Burlakova S.V., Egorycheva M.T., Fomenko V.V., Salakhutdinov N.F., Shcherban A.B. Biological justification of the use of Novohisol with natural fungicides in the cultivation of bread wheat. Chemistry for Sustainable Development. 2025;3:303-314 (in Russian)

6. Chakraborty M., Hasanuzzaman M., Rahman M., Khan Md.A.R., Bhowmik P., Mahmud N.U., Tanveer M., Islam T. Mechanism of plant growth promotion and disease suppression by chitosan biopolymer. Agriculture. 2020;10:624. doi 10.3390/agriculture10120624

7. Chandler D., Bailey A.S., Tatchell G.M., Davidson G., Greaves J., Grant W.P. The development, regulation and use of biopesticides for integrated pest management. Philos Trans R Soc Lond B Biol Sci. 2011;366:1987-1998. doi 10.1098/rstb.2010.0390

8. Elsharkawy M.M., Omara R.I., Mostafa Y.S., Alamri S.A., Hashem M., Alrumman S.A., Ahmad A.A. Mechanism of wheat leaf rust control using chitosan nanoparticles and salicylic acid. J Fungi. 2022; 8(3):304. doi 10.3390/jof8030304

9. Faoro F., Maffi D., Cantu D., Iriti M. Chemical-induced resistance against powdery mildew in barley: the effects of chitosan and benzothiadiazole. BioControl. 2008;53(2):387-401. doi 10.1007/s10526-007-9091-3

10. Ghauoth A., Arul J., Grenier J., Benhamou N., Asselin A., Belanger G. Effect of chitosan on cucumber plants: suppression of Pythium aphanidermatum and induction of defense reaction. Phytopathology. 1994;84(3):313-320. doi 10.1094/PHYTO-84-31

11. Gill U.S., Lee S., Mysore K.S. Host versus nonhost resistance: distinct wars with similar arsenals. Phytopathology. 2015;105(5):580-587. doi 10.1094/PHYTO-11-14-0298-RVW

12. Haggag W.M.W., Hussein M.M., Medhat M.T., El Habbasha S.F. Enhancement of wheat resistant to diseases by elicitors. Int J Sci Res. 2014;3(11):1526-1530

13. Japaridze L.I. Practicum on Microscopic Chemistry of Plants. Moscow: Sovetskaya Nauka Publ., 1953 (in Russian)

14. Lopez-Moya F., Martin-Urdiroz M., Oses-Ruiz M., Were V.M., Fricker M.D., Littlejohn G.R., Lopez-Llorca L.V., Talbot N.J. Chitosan inhibits septin-mediated plant infection by the rice blast fungus Magnaporthe oryzae in a protein kinase C and Nox1 NADPH oxidase-dependent manner. New Phytol. 2021;230(4):1578-1593. doi 10.1111/nph.17268

15. Maksimov I.V., Cherepanova E.A. Pro-/antioxidant system and resistance of plants to pathogens. Uspehi Sovremennoy Biologii = Advances in Current Biology. 2006;126(3):250-261 (in Russian)

16. Malerba M., Cerana R. Chitosan effects on plant systems. Int J Mol Sci. 2016;17(7):996. doi 10.3390/ijms17070996

17. Manjunatha G., Roopa K.S., Prashanth G.N., Shetty H.S. Chitosan enhances disease resistance in pearl millet against downy mildew caused by Sclerospora graminicola and defence-related enzyme activation. Pest Manag Sci. 2008;64:1250-1257. doi 10.1002/ps.1626

18. Manjunatha G., Niranjan-Raj S., Prashanth G.N., Deepak S., Amruthesh K.N., Shetty H.S. Nitric oxide is involved in chitosaninduced systemic resistance in pearl millet against downy mildew disease. Pest Manag Sci. 2009;65(7):737-743. doi 10.1002/ps.1710

19. Meng X., Yang L., Kennedy J.F., Tian S. Effects of chitosan and oligochitosan on growth of two fungal pathogens and physiological properties in pear fruit. Carbohydr Polym. 2010;81(1):70-75. doi 10.1016/j.carbpol.2010.01.057

20. Moldenhauer J., Moerschbacher B.M., van der Westhuizen A.J. Histological investigation of stripe rust (Puccinia striiformis f. sp. tritici) development in resistant and susceptible wheat cultivars. Plant Pathology. 2006;55:469-474. doi 10.1111/j.1365-3059.2006.01385.x

21. Nandeeshkumar P., Sudisha J., Ramachandra K.K., Prakash H., Niranjana S., Shekar S.H. Chitosan induced resistance to downy mildew in sunflower caused by Plasmopara halstedii. Physiol Mol Plant Pathol. 2008;72(4-6):188-194. doi 10.1016/j.pmpp.2008.09.001

22. Orzali L., Forni C., Riccioni L. Effect of chitosan seed treatment as elicitor of resistance to Fusarium graminearum in wheat. Seed Sci Technol. 2014;42(2):132-149. doi 10.15258/sst.2014.42.2.03

23. Orzali L., Corsi B., Forni C., Riccioni L. Chitosan in agriculture: a new challenge for managing plant disease. In: Shalaby E.A. (Ed.) Biological Activities and Application of Marine Polysaccharides. InTech. 2017;87-96. doi 10.5772/66840

24. Plotnikova L.Ya. Influence of the surface features and physiological reactions of non-host species on the development of cellular structures of rust fungi. Tsitologiia. 2008;50(5):439-446 (in Russian)

25. Plotnikova L.Ya. Effect of benzothiadiazole, an inducer of systemic acquired resistance, on the pathogenesis of wheat brown rust. Russian Journal of Plant Physiology. 2009;56(4):517-526. doi 10.1134/S1021443709040116

26. Plotnikova L., Knaub V. Exploitation of the genetic potential of Thinopyrum and Agropyron genera to protect wheat from diseases and environmental stresses. Vavilovskii Zhurnal Genetiki i Selektsii = Vavilov J Genet Breed. 2024;28(5):536-553. doi 10.18699/vjgb-24-60

27. Plotnikova L.Y., Meshkova L.V. Evolution of cytophysiological relationships between leaf rust causal agent and common wheat in the process of overcoming of resistance determined by the gene Lr19. Mikologiya i Fitopatologiya = Mycology and Phytopathology. 2009; 43(4):343-357 (in Russian)

28. Plotnikova L., Pozherukova V., Knaub V., Kashuba Y. What was the reason for the durable effect of Sr31 against wheat stem rust? Agriculture. 2022;12:2116. doi 10.3390/agriculture12122116

29. Plotnikova L., Knaub V., Pozherukova V. Nonhost resistance of Thinopyrum ponticum to Puccinia graminis f. sp. tritici and the effects of the Sr24, Sr25, and Sr26 genes introgressed to wheat. Int J Plant Biol. 2023;14:435-457. doi 10.3390/ijpb14020034

30. Popova E.V., Domnina N.S., Kovalenko N.M., Sokornova S.V., Tyuterev S.L. Influence of chitosan hybrid derivatives on induced wheat resistance to pathogens with different nutrition strategies. Applied Biochemistry and Microbiology. 2018;54(5):535-539. doi 10.1134/S0003683818050150

31. Rabea E.I., Badawy M.E., Rogge T.M., Stevens C.V., Höfte M., Steurbaut W., Smagghe G. Insecticidal and fungicidal activity of new synthesized chitosan derivatives. Pest Manag Sci. 2005;61(10):951- 960. doi 10.1002/ps.1085

32. Richter T., Gulich M., Richter K. Quality control and good manufacturing practice (GMP) for chitosan-based biopharmaceutical products. In: Sarmento B., das Neves J. (Eds) Chitosan-Based Systems for Biopharmaceuticals: Delivery, Targeting and Polymer Therapeutics. John Wiley & Sons, 2012;503-542. doi 10.1002/9781119962977.ch26

33. Rkhaila A., Chtouki T., Erguig H., El Haloui N., Ounine K. Chemical proprieties of biopolymers (chitin/chitosan) and their synergic effects with endophytic bacillus species: unlimited applications in agriculture. Molecules. 2021;26(4):1117. doi 10.3390/molecules26041117

34. Roelfs A.P., Singh R.P., Saari E.E. Rust diseases of wheat: concepts and methods of disease management. Cimmyt, Mexico DF, Mexico, 1992.

35. Rsaliyev A.S., Rsaliyev Sh.S. Principal approaches and achievements in studying race composition of wheat stem rust. Vavilovskii Zhurnal Genetiki i Selektsii = Vavilov J Genet Breed. 2018;22(8): 967-977. doi 10.18699/VJ18.439 (in Russian)

36. Shcherban A.B. Chitosan and its derivatives as promising plant protection tools. Vavilovskii Zhurnal Genetiki i Selektsii = Vavilov J Genet Breed. 2023;27(8):1010-1021. doi 10.18699/VJGB-23-116

37. Singh Y., Nair A.M., Verma P.K. Surviving the odds: from perception to survival of fungal phytopathogens under host-generated oxidative burst. Plant Commun. 2021;2:100142. doi 10.1016/j.xplc.2021.100142

38. Sørensen C.K., Thach T., Hovmøller M.S. Evaluation of spray and point inoculation methods for the phenotyping of Puccinia striiformis on wheat. Plant Disease. 2016;100(6):1064-1070. doi 10.1094/PDIS-12-15-1477-RE

39. Sternshis M.V., Belyaev A.A., Tsvetkova V.P., Shpatova T.V., Lelyak A.A., Bakhvalov S.A. Biopreparations Based on Bacteria of the Genus Bacillus for Plant Health Management. Novosibirsk: Publishing House of SB RAS, 2016 (in Russian)

40. Tarchevsky I.A. Elicitor-induced signaling pathways and their interaction. Russian Journal of Plant Physiology. 2000;47(2):285-294

41. Teplyakova O.I., Fomenko V.V., Salakhutdinov N.F., Vlasenko N.G. Novochizol™ seed treatment: effects on germination, growth and development in soft spring wheat. Nat Prod Chem Res. 2022;10(5): 1-4. doi 10.35248/naturalproducts.10.5.1-04

42. Tyuterev S.L. Ecologically safe inducers of plant resistance to diseases and physiological stresses. Vestnik Zashchity Rasteniy = Plant Protection News. 2015;1(83):3-13 (in Russian)

43. USDA. World Agricultural Production; USDA Foreign Agricultural Service. Washington, DC, USA, 2016.

44. Vander P., Vårum K.M., Domard A., El Gueddari N.E., Moerschbacher B.M. Comparison of the ability of partially N-acetylated chitosans and chitooligosaccharides to elicit resistance reactions in wheat leaves. Plant Physiol. 1998;118(4):1353-1359. doi 10.1104/pp.118.4.1353

45. Varlamov V.P., Ilyina A.V., Shagdarova B.T., Lunkov A.P., Mysyakina I.S. Chitin/chitosan and its derivatives: fundamental problems and practical approaches. Biochemistry. 2020;85:154-176. doi 10.1134/S0006297920140084

46. Woldeab G., Hailu E., Bacha N. Protocols for race analysis of wheat stem rust (Puccinia graminis f. sp. tritici). Ambo, Ethiopia: EIAR, 2017.

47. Yarullina L.G., Kalatskaja J.N., Cherepanova E.A., Yalouskaya N.A., Tsvetkov V.O., Ovchinnikov I.A., Burkhanova G.F., Rybinskaya K.I., Sorokan A.V., Herasimovich K.M., Zaikina E.A., Nikolaichuk V.V., Hileuskaya K.S., Mardanshin I.S. Approaches to improving biological activity of agricultural formulations based on bacteria of the genus Bacillus and chitosan nanocomposites (review). Applied Biochemistry and Microbiology. 2023;59(5)549-560. doi 10.1134/s0003683823050186

48. Yarullina L.G., Burkhanova G.F., Tsvetkov V.O., Cherepanova E.A., Sorokan A.V., Zaikina E.A., Mardanshin I.S., Fatkullin I.Y., Maksimov I.V., Kalatskaja J.N., Yalouskaya N.A., Rybinskay E.I. The effect of chitosan conjugates with hydroxycinnamic acids and Bacillus subtilis bacteria on the activity of protective proteins and resistance of potato plants to Phytophthora infestans. Appl Biochem Microbiol. 2024a;60(2):231-240. doi 10.1134/S0003683824020194

49. Yarullina L., Kalatskaja J., Tsvetkov V., Burkhanova G., Yalouskaya N., Rybinskaya K., Zaikina E., Cherepanova E., Hileuskaya K., Nikalaichuk V. The influence of chitosan derivatives in combination with Bacillus subtilis bacteria on the development of systemic resistance in potato plants with viral infection and drought. Plants. 2024b;13: 2210. doi 10.3390/plants13162210

50. Yuan M., Pok B., Ngou M., Ding P., Xin X.-F. PTI-ETI crosstalk: an integrative view of plant immunity. Curr Opin Plant Biol. 2021;62: 102030. doi 10.1016/j.pbi.2021.102030

51. Yuan P., Qian W., Jiang L., Jia C., Ma X., Kang Z., Liu J. A secreted catalase contributes to Puccinia striiformis resistance to host-derived oxidative stress. Stress Biol. 2021;1(1):22. doi 10.1007/s44154-021-00021-2

52. Zhao Y., Zhu X., Chen X., Zhou J. From plant immunity to crop disease resistance. J Genet Genom. 2022;49(8):693-703. doi 10.1016/j.jgg.2022.06.003

53. Zheng K., Lu J., Li J., Yu Y., Zhang J., He Z., Ismail O.M., Wu J., Xie X., Li X., Xu G., Dou D., Wang X. Efficiency of chitosan application against Phytophthora infestans and the activation of defence mechanisms in potato. Int J Biol Macromol. 2021;182:1670-1680. doi 10.1016/j.ijbiomac.2021.05.097