Monitoring of the leaf biochemical compositions in Mexican lime (Citrus aurantifolia Swingle) during the progression of Witches’ Broom Disease of Lime (WBDL)

Document Type : Full Paper

Authors

Assistant professor, Research Center of Citrus and Subtropical Fruit, Horticultural Science Research Institute, Agricultural Research and Education Organization (AREO), Ramsar, Iran

Abstract

Witches’ broom disease of lime (WBDL), caused by ‘Candidatus Phytoplasma aurantifolia’, is major limiting factor for Mexican lime (Citrus aurantifolia Swingle) production in southern Iran. In this investigation, biochemical composition profiling of the healthy and infected rooted Mexican lime wasmonitored during WBDL progression (90-330 days) as well as to distinguish infected from healthy Mexican limes under greenhouse conditions. For this aim, stem cuttings is taken from healthy Mexican lime. After three months, half of Mexican lime cutting were inoculated by bark-grafting from WBDL-affected Mexican. Six healthy and infected plants were sampled 90, 150, 210, 270 and 330 days after inoculation with Phytoplasma. Pigments, total phenol, antioxidant capacity, soluble carbohydrate and starch content in leaves were measured. Results showed Phytoplasmal infection caused growth inhibition and reduced stem and root dry mater. Moreover, infection mostly caused to a marked decrease in pigments and significantly increased soluble carbohydrates and starch. In Mexican lime leaves, phenol contents and antioxidant capacity were increased after inoculation with Phytoplasma. In addition, the principal components analysis (PCA) revealed a clear distinction between the leaf biochemical profiles of healthy and infected plants during the progression of WBDL. In conclusion, this study provides new insights into the lime response to phytoplasma infection during the progression of WBDL. Finally, we showed that healthy plants can be isolated from infected plants using biochemical profiles.

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  1. Bertamini, M., Grando, M. S. & Nedunchezhian, N. (2003). Effects of phytoplasma infection on pigments, chlorophyll-protein complex and photosynthetic activities in field grown apple leaves. Biologia Plantarum, 47, 237-242.
  2. Bertamini, M., Nedunchezhian, N., Tomasi, F. & Grando, S. (2002). Phytoplasma [Stolbur subgroup (Bois Noir-BN)] infection inhibits photosynthetic pigments, ribulose-1, 5-biphosphate carboxylase and photosynthetic activities in field grown grapevine (Vitis vinifera L. cv. Chardonnay) leaves. Physiological and Molecular Plant Pathology, 61, 357-366.
  3. Bertamini, M. & Nedunchezhian, N. (2001). Effects of phytoplasma [Stolbur-Subgroup (Bois Noir-BN)] on photosynthetic pigments, saccharides, ribulose 1,5-bisphosphate carboxylase, nitrate and nitrite reductases, and photosynthetic activities in field-grown grapevine (Vitis vinifera L. cv. Chardonnay) leaves. Photosynthetica, 39, 119-122.
  4. Boeing, J., Barizao, E. O., Silva, B. C., Montanher, P. F., Almeida, V. & Visentainer J. (2014). Evaluation of solvent effect on the extraction of phenolic compounds and antioxidant capacities from the berries: application of principal component analysis. Chemistry Central Journal, 8, 1-9. 
  5. Brand-Williams, W., Cuvelier, M. E. & Berset, C. (1995). Use of a free radical method to evaluate antioxidant activity. LWT – Food Science and Technology, 28, 25-30.
  6. Chung, K. R., Khan, I. A. & Brlansky, R. H. (2006). Citrus diseases exotic to Florida: witches' broom disease of lime (WBDL), University of Florida (Electronic Data Information Systems) Database, Fact Sheet.
  7. Deng, S. J. & Hiruki, C. (1991). Amplification of 16S ribosomal-RNA genes from culturable and nonculturable mollicutes. Journal of Microbiological Methods, 14, 53-61.
  8. Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A. & Smith, F. (1956). Colorimetric method for determination of sugars and related substances. Analytical Chemistry, 28(3), 350-356.
  9. Friend, J. (1979). Phenolic substances and plant disease. In: T. Swain, J.B. Harborne, and C.F. Van Sumere (Eds), Biochemistry of plant phenolics. (pp. 557-558). Plenum Press, New York.
  10. Foyer, C. H., Lelandais, M. & Kunert, K. J. (1994). Photooxidative stress in plants. Plant Physiology, 92, 696-717.
  11. Garnier, M., Zreik, L. & Bov, J. M. (1991). Witches' broom disease of lime trees in Oman: Transmission of a mycoplasma-like organism (MLO) to periwinkle and citrus and the production of monoclonal antibodies against the MLO. In: Proceedings of 11th conference of the International Organization of Citrus Virologists, 6-10 Nov., c/o Department of Plant Pathology, Riverside, California, pp. 448-453.
  12. Giorno, F., Guerriero, G., Biagetti, M., Ciccotti, A. M. & Baric, S. (2013). Gene expression and biochemical changes of carbohydrate metabolism in vitro micro-propagated apple plantlets infected by ‘Candidatus Phytoplasma mali’. Plant Physiology and Biochemistry, 70, 311-317.
  13. Gundersen, D. E. & Lee, I. M. (1996). Ultrasensitive detection of phytoplasmas by nested-PCR assays using two universal primer pairs. Phytopathologia Mediterranea, 35, 144-151.
  14. Junqueira, A. C., Bedendo, I. P. & Pascholati, S. F. (2011). Effect of phytoplasma infection on the activity of peroxidase, β-1,3 glucanase and chitinase in corn plants. Summa Phytopathologica, 37, 194-198.
  15. Kartte, S. & Seemuller, E. (1991). Histopathology of apple proliferation in Malus taxa and hybrids of different susceptibility. Journal of Phytopathology, 131, 149-160.
  16. Lepka, P., Stitt, M., Moll, E. & Seemüller, E. (1999). Effect of phytoplasmal infection on concentration and translocation of carbohydrates and amino acids in periwinkle and tobacco. Physiological and Molecular Plant Pathology, 55, 59-68.
  17. McCready, R. M., Guggolz, J., Silviera, V. & Owens, H. S. (1950). Determination of starch and amylose in vegetables. Analytical Chemistry, 22(9), 1156-1158
  18. Mollayi, S., Zadali, R., Farzaneh, M. & Ghassempour, A. (2015). Metabolite profiling of Mexican lime (Citrus aurantifolia) leaves during the progression of witches’ broom disease. Phytochemistry Letters, 13, 290-296.
  19. Murray, M. G. & Thomson, W. F. (1980). Rapid isolation of high molecular weight plant DNA. Nucleic Acids Research, 8, 4321-4325.
  20. Salehi, M., Izadpanah, K. & Rahimian, H. (1997). Witches’ broom disease of lime in Sistan, Baluchistan. Iranian Journal of Plant Pathology, 33, 76.
  21. Sestak, Z. (1966). Limitation for finding linear relationship between chlorophyll content and photosynthetic activity. Biologia Plantarum, 8, 336-346.
  22. Schneider, B., Seemuller, E., Smart, C. D. & Kirkpatrick, B. C. (1995). Phylogenetic classification of plant pathogenic mycoplasmalike organisms or phytoplasmas. In S. Razin (Ed.) Molecular and Diagnostic Procedures in Mycoplasmology. (pp. 369-380). Academic Press, San Diego.
  23. Singleton, V. L. & Rossi, J. A. (1965). Colorimetry of total phenolics with phosphomolybdic–phosphotungstic acid reagents. American Journal of Enology and Viticulture, 16, 144-158.
  24. Tan, Y., Wei, H. R., Wang, J. W., Zong, X. J., Zhu, D. Z. & Liu, Q. Z. (2015). Phytoplasmas change the source-sink relationship of field-grown sweet cherry by disturbing leaf function. Physiological and Molecular Plant Pathology, 92, 22-27.
  25. Zafari, S., Niknam, V., Musetti, R. & Noorbakhsh, S. N. (2012). Effect of phytoplasma infection on metabolite content and antioxidant enzyme activity in lime (Citrus aurantifolia). Acta Physiologiae Plantarum, 34, 561-568.
  26. Zimmermann, M. R., Schneider, B., Mithofer, A., Reichelt, M., Seemuller, E. & Furch, A. (2015). Implications of "Candidatus Phytoplasma mali" infection on phloem function of apple trees. Journal of Endocytobiosis and Cell Research, 26, 67-75.