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Phytoplasma Cultivation

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Phytoplasmas: Plant Pathogenic Bacteria - III

Abstract

The possibility to grow phytoplasmas in complex media as the mycoplasmas was ruled out for more than 40 years due to the inconsistency of the first isolation trials. The use of micropropagated infected periwinkle shoots first and tissues infected directly from the fields on artificial media was recently confirmed. Isolation and cultivation together with the first biochemical and biological characterization of some phytoplasmas were consistently obtained. Phytoplasmas were successfully grown from grapevine, cassava and coconut palm tissues severely infected and in some cases also from asymptomatic tissues from trees or plants growing in severely infected areas. Phytoplasmas from diverse ribosomal groups were isolated in particular from cassava with frog skin the detected prokaryotes in culture were molecularly identified as belonging to the 16SrIII group as in the original plants. Aster yellows and ‘stolbur’ phytoplasmas, group 16SrI and 16SrXII, respectively, were consistently grown from diverse host plants such as grapevine and coconut palms. Seed transmission in corn of aster yellows phytoplasmas was also confirmed by isolation from seedlings of viable cells. The growth and biochemical and biological characterization of these bacteria is therefore the most important recent step in the study of phytoplasmas that will allow to improve their knowledge and to carry out focused management in the field in order to reduce their impact on cultivated and wild crops worldwide.

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References

  • Alma A, Bosco D, Danielli A, Bertaccini A, Vibio M, Arzone A (1997) Identification of phytoplasmas in eggs, nymphs of Scaphoideus titanus Ball reared on healthy plants. Insect Molecular Biology 2, 115–121.

    Article  CAS  PubMed  Google Scholar 

  • Alma A, Palermo S, Boccardo G, Conti M (2001) Transmission of chrysanthemum yellows, a subgroup 16SrI-B phytoplasma, to grapevine by four leafhopper species. Journal of Plant Pathology 83, 181–187.

    Google Scholar 

  • Alvarez E, Mejía JF, Llano GA, Loke JB, Calari A, Duduk B, Bertaccini A (2009) Detection and molecular characterization of a phytoplasma associated with frogskin disease in Cassava. Plant Disease 93, 1139–1145.

    Article  CAS  PubMed  Google Scholar 

  • Alvarez E, Mejía JF, Pardo JM (2010) Development of a real-time PCR assay, to detect and quantify a 16SrIII-L phytoplasma associated with cassava frogskin disease (CFSD). Phytopathology 100, S5.

    Google Scholar 

  • Alvarez E, Betancourth C, Muñoz J (2017) Pathogenicity of a 16SrIII-L phytoplasma associated with frogskin disease of cassava (Manihot esculenta Crantz) in Colombia. Phytopathology 107, S2.5.

    Article  Google Scholar 

  • Angelini E, Constable F, Duduk B, Fiore N, Quaglino F, Bertaccini A (2018) Grapevine phytoplasmas. In: Phytoplasmas: Plant Pathogenic Bacteria-I. Characterization and Epidemiology of Phytoplasma-Associated Diseases. Eds Rao GP, Bertaccini A, Fiore N, Liefting L. Springer, Singapore, 123–151 pp.

    Chapter  Google Scholar 

  • Arocha Rosete Y, Atta Diallo H, Konan Konan JL, Yankey N, Saleh M, Pilet F, Contaldo N, Paltrinieri S, Bertaccini A, Scott JA (2017) Improving sampling and detection for the coconut lethal yellowing phytoplasma in coconut-growing villages of Grand-Lahou, Côte d’Ivoire. Annals of Applied Biology 170, 333–347.

    Article  CAS  Google Scholar 

  • Bertaccini A, Davis RE, Lee I-M (1992) In vitro micropropagation for maintenance of mycoplasmalike organisms in infected plant tissues. HortScience 27, 1041–1043.

    Article  Google Scholar 

  • Bertaccini A, Contaldo N, Calari A, Paltrinieri S, Windsor HM, Windsor DG (2010) Preliminary results of axenic growth of phytoplasmas from micropropagated infected periwinkle shoots. 18th Congress of the International Organization for Mycoplasmology, Chianciano Terme, Italy, July 11–16 147, 153.

    Google Scholar 

  • Brown DR, Whitcomb RF, Bradbury JM (2007) Revised minimal standards for description of new species of the class Mollicutes (division Tenericutes). International Journal of Systematic and Evolutionary Microbiology 57, 2703–2719.

    Article  CAS  PubMed  Google Scholar 

  • Contaldo N, Bertaccini A, Paltrinieri S, Windsor HM, Windsor DG (2012) Axenic culture of plant pathogenic phytoplasmas. Phytopathologia Mediterranea 51, 607–617.

    CAS  Google Scholar 

  • Contaldo N, Bertaccini A, Paltrinieri S, Windsor DG, Windsor HM (2013) Cultivation of several phytoplasmas from a micropropagated plant collection. Petria 23, 13–18.

    Google Scholar 

  • Contaldo N, Windsor DG, Windsor HM, Paltrinieri S, Satta E, Bertaccini A (2014a) Phytoplasma cultivation from a micropropagated plant collection. IOM 2014, June 1-6, Blumenau, Brazil 33, 40.

    Google Scholar 

  • Contaldo N, Satta E, Bertaccini A, Windsor DG (2014b) Methods for isolation by culture, and subsequent molecular identification, of phytoplasmas from plants sourced in the field. IOM 2014, June 1-6, Blumenau, Brazil 106, 56.

    Google Scholar 

  • Contaldo N, Satta E, Zambon Y, Canel A, Paltrinieri S, Bertaccini A (2014c) Morphological and molecular identification of phytoplasmas in culture from plants sourced in the field. Journal of Plant Pathology 96, S.4.39.

    Google Scholar 

  • Contaldo N, Satta E, Paltrinieri S, Gherardi M, Laurita R, Stancampiano A, Zambon Y, Colombo V, Bertaccini A (2015a) Interaction between cold atmospheric plasma and phytoplasmas in micropropagated infected periwinkle shoots. Phytopathogenic Mollicutes 5(1-Supplement), S127-S128.

    Article  Google Scholar 

  • Contaldo N, Zambon Y, Satta E, Paltrinieri S, Bertaccini A (2015b) Cultivation in chemically defined media of phytoplasmas from field-infected grapevine plants showing yellows symptoms. 18th Congress of the International Council for the Study of Virus and Virus-like Diseases of the Grapevine. September 7-11, Ankara, Turkey, 121–123.

    Google Scholar 

  • Contaldo N, Satta E, Zambon Y, Paltrinieri S, Bertaccini A (2016a) Development and evaluation of different complex media for phytoplasma isolation and growth. Journal of Microbiological Methods 127, 105–110.

    Article  CAS  PubMed  Google Scholar 

  • Contaldo N, Zambon Y, Paltrinieri S, Mori N, Mitrovic J, Duduk B, Bertaccini A (2016b) Characterization of ‘Candidatus Phytoplasma solani’ strains from grapevines, Hyalesthes obsoletus, reference strains in periwinkle and in colonies. Mitteilungen Klosterneuburg 66, 63–69.

    Google Scholar 

  • Contaldo N., D. Dolanc, A. Bertaccini, M. Dermastia. 2018a. Isolation of diverse phytoplasmas from symptomatic grapevine samples. “Bois noir” Fifth Workshop, Liubljana September 18 and 19, a16.

    Google Scholar 

  • Contaldo N., G. D’Amico, Y. Zambon, A. Bertaccini. 2018b. ‘Candidatus Phytoplasma asteris’ isolated from grapevine: preliminary metabolic features. 22th Congress of the International Organization for Mycoplasmology (IOM) July 9–12, Portsmouth, NH, USA 54, 40.

    Google Scholar 

  • Contaldo N, D’Amico G, Paltrinieri S, Diallo HA, Bertaccini A, Arocha Rosete Y (2019) Molecular and biological characterization of phytoplasmas from coconut palms affected by the lethal yellowing disease in Africa. Microbiological Research 223–225, 51–57.

    Article  PubMed  CAS  Google Scholar 

  • Danielli A, Bertaccini A, Bosco D, Alma A, Vibio M, Arzone A (1996) May evidence of 16SrI-group-related phytoplasmas in eggs, nymphs and adults of Scaphoideus titanus Ball suggest their transovarial transmission? IOM Letters 4, 190–191.

    Google Scholar 

  • Davis RE, Worley JF, Whitcomb RF, Ishijima T, Steere RL (1972) Helical filaments produced by a mycoplasma-like organism associated with corn stunt disease. Science 176, 521–523.

    Article  PubMed  Google Scholar 

  • Doi YM, Teranaka M, Yora K, Asuyama H (1967) Mycoplasma or PLT-group-like microorganisms found in the phloem elements of plants infected with mulberry dwarf, potato witches’ broom, aster yellows, or paulownia witches’ broom. Annals of the Phytopathological Society of Japan 33, 259–266.

    Article  Google Scholar 

  • Elmendorf E (1977) Newly discovered prokaryotic plant pathogens. A bibliography. R 2901 Research Bulletin College Agriculture University of Wisconsin, Madison, United States of America, 1–85 pp.

    Google Scholar 

  • Freundt EA, Erno H, Lemcke RM (1979) Identification of mycoplasma. In: Methods in Microbiology. Ed Bergan N, Academic Press, London, United Kingdom, 396 pp.

    Google Scholar 

  • Gasparich GE, Whitcomb RF, Dodge D, French FE, Glass J, Williamson DL (2004) The genus Spiroplasma and its non-helical descendants: phylogenetic classification, correlation with phenotype and roots of the Mycoplasma mycoides clade. International Journal of Systematic and Evolutionary Microbiology 54, 893–918.

    Article  CAS  PubMed  Google Scholar 

  • Ghosh SK, Raychaudhuri SP, Chenulu VV, Varma A (1975) Isolation, cultivation and characterization of mycoplasma-like organisms from plants. Proceedings of the Indian National Science Academy 41B, 362–366.

    Google Scholar 

  • Giannotti J, Vago C (1971) Role des mycoplasmes dans l’étiologie de la phyllodie du trèfle: culture et transmission expérimentale de la maladie. Physiologie Vegetale 9, 541–553.

    Google Scholar 

  • Jones AL, Whitcomb RF, Williamson DL, Coan ME (1977) Comparative growth and isolation of spiroplasmas in media beside on insect tissue culture formulation. Phytopathology 67, 738–746.

    Article  Google Scholar 

  • Ishiie T, Doi Y, Yora K, Asuyama H (1967) Suppressive effects of antibiotics of tetracycline group on symptom development of mulberry dwarf disease. Annals of the Phytopathological Society of Japan 33, 267–275.

    Article  CAS  Google Scholar 

  • Kunkel LO (1926) Studies on aster yellows. American Journal of Botany 23, 646–705.

    Article  Google Scholar 

  • Kunkel LO (1931) Celery yellows of California not identical with the aster yellows of New York. Boyce Thompson Institute 4, 405–414.

    Google Scholar 

  • Kunkel LO (1955) Cross protection between strains of aster yellow-type viruses. Advances in Virus Research 3, 251–273.

    Article  CAS  PubMed  Google Scholar 

  • Lee I-M, Davis RE (1992) Mycoplasmas which infect plants and insects. In: Mycoplasmas: Molecular Biology and Pathogenesis. Eds Maniloff J, McElhansey RN, Finch LR, Baseman JB. American Society for Microbiology. Washington DC, United States of America, 379–390 pp.

    Google Scholar 

  • Lim PO, Sears BB (1989) 16S rRNA sequence indicates that plant-pathogenic mycoplasmalike organisms are evolutionarily distinct from animal mycoplasmas. Journal of Bacteriology 171, 5901–5906.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lin SC, Lee CS, Chin RJ (1970) Isolation and cultivation of, and inoculation with a mycoplasma causing white leaf disease of sugarcane. Phytopathology 60, 795–797.

    Article  Google Scholar 

  • Lombardo G, Pignattelli P (1970) Cultivation in a cell-free medium of a mycoplasma-like organism from Vinca rosea with phyllody symptoms on the flowers. Annals of Microbiology 20, 83–88.

    Google Scholar 

  • Maramorosch K (2011) Historical reminiscences of phytoplasma discovery. Bulletin of Insectology 64(Supplement), S5-S8.

    Google Scholar 

  • Mc Coy RE, Caudwell A, Chang CJ, Chen TA, Chiykowskyi LN, Cousin MT, Dale de Leeuw GTN, Golino DA, Hackett KJ, Kirkptrick BC, Marwitz R, Petzold H, Shina RH, Sugiura M, Whitcomb RF, Yang IL, Zhu BM, Seemüller E (1989) Plant diseases associated with mycoplasmalike organisms. In: The Mycoplasmas, volume 5. Eds Whitcomb RF, Tully JG. Academic Press, New York, United States of America, 545–640 pp.

    Google Scholar 

  • Mori N, Martini M, Bressan A, Guadagnini M, Girolami V, Bertaccini A (2002) Experimental transmission by Scaphoideus titanus Ball of two molecularly distinct “flavescence dorée” type phytoplasmas. Vitis 41, 99–102.

    Google Scholar 

  • Müller HM, Kleinhempel H, Rabitschuch AW, Spaar D, Müller JH (1975) Mykoplasmen in der Phytopathologie, Bibliographisches Verzeichnis der Literatur fur die Jahre 1967–1974. Institut fur Landwritschaftliche Information und Dokumentation, Berlin.

    Google Scholar 

  • Nasu S, Sugiura M, Wakimoto T, Iida TT (1967) On the pathogen of rice yellow dwarf virus. Annals of Phytopathological Society Japan 33, 343–344.

    Google Scholar 

  • Nocard E, Roux ER (1898) Le microbe de la péripneumonie. Annales de l’Institute Pasteur 12, 240–262.

    Google Scholar 

  • Poghosyan A, Lebsky V (2004) Aislamiento y estudio ultrastructural de tres cepas de fitoplasmas causantes de enfermedades tipo “stolbur” en solanaceae. Fitopatologia Colombiana 8, 21–30.

    Google Scholar 

  • Rao GP, Bertaccini A, Fiore N, Liefting L (2018) Phytoplasmas: Plant Pathogenic Bacteria-I. Characterization and Epidemiology of Phytoplasma-Associated Diseases. Springer, Singapore, 345 pp.

    Google Scholar 

  • Rid M, Mesca C, Ayasse M, Gross J (2016) Apple proliferation phytoplasma influences the pattern of plant volatiles emitted depending on pathogen virulence. Frontieres in Ecological Evolution 3, 152.

    Google Scholar 

  • Saccardo F, Martini M, Palmano S, Ermacora P, Scortichini M, Loi N, Firrao G (2012) Genome drafts of four phytoplasma strains of the ribosomal group 16SrIII. Microbiology 158, 2805–2814.

    Article  CAS  PubMed  Google Scholar 

  • Saglio P, L’Hospital M, Lafleche D, Dupont G, Bové J-M, Tully JG, Freundt EA (1973) Spiroplasma citri gen. and sp. n.: a mycoplasma-like organism associated with “stubborn” disease of citrus. International Journal of Systematic Bacteriology 23, 191–204.

    Article  CAS  Google Scholar 

  • Satta E, Contaldo N, Paltrinieri S, Bertaccini A (2016) Biological and molecular proof of phytoplasma seed transmission in corn. 21th Congress of the IOM, Brisbane, Australia, July 3-7 61, 65–66.

    Google Scholar 

  • Satta E, Paltrinieri S, Bertaccini A (2019) Phytoplasma transmission by seed. In: Phytoplasmas: Plant Pathogenic Bacteria-II. Transmission and Management of Phytoplasma Associated Diseases. Chapter 6. Eds. Bertaccini A, Weintraub PG, Rao GP, Springer, Singapore 131–147 pp.

    Google Scholar 

  • Seemüller E, Schneider B (2007) Differences in virulence and genomic features of strains of ‘Candidatus Phytoplasma mali’, the apple proliferation agent. Phytopathology 97, 964–970.

    Article  CAS  PubMed  Google Scholar 

  • Skripal IG, Malinivskaya LP (1984) Medium SMIMB-72 for isolation and cultivation of phytopathogenic mycoplasmas. Microbiologiceskii Zhurnal 46, 71–75.

    Google Scholar 

  • Skripal IG, Malinovskaya LP, Onishchenko AN (1984) A method for isolation of mycoplasmas from plants affected by yellows diseases. Microbiologiceskii Zhurnal 46, 93–96.

    Google Scholar 

  • Whitcomb RF, Tully GJ (1975) The Mycoplasmas III. The Academic Press, New York, United States of America, 351 pp.

    Google Scholar 

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Authors and Affiliations

  1. Department of Agricultural and Food Sciences, Alma Mater Studiorum – University of Bologna, Bologna, Italy

    Nicoletta Contaldo & Assunta Bertaccini

Authors
  1. Nicoletta Contaldo
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  2. Assunta Bertaccini
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Editors and Affiliations

  1. Department of Agricultural and Food Sciences, Alma Mater Studiorum – University of Bologna, Bologna, Italy

    Assunta Bertaccini

  2. Department of Clinical Plant Science, Faculty of Bioscience and Applied Chemistry, Hosei University, Tokyo, Japan

    Kenro Oshima

  3. Department of Integrative Infection Biology Crops-Livestock, University of Hohenheim, Stuttgart, Germany

    Michael Kube

  4. Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, Delhi, India

    Govind Pratap Rao

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Contaldo, N., Bertaccini, A. (2019). Phytoplasma Cultivation. In: Bertaccini, A., Oshima, K., Kube, M., Rao, G. (eds) Phytoplasmas: Plant Pathogenic Bacteria - III. Springer, Singapore. https://doi.org/10.1007/978-981-13-9632-8_6

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