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Journal of Bacteriology, November 2003, p. 6513-6521, Vol. 185, No. 22
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.22.6513-6521.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification and Characterization of Phytoplasmal Genes, Employing a Novel Method of Isolating Phytoplasmal Genomic DNA

Sharon Melamed,¹ Edna Tanne,² Raz Ben-Haim,¹ Orit Edelbaum,¹ David Yogev,³ and Ilan Sela^1*

Institute for Plant Sciences, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot 76100,¹ The Volcani Center, Agricultural Research Organization, Bet Dagan 50250,² Department of Membrane Research, The Institute of Microbiology, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel³

Received 14 July 2003/ Accepted 18 August 2003

Phytoplasmas are unculturable, insect-transmissible plant pathogens belonging to the class Mollicutes. To be transmitted, the phytoplasmas replicate in the insect body and are delivered to the insect's salivary glands, from where they are injected into the recipient plant. Because phytoplasmas cannot be cultured, any attempt to recover phytoplasmal DNA from infected plants or insects has resulted in preparations with a large background of host DNA. Thus, studies of the phytoplasmal genome have been greatly hampered, and aside from the rRNA genes, only a few genes have hitherto been isolated and characterized. We developed a unique method to obtain host-free phytoplasmal genomic DNA from the insect vector's saliva, and we demonstrated the feasibility of this method by isolating and characterizing 78 new putative phytoplasmal open reading frames and their deduced proteins. Based on the newly accumulated information on phytoplasmal genes, preliminary characteristics of the phytoplasmal genome are discussed.

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* Corresponding author. Mailing address: The Hebrew University, Faculty of Agricultural, Food and Environmental Quality Sciences, Virus Laboratory, Rehovot 76100, Israel. Phone: 972-8-9489377. Fax: 972-8-9473402. E-mail: sela{at}agri.huji.ac.il.

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Journal of Bacteriology, November 2003, p. 6513-6521, Vol. 185, No. 22
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.22.6513-6521.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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