Detection of Citrus psorosis ophiovirus in Citrus CV. Navel Orange in Egypt

Authors

  • R. E. SALEM Agricultural Genetic Engineering Research Institute (AGERI), ARC, Giza, Egypt
  • FATMA BADAWY Dept. Genetics, Faculty of Agric., Ain Shams University, Shoubra El-Kheima, Egypt
  • M. I. SALAMA Agricultural Genetic Engineering Research Institute (AGERI), ARC, Giza, Egypt
  • HANAN NOUR EL- DIN Agricultural Genetic Engineering Research Institute (AGERI), ARC, Giza, Egypt
  • KH. A. EL-DOUGDOUG Dept. Agric. Microbiol., Faculty of Agric., Ain Shams University, Shoubra El-Kheima, Egypt
  • EMAN M. FAHMY Dept. Genetics, Faculty of Agric., Ain Shams University, Shoubra El-Kheima, Egypt

Abstract

Citrus is one of the most important fruit crops worldwide. Most of the citrus cultivars are grown as grafted plants. Virus pathogens transmitted by grafting as well as insect vectors could cause economic problems. Among graft transmissible diseases that have been reported from Egypt as well as in the rest of the Mediterranean countries; citrus psorosis disease that is of the most serious and remain the most spread disease in Egypt (Roistacher, 1991; Sofy, 2008).
In infected trees, scaly bark symptoms appears on the trunk, staining of interior wood of branch and gummy as well as shortened leaf internodes and mottling patterns on leaves (Ghazal et al., 2008; El-Dougdoug et al., 2009).
Citrus psorosis virus (CPsV), the type species of the genus Ophiovirus (Vaira et al., 2005), is the putative causal agent of psorosis, a major graft-transmissible disease of citrus (Roistacher, 1993).
Virus particles are formed by three single-stranded, negative sense RNAs and a coat protein (CP) of 48 kDa (Derrick et al., 1988; Martı´n et al., 2005). RNA1 contains 8184 bp and its complementary strand has two ORFs potentially encoding a 24 kDa protein of unknown function and a 280 kDa protein with motifs characteristic of RNA-dependent RNA polymerases (RdRp). RNA2 contains 1644 bp and its complementary strand encodes a 54 kDa protein without similarities to other known proteins and RNA3 contains 1454 bp and its complementary strand encodes the CP (Barthe et al., 1998; Sa´nchez de la Torre et al., 1998 & 2002; Naum-Onganı´a et al., 2003; Martı´n et al., 2005).
The sensitive, reliable and rapid identification of plant viruses is essential for effective disease control. For many years, laborious and costly indexing on citrus indicators was the only diagnostic method available (Roistacher, 1993).
The first breakthrough in the development of a laboratory diagnostic testing technique came in 1977 with the development of the ELISA (Clark and Adams, 1977).
DAS-ELISA (Garcia et al., 1997) and TAS-ELISA (Alioto et al., 1999) are methods developed and applied for psorosis diagnosis in field trees.
Polymerase chain reaction (PCR) was developed in the early 1990’s and has the ability to selectively amplify a part of the target deoxyribonucleic acid (DNA). Reverse transcription-polymerase chain reaction (RT-PCR) (Garcia et al., 1997; Legarreta et al., 2000; D’Onghia et al., 2001), are now being utilized and several primers have been designed for the CPsV detection by the RT-PCR providing an alternative method for diagnosis (Barthe et al., 1998; Legarreta et al., 2000).
This study aimed to detect CPsV in citrus trees cv. Navel orange via biological, serological and molecular methods.

References

Alioto, D., M. Gangemi, S. Deaglio, P. Sposato, E. Noris, E. Luisoni and R. G. Milne (1999). Improved detection of citrus psorosis virus using polyclonal and monoclonal antibodies. Plant Pathol., 48: 735-741.

Barthe, G. A., T. L. Ceccardi, K. L. Manjunath and K. S. Derrick (1998). Citrus psorosis virus: nucleotide sequencing of the coat protein gene and detection by hybridization and RT-PCR. J. Gen. Virol. 79: 1531-1537.

Clark, M. F. and A. N. Adams (1977). Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. Journal of General Virology, 34: 475-483.

Derrick, K. S., K. S. Brlansky, R. F. Lee, L. W. Timmer and T. K. Nguyen (1988). Two components associated with citrus ringspot virus. In Proc. 10th Conf. International Organization of Citrus Virologists, p. 340-342. Edited by L. W. Timmer, S. M. Garnsey and L. Navarro. Riverside: Univ. of California IOCV.

Djelouah, K., O. Potere, D. Boscia, A.M. D’Onghia and V. Savino (2000). Production of monoclonal antibodies to Citrus psorosis associated virus. Proc. 14th Conf. International Organization of Citrus Virologists, Campinas, 1998: 152-158.

D'Onghia, A. M., K. Djelouah, D. Alioto, M. A. Castellano and V. Savino (1998). ELISA correlates with biological indexing for the detection of citrus psorosis-associated virus. J. Plant Pathol., 80: 157-163.

D’Onghia, A. M., K. Djelouah, D. Frasheri, and O. Potere (2001). Citrus psorosis virus detection by direct tissue blot immunoassay. J. Plant Pathol., 83: 139-142.

El-Dougdoug, Kh. A., S. A. Ghazal, A. A. Mousa, H. Fahmy and A. R. Sofy (2009). Differentiation among three Egyptian isolates of Citrus psorosis virus. International Journal of Virology, 5: 49-63.

Garcia, M. L., M. E. De La Torre, E. Dal Bo, K. Djelouah, N. Rouag, E. Luisoni, R. G. Milne and O. Grau (1997). Detection of Citrus psorosis ringspot virus using RT-PCR and DAS ELISA. Plant Pathology, 46: 830-836.

Garnsey, S. M. and M. Cambra (1991). Enzyme-linked immunosorbent assay (ELISA) for citrus pathogens. Pages 193-216 in: Grafttransmissible diseases of citrus-A handbook for detection and diagnosis. C.N Roistacher, ed. Food and Agriculture Organization of the United Nations, Rome.

Ghazal, S. A., Kh. A. El-Dougdoug, A. A. Mousa, H. Fahmy and A. R. Sofy (2008). Isolation and identification of Citrus psorosis virus Egyptian isolate (CPsV-EG). Commun. Agric. Appl. Biol. Sci., 73: 285-295.

Legarreta, G. G., M. L. Garcia, N. Costa and O. Grau (2000). A highly sensitive heminested RT PCR assay for the detection of Citrus psorosis virus targeted to a conserved region of the genome. Journal of Virological Methods, 84: 15-22.

Martı´n, S., C. Lo´ pez, M. L. Garcı´a, G. Naum-Onganı´a, O. Grau, R. Flores, P. Moreno and J. Guerri (2005). The complete nucleotide sequence of a Spanish isolate of Citrus psorosis virus: comparative analysis with other ophioviruses. Arch. Virol., 150: 167-176.

Naum-Onganı´a, G., S. Gago-Zachert, E. Pen˜ a, O. Grau and M. L. Garcı´a (2003). Citrus psorosis virus RNA 1 is of negative polarity and potentially encodes in its complementary strand a 24 K protein of unknown function and 280 K putative RNA dependent RNA polymerase. Virus Res., 96: 49-61.

Potere, O., D. Boscia, K. Djelouah, V. Elicio and V. Savino (1999). Use of monoclonal antibodies to citrus psorosis virus for diagnosis. J. Plant Pathol., 81: 209-212.

Roistacher, C. N. (1991). Graft transmissible Diseases of Citrus. Handbook for Detection and Diagnosis. FAO, Rome, Italy.

Roistacher, C. N. (1993). Psorosis: A review. In: Moreno P., da Graca J.V. and Timmer L.W. (eds). Proc. 12th Conf. International Organization of Citrus Virologists IOCV, Riverside 1993: 134-139

Sambrook, J., E. F. Fritsch and T. Maniatis (1989). Molecular Cloning: A Laboratory Manual Cold Spring Harbor Laboratory, Cold Spring Harbor New York.

Sa´nchez, de la Torre, E., O. Riva, R. Zandomeni, O. Grau and M. L. Garcı´a (1998). The top component of citrus psorosis virus contains two ssRNAs, the smaller encodes the coat protein. Mol Plant Pathol On-Line. http://www.bspp.org.uk/ mppol/1998/1019sanchez/index.htm

Sa´nchez, de la Torre, M. E., C. Lo´ pez, O. Grau and M. L. Garcı´a (2002). RNA 2 of Citrus psorosis virus is of negative polarity and has a single open reading frame in its complementary strand. J. Gen. Virol., 83: 1777-1781.

Sofy, A. R. (2008). Biological and molecular studies on Citrus psorosis virus in Egypt. M. Sc., Fac. of Science, Al-Azhar Univ., PP. 195, Egypt.

Vaira, A. M., G. P. Accotto, S. Gago-Zachert, M. L. Garcia, O. Grau, R. G. Milne, T. Morikawa, T. Natsuaki, V. Torok, M. Verbeek and H. J. Vetten (2005). Genus Ophiovirus. In: Fauquet C. M., Mayo M. A., Maniloff J., Desselberger U., Ball L. A. (eds.). Virus Taxonomy. Eight Report of the International Committee on Taxonomy of Viruses, p 673-679. Elsevier Academic Press, San Diego, USA.

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2016-01-11

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