Citrus rootstock breeding: response of four allotetraploid somatic hybrids to Citrus tristeza virus induced infections
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Four allotetraploid somatic hybrids of citrus, with potential for rootstock improvement, have been evaluated for their response to Citrus tristeza virus (CTV) infection. CTV is the most important viral pathogen affecting citrus production worldwide. Somatic combinations of ‘Milam’ lemon (Citrus jambhiri Lush.) + Sour orange (C. aurantium L Osb.), Calamondin (C. madurensis Lour.) + ‘Keen’ sour orange (C. aurantium L.), Calamondin + ‘Femminello‘ lemon (C. limon L. Burm. F.) and Cleopatra mandarin (C. reshni Hort. ex Tan.) + ‘Femminello’ lemon, were studied. Plants were grafted with CTV-infected “Valencia” sweet orange budwood. Two different CTV strains collected in Sicily, considered as “mild” and “severe”, were used to inoculate candidate rootstocks. The goal of this work is to select a rootstock alternative to CTV susceptible Sour orange, still the prevalent rootstock in the Mediterranean basin. DAS-ELISA and real-time PCR assays confirmed a decreased level of viral replication in tested somatic hybrids, as compared to the susceptible genotypes sour orange and Citrus alemow. The Calamondin+‘Keen’ sour orange genotype did not support any replication in either CTV strain. Somatic hybridization is confirmed to be an effective tool to obtain functionally new rootstocks. Our results can be considered a starting point to open new approaches for the Mediterranean citrus industry. Indeed, the four somatic hybrids have been propagated for agronomical multisite evaluation trials to further assess if their horticultural performance, fruit holding capacity, and soil adaptation are adequate to replace sour orange.
KeywordsCTV Protoplast fusion Rootstocks DAS-ELISA RT-PCR Real-time qRT-PCR
The authors wish to thank Mr. Antonio Motisi (IBBR-CNR, Palermo) for skillful technical assistance.
All the authors conceived and designed the experiments. LA and SP carried out the experiments. LA developed the plant materials. SP performed the molecular analysis. LA, FM, SD and SFDB commented the results. FM and SFDB drafted the manuscript. All the authors critically read and approved the final manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
The experiments were performed in compliance with the European current laws.
- Allario, T., Brumos, J., Colmenero, J. M., Domingo, I., José, J., Antonio, P. J., et al. (2009). Autotetraploid Citrus limonia rootstocks are more tolerant to water deficit than parental diploids. Intern Confon Polyploidy, Hybrid and Biodivers, May 17–20, 2009, Saint-Malo France, program and abstracts. Rennes: Universite de Rennes, 1, 98.Google Scholar
- Cambra, M., Garnsey, S. M., Permar, T. A., Henderson, C. T., Gumpf, D. J., & Vela, C. (1990). Detection of Citrus tristeza virus (CTV) with a mixture of monoclonal antibodies. Phytopathol, 80, 1034.Google Scholar
- Cambra, M., Gorris, M. T., Olmos, A., Martínez, M. C., Romàn, M. P., Bertolini, E., et al. (2002). European diagnostic protocols (DIAGPRO) for Citrus tristeza virus in adult trees. In N. Duran-Vila, R. G. Milne, & J. V. da Graça (Eds.), Proc of the 15th Conf of the Intern Organ of Citrus Virol IOCV (pp. 69–77). CA: Riverside.Google Scholar
- Castle, W. S. (1987). Citrus rootstocks. In R. C. Rom & R. F. Carlson (Eds.), Rootstocks for fruit crops (pp. 361–399). NY: Wiley.Google Scholar
- Coletta-Filho, H. D., Carlos, E. F., Alves, K. C. S., Pereira, M. A. R., Boscariol-Camargo, R. L., de Souza, A. A., & Machado, M. A. (2010). In planta multiplication and graft transmission of “Candidatus Liberibacter asiaticus” revealed by real-time PCR. European Journal of Plant Pathology, 126(1), 53–60.CrossRefGoogle Scholar
- Dambier, D., Benyahia, H., Pensabene-Bellavia, G., Kacar, Y. A., Froelicher, Y., Belfalah, Z., et al. (2011). Somatic hybridization for citrus rootstock breeding: An effective tool to solve some important issues of the Mediterranean citrus industry. Plant Cell Reports, 30, 883–900.CrossRefGoogle Scholar
- EPPO (2009). EPPO Reporting Service n. 1, 2, 3, 6, 8, 9, 10, 11. http://www.eppo.org/PUBLICATIONS/reporting/reporting_service.htm [accessed on 10 February 2012].Google Scholar
- Etebu, E., & Nwauzoma, A. B. (2014). A review on sweet orange (Citrus sinensis L. Osb.): Health, diseases and management. Am J of Res Comm, 2(2), 33–70.Google Scholar
- Fatta Del Bosco, S., Napoli, E., Mercati, F., Abbate, L., Carimi, F., & Ruberto, G. (2017). Somatic cybridization for Citrus: Polyphenols distribution in juices and peel essential oil composition of a diploid cybrid from Cleopatra mandarin (Citrus reshni Hort. Ex tan.) and sour orange (Citrus aurantium L.). Genet Resour Crop Ev, 64(2), 261–275.CrossRefGoogle Scholar
- Garnsey, M., Gumpf, S. M., Roistacher, D. J., Civerolo, C. N., Lee, E. L., Yokomi, R. F., et al. (1987). Toward a standardized evaluation of the biological properties of citrus tristeza virus. Phytophylactica, 19, 151–158.Google Scholar
- Grosser, J. W., Chandler, J. L. (2003) New Citrus rootstock via protoplast fusion. Acta Hort ic 622, 491-497.Google Scholar
- Grosser, J. W., & Gmitter Jr., F. G. (1990). Protoplast fusion and citrus improvement. Plant Breed Rev 8. Timber Press Inc, 8, 339–374.Google Scholar
- Grosser, J. W., Gmitter Jr., F. G., & Castle, W. S. (1995). Production and evaluation of citrus somatic hybrid rootstocks. Prog Rep Proc Fla State Hort Soc, 108, 140–143.Google Scholar
- Grosser, J. W., Chandler, J. L. (2003) New Citrus rootstock via protoplast fusion. Acta Hortic, 622, 491–497.Google Scholar
- Hodgson, R. W. (1967). Horticultural varieties of citrus. In W. Reuther, H. J. Webber, & L. D. Batchelor (Eds.), The citrus ind 1 (pp. 431–591). Riverside: University of California.Google Scholar
- Husband, B. C., Baldwin, S. J., Suda, J. (2013). The incidence of polyploidy in natural plant populations: Major patterns and evolutionary processes. In: Greilhuber J., Dolezel J., Wendel J.F. (Eds.), Plant Genome Divers, Volume 2. Springer, Vienna. Google Scholar
- Hussain, S., Curk, F., Dhuique-Mayer, C., Urban, L., Ollitrault, P., Luro, F., & Morillon, R. (2012). Autotetraploid trifoliate orange (Poncirus trifoliata) rootstocks do not impact clementine quality but reduce fruit yields and highly modify rootstock/scion physiology. Scientia Horticulturae, 134, 100–107.CrossRefGoogle Scholar
- Jannati, M., Fotouhi, R., Abad, A. P., & Salehi, Z. (2009). Genetic diversity analysis of Iranian citrus varieties using micro satellite (SSR) based markers. J Hortic For, 1(7), 120–125.Google Scholar
- Johnson, A. A. T., & Veilleux, R. E. (2001). Somatic hybridization and application in plant breeding. Plant Breed Rev, 20, 167–225.Google Scholar
- Louzada, E. S., Grosser, J. W., Gmitter Jr., F. G., Nielsen, B., Chandler, J. L., Deng, X. X., et al. (1992). Eight new somatic hybrid Citrus with potential for improved disease resistance. Horticultural Science, 27(9), 1033–1036.Google Scholar
- Mourao Filho, F. A. A., Pio, R., Jannuzzi Mendes, B. M., de Azavedo, F. A., Schinor, E. H., Albuquerque Entelmann, F., et al. (2008). Evaluation of citrus somatic hybrids for tolerance to Phytophthora nicotianae and to citrus tristeza virus. Scientia Horticulturae, 115, 301–308.CrossRefGoogle Scholar
- Napoli, E., Ruberto, G., Abbate, L., Mercati, F., Del Bosco, S. F. (2016). Citrus genetic improvement: New citrus hybrids from breeding procedures and evaluation of their genetic and phytochemical aspects. In: Simmons D. (Ed), Citrus fruits: production, consumption and health benefits. Nova Science Publishers, Incorporated. Google Scholar
- Ollitrault, P., & Navarro, L. (2012). Citrus. In M. Badenes & D. Byrne (Eds.), Fruit breeding. Handbook of plant breeding (Vol. 8, pp. 623–662). Boston: Springer.Google Scholar
- Ollitrault, P., Froelicher, Y., Dambier, D., Seker, M. (2000). Rootstock Breeding by somatic hybridization for the Mediterranean Citrus industry. ISHS Acta Hort 535: First International Citrus Biotechnology Symposium.Google Scholar
- Osman, F., Hodzic, E., Kwon, S. J., Wang, J., & Vidalakis, G. (2015). Development and validation of a multiplex reverse transcription quantitative PCR (RT-qPCR) assay for the rapid detection of Citrus tristeza virus, Citrus psorosis virus, and Citrus leaf blotch virus. Journal of Virological Methods, 220, 64–75.CrossRefGoogle Scholar
- Pereira de Carvalho Costa, M. A., Mendes, B. M. J., & Mourao Filho, F. A. A. (2003). Somatic hybridization for improvement of citrus rootstock: Production of five new combinations with potential for improved disease resistance. Australian Journal of Experimental Agriculture, 43, 1151–1156.CrossRefGoogle Scholar
- Roistacher, C. N. (1993). Psorosis a review. In: Proocedings of the 12 th Conference of the International Organization of Citrus Virologists, 1192(1), 139–154.Google Scholar
- Shafieizargar, A., Awang, Y., Juraimi, A. S., & Othman, R. (2013). Comparative studies between diploid and tetraploid Dez orange (Citrus sinensis (L.) Osb:) under salinity stress. Australian Journal of Crop Science, 7, 1436–1441.Google Scholar
- Zanutto, C. A., Corazza, M. J., Nunes, W. M. C., & Muller, G. W. (2013). Evaluation of the protective capacity of new mild Citrus tristeza virus (CTV) isolates selected for a preimmunization program. Science in Agriculture, 70, 116–124.Google Scholar