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Anther culture in Citrus clementina: a way to regenerate tri-haploids

M. A. Germanà A C , B. Chiancone A , O. Lain B and R. Testolin B
+ Author Affiliations
- Author Affiliations

A Dipartimento SENFIMIZO, Facoltà di Agraria, Viale delle Scienze 11, 90128 Palermo, Italy.

B Dipartimento di Scienze Agrarie e Ambientali, Viale delle Scienze 208, 33100 Udine, Italy.

C Corresponding author. Email:

Australian Journal of Agricultural Research 56(8) 839-845
Submitted: 27 January 2005  Accepted: 20 May 2005   Published: 25 August 2005


Abstract. Regenerants from anther culture of Citrus clementina Hort. ex Tan. cvv. Nules, SRA 63, and Monreal were obtained in different experiments from 1994 to 2002. Genetic analysis of 37 such regenerants was carried out using 4 microsatellite markers that were heterozygous in the parental genotypes. The results showed that in all cases but one the regenerants carried only one or the other allele of the parental genotype, and were therefore homozygous and produced through a process of gametophytic embryogenesis. Ploidy analysis by flow cytometry of 94 regenerants showed that as many as 82% of them were tri-haploids, rather than haploids or doubled-haploids as expected, with other ploidy levels such as n, 2n, 4n, and 6n occurring at very low percentages (2, 10, 3, and 3%, respectively). Regeneration from anther culture is therefore proposed as a rapid, and attractive method of obtaining new triploid varieties in clementine, which could be of great interest for the fresh fruit market that now requires fruit to be seedless.

Additional keywords: Citrus, tissue culture, haploids, flow cytometry, microsatellites, simple sequence repeat.


This work was partially supported by the National Project ‘Plant Biotechnology’ of the Italian Ministry of Agriculture. The authors thank M. Hofer, L. Navarro, and O. Olivares-Fuster for the flow cytometric analyses, and A. Ross Ferguson for the manuscript review.


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