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 BA 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: agermana@unipa.it
Australian Journal of Agricultural Research 56(8) 839-845 https://doi.org/10.1071/AR05025
Submitted: 27 January 2005 Accepted: 20 May 2005 Published: 25 August 2005
Abstract
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.
Acknowledgments
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.
Chen Z,
Wang H, Liao H
(1980) The induction of Citrus pollen plants in artificial media. Acta Genetica Sinica 7, 189–192.
Esen A, Soost RK
(1972) Unexpected triploids in citrus: their origin, identification, and possible use. Journal of Heredity 62, 329–333.
FAOSTAT (2002).
(
:
)
https://www.FAO.org
Froelicher Y, Ollitrault P
(2000) Effects of the hormonal balance on Clausena excavata androgenesis. Acta Horticulturae 535, 139–146.
Geraci G,
Esen A, Soost RK
(1975) Triploid progenies from 2x × 2x crosses of Citrus cultivars. Journal of Heredity 66, 177–178.
| PubMed |
Germanà MA
(1997) Haploidy in Citrus.‘ ‘ haploid production in higher plants’. Vol. 5. 5, pp. 195–217.,(Eds SM Jain, SK Sopory, RE Veilleux)
pp. 195–217. (Kluwer Academic Publisher: Dordrecht, The Netherlands)
Germanà MA
(2003) Haploids and doubled haploids in Citrus spp. ‘Doubled haploid production in crop plants. A manual’. FAO-IAEA.,(Eds M Maluszynsky, KJ Kasha, BP Forster, I Szaejko)
pp. 303–308. (Kluwer Academic Publisher: Dordrecht, The Netherlands)
Germanà MA, Chiancone B
(2003) Improvement of the anther culture protocol in Citrus clementina Hort. ex Tan. Plant Cell Reports 22, 181–187.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Germanà MA,
Crescimanno FG, Motisi A
(2000a) Factors affecting androgenesis in Citrus clementina Hort. ex Tan. Advances in Horticultural Science 14, 49–58.
Germanà MA,
Crescimanno FG,
de Pasquale F, Wang YY
(1991) Androgenesis in 5 cultivars of Citrus limon L. Burm. Acta Horticulturae 300, 315–324.
Germanà MA,
Crescimanno FG,
Reforgiato G, Russo MP
(2000b) Preliminary characterization of several doubled haploids of Citrus clementina cv. Nules. Acta Horticulturae 535, 183–190.
Germanà MA, Reforgiato G
(1997) Haploid embryos regeneration from anther culture of ‘Mapo’ tangelo (Citrus deliciosa × C. paradisi). Advances in Horticultural Science 11, 147–152.
Germanà MA,
Wang YY,
Barbagallo MG,
Iannolino G, Crescimanno FG
(1994) Recovery of haploid and diploid plantlets from anther culture of Citrus clementina Hort. ex Tan. and Citrus reticulata Blanco. Journal of Horticultural Science 69, 473–480.
Gmitter FG,
Ling XB, Deng XX
(1990) Induction of triploid Citrus plants from endosperm calli in vitro.
Theoretical and Applied Genetics 80, 785–790.
| Crossref | GoogleScholarGoogle Scholar |
Grosser JW,
Ollitrault P, Olivares Fuster O
(2000) Somatic hybridization in citrus: an effective tool to facilitate variety improvement. In Vitro Cellular and Developmental Biology-Plant 36, 434–449.
Hidaka T,
Yamada Y, Shichijo T
(1979)
In vitro differentiation of haploid plants by anther culture in Poncirus trifoliata (L.) Raf. Japanese Journal of Breeding 29, 248–254.
Hofer M,
Gomez A,
Aguiriano E,
Manzanera JA, Bueno MA
(2002) Analysis of simple sequence repeat markers in homozygous lines of apple. Plant Breeding 121, 159–162.
| Crossref | GoogleScholarGoogle Scholar |
Kadota M, Niimi Y
(2004) Production of triploid plants of Japanese pear (Pyrus pyrifolia Nakai). Euphytica 138, 141–147.
| Crossref | GoogleScholarGoogle Scholar |
Kijas JMH,
Fowler JCS, Thomas MR
(1995) An evaluation of sequence tagged microsatellite site markers for genetic analysis within Citrus and related species. Genome 38, 349–355.
| PubMed |
Kijas JMH,
Thomas MR,
Fowler JCS, Roose ML
(1997) Integration of trinucleotide microsatellites into a linkage map of Citrus. Theoretical and Applied Genetics 94, 701–706.
| Crossref | GoogleScholarGoogle Scholar |
Kobayashi S,
Ohgawara T,
Saito W,
Nakamura Y, Omura M
(1997) Production of triploid somatic hybrids in Citrus. Journal of Japanese Society for Horticultural Science 66, 453–458.
Murashige T, Skoog FA
(1962) Revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15, 473–497.
Ollitrault P,
Allent V, Luro F
(1996) Production of haploid plants and embryogenic calli of clementine (Citrus reticulata Blanco) after in situ parthenogenesis induced by irradiated pollen. Proceedings International Society of Citriculture South Africa 2, 913–917.
Ollitrault P,
Vanel F,
Froelicher Y, Dambier D
(2000) Creation of triploid Citrus hybrids by electrofusion of haploid and diploid protoplasts. Acta Horticulturae 535, 191–198.
Raquin C, Pilet V
(1972) Production de plantes a partir d’antheres de Petunia cultiveés in vitro. Comptes Rendus D 274, 1019–1022.
Soost RK, Cameron JW
(1980) ‘Oroblanco’, a triploid pummelo-grapefruit hybrid. HortScience 15, 667–669.
Soost RK, Cameron JW
(1985) ‘Melogold’, a triploid pummelo-grapefruit hybrid. HortScience 29, 1134–1135.
Sunderland N,
Collins GB, Dunwell JM
(1974) The role of nuclear fusion in pollen embryogenesis of Datura innoxia Mill. Planta 117, 227–241.
| Crossref | GoogleScholarGoogle Scholar |
Trabut L
(1902) L’hybridation des Citrus: una nouvelle tangerine «La Clémentiné» Revue Horticole 74, 232–234.
