Citrus somatic hybridization with potential for direct tetraploid scion cultivar development

Summary Protoplast fusion has been an effective tool with several applications for citrus cultivar improvement. The evaluation of fruit characteristics of certain previously produced citrus somatic hybrids with ‘Page’ tangelo or ‘Murcott’ tangor as parents, has shown that some allotetraploids produce high quality fruit with direct fresh-fruit cultivar potential. Based on this information, additional somatic hybrids combining these parents with other high quality scions were produced as follows: ‘Murcott’ tangor + ‘Dancy’ tangerine, ‘Murcott’ + LB8-8 (‘Clementine’ ‘Minneola’), ‘Page’ tangelo + ‘Murcott’, ‘Page’ + LB8-9 (‘Clementine’ ‘Minneola’), ‘Page’+ (‘Clementine’ ‘Satsuma’), ‘Page’ + Orie Lee hybrid (‘Clementine’ ‘Murcott’), and ‘Page’ + ‘Ortanique’ tangor, respectively. Their tetraploid level was confirmed by flow cytometry, and their hybridity was verified by random amplified polymorphic DNA (RAPD) or simple sequence repeat (SSR) analysis. Polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) analysis from mitochondria (mt) and chloroplast (cp) DNA, and cpSSR analysis, were also applied to analyse their cytoplasmic genomes. These new hybrids have potential for good quality fruit with reduced seed numbers, and can also serve as tetraploid breeding parents for subsequent triploid production.

[1]  T. Moriguchi,et al.  Somatic hybrids obtained by fusion betweenPoncirus trifoliata (2x) andFortunella hindsii (4x) protoplasts , 1997, Plant Cell Reports.

[2]  W. Guo,et al.  Wide somatic hybrids of Citrus with its related genera and their potential in genetic improvement , 2001, Euphytica.

[3]  J. Grosser,et al.  Transmission of organelle genomes in citrus somatic hybrids , 2000, Plant Cell, Tissue and Organ Culture.

[4]  J. Grosser,et al.  Further evidence of a cybridization requirement for plant regeneration from citrus leaf protoplasts following somatic fusion , 1996, Plant Cell Reports.

[5]  S. Kobayashi,et al.  Analysis of cytoplasmic genomes in somatic hybrids between navel orange (Citrus sinensis Osb.) and ‘Murcott’ tangor , 1991, Theoretical and Applied Genetics.

[6]  P. Ollitrault,et al.  Application of Cleaved Amplified Polymorphic Sequence Method for Analysis of Cytoplasmic Genome among Aurantioideae Intergeneric Somatic Hybrids , 2003 .

[7]  C. Yun cpSSR: a New Tool to Analyze Chloroplast Genome of Citrus Somatic Hybrids , 2003 .

[8]  W. Guo,et al.  Molecular characterization of cytoplasmic and nuclear genomes in phenotypically abnormal Valencia orange (Citrus sinensis) + Meiwa kumquat (Fortunella crassifolia) intergeneric somatic hybrids , 2002, Plant Cell Reports.

[9]  Yunjiang Cheng,et al.  Regeneration and molecular characterization of intergeneric somatic hybrids between Citrus reticulata and Poncirus trifoliata , 2002, Plant Cell Reports.

[10]  T. Cardi,et al.  Organelle DNA analysis of Solanum and Brassica somatic hybrids by PCR with ’universal primers’ , 2001, Theoretical and Applied Genetics.

[11]  W. Powell,et al.  Chloroplast microsatellites: new tools for studies in plant ecology and evolution. , 2001, Trends in ecology & evolution.

[12]  P. Ollitrault,et al.  Somatic hybridization in citrus: An effective tool to facilitate variety improvement , 2000, In Vitro Cellular & Developmental Biology - Plant.

[13]  G. Bryan,et al.  Polymorphic simple sequence repeat markers in chloroplast genomes of Solanaceous plants , 1999, Theoretical and Applied Genetics.

[14]  J. Grosser,et al.  Somatic Hybridization, an Integral Component of Citrus Cultivar Improvement: II. Rootstock Improvement , 1998 .

[15]  R. M. Pérez,et al.  Embryogenesis in vitro of several Citrus species and cultivars , 1998 .

[16]  M. Thomas,et al.  Integration of trinucleotide microsatellites into a linkage map of Citrus , 1997, Theoretical and Applied Genetics.

[17]  W. Saito,et al.  Fruit Characteristics and Pollen Fertility of Citrus Somatic Hybrids , 1995 .

[18]  J. Saunt Citrus varieties of the world : an illustrated guide , 1990 .

[19]  J. Grosser,et al.  Protoplast Fusion and Citrus Improvement , 1990 .

[20]  T. Murashige,et al.  Growth factor requirements of Citrus tissue culture , 1969 .