scholarly journals Production of Somatic Hybrid and Autotetraploid Breeding Parents for Seedless Citrus Development

HortScience ◽  
1992 ◽  
Vol 27 (10) ◽  
pp. 1125-1127 ◽  
Author(s):  
Jude W. Grosser ◽  
Frederick G. Gmitter ◽  
E.S. Louzada ◽  
J.L. Chandler
Keyword(s):  
Somatic Hybrid ◽  
Somatic Hybrids ◽  
Sweet Orange ◽  
Electrophoretic Analysis ◽  
Root Tip ◽  
Banding Patterns ◽  
Hybrid Plants ◽  
Sexual Hybrid ◽  
Seedling Leaf ◽  
Cell Chromosome

Allotetraploid somatic hybrid plants of `Nova' tangelo [a sexual hybrid of `Clementine mandarin (C. reticulata Blanco) × `Orlando' tangelo (C. reticulata × C. paradisi Macf.)] + `Succari' sweet orange (C. sinensis L. Osbeck), and `Hamlin' sweet orange (C. sinensis L. Osbeck) + `Dancy' tangerine (C. reticulata) were regenerated following protoplast fusion. `Nova' and `Hamlin' protoplasts were isolated from ovule-derived embryogenic callus and suspension cultures, respectively, and fused using a polyethylene glycol method with seedling leaf-derived protoplasts of `Succari' and `Dancy', respectively. Plants were regenerated via somatic embryogenesis, and somatic hybrids were identified on the basis of leaf morphology, root-tip cell chromosome number, and electrophoretic analysis of peroxidase and phosphoglucose mutase isozyme banding patterns. Diploid plants were regenerated from unfused protoplasts of `Hamlin', `Nova', and `Succari'. Tetraploid plants of `Hamlin' and `Succari' were also recovered, apparently resulting from homokaryotic fusions. No `Dancy' plants were recovered. The somatic hybrid and autotetraploid plants can be used for interploid hybridization with selected monoembryonic scions to generate improved seedless triploid tangor/tangelo cultivars. The lack of suitable tetraploid breeding parents has previously inhibited the development of quality seedless cultivars by this method.

scholarly journals Production of Tetraploid Somatic Hybrid Breeding Parents for Use in Lemon Cultivar Improvement

HortScience ◽  
1992 ◽  
Vol 27 (5) ◽  
pp. 445-447 ◽  
Author(s):  
N. Tusa ◽  
J.W. Grosser ◽  
F.G. Gmitter ◽  
E.S. Louzada
Keyword(s):  
Somatic Hybrid ◽  
Somatic Hybrids ◽  
Sweet Orange ◽  
Callus Cultures ◽  
Hybrid Breeding ◽  
Root Tip ◽  
Phosphogluconate Dehydrogenase ◽  
Hybrid Plants ◽  
Seedling Leaf ◽  
Cell Chromosome

Allotetraploid somatic hybrid plants of `Hamlin' sweet orange (Citrus sinensis L. Osbeck) + `Femminello' lemon (C. limon L. Burm. f.), and Milam lemon (purported hybrid of C. jambhiri Lush) + `Femminello' lemon were regenerated via somatic embryogenesis following protoplast fusion. `Hamlin' and Milam protoplasts were isolated from undeveloped ovule-derived embryogenic callus cultures and fused using a polyethylene glycol method with seedling leaf-derived protoplasts of `Femminello' lemon. Somatic hybrids were identified on the basis of leaf morphology, root-tip cell chromosome number, and electrophoretic analyses of phosphoglucose isomerase, phosphoglucose mutase, and 6-phosphogluconate dehydrogenase leaf isozymes. The somatic hybrids will be used in interploid crosses with lemon in an effort to generate seedless triploid lemon types with improved tolerance to mal secco disease.

scholarly journals 362 PRODUCTION OF SEVEN NEW INTERGENERIC SOMATIC HYBRIDS FOR CITRUS ROOTSTOCK IMPROVEMENT

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 482g-483
Author(s):  
Francisco A. A. Mourão Fo ◽  
Jude W. Grosser ◽  
Frederick G. Gmitter
Keyword(s):  
Somatic Hybrid ◽  
Protoplast Culture ◽  
Somatic Hybrids ◽  
Sweet Orange ◽  
Electrophoretic Analysis ◽  
Root Tip ◽  
Chromosome Counts ◽  
Banding Patterns ◽  
Hybrid Plants ◽  
Citrus Rootstock

Protoplast culture following polyethylene glycol (PEG)-induced fusion resulted in the regeneration of somatic hybrid plants from the following combinations: `Succari' sweet orange (C. sinensis L. Osbeck) + Severinia disticha; `Hamlin' sweet orange (C. sinensisj + S. disticha: `Valencia' sweet orange (C. sinesis) + S. disticha; `Nova' tangelo (C. reticulata hybrid) + S. disticha; `Succari' sweet orange + S. buxifolia; `Nova' tangelo + Citropsis gilletiana; and `Succari' sweet orange + Atlantia ceylanica. `Succari', `Hamlin', `Valencia', and `Nova' protoplasts were Isolated from ovule-derived embryogenic callus and/or suspension cultures whereas protoplasts of S. disticha, S. buxifolia, C. gilletiana, and A. ceylanica were isolated from leaves of potted trees in a greenhouse. Plants were regenerated via somatic embryogenesis and somatic hybrids were identified on the basis of leaf morphology. Electrophoretic analysis of isozyme banding patterns and root tip chromosome counts are being performed. Somatic hybrids with S. disticha are apparently weak whereas the other somatic hybrid plants with S. buxifolia, C. gilletiana, and A. ceylanica exhibit adequate vigor. These are more examples that the the techique of protoplast fusion can be an important tool in overcoming barriers to hybridization of sexually incompatible species.

scholarly journals 361 IN VITRO CITRUS BREEDING FOR SCION IMPROVEMENT

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 482f-482
Author(s):  
Francisco A. A. Mourão Fo ◽  
Jude W. Grosser ◽  
Frederick G. Gmitter
Keyword(s):  
Somatic Hybrid ◽  
Somatic Hybrids ◽  
Sweet Orange ◽  
Cell Suspension Cultures ◽  
Electrophoretic Analysis ◽  
Root Tip ◽  
Embryogenic Cell ◽  
Citrus Breeding ◽  
Hybrid Plants

Protoplast culture following polyethylene glycol (PEG)-induced fusion resulted in the regeneration of somatic hybrid plants from the following combinations: `Succari' sweet orange (C. sinensis L. Osbeck) + `Ponkan' mandarin (C. reticulata Blanco), `Succari' sweet orange + `Dancy' mandarin (C. reticulata), `Succari' sweet orange + `Page' tangelo [a sexual hybrid between `Minneola' tangelo (C. reticulata × C. paradisi Mcf.) × `Clementine' mandarin (C. reticulata)], `Valencia' sweet orange (C. sinensis) + `Page' tangelo. `Succari' and `Valencia' protoplasts were isolated -from ovule-derived embryogenic cell suspension cultures and from seedling leaves for the other parents. Somatic hybrid plants were Identified on the basis of leaf morphology and electrophoretic analysis of isozyme banding patterns. Root tip cell chromosome counting is being performed on all plants. Other putative somatic hybrids Include: `Succari' sweet orange + `Minneola' tangelo; `Succari' sweet orange + `Murcott' tangos (C. sinensis × C. reticulata); `Valencia' sweet orange + `Murcott' tangor; and `Valencia' sweet orange + `Dancy' mandarin. These plants may have direct cultivar potential, but there primary use will be for interploid hybridization with selected monoembryonic scions to produce improved seedless triploids.

scholarly journals Caipira sweet orange + Rangpur lime: a somatic hybrid with potential for use as rootstock in the Brazilian citrus industry

2000 ◽  
Vol 23 (3) ◽  
pp. 661-665 ◽  
Author(s):  
Fernanda Januzzi Mendes-da-Glória ◽  
Francisco de Assis Alves Mourão Filho ◽  
Luis Eduardo Aranha Camargo ◽  
Beatriz Madalena Januzzi Mendes
Keyword(s):  
Somatic Hybrid ◽  
Leaf Morphology ◽  
Somatic Hybrids ◽  
Sweet Orange ◽  
Southeastern Brazil ◽  
Citrus Industry ◽  
Rangpur Lime ◽  
Rapd Profile ◽  
Hybrid Plants ◽  
Regenerated Plants

Protoplast culture following polyethylene glycol-induced fusion resulted in the regeneration of somatic hybrid plants between Caipira sweet orange (Citrus sinensis L. Osbeck) and Rangpur lime (C. limonia L. Osbeck). The plants were confirmed as somatic hybrids by leaf morphology, chromosome number and RAPD profile. All regenerated plants were tetraploid (2n = 4x = 36), with intermediate leaf morphology and complementary RAPD banding profile of both parents. This combination may be useful as a rootstock for the citrus industry in Southeastern Brazil since this somatic hybrid could combine the drought tolerance and vigor of Rangpur lime with the blight tolerance of Caipira sweet orange.

scholarly journals Eight New Somatic Hybrid Citrus Rootstocks with Potential for Improved Disease Resistance

HortScience ◽  
1992 ◽  
Vol 27 (9) ◽  
pp. 1033-1036 ◽  
Author(s):  
Eliezer S. Louzada ◽  
Jude W. Grosseti ◽  
Frederick G. Gmitter ◽  
Beatriz Nielsen ◽  
J.L. Chandler ◽  
...  
Keyword(s):  
Somatic Hybrid ◽  
Somatic Hybrids ◽  
Sweet Orange ◽  
Field Trials ◽  
Poncirus Trifoliata ◽  
Sour Orange ◽  
Rough Lemon ◽  
Hybrid Plants ◽  
Volkamer Lemon ◽  
Cleopatra Mandarin

Protoplast culture following polyethylene glycol-induced fusion resulted in the regeneration of vigorous tetraploid somatic hybrid plants from eight complementary parental rootstock combinations: Citrus reticulata Blanco (Cleopatra mandarin) + C. aurantium L. (sour orange), C. reticulata (Cleopatra mandarin) + C. jambhiri Lush (rough lemon), C. reticulata (Cleopatra mandarin) + C. volkameriana Ten. & Pasq. (Volkamer lemon), C. reticulata (Cleopatra mandarin) + C. limonia Osb. (Rang-pur), C. sinensis (L.) Osb. (Hamlin sweet orange) + C. limonia (Rangpur), C. aurantium (sour orange) + C. volkameriana (Volkamer lemon) zygotic seedling, C. auruntium hybrid (Smooth Flat Seville) + C. jambhiri (rough lemon), and C. sinensis (Valencia sweet orange) + Carrizo citrange [C. paradisi Macf. × Poncirus trifoliata (L.) Raf.]. Diploid plants were regenerated from nonfused callus-derived protoplasts of Valencia sweet orange and Smooth Flat Seville and from nonfused leaf protoplasts of sour orange, Rangpur, rough lemon, and Volkamer lemon. Regenerated plants were classified according to leaf morphology, chromosome number, and leaf isozyme profiles. All somatic hybrid plants were tetraploid (2n = 4× = 36). One autotetraploid plant of the Volkamer lemon zygotic was recovered, apparently resulting from a homokaryotic fusion. These eight new citrus somatic hybrids have been propagated and entered into field trials.

scholarly journals ASYMMETRIC PROTOPLASM FUSION IN TOMATO

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1130b-1130
Author(s):  
P.F. McCabe ◽  
L.J. Dunbar ◽  
A. Guri ◽  
K.C. Sink
Keyword(s):  
Somatic Hybrid ◽  
Morphological Characteristics ◽  
Regeneration Medium ◽  
Chromosome Counts ◽  
Donor Genome ◽  
Hybrid Plants ◽  
Sexual Hybrid ◽  
Gene Nptii ◽  
Exact Quantification ◽  
Selection Of

Sexual hybrid plants of Lycopersicon esculentum × L. pennellii (E × P) have been transformed and the T-DNA inserts genetically mapped. Donor protoplasts of E × P were isolated from leaves, and subsequently irradiated with 0, 5, 10 and 20 krad of a 60Co. They were then fused with suspension-derived protoplasts of S. lycopersicoides using the PEG-CA++-high pH technique. The protoplasts were cultured in medium 8E at 1.5 × 106 protoplasm/ml. Selection of heterokaryon-derived macrocalli was facilitated by the inability of irradiated donor protoplasts to divide and by plating p-calli on regeneration medium containing kanamycin - an antibiotic for which the resistance gene NPTII is part of the T-DNA. Morphological characteristics of the resulting asymmetric somatic hybrid plants indicate that 10 and 20 krad irradiation eliminates a larger port ion of the genome than does 5 krad. This has been confirmed with isozyme analysis and chromosome counts which generally show the 5 krad asymmetric somatic hybrid plants to differ little from symmetric hybrids although they contain some significant exceptions. Isozyme data reveals the 10 and 20 krad plants to have received much less of the donor genome. Exact quantification is continuing using isozyme markers, chromosome counts and cDNA probes.

scholarly journals SOMATIC HYBRIDIZATION BETWEEN CITRUS RETICULATA AND CITROPSIS GABUNENSIS THROUGH ELECTROFUSION

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1155d-1155
Author(s):  
Jinq-Tian Ling ◽  
N. Nito ◽  
M. Iwamasa
Keyword(s):  
Electric Fields ◽  
Somatic Hybrid ◽  
Somatic Hybrids ◽  
Morphological Characteristics ◽  
Chromosome Count ◽  
Citrus Reticulata ◽  
Embryogenic Calli ◽  
Isoenzyme Analysis ◽  
Hybrid Plants ◽  
Agar Selection

Protoplasts were isolated from embryogenic calli of Citrus reticulata cv. Ponkan and Citropsis gabunensis, and fused in electric fields. The maximal fusion efficiency was obtained by application of AC at 75 V/cm (1.0 MHz) and DC square-wave pulse at 1.125 KV/cm for 40 usec. Fusion-treated protoplasts were cultured on MT medium without phytohormone, solidified with 0.6% agar. Colonies from the protoplasts were proliferated on MT medium with zeatin 1 mg/l and 0.9% agar. Selection of somatic hybrid callus was based on chromosome count and isoenzyme analysis. The somatic hybrids were tetraploid (2n=36). C. reticulata and C. gabunensis were both homozygous at Got-1 locus, but distinguishable easily because band of the latter migrated faster than that of the former. In zymogram of somatic hybrid, both parent bands were retained and a new hybrid band was also evident between them. Embryos from somatic-hybrid callus regenerated intact plant. The hybrid plants showed intermediate morphological characteristics of the parents.

scholarly journals Somatic Hybridization of Complementary Citrus Rootstock:Five New Hybrids

HortScience ◽  
1994 ◽  
Vol 29 (7) ◽  
pp. 812-813 ◽  
Author(s):  
Jude W. Grosser ◽  
Frederick G. Gmitter ◽  
J.L. Chandler ◽  
Eliezer S. Louzada
Keyword(s):  
Somatic Hybrid ◽  
Somatic Hybridization ◽  
Poncirus Trifoliata ◽  
Sour Orange ◽  
Trifoliate Orange ◽  
Hybrid Plants ◽  
Regenerated Plants ◽  
Citrus Rootstock ◽  
Sexual Hybrid ◽  
Cleopatra Mandarin

Protoplasm culture following polyethylene glycol-induced fusion resulted in the regeneration of tetraploid somatic hybrid plants from the following attempted parental combinations: Cleopatra mandarin (Citrus reticulata Blanco) + Argentine trifoliate orange [Poncirus trifoliata (L.) Raf.]; `Succari' sweet orange [C. sinensis (L.) Osb.] + Argentine trifoliate orange; sour orange (C. aurantium L.) + Flying Dragon trifoliate orange (P. trifolita); sour orange + Rangpur (C. limonia Osb.); and Milam lemon (purported sexual hybrid of C. jambhiri Lush × C. sinensis) + Sun Chu Sha mandarin (C. reticulate Blanco). Protoplasm isolation, fusion, and culture were conducted according to previously published methods. Regenerated plants were classified according to leaf morphology, chromosome number, and peroxidase, phosphoglucomutase, and phosphoglucose isomerase leaf isozyme profiles. All of the somatic hybrid plants were tetraploid, as expected (2n = 4x = 36), and all five selections have been propagated and entered into commercial citrus rootstock trials.

scholarly journals New Intergeneric Somatic Hybrids Combining Amblycarpa Mandarin with Six Trifoliate/Trifoliate Hybrid Selections for Lime Rootstock Improvement

HortScience ◽  
2004 ◽  
Vol 39 (2) ◽  
pp. 355-360 ◽  
Author(s):  
Victor Medina-Urrutia ◽  
Karla Fabiola ◽  
Lopez Madera ◽  
Patricia Serrano ◽  
G. Ananthakrishnan ◽  
...  
Keyword(s):  
Somatic Hybrid ◽  
Somatic Hybridization ◽  
Somatic Hybrids ◽  
Field Evaluation ◽  
Randomly Amplified Polymorphic Dna ◽  
Hybrid Plants ◽  
Commercially Important ◽  
Size Controlling ◽  
Commercial Field

No presently available rootstock combines all the available rootstock attributes necessary for efficient long-term citriculture (production and harvesting) of Mexican limes and other commercially important scions. In the present study, somatic hybridization techniques were used to combine the widely adapted Amblycarpa mandarin (also known as Nasnaran mandarin) with six different trifoliate/trifoliate hybrid selections: Benton, Carrizo, and C-35 citranges; Flying Dragon and Rubidoux trifoliate oranges; and a somatic hybrid of sour orange + Flying Dragon. The ultimate goal of this research is to generate polyploid somatic hybrids that express the complementary horticultural and disease resistance attributes of the corresponding parents, and have direct potential as improved tree-size controlling rootstocks. Somatic hybrids from all six parental combinations were confirmed by a combination of leaf morphology, flow cytometry, and randomly amplified polymorphic DNA (RAPD) (for nuclear hybridity) and cleaved amplified polymorphic sequence (CAPS) analyses (for mtDNA and cpDNA). This is the first report of citrus somatic hybridization using Amblycarpa mandarin. Unexpected hexaploid somatic hybrid plants were recovered from the fusion of Amblycarpa mandarin + C-35 citrange. Hexaploid hybrids should be very dwarfing and may have potential for producing potted ornamental citrus. Resulting somatic hybrid plants from all six combinations have been propagated by tissue culture and/or rooted cuttings and are being prepared for commercial field evaluation for their potential as improved rootstocks for Mexican lime and other important scions.

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