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Jude W Grosser - One of the best experts on this subject based on the ideXlab platform.
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protoplast fusion for production of tetraploids and triploids applications for scion and rootstock breeding in citrus
Plant Cell Tissue and Organ Culture, 2011Co-Authors: Jude W Grosser, Frederick G. GmitterAbstract:Protoplast fusion technology has been utilized in many crops to generate allotetraploid Somatic hybrids, and sometimes autotetraploids as a byproduct of the process. A brief history of this technology development is provided, along with a simple protocol developed for citrus, which can be easily adapted to other plants. Protoplast fusion has become a significant tool in ploidy manipulation that can be applied in various cultivar improvement schemes. In rare cases, a new Somatic hybrid may have direct utility as an improved cultivar; however, the most important application of Somatic Hybridization is the building of novel germplasm as a source of elite breeding parents for various types of conventional crosses for both scion and rootstock improvement. Somatic Hybridization is generating superior allotetraploid breeding parents for use in interploid crosses to generate seedless triploids. Seedlessness is a primary breeding objective for new fresh fruit citrus varieties, and several thousand triploid hybrids have been produced using Somatic hybrids as the tetraploid parent. Protoplast fusion is also being utilized to produce Somatic hybrids that combine complementary diploid rootstocks, which have shown good potential for tree size control. Tree size control has gained importance as a means of reducing harvesting costs, maximizing the efficiency of modern cold protection methodology, and facilitating the adaptation of new fruit production systems. Successful Somatic Hybridization in citrus rootstock improvement has enabled rootstock breeding at the tetraploid level via sexual Hybridization, which can yield maximum genetic diversity in zygotic progeny upon which to impose selection for the many traits required in improved rootstock cultivars, including disease and insect resistance, broad adaptation, tree size control, and the ability to consistently produce high yields of quality fruit. Recent progress and successful examples of these applications are discussed. Finally, a discussion of the genetic potential of Somatic hybrids as breeding parents, including meiotic behavior and inheritance is provided.
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production of mandarin pummelo Somatic hybrid citrus rootstocks with potential for improved tolerance resistance to sting nematode
Scientia Horticulturae, 2007Co-Authors: Jude W Grosser, J.l. Chandler, Larry W. DuncanAbstract:Abstract Sting nematode (Belonolaimus longicaudatus Rau) has become a primary factor limiting citrus production in localized regions of the central Florida sandridge citrus production area, making the development of resistant rootstocks a new breeding objective. In efforts to develop a replacement rootstock for the widely adapted sour orange, our focus has been on Somatic Hybridization of selected mandarin + pummelo combinations [Grosser, J.W., Gmitter, Jr., F.G., 1990. Protoplast fusion and citrus improvement. Plant Breed. Rev. 8, 339–374; Ananthakrishnan, G., Calovic, M., Serrano, P., Grosser, J.W., 2006. Production of additional allotetraploid Somatic hybrids combining mandarins and sweet oranges with pre-selected pummelos as potential candidates to replace sour orange rootstock. In Vitro Cell. Dev.: Plant 42, 367–371], since sour orange is probably an introgression hybrid of mandarin and pummelo as suggested by molecular marker analyses [Nicolosi, E., Deng, Z.N., Gentile, A., La Malfa, S., Tribulato, E., 2000. Citrus phylogeny and genetic origin of important species as investigated by molecular markers. Theor. Appl. Genet. 100, 1155–1166; Gulsen, O., Roose, M.L., 2001. Lemons: diversity and relationships with selected Citrus genotypes as measured with nuclear genome markers. J. Am. Soc. Hort. Sci. 126, 309–317]. Somatic hybrid plants were produced from four new mandarin (C. reticulata Blanco) + pummelo (C. grandis L. Osbeck) parental combinations by fusing embryogenic suspension culture-derived protoplasts isolated from selected mandarins with leaf protoplasts of pummelo seedlings previously selected for tolerance/resistance to the sting nematode (B. longicaudatus Rau) as follows: Amblycarpa mandarin + ‘Liang Ping Yau’ (seedling) pummelo seedling SN7; Amblycarpa mandarin + ‘Hirado Buntan Pink’ (HBP) pummelo seedling SN3; Murcott tangor + pummelo seedling SN3; and Shekwasha mandarin + pummelo seedling SN3. Somatic Hybridization was verified by ploidy analysis (via flow cytometry) and RAPD analyses. Mandarin parents were selected for wide soil-adaptation and ability to produce friable embryogenic callus lines. Pummelo seedlings used as leaf parents were identified from a previous screen of large seed populations (200 each) from four pummelos for resistance to sting nematode as follows: ‘Hirado Buntan Pink‘; ‘Red Shaddock‘; ‘Large Pink Pummelo’ and a seedling pummelo of ‘Liang Ping Yau‘. Ten resistant/tolerant pummelo seedlings were selected from the 800 pummelo seeds planted in the screen for further study. The four new Somatic hybrids have been propagated to evaluate their horticultural performance and resistance to the sting nematode. These potential Somatic hybrid rootstocks should also have potential to control tree size due to polyploidy.
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PRODUCTION OF ADDITIONAL ALLOTETRAPLOID Somatic HYBRIDS COMBINING MANDARINS AND SWEET ORANGE WITH PRE-SELECTED PUMMELOS AS POTENTIAL CANDIDATES TO REPLACE SOUR ORANGE ROOTSTOCK
In Vitro Cellular & Developmental Biology – Plant, 2006Co-Authors: G Ananthakrishnan, Milica Ćalović, Patricia Serrano, Jude W GrosserAbstract:Sour orange (Citrus aurantium L.) rootstock has historically been a widely utilized eitrus rootstock throughout the world due to its wide soil adaptability and superior horticultural performance. However, quick-decline isolates of citrus tristeza virus (CTV) have demolished entire industries of sour orange rootstock in some countries, including Brazil and Venezuela. CTV is presently destroying millions of trees of sour orange rootstock in Florida and threatens the citrus industries of Texas and Mexico, where sour orange is the predominant rootstock. Efforts to replace sour orange rootstock are combining traditional breeding and biotechnology approaches, including Somatic Hybridization and transformation. Molecular techniques have confirmed that sour orange is probably a hybrid of mandarin and pummelo. A major focus of our program continues to be the Somatic Hybridization of superior mandarins with pre-selected pummelo parents. Here, we report the regeneration of allotetraploid Somatic hybrid plants from seven new mandarin+pummelo combinations and one new sweet orange+pummelo combination. All new Somatic hybrids were confirmed by leaf morphology, ploidy analysis via flow cytometry, and random amplified polymorphic DNA analysis to show nuclear contributions from both parents in corresponding hybrids. These new Somatic hybrids are being propagated by tissue culture and/or rooted cuttings for further evaluation of disease resistance and horticultural performance in field trials.
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building a replacement sour orange rootstock Somatic Hybridization of selected mandarin pummelo combinations
Journal of the American Society for Horticultural Science, 2004Co-Authors: Jude W Grosser, G Ananthakrishnan, Victor Medinaurrutia, Patricia SerranoAbstract:Sour orange has been a premier citrus rootstock worldwide due to its ability to perform on challenging soils and to produce and hold high-quality fruit. However, increasingly widespread quick-decline isolates of citrus tristeza virus (CTV) have destroyed entire industries on sour orange in some countries, and are in the process of destroying millions of trees on sour orange in Florida. CTV also threatens other citrus locations planted heavy to sour orange, including Texas and Mexico. An acceptable alternative rootstock to replace sour orange is in high demand but has yet to be developed. Molecular analyses have recently determined that sour orange is probably a hybrid of pummelo and mandarin. We report the production of 12 new mandarin + pummelo Somatic hybrids produced by protoplast fusion from selected superior mandarin and pummelo parents, in efforts to develop a suitable replacement sour-orange-like rootstock that is resistant to CTV-induced quick decline. Somatic hybrids from all 12 parental combinations were con- fi rmed by a combination of leaf morphology, fl ow cytometry, and RAPD analyses (for nuclear hybridity). These new mandarin + pummelo Somatic hybrids are being propagated by rooted cuttings as necessary to conduct quick-decline resistance assays and to assess horticultural performance in replicated fi eld trials.
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observations and suggestions for improving Somatic Hybridization by plant protoplast isolation fusion and culture
Hortscience, 1994Co-Authors: Jude W GrosserAbstract:De (Triticum aestivum): In vitro production and utilization of doubled haploids, p. 101–124. In: Y.P.S. Bajaj (ed.). Biotechnology in agriculture and forestry. vol. 12: Haploids in crop improvement I. Springer-Verlag, Berlin. Powell, W., P.D.S. Caligari, and J.M. Dunwell. 1986. Field performance of lines derived from haploid and diploid tissues of Hordeum vulgare. Theor. Applied Genet. 72:458–465. Reed, S.M. and E.A. Wernsman. 1989. DNA amplification among antherderived doubled haploid lines of tobacco and its relationship to agronomic performance. Crop Sci. 29:1072–1076. 401 Research Group, Laboratory of Plant Cell and Tissue Culture, Institute of Genetics, Academia Sinica. 1975. Primary study on induction of pollen plants of Zea mays. Acta Genet. Sin. 2:143. Research Laboratory of Breeding. 1981. A preliminary study on the heredity and vitality of the progenies of tobacco pollen plants, p. 223–225. In: H. Hul (ed.). Plant tissue culture. Proc. Symp. Plant Tissue Culture, Peking, 1978. Pitman Publishing, London. Rufty, R.C., E.A. Wernsman, C.E. Main, and G.V. Gooding, Jr. 1990. Registration of NC-BMR 42 and NC-BMR 90 germplasm lines of tobacco. Crop Sci. 30:241–242. Schaeffer, G.W., F.T. Sharpe, Jr., and P.B. Cregan. 1984. Variation for improved protein and yield from rice anther culture. Theor. Applied Genet. 67:383–389. Simmonds, N.W. 1979. Principles of crop improvement. Longman, London. Thomas, E., F. Hoffman, and G. Wenzel. 1975. Haploid plantlets from microspores of rye. Z. Pflanzenzuchtg. 75:106–113. Wang, Y., C. Sun, C. Wang, and N. Chien. 1973. The induction of the pollen plantlets of Triticale and Capsicum annuum from anther culture. Sci. Sinica 16:147–151. Wenzel, G., F. Hoffmann, and E. Thomas. 1977. Anther culture as a breeding tool in rape. I. Ploidy level and phenotype of androgenetic plants. Z. Pflanzenzuchtg. 78:149–155. Wenzel, G., O. Schieder, T. Przewozny, S.K. Sopory, and G. Melchers. 1979. Comparison of single cell culture derived Solanum tuberosum L. plants and a model for their application in breeding programs. Theor. Applied Genet. 55:49–55. Winzeler, H., J. Schmid, and P.M. Fried. 1987. Field performance of androgenetic, doubled haploid spring wheat lines in comparison with lines selected by the pedigree system. Plant Breeding 99:41–48. Wu, J. 1986. Breeding haploid corn by anther culture, p. 149–161. In: H. Hu and H. Yang (eds.). Haploids of higher plants in vitro. Springer-Verlag, Berlin. Yang, X. and H. Fu. 1989. Hua-03—A high protein indica rice. Intl. Rice Res. Nwsl. 14(3):14–15. Zhu, D. and X. Pan. 1990. Rice (Oryza sativa L.): Guan 18—An improved variety through anther culture, p. 204–211. In: Y.P.S. Bajaj (ed.). Biotechnology in agriculture and forestry 2: Haploids in crop improvement I. Springer-Verlag, Berlin. Zhao, Y., X. He, J. Wang, and W. Liu. 1990. Anther culture 28—A new diseaseresistant and high-yielding variety of winter wheat, p. 353–362. In: Y.P.S. Bajaj (ed.). Biotechnology in agriculture and forestry 13: Wheat. SpringerVerlag, Berlin.
Huimin Chen - One of the best experts on this subject based on the ideXlab platform.
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transfer of small chromosome fragments of agropyron elongatum to wheat chromosome via asymmetric Somatic Hybridization
Science China-life Sciences, 2004Co-Authors: Jing Wang, Fengning Xiang, Huimin ChenAbstract:The chromosome constitution of hybrids and chromatin patterns of Agropyron elongatum (Host)Neviski in F5 Somatic hybrid lines -1–3 and I-1-9 between Triticum aestivum L. and A. elongatum were analyzed. Based on the statistic data of pollen mother cells, F5 I-1-9 and-1-3 had 20–21 bivalents with a frequency of 84.66% and 85.28%, of which, 89.83% and 89.57% were ring bivalents. The result indicated that both hybrid lines were basically stable in the chromosome constitution and behavior. RAPD analysis showed that the two hybrids contained biparental and integrated DNA. GISH (Genome in situ Hybridization) revealed that in the form of small chromosome segments, A. elongatum chromatin was scattered on 4–6 wheat chromosomes near by the region of centromere and telomere in the two hybrid lines. SSR analysis indicated that A. elongatum DNA segments were distributed on the 2A, 5B, 6B and 2D wheat chromosomes in the hybrids, which was in accordance with the GISH results that small-segments intercalated poly-site.
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asymmetric Somatic Hybridization between wheat triticum aestivum and avena sativa l
Science China-life Sciences, 2003Co-Authors: Fengning Xiang, Huimin ChenAbstract:Protoplasts from cell suspensions ofyoung-embryo-derived calli, which were nonregenerable for long-term subculture and protoplasts from embryogenic calli with the regeneration capacity of 75% ofthe same wheat Jinan 177, were mixed as recipient. Protoplasts from embryogenic calli of Avena sativa (with the regeneration capacity ofless than 10%) irradiated with UV at an intensity of 300 μW/cm2 for 30 s, 1 min, 2 min, 3 min, 5 min were used as the donor. Protoplasts ofthe recipient and the donor were fused by PEG method. Many calli and normal green plants were regenerated at high frequency, and were verified as Somatic hybrids by chromosome counting, isozyme, 5S rDNA spacer sequence analysis and GISH (genomic in situ Hybridization). Fusion combination between protoplasts either from the cell suspensions or from the calli and UV-treated Avena sativa protoplasts could not regenerate green plants.
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asymmetric Somatic Hybridization between wheat triticum aestivum l and agropyron elongatum host nevishi
Theoretical and Applied Genetics, 2003Co-Authors: Fengning Xiang, Aifen Zhou, Huai Wang, Huimin ChenAbstract:Suspension-derived protoplasts of Agropyron elongatum irradiated by ultra-violet light (UV) were fused with the suspension-derived protoplasts of Triticum astivum using PEG. Fertile intergeneric Somatic hybrid plants were produced and various hybrid lines have been selected and propagated in successive generations. Their hybrid nature was confirmed by analysis of profiles of isozymes, RAPDs, and 5S rDNA spacer sequences, and via GISH analysis. By the procedure described, the phenotype and chromosome number of wheat could be maintained besides transfer of a few chromosomes and chromosomal fragments from the donor A. elongatum. The results above indicated that highly asymmetric fertile hybrid plants and hybrid progenies of wheat were produced via Somatic Hybridization.
Frederick G. Gmitter - One of the best experts on this subject based on the ideXlab platform.
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protoplast fusion for production of tetraploids and triploids applications for scion and rootstock breeding in citrus
Plant Cell Tissue and Organ Culture, 2011Co-Authors: Jude W Grosser, Frederick G. GmitterAbstract:Protoplast fusion technology has been utilized in many crops to generate allotetraploid Somatic hybrids, and sometimes autotetraploids as a byproduct of the process. A brief history of this technology development is provided, along with a simple protocol developed for citrus, which can be easily adapted to other plants. Protoplast fusion has become a significant tool in ploidy manipulation that can be applied in various cultivar improvement schemes. In rare cases, a new Somatic hybrid may have direct utility as an improved cultivar; however, the most important application of Somatic Hybridization is the building of novel germplasm as a source of elite breeding parents for various types of conventional crosses for both scion and rootstock improvement. Somatic Hybridization is generating superior allotetraploid breeding parents for use in interploid crosses to generate seedless triploids. Seedlessness is a primary breeding objective for new fresh fruit citrus varieties, and several thousand triploid hybrids have been produced using Somatic hybrids as the tetraploid parent. Protoplast fusion is also being utilized to produce Somatic hybrids that combine complementary diploid rootstocks, which have shown good potential for tree size control. Tree size control has gained importance as a means of reducing harvesting costs, maximizing the efficiency of modern cold protection methodology, and facilitating the adaptation of new fruit production systems. Successful Somatic Hybridization in citrus rootstock improvement has enabled rootstock breeding at the tetraploid level via sexual Hybridization, which can yield maximum genetic diversity in zygotic progeny upon which to impose selection for the many traits required in improved rootstock cultivars, including disease and insect resistance, broad adaptation, tree size control, and the ability to consistently produce high yields of quality fruit. Recent progress and successful examples of these applications are discussed. Finally, a discussion of the genetic potential of Somatic hybrids as breeding parents, including meiotic behavior and inheritance is provided.
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intergeneric Somatic Hybridization of sexually incompatible parents citrus sinensis and atalantia ceylanica
Plant Cell Reports, 1993Co-Authors: E S Louzada, Jude W Grosser, Frederick G. GmitterAbstract:Protoplast fusion using polyethylene glycol (PEG) was conducted to combine Citrus sinensis (L.) Osbeck cv. ‘Hamlin’ sweet orange protoplasts, isolated from nucellus-derived embryogenic callus with Atalantia ceylanica (Arn.) Oliv, leaf protoplasts. Five plants regenerated from independent fusion events following protoplast culture were identified as intergeneric allotetraploid Somatic hybrids of ‘Hamlin’ sweet orange and A. ceylanica, and confirmed by isozyme analysis and chromosome number determination in root tip cells (2n=4x=36). Two different types of leaf morphology were observed among the hybrids (normal and narrow), although no differences in chromosome number nor isozyme banding patterns were observed. This is the first report of the production of hybrid plants between these sexually incompatible genera.
Sergio Fatta Del Bosco - One of the best experts on this subject based on the ideXlab platform.
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genetic improvement of citrus fruits the essential oil profiles in a citrus limon backcross progeny derived from Somatic Hybridization
Food Research International, 2013Co-Authors: Sergio Fatta Del Bosco, Loredana Abbate, Nicasio Tusa, Tonia Strano, Agatino Renda, Giuseppe RubertoAbstract:Abstract The peel essential oil profiles of an interspecific allotetraploid Somatic hybrid, achieved by protoplast fusion of the ‘Valencia’ sweet orange ( Citrus sinensis L. Osbeck) and ‘Femminello’ lemon ( Citrus limon L. Burm), and three sexual hybrids obtained by back crosses between Femminello lemon and the allotetraploid Somatic hybrid (Valencia + Femminello), have been studied by gas chromatography (GC) combined with a flame ionization detector (FID) and a mass spectrometry (MS). In total, 83 components were fully characterized and grouped in four classes (monoterpene hydrocarbons, oxygenated monoterpenes, sesquiterpenes, and others) for an easier comparison of all oils. A statistical treatment by linear discriminant analysis (LDA) of the compositional data from the allotetraploid hybrid and the three sexual hybrids show an intermediate essential oil profile with respect to those of both parents. The contribution of ‘Femminello’ lemon parent is in all cases predominant in the production of the volatile profiles of the new hybrids; however, a different behavior in the peel essential accumulation between the allotetrapolid hybrid and the three hybrids is observed.
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Citrus cybrid leaf essential oil.
Flavour and Fragrance Journal, 2000Co-Authors: Giuseppe Alonzo, Sergio Fatta Del Bosco, Eristanna Palazzolo, Filippo Saiano, N TusaAbstract:The essential oil from the leaves of a Citrus cybrid plant, obtained by Somatic Hybridization between ‘Valencia’ sweet orange (Citrus sinensis L. Osbeck), embryogenic parent, and ‘Femminello’ lemon (C. limon L. Burm f.), leaf parent, has been studied by steam distillation and GC–MS analysis and compared with the correspondent oils from the parent plants. Although the overall composition of the cybrid essential oil appears much closer to that of ‘Femminello’, the relatively higher percentage of β-caryophyllene, as well as the qualitative and quantitative modifications of the esteric fraction, are characteristic of the new hybrid. Since cybrid cells possess the nuclear genome of the mesophyll parent, ‘Femminello’ lemon, and the mitochondrial genome of the nucellar parent, ‘Valencia’ sweet orange, it seems possible to conclude that the genetic information coding for the biosynthesis of the essential oils is mainly embedded in the nucleus, although interactions between nuclear genome and cytoplasmic genome can not be excluded. Copyright © 2000 John Wiley & Sons, Ltd.
Chen Huimin - One of the best experts on this subject based on the ideXlab platform.
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agronomic trait and protein component of f_2 hybrid originated from intergeneric Somatic Hybridization between triticum aestivum and agropyron elongatum
Acta Botanica Sinica, 2001Co-Authors: Xiang Fengning, Feng Baomin, Xia Guangmin, Chen HuiminAbstract:Protoplasts derived from common wheat ( Triticum aestivum L. cv. Jinan 177) were fused with UV_treated protoplasts of Agropyron elongatum (Host) Nevski by PEG method, and fertile asymmetric Somatic hybrid plants resembling wheat morphology were obtained. The F 2 hybrid plants could be divided into 3 types according to their morphology. Type Ⅰ hybrids had high and loosely standing stalks with big spikes and grains. TypeⅡ hybrids were dwarf and compact in shape with high tillering ability and smaller spikes. Type Ⅲ hybrids were similar to typeⅠas a whole but had more compact and erect spikes. All the F 2 hybrid lines were superior to wheat in seed protein content, although some difference existed between themselves. Protein analysis of immature embryos and flag leaves from hybrids by two_dimensional electrophoresis showed that they possessed characteristic proteins of both parents and some new proteins. There existed also some different kinds of proteins in different lines.
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asymmetric Somatic Hybridization between wheat triticum aestivum and bromus inermis
Acta Botanica Sinica, 1999Co-Authors: Xiang Fengning, Xia Guangmin, Chen Huimin, Zhou Aifen, Huang Yue, Zhai XiaolingAbstract:Asymmetric Hybridization was conducted between wheat and Bromus inermis Leyss which is a distantly related intergeneric plant (belonging to different tribe) of wheat and possesses some favorable traits, such as resistant to cold, drought and disease. Protoplasts isolated from young embryo derived calli of common wheat ( Triticum aestivum L., cv. 99P, (AABBDD), 2n=42) were fused with UV treated protoplasts isolated from young embryo derived calli of Bromus inermis by PEG method. Three clones (No.1～No.3) were regenerated from the fusion products and differentiated into albino seedlings. The clones and the seedlings were all verified as hybrids by chromosome counting, isozyme and RAPD analysis. Their isozyme and RAPD pattern contained the characteristic bands of both parents as well as new band(s). The chromosome numbers of albino were in the range of 42～54 with small chromosomes of Bromus inermis and chromosome fragments. The above results confirmed that hybrid albinos were obtained.
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plant regeneration from intergeneric Somatic Hybridization between triticum aestivum l and leymus chinensis trin tzvel
Plant Science, 1996Co-Authors: Xia Guangmin, Chen HuiminAbstract:Abstract The suspension derived protoplasts of wheat (Triticum aestivum) cv. Jinan 177 were used as a recipient to fuse with the protoplasts of the 60Co gamma-ray irradiated calli of Legmus chinensis. The wheat suspension cells and their protoplasts were not capable of differentiating to whole plants. The irradiated calli of L. chinensis were also the same. The protoplasts originated from the treated or untreated calli were both unable to divide under the conditions of this experiment. However, the fusion products grew and developed to whole plants which were identified as hybrids according to the analysis of chromosome, isozyme and morphology. The above result revealed that the lost regeneration capacity of both parents could be complementarily restored through Somatic Hybridization. This phenomenon also occurred with our work on Triticum aestivum (+) Haynaldia villosa, T. aestivum (+) Agropyron elongatum and T. aestivum (+) Psathyrostachys juncea.
