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Production and characterization of somatic hybrids between rice (Oryza sativa L.) and reed (Phragmites communis Trin.) obtained by protoplast fusion

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Abstract

Protoplasts were isolated from cell suspension cultures of rice (Oryza sativa L.) and reed (Phragmites communis Trin.), fused by a PEG–high pH-high Ca2+ method and cultured on Ry-2 medium to obtain minicalli. Hybrids were selected using lack of dividing ability of reed protoplast and plant regeneration ability of rice protoplast. Regenerated plantlets showed intermediate traits of both or either of parents and the leaves showed variant traits in the shape. Especially, mature plants showed intermediate traits of parents or variant traits in reproductive organs. Selected hybrid plants did not have fertility because pollen and egg cells developed abnormally. Chromosome counting of hybrid plants revealed their cells were tetraploid (2n = 4x = 72) and aneuploidy. The numbers of the chromosome were different not only between the hybrid plants but also within the individual one. However, the peroxidase, the random amplified polymorphic DNA and the simple sequence repeat analysis indicated that regenerated plants were somatic hybrids. The somatic hybrids obtained in this study suggested the useful information for the production of intergeneric hybrids of rice and other plants, and for the characterization of the traits in hybrids.

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Abbreviations

2, 4-D:

2, 4-Dichlorophenoxyacetic acid

NAA:

α-Naphthaleneacetic acid

BA:

6-Benzyladenine

PEG:

Polyethylene glycol

MS:

Murashige and Skoog (1962)

B5:

Gamborg et al. (1968)

Ry-2:

Yamada and Zhi-qi (1986)

RAPD:

Random amplified polymorphic DNA

SSR:

Simple sequence repeat

References

  • Aixia C, Guangmin X (2004) Somatic hybridization between wheat and Italian ryeglass. Plant Sci 166:1219–1226

    Article  CAS  Google Scholar 

  • Aleza P, Garcia-Lor A, Juárez J, Navarro L (2016) Recovery of Citrus cybrid plants with diverse mitochondrial and chloroplast genome combinations by protoplast fusion followed by in vitro shoot, root, or embryo micrografting. Plant Cell, Tissue Organ Cult 126:205–217

    Article  CAS  Google Scholar 

  • Cappelle C, Mörchen M, Hilbert JL, Rambaud C (2007) Regeneration and molecular characterization of a male sterile interspecific somatic hybrid between Cichorium intybus and C. endivia. Plant Sci 172:596–603

    Article  CAS  Google Scholar 

  • Carlson P, Smith H, Dearsing R (1972) Parasexual interspecific plant hybridization. Proc Nat Acad Sci USA 69:2292–2294

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chunhui X, Guangmin X, Daying Z, Fengning X, Huimin C (2003) Integration of maize nuclear and mitocondrial DNA into the wheat genome through somatic hybridization. Plant Sci 165:1001–1008

    Article  CAS  Google Scholar 

  • Collonnier C, Fock I, Mariska I, Servaes A, Vedel F, Siljak-Yakovlev S, Souvannavong V, Sihachakr D (2003) GISH confirmation of somatic hybrids between Salanum melongena and S. torvum and assessment of resistance to both the fungal and the bacterial wilts. Plant Physiol Biochem 41:459–470

    Article  CAS  Google Scholar 

  • Collonnier C, Mulya K, Fock I, Mariska I, Servaes A, Vedel F, Siljak-Yakovlev S, Souvannavong V, Ducreux G, Sihachakr D (2001) Source of resistance against Raistonia solanacearum in fertile somatic hybrids of eggplant (S. melongena L. with S. aethiopcum L.). Plant Sci 160:301–313

    Article  CAS  PubMed  Google Scholar 

  • Dambier D, Benyahia H, Pensabene-Bellavia G, Kaçar YA, Froelicher Y, Belfalah Z, Lhou B, Handaji N, Printz B, Morillon R, Yesiloglu T, Navarro L, Ollitrault P (2011) Somatic hybridization for citrus rootstock breeding: An effective tool to solve some important issues of the Mediterranean citrus industry. Plant Cell Rep 30:883–900

    Article  CAS  PubMed  Google Scholar 

  • Feng YY, Takahashi H, Akagi H, Mori K (2006) New interspecific rice genome constructions obtained by symmetric cell fusion. Plant Breeding 125:403–404

    Article  CAS  Google Scholar 

  • Fengning X, Guangmin X, Huimin C (2003) Effect of UV dosage on somatic hybridization between common wheat and Avena sativa L. Plant Sci 164:697–707

    Article  CAS  Google Scholar 

  • Fiuk A, Rybczyński JJ (2008) Factors influencing efficiency of somatic embryogenesis of Gentiana kurroo (Royle) cell suspension. Plant Biotechnol Rep 2:33–39

    Article  Google Scholar 

  • Fork I et al (2001) Use of Solanum stenotomum for introduction of resistance to bacterial wilt in somatic hybrids of potato. Plant Physiol Biochem 39:899–908

    Article  Google Scholar 

  • Forsberg J et al (1998) Comparison of UV light, X-ray and restriction enzyme treatment as tools in production of asymmetric somatic hybrids between Brassia napus and Arabidopsis thaliana. Theoret Appl Genet 96:1178–1185

    Article  CAS  Google Scholar 

  • Fu CH, Chen CL, Guo WW, Deng XX (2004) GISH, AFLP and PCR-RFLP analysis of an intergeneric somatic hybrid combining Goutou sour orange and Poncirus trifoliata. Plant Cell Rep 23:391–396

    Article  CAS  PubMed  Google Scholar 

  • Fukui K, Iijima K (1991) Somatic chromosome map of rice by imaging methods. Theor Appl Genet 81:589–596

    Article  CAS  PubMed  Google Scholar 

  • Gleba YY, Hoffmann F (1989) ‘Arabidobrassica’: a novel plant obtained by protoplast fusion. Planta 149:112–117

    Article  Google Scholar 

  • Guan Q, Guo Y, Wei Y, Meng F, Zhang Z (2010) Regeneration of somatic hybrids of ginger via chemical protoplast fusion. Plant Cell Tissue Organ Cult 102:279–284

    Article  CAS  Google Scholar 

  • Guo WW, Wu RC, Cheng YJ, Deng XX (2007) Production and molecular characterization of Citrus intergeneric somatic hybrids between red tangerine and citrange. Plant Breeding 126:72–76

    Article  CAS  Google Scholar 

  • Guo X, Xie C, Cai X, Song B, He L, Liu J (2010) Meiotic behavior of pollen mother cells in relation to ploidy level of somatic hybrids between Solanum tuberosum and S. chacoense. Plant Cell Rep 29:1277–1285

    Article  CAS  PubMed  Google Scholar 

  • Guri A, Levi A, Sink KC (1988) Morphological and molecular characterization of somatic hybrid plants between Lycopersicon esculentum and Solanum nigrum. Mol Gen Genet 212:191–198

    Article  CAS  Google Scholar 

  • Hashizume T, Shimamoto I, Harushima Yui M, Sato T, Imai T, Hirai M (1996) Construction of a linkage map for watermelon (Citrullus lanatus) using random amplified polymorphic DNA (RAPD). Euphytica 90:265–273

    Article  CAS  Google Scholar 

  • Hayashi Y et al (1988) Hybrids of rice (Oryza sativa L.) and wild Oryza species obtained by cell fusion. Plantech Mol Gen Genet 214:6–10

    Article  Google Scholar 

  • Hayashi Y et al (1989) Production of cytoplasmic male sterile rice (Oryza Sativa L.) by cell fusion. Nat Biotechnol 7:1171–1174

    Article  Google Scholar 

  • Ishikawa S, Bang SW, Kaneko Y, Matsuzawa Y (2003) Production and characterization of intergeneric somatic hybrids between Moricandia arvensis and Brassica oleracea. Plant Breeding 122:233–238

    Article  Google Scholar 

  • Iwamoto Y, Hirai M, Ohmido N, Fukui K, Ezura H (2007) Fertile somatic hybrids between Solanum integrifolium and S. sanitwongsei (syn. S. kurzii) as candidates for bacterial wilt-resistant rootstock of eggplant. Plant Biotechnology 24:179–184

    Article  Google Scholar 

  • Jarl CI, Bokelmann GS, Haas JM (1995) Protoplast regeneration and fusion in Cucumis: melon×cucumber. Plant Cell Tiss Org Cult 43:259–265

    Article  Google Scholar 

  • Kaneko Y, Bang SW, Matsuzawa Y (2000) Early-bolting trait and RAPD markers in the specific monosomic addition line of radish carrying the e-chromosome of Brassica oleracea. Plant Breeding 119:137–140

    Article  CAS  Google Scholar 

  • Kirti PB, Narasimhulu SB, Prakash S, Chopra VL (1992) Somatic hybridization between Brassica juncea and Moricandia arvensis by protoplast fusion. Plant Cell Rep 11:318–321

    CAS  PubMed  Google Scholar 

  • Kumar MK, Sandeep BV, Rao PS (2018) Development of salt tolerant callus cultures by somatic hybridization between Oryza sativa and mangrove grass Myriostschya wightiana. Annals of Agrarian Science 16:396–404

    Article  Google Scholar 

  • Laiqur R, Ahuja PS, Banerjee S, Bhargava SC (1998) Analysis of asymmetric inter-genetic somatic hybrid plants produced between Hyoscymus maticus L. and double mutant of Nicotiana tabacum L. J Genet Breed 52:333–337

    Google Scholar 

  • Luthra SK, Tiwari JK, Kumar V, Lal M (2019) Evaluation of interspecific somatic hybrids of potato (Solanum tuberosum) and Wild S. cardiophyllum for adaptability, tuber dry matter, keeping quality and late blight resistance. Agricult Res 8:158–164

    Article  Google Scholar 

  • Mizuhiro N, Ito K, Mii M (2001) Production and characterization of interspecfic somatic hybrids between Primula malaeoides and P. obconica. Plant Sci 161:489–496

    Article  CAS  Google Scholar 

  • O’Connel MA, Hanson MR (1987) Regeneration of somatic hybrid plants formed between Lycopersicon esculentum and L. pennellii. Theor Appl Genet 75:83–89

    Article  Google Scholar 

  • Rakosy-Tican E, Thieme R, Nachtigall M, Molnar I, Tunde-Eva D (2015) The recipient potato cultivar influences the genetic makeup of the somatic hybrids between five potato cultivars and one cloned accession of sexually incompatible species Solanum bulbocastanum Dun. Plant Cell, Tissue Organ Cult 122(10):395–407

    Article  CAS  Google Scholar 

  • Smyda-Dajmund P, Śliwka J, Wasilewicz-Flis I, Jakuczun H, Zimnoch-Guzowska E (2016) Genetic composition of interspecific potato somatic hybrids and autofused 4×plants evaluated by DArT and cytoplasmic DNA markers. Plant Cell Rep 35:1345–1358

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Suiyun C, Guangmin X, Taiyong Q, Fengmin X (2004) Introgression of salt-tolerance from somatic hybrids between common wheat and Thinopyrum ponticum. Plant Sci 167:773–779

    Article  CAS  Google Scholar 

  • Terada R et al (1987) Plantlet regeneration from somatic hybrids of rice (Oryza sativa L.) and barnyard grass (Echinochloa oryzicola Vasing). Mol Gen Genet 210:39–43

    Article  Google Scholar 

  • Teresa F et al (2018) An integrated proteomic and metabolomic study to evaluate the effect of nucleus-cytoplasm interaction in a diploid citrus hybrid between sweet orange and lemon. Plant Mol Biol 98:407–425

    Article  CAS  Google Scholar 

  • Tomiczak K, Mikuła A, Sliwinska E, Rybczyński JJ (2015) Autotetraploid plant regeneration by indirect somatic embryogenesis from leaf mesophyll protoplasts of diploid Gentiana decumbens Lf. Vitro Cell Dev Biol Plant 51:350–359

    Article  CAS  PubMed  Google Scholar 

  • Tomiczak K, Sliwinska E, Rybczyński JJ (2017) Protoplast fusion in the genus Gentiana: genomic composition and genetic stability of somatic hybrids between Gentiana kurroo Royle and G. cruciata L. Plant Cell, Tissue Organ Cult 131:1–14

    Article  CAS  Google Scholar 

  • Trabace T, Fiore MD, Ambrosio C, Vanadia S, Sunseri F (1996) Sunflower cytoplasmic hybrids revealed by PCR assay using male sterility as selectable marker. J Genet Breed 50:29–34

    Google Scholar 

  • Waara S, Glimelius K (1995) The potential of somatic hybridization in crop breeding. Euphytica 85:217–255

    Article  Google Scholar 

  • Wang W, Cui SX, Zhang CL (2001) Plant regeneration from embryogenic suspension cultures of dune reed. Plant Cell, Tissue Organ Cult 67:11–17

    Article  CAS  Google Scholar 

  • Wei Y, Guangmin X, Daying Z, Huimin C (2001) Transfer of salt tolerance from Aeluropus littorulis sinensis to wheat via asymetric somatic hybridization. Plant Sci 161:259–266

    Article  CAS  PubMed  Google Scholar 

  • Wójcik AI, Rybczyński JJ (2015) Electroporation and morphogenic potential of Gentiana kurroo (Royle) embryogenic cell suspension protoplasts. Biotechnologia 96:19–29

    Article  Google Scholar 

  • Yamada Y, Zhi-qi Y (1986) Plant regeneration from protoplast-derived callus of rice (O. sativa L.). Plant Cell Rep 5:85–88

    Article  CAS  PubMed  Google Scholar 

  • Yamaguchi J, Shiga T (1993) Characteristics of regenerated plants via protoplast electrofusion between melon (Cucumis melo) and pumpkin (interspecific hybrid, Cucurbita maxima×C. Moschata). Jpn J Breed 43:173–182

    Article  CAS  Google Scholar 

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Acknowledgements

We appreciate Prof. Hyong Bom Pak, Faculty of Life Sciences, Kim Il Sung University, DPR Korea for invaluable advice. This study was supported by the DPR Korea State Commission of Science and Technology.

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  1. Department of Genetics, Faculty of Life Science, KIM IL SUNG University, Pyongyang, Democratic People’s Republic of Korea

    Kang Kim, Tong Su Ho & Un Suk Hwang

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  1. Kang Kim
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Correspondence to Kang Kim.

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Kim, K., Ho, T.S. & Hwang, U.S. Production and characterization of somatic hybrids between rice (Oryza sativa L.) and reed (Phragmites communis Trin.) obtained by protoplast fusion. J. Plant Biochem. Biotechnol. 31, 370–379 (2022). https://doi.org/10.1007/s13562-021-00689-7

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