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Molecular and morphological characterization of somatic hybrids between Solanum tuberosum L. and S. etuberosum Lindl.

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Abstract

Interspecific potato somatic hybrids between Solanum tuberosum L. (di)haploid C-13 and 1 endosperm balance number non-tuberous wild species S. etuberosum Lindl. were produced by protoplasts electrofusion. The objective was to transfer virus resistance from this wild species into the cultivated potatoes. Post-fusion products were cultured in VKM medium followed by regeneration of calli in MS13 K medium at 20°C under a 16-h photoperiod, and regenerants were multiplied on MS medium. Twenty-one somatic hybrids were confirmed by RAPD, SSR and cytoplasm (chloroplast/mitochondria) type analysis possessing species-specific diagnostic bands of corresponding parents. Tetraploid nature of these somatic hybrids was determined through flow cytometry analysis. Somatic hybrids showed intermediate phenotypes (plant, leaves and floral morphology) to their parents in glass-house grown plants. All the somatic hybrids were male-fertile. ELISA assay of somatic hybrids after artificial inoculation of Potato virus Y (PVY) infection reveals high PVY resistance.

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Abbreviations

CRBC:

Chicken red blood cell

CTAB:

Cetyltrimethylammonium bromide

nDNA:

Nuclear DNA

DUS:

Distinctness, uniformity and stability

EBN:

Endosperm balance number

FC:

Flow cytometry

HEPES:

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

PVY:

Potato virus Y

RAPD:

Random amplified polymorphic DNA

RCBD:

Randomized complete block design

SSR:

Simple sequence repeat

TPS:

True potato seed

References

  • Anonymous (2007) Procedures for standard evaluation trials of advanced potato clones. An International Cooperators’ Guide. International Potato Center (CIP), Lima-Peru, pp 75–94

  • Arumuganathan K, Earle ED (1991a) Nuclear DNA content of some important plant species. Plant Mol Biol Rep 9:208–218

    Article  CAS  Google Scholar 

  • Arumuganathan K, Earle ED (1991b) Estimation of nuclear DNA content of plants by flow cytometry. Plant Mol Biol Rep 9:229–241

    Article  CAS  Google Scholar 

  • Barone A, Li J, Sebastiano A, Cardi T, Frusciante L (2002) Evidence for tetrasomic inheritance in a tetraploid Solanum commersonii (+) S. tuberosum somatic hybrid through the use of molecular markers. Theor Appl Genet 104:539–546

    Article  CAS  PubMed  Google Scholar 

  • Behnke M (1975) Regeneration in Gewebekulturen einiger dihaplider Solanum tuberosum-Klone. Z Pflanzenziicht 75:262–265

    Google Scholar 

  • Bidani A, Nouri-Ellouz O, Lakhoua L, Sihachakr D, Cheniclet C, Mahjoub A, Drira N, Gargouri-Bouzid R (2007) Interspecific potato somatic hybrids between Solanum berthaultii and Solanum tuberosum L. showed recombinant plastome and improved tolerance to salinity. Plant Cell Tiss Organ Cult 91:179–189

    Article  CAS  Google Scholar 

  • Binding H, Nehls R (1977) Regeneration of isolated protoplast to plant Solanum dulcamara L. Z Pflanzenphysiol 85:279–280

    Google Scholar 

  • Binding H, Nehls R, Schieder O, Sopory SK, Wenzel C (1978) Regeneration of mesophyll protoplast isolated from dihaploid clones of Solanum tuberosum L. Plant Physiol 43:52–54

    Article  Google Scholar 

  • Caetano-Anolles G, Bassam BJ, Gresshoff PM (1992) Primer template interactions during DNA amplification fingerprinting with single arbitrary oligonucleotides. Mol Gen Genet 235:157–165

    Article  CAS  PubMed  Google Scholar 

  • Chanemougasoundharam A, Sharma S, Sarkar S, Pandey SK, Khurana SMP, Thakur KC, Manivel P (2004) Induction of andrognesis and regeneration of androgenic plants from tetraploid Indian potato (Solanum tuberosum L. subsp. tuberosum) cultivars. Potato J 31:59–65

    Google Scholar 

  • Chavez R, Brown CR, Iwanaga M (1988) Application of interspecific sesquiploidy to introgression of PLRV resistance from non-tuber-bearing Solanum etuberosum to cultivated potato germplasm. Theor Appl Genet 76:497–500

    Google Scholar 

  • Devos KM, Gale MD (1992) The use of random amplified polymorphic DNA markers in wheat. Theor Appl Genet 84:567–572

    Article  Google Scholar 

  • Fish N, Karp A, Jones MGK (1988) Production of somatic hybrids by electrofusion in Solanum. Theor Appl Genet 76:260–266

    Google Scholar 

  • Galbraith DW, Harkins KR, Maddox JM, Ayres NM, Sharma DP, Firoozabady E (1983) Rapid flow cytometric analysis of the cell cycle in intact plant tissues. Science 220:1049–1051

    Article  CAS  PubMed  Google Scholar 

  • Gavrilenko T, Thieme R, Heimbach U, Thieme T (2003) Fertile somatic hybrids of Solanum etuberosum + dihaploid Solanum tuberosum and their backcrossing progenies: relationships of genome dosage with tuber development and resistance to potato virus Y. Euphytica 131:323–332

    Article  CAS  Google Scholar 

  • Ghislain M, Jorge N, Herrera MR, Pignataro J, Guzman F, Bonierbale M, Spooner DM (2009) Robust and highly informative microsatellite-based genetic identity kit for potato. Mol Breed 23:377–388

    Article  CAS  Google Scholar 

  • Gibson RW, Jones MGK, Fish N (1988) Resistance to potato leaf roll virus and potato virus Y in somatic hybrids between dihaploid Solanum tuberosum and S. brevidens. Theor Appl Genet 76:113–117

    Google Scholar 

  • Gillen AM, Novy RG (2007) Molecular characterization of the progeny of Solanum etuberosum identifies a genomic region associated with resistance to potato leafroll virus. Euphytica 155:403–415

    Article  CAS  Google Scholar 

  • Gopal J, Kumar V, Pandey SK, Kumar R, Pandey PC, Singh SV (2008) Morphological descriptors for DUS testing of potato varieties. Plant Genet Res News 154:40–47

    Google Scholar 

  • Greplova M, Polzerova H, Vlastnkova H (2008) Electrofusion of protoplasts from Solanum tuberosum, S. bulbocastanum and S. pinnatisectum. Acta Physiol Plant 30:787–796

    Article  CAS  Google Scholar 

  • Helgeson JP, Haberlach GT (1999) Somatic hybrids of Solanum tuberosum and related species. In: Altman A, Ziv M, Izhar S (eds) Plant biotechnology and in vitro biology in the 21st century. Kluwer, Dordrecht, pp 151–154

    Google Scholar 

  • Horsman K, Gavrilenko T, Bergervoet JEM, Huigen DJ, Joe ATW, Jacobsen E (2001) Alteration of the genomic composition of Solanum nigrum (+) potato backcross derivatives by somatic hybridization: selection of fusion hybrids by DNA measurements and GISH. Plant Breed 120:201–207

    Article  CAS  Google Scholar 

  • Jackson SA, Hanneman RE Jr (1999) Crossability between cultivated and wild tuber- and non-tuber-bearing Solanums. Euphytica 109:51–67

    Article  Google Scholar 

  • Johnston SA, den Nijs APM, Peloquin SJ, Hannemann RE (1980) The significance of genetic balance to endosperm development in interspecific crosses. Theor Appl Genet 57:5–9

    Google Scholar 

  • Lössl A, Götz M, Braun A, Wenzel G (2000) Molecular markers for cytoplasm in potato: male sterility and contribution of different plastid-mitochondrial configurations to starch production. Euphytica 116:221–230

    Article  Google Scholar 

  • Maciejewska U, Skierski JS, Szczerbakowa A (1999) Nuclear DNA content of Solanum species grown in vitro as determined by flow cytometry. Acta Physiol Plant 21:37–43

    Article  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Naess SK, Braden JM, Wielgus SM, Haberlach GT, McGrath JM, Helgeson JP (2001) Analysis of introgression of Solanum bulbocastanum DNA into potato breeding lines. Mol Genet Genomics 265:694–704

    Article  CAS  PubMed  Google Scholar 

  • Nouri-Ellouza O, Gargouri-Bouzid R, Sihachakr D, Triki MA, Ducreux G, Drira N, Lakhoua L (2006) Production of intraspecific somatic hybrids with improved tolerance to PVY and Pythium aphanidermatum. J Plant Physiol 163:1321–1332

    Article  Google Scholar 

  • Novy RG, Helgeson JP (1994a) Resistance to potato virus Y in somatic hybrids between Solanum etuberosum and S. tuberosum × S. berthaultii hybrid. Theor Appl Genet 89:783–786

    Google Scholar 

  • Novy RG, Helgeson JP (1994b) Somatic hybrids between Solanum etuberosum and diploid, tuber bearing Solanum clones. Theor Appl Genet 89:775–782

    Google Scholar 

  • Novy RG, Nasruddin A, Ragsdale DW, Radcliffe EB (2002) Genetic Resistances to Potato Leafroll Virus, Potato Virus Y, and Green Peach Aphid in Progeny of Solanum etuberosum. Am J Potato Res 79:9–18

    Article  Google Scholar 

  • Novy RG, Gillen AM, Whitworth JL (2007) Characterization of the expression and inheritance of potato leafroll virus (PLRV) and potato virus Y (PVY) resistance in three generations of germplasm derived from Solanum etuberosum. Theor Appl Genet 114:1161–1172

    Article  CAS  PubMed  Google Scholar 

  • Oberwalder B, Schilde-Rentschler L, Ruoβ B, Wittemann S, Ninnemann H (1998) Asymmetric protoplast fusions between wild species and breeding lines of potato–effect of recipients and genome stability. Theor Appl Genet 97:134–1354

    Article  Google Scholar 

  • Oberwalder B, Schilde-Rentschler L, Loffelhardt-Ruoβ B, Ninnemann H (2000) Differences between hybrids of Solanum tuberosum L. and Solanum circaeifolium Bitt. obtained from symmetric and asymmetric fusion experiments. Potato Res 43:71–82

    Article  Google Scholar 

  • Pehu E, Karp A, Moore K, Steele S, Dunckley R, Jones MGK (1989) Molecular, cytogenetic and morphological characterization of somatic hybrids of dihaploid Solanum tuberosum and diploid S. brevidens. Theor Appl Genet 78:696–704

    Article  CAS  Google Scholar 

  • Polgar Z, Wielgus SM, Horvath S, Helgeson JP (1999) DNA analysis of potato + Solanum brevidens somatic hybrid lines. Euphytica 105:103–107

    Article  Google Scholar 

  • Rokka VM, Xu YS, Kankila J, Kuusela A, Pulli S, Pehu E (1994) Identification of somatic hybrids of dihaploid Solanum tuberosum lines and S. brevidens by species specific RAPD patterns and assessment of disease resistance of the hybrids. Euphytica 80:207–217

    Article  CAS  Google Scholar 

  • Rokka VM, Tauriainen A, Pietilä L, Pehu E (1998) Interspecific somatic hybrids between wild potato Solanum acaule Bitt. and anther-derived dihaploid potato (Solanum tuberosum L.). Plant Cell Rep 18:82–88

    Article  CAS  Google Scholar 

  • Saghai-Maroof MA, Soliman KM, Jorgensen RA, Allard RW (1984) Ribosomal DNA spacer-length polymorphisms in barley: mendelian inheritance, chromosomal locations, and population dynamics. Proc Natl Acad Sci USA 81:8014–8018

    Article  CAS  PubMed  Google Scholar 

  • Sharma S, Sarkar D, Pandey SK (2010) Phenotypic characterization and nuclear microsatellite analysis reveal genomic changes and rearrangements underlying androgenesis in tetraploid potatoes (Solanum tuberosum L.). Euphytica 171:313–326

    Article  CAS  Google Scholar 

  • Spooner DM, Hijmans R (2001) Potato systematics and germplasm collecting, 1989–2000. Am J Potato Res 78:237–268

    Article  Google Scholar 

  • Sutula CL, Gillett JM, Morrissey SM, Ramsdell DC (1986) Interpreting ELISA data and establishing the positive-negative threshold. Plant Dis 70:722–726

    Article  Google Scholar 

  • Szczerbakowa A, Maciejewska U, Pawlowski P, Skierski JS, Wielgat B (2001) Electrofusion of protoplasts from Solanum tuberosum, S. nigrum and S. bulbocastanum. Acta Physiol Plant 2:169–179

    Article  Google Scholar 

  • Szczerbakowa AU, Maciejewska E, Zimnoch-Guzowska E, Wielgat B (2003) Somatic hybrids Solanum nigrum (+) S. tuberosum: morphological assessment and verification of hybridity. Plant Cell Rep 21:577–584

    CAS  PubMed  Google Scholar 

  • Szczerbakowa A, Boltowicz D, Lebecka R, Radomski P, Wielgat B (2005) Characteristics of the interspecific somatic hybrids Solanum pinnatisectum (+) S. tuberosum H-8105. Acta Physiol Plant 3:265–273

    Article  Google Scholar 

  • Thieme R, Darsow U, Gavrilenko T, Dorokhov D, Tiemann H (1997) Production of somatic hybrids between S. tuberosum L. and late blight resistant Mexican wild potato species. Euphytica 97:189–200

    Article  Google Scholar 

  • Thieme R, Darsow U, Rakosy-Tican L, Kang Z, Gavrilenko T, Antonova O, Heimbach U, Thieme T (2004) Use of somatic hybridization to transfer resistance to late blight and potato virus Y (PVY) into cultivated potato. Plant Breed Seed Sci 50:113–118

    Google Scholar 

  • Thieme R, Rakosy-Tican E, Gavrilenko T, Antonova O, Schubert J, Nachtigall M, Heimbach U, Thieme T (2008) Novel somatic hybrids (Solanum tuberosum L. + Solanum tarnii) and their fertile BC1 progenies express extreme resistance to potato virus Y and late blight. Theor Appl Genet 116:691–700

    Article  CAS  PubMed  Google Scholar 

  • Thompson AL, Taylor RJ, Pasche JS, Novy RG, Gudmestad NC (2007) Resistance to Phytophthora erythroseptica and Pythium ultimum in a Potato Clone Derived from S. berthaultii and S. etuberosum. Am J Potato Res 84:149–160

    Article  Google Scholar 

  • USDA ARS National Genetic Resources Program (2003) Germplasm Resources Information Network-(GRIN). [Online Database] http://www.ars-grin.gov/cgi-bin/npgs/html/obs.pl?1189587 National Germplasm Resources Laboratory, Cited 23 Sept 2003

  • Valkonen JPT, Rokka V-M (1998) Combination and expression of two virus resistance mechanisms in interspecific somatic hybrids of potato. Plant Sci 131:85–94

    Google Scholar 

  • Valkonen JPT, Brigneti G, Salazar LF, Pehu LF, Gibson RW (1992a) Interactions of the Solanum spp. of the Etuberosa group and nine potato-infecting viruses and viroid. Ann Appl Biol 120:301–313

    Article  Google Scholar 

  • Valkonen JPT, Brigneti G, Pehu E (1992b) Resistance to Myzus persicae (Suls.) in wild potatoes of the series Etuberosa. Acta Agric Scand, Sectt B. Soil Plant Sci 42:118–127

    Google Scholar 

  • Valkonen JPT, Slack SA, Plaisted RL (1994) Use of the virus strain group concept to characterize the resistance to PVX and PVYo in the potato cv. “Allegany.” Am Potato J 71:507–516

    Google Scholar 

  • Van der Hoeven R, Fulton T, Ilut DC, Tanksley SD (2001) Development of a Solanaceae genome database: SGN. Plant and animal genome IX conference, 2001, San Diego, CA. Abstract #W54_06. Available: http://sgn.cornell.edu

  • Zhou A, Xia G, Zhang X, Chen H, Hu H (2001) Analysis of chromosomal and organellar DNA of somatic hybrids between Triticum aestivum and Haynaldia villosa Schur. Mol Genet Genomics 265:387–393

    Article  CAS  PubMed  Google Scholar 

  • Zimnoch-Guzowska E, Lebecka R, Kryszczuk A, Maciejewska U, Szczerbakowa A, Wielgat B (2003) Resistance to Phytophthora infestans in somatic hybrids of Solanum nigrum L. and diploid potato. Theor Appl Genet 107:43–48

    CAS  PubMed  Google Scholar 

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Acknowledgments

Authors are indebted to Dr. PS Ahuja, Director, Institute of Himalayan Bioresource and Technology (IHBT), Palampur, Himachal Pradesh, for the facility provided for FC analysis. The authors thank Mr. Sheeshram Thakur for the in vitro multiplication and field management. Hearty thanks are also extended to Mr. Akshay and Sanjay, IHBT, for help rendered in FC analysis. The financial assistance received in the form of an ad-hoc research project (F. No. 8-45/2004-Hort. II) from the Indian Council of Agricultural Research (ICAR), New Delhi, is gratefully acknowledged.

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  1. D. Sarkar

    Present address: Division of Crop Improvement, Central Research Institute for Jute and Allied Fibres (CRIJAF), Barrackpore, Kolkata, 700 120, West Bengal, India

Authors and Affiliations

  1. Cell and Molecular Biology Laboratory, Division of Crop Improvement, Central Potato Research Institute (CPRI), Shimla, 171 001, Himachal Pradesh, India

    Jagesh K. Tiwari,  Poonam, D. Sarkar, SK. Pandey & Jai Gopal

  2. Institute of Himalayan Bioresource and Technology (IHBT), Palampur, 176 061, Himachal Pradesh, India

    S. Raj Kumar

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  1. Jagesh K. Tiwari
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Correspondence to Jagesh K. Tiwari.

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Tiwari, J.K., Poonam, Sarkar, D. et al. Molecular and morphological characterization of somatic hybrids between Solanum tuberosum L. and S. etuberosum Lindl.. Plant Cell Tiss Organ Cult 103, 175–187 (2010). https://doi.org/10.1007/s11240-010-9765-x

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