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Interspecific potato somatic hybrids between Solanum tuberosum and S. cardiophyllum, potential sources of late blight resistance breeding

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Abstract

Solanum cardiophyllum Lindl. (PI 341233) (2n = 2x = 24, 1EBN) is a diploid wild potato species highly resistant to late blight, the most serious disease of potato. However, it poses a problem of sexual incompatibility with common potato. So to circumvent this problem, in this study, we developed interspecific potato somatic hybrids between cultivated S. tuberosum dihaploid C-13 and wild species S. cardiophyllum via protoplast fusion. Out of 26 regenerants, only 4 were confirmed as true somatic hybrids containing both parental genomes based on molecular markers and phenotypes. Somatic hybrids were identified by RAPD, ISSR, SSR, AFLP and cytoplasm (chloroplast and mitochondrial genomes) type molecular markers. Intermediate phenotypes of somatic hybrids were confirmed by leaf, flower and tuber traits. Late blight resistance of the hybrids was assessed by challenge inoculation of P. infestans under controlled conditions. Somatic hybrids were found tetraploid by flow cytometry, exhibited high pollen stainability by acetocarmine staining and formed berries and viable seeds after crossing with a common potato variety. Somatic hybrids possessed diverse cytoplasm types (W/α, W/γ and T/β) as assessed by chloroplast and mitochondrial genome-specific markers. Further, cluster analysis based on the Jaccard’s coefficient of molecular profiles generated by all above markers showed genetic distinctness in somatic hybrids and parents. Taken together, these interspecific somatic hybrids with diverse cytoplasm background have potential to be employed in potato breeding programmes to widen the cultivated gene pool through conventional and molecular methods.

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Abbreviations

AFLP:

Amplified fragment length polymorphism

AUDPC:

Area under the disease progress curve

DAP:

Days after planting

FC:

Flow cytometer

EBN:

Endosperm balance number

ISSR:

Inter simple sequence repeat

MS:

Murashige and Skoog

RAPD:

Random amplified polymorphic DNA

SSR:

Simple sequence repeat

TPS:

True potato seed

References

  • Ahn YK, Park T-H (2013) Resistance to common scab developed by somatic hybrids between Solanum brevidens and Solanum tuberosum. Acta Agric Scand Sect B 63:595–603

    CAS  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, Tissue 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

    Article  Google Scholar 

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

    Article  Google Scholar 

  • Bradshaw JE, Bryan GJ, Ramsay G (2006) Genetic resources (including wild and cultivated Solanum species) and progress in their utilisation in potato breeding. Potato Res 49:49–65

    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 

  • Chen L, Guo X, Xie C, He L, Cai X, Tian L, Song B, Liu J (2013) Nuclear and cytoplasmic genome components of Solanum tuberosum + S. chacoense somatic hybrids and three SSR alleles related to bacterial wilt resistance. Theor Appl Genet 126:1861–1872

    Article  CAS  PubMed  Google Scholar 

  • Chimote VP, Chakrabarti SK, Pattanayak D, Pandey SK, Naik PS (2008) Molecular analysis of cytoplasm type in Indian potato varieties. Euphytica 162:69–80

    Article  CAS  Google Scholar 

  • Eeckhaut T, Lakshmanan PS, Deryckere D, Von Bockstaele E, Van Huylenbroeck J (2013) Progress in plant protoplast research. Planta 238:991–1003

    Article  CAS  Google Scholar 

  • Ghislain M, Núñez J, 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 

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

    Google Scholar 

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

    Article  Google Scholar 

  • Hein I, Birch PRJ, Danan S, Lefebvre V, Odeny DA, Gebhardt C, Trognitz F, Bryan GJ (2009) Progress in mapping and cloning qualitative and quantitative resistance against Phytophthora infestans in potato and its wild relatives. Potato Res 52:215–227

    Article  Google Scholar 

  • Helgeson JP, Pohlman JD, Austin S, Haberlach GT, Wielgus SM, Ronis D, Zambolim L, Tooley P, McGrath JM, James RV, Stevenson WR (1998) Somatic hybrids between Solanum bulbocastanum and potato: a new source of resistance to late blight. Theor Appl Genet 96:738–742

    Article  Google Scholar 

  • Iovene M, Aversano R, Savarese S, Caruso I, Dimatteo A, Cardi T, Frusciante L, Carputo D (2012) Interspecific somatic hybrids between Solanum bulbocastanum and S. tuberosum and their haploidization for potato breeding. Biol Plant 56:1–8

    Article  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 

  • Lössl A, Adler N, Horn R, Frei U, Wenzel G (1999) Chondriome-type characterization of potato: mt α, β, γ, δ, ε and novel plastid-mitochondrial configurations in somatic hybrids. Theor Appl Genet 98:1–10

    Article  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 

  • 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 

  • Polzerová H, Patzak J, Greplová M (2011) Early characterization of somatic hybrids from symmetric protoplast electrofusion of Solanum pinnatisectum Dun. and Solanum tuberosum L. Plant Cell, Tissue Organ Cult 104:163–170

    Article  Google Scholar 

  • Rohlf F (2006) NTSYSpc: numerical taxonomy system (ver. 2.2). Exeter Publishing, Ltd, Setauket

    Google Scholar 

  • Sarkar D, Tiwari JK, Shushruti Sharma, Poonam Sharma Sanjeev, Gopal J, Singh BP, Luthra SK, Pandey SK, Pattanayak D (2011) Production and characterization of somatic hybrids between Solanum tuberosum L. and S. pinnatisectum Dun. Plant Cell, Tissue Organ Cult 107:427–440

    Article  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 

  • Sharma S, Sarkar D, Pandey SK, Chandel P, Tiwari JK (2011) Stoloniferous shoot protoplast, an efficient cell system in potato for somatic cell genetic manipulations. Sci Hortic 128:84–91

    Article  CAS  Google Scholar 

  • Smyda P, Jakuczun H, Debski K, Śliwka J, Thieme R, Nachtigall M, Wasilewicz-Flis I, Zimnoch-Guzowska E (2013) Development of somatic hybrids Solanum × michoacanum Bitter. (Rydb.) (+) S. tuberosum L. and autofused 4x S. × michoacanum plants as potential sources of late blight resistance for potato breeding. Plant Cell Rep 32:1231–1241

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Song J, Bradeen JM, Naess SK, Raasch JA, Wielgus SM, Haberlach GT, Liu J, Kuang H, Austin-Phillips S, Buell CR, Helgeson JP, Jiang J (2003) Gene RB cloned from Solanum bulbocastanum confers broad spectrum resistance to potato late blight. Proc Natl Acad Sci USA 100:9128–9133

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Tarwacka J, Polkowska-Kowalczyk L, Kolano B, Śliwka J, Wielgat B (2013) Interspecific somatic hybrids Solanum villosum (+) S. tuberosum, resistant to Phytophthora infestans. J Plant Physiol 170:1541–1548

    Article  CAS  PubMed  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 

  • Thieme R, Rakosy-Tican E, Nachtigall M, Schubert J, Hammann T, Antonova O, Gavrilenko T, Heimbach U, Thieme T (2010) Characterization of the multiple resistance traits of somatic hybrids between Solanum cardiophyllum Lindl. and two commercial potato cultivars. Plant Cell Rep 29:1187–1201

    Article  CAS  PubMed  Google Scholar 

  • Tiwari JK, Poonam Sarkar D, Pandey SK, Gopal J, Kumar SR (2010) Molecular and morphological characterization of somatic hybrids between Solanum tuberosum L. and S. etuberosum Lindl. Plant Cell, Tissue Organ Cult 103:175–187

    Article  CAS  Google Scholar 

  • Tiwari JK, Poonam Kumar V, Singh BP, Sharma S, Luthra SK, Bhardwaj V (2013a) Evaluation of potato somatic hybrids of dihaploid S. tuberosum (+) S. pinnatisectum for late blight resistance. Potato J 40:176–179

    Google Scholar 

  • Tiwari JK, Saurabh S, Chandel P, Singh BP, Bhardwaj V (2013b) Analysis of genetic and epigenetic variation in potato somatic hybrid by AFLP and MASP markers. Electr J Biotech. doi:10.2225/vol16-issue 6-full text-9)

  • Tiwari JK, Sundaresha S, Singh BP, Kaushik SK, Chakrabarti SK, Bhardwaj V, Chandel P (2013c) Molecular markers for late blight resistance breeding of potato: an update. Plant Breed 132:237–245

    Article  CAS  Google Scholar 

  • Tiwari JK, Chandel P, Singh BP, Bhardwaj V (2014) Analysis of plastome and chondriome genome types in potato somatic hybrids from Solanum tuberosum × Solanum etuberosum. Genome 57:29–35

    Article  CAS  PubMed  Google Scholar 

  • Tiwari JK, Devi S, Sharma S, Chandel P, Rawat S, Singh BP (2015) Allele mining in Solanum germplasm: cloning and characterization of RB-homologous gene fragments from late blight resistant wild potato species. Plant Mol Biol Rep 33:1584–1598

    Article  CAS  Google Scholar 

  • Vos P, Hogers R, Bleeker M, Reijans M, Vandelee T, Hornes M, Frijters A, Pot J, Paleman J, Kuiper M, Zabeau M (1995) AFLP: a new technique for DNA fingerprinting. Nucleic Acid Res 23:4407–4414

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Wang M, Allefs S, van den Berg RG, Vleeshouwers VGAA, van der Vossen EAG, Vosman B (2008) Allele mining in Solanum: conserved homologues of Rpi-blb1 are identified in Solanum stoloniferum. Theor Appl Genet 116:933–943

    Article  CAS  PubMed  Google Scholar 

  • Wang J, Jiang J, Wang Y (2013) Protoplast fusion for crop improvement and breeding in China. Plant Cell, Tissue Organ Cult 112:131–142

    Article  Google Scholar 

  • Yu Y, Ye W, He L, Cai X, Liu T, Liu J (2013) Introgression of bacterial wilt resistance from eggplant to potato via protoplast fusion and genome components of the hybrids. Plant Cell Rep 32:1687–1701

    Article  CAS  PubMed  Google Scholar 

  • 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 

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Acknowledgments

The financial and manpower assistances received by JKT from the DBT, Govt. of India (Project No.BT/PR4291/AGR/2/838/2011) and ICAR-CPRI, Shimla (P1-2010/3-IPR-F30/0210) is gratefully acknowledged. The authors are thankful to Dr. Geetha, ICAR-DAMPR, Boriavi, Gujarat, India for FC facility and Mr. Sheeshram Thakur for in vitro multiplication.

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Authors and Affiliations

  1. Central Potato Research Institute, Shimla, Himachal Pradesh, 171 001, India

    Poonam Chandel, Jagesh Kumar Tiwari, Nilofer Ali, Sapna Devi, Sanjeev Sharma & Bir Pal Singh

  2. Central Potato Research Institute Campus, Modipuram, Meerut, Uttar Pradesh, 250 110, India

    Satish Kumar Luthra

  3. Himachal Pradesh University, Shimla, Himachal Pradesh, 171 005, India

    Shashi Sharma

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  1. Poonam Chandel
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  2. Jagesh Kumar Tiwari
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  8. Bir Pal Singh
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Correspondence to Jagesh Kumar Tiwari.

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Chandel, P., Tiwari, J.K., Ali, N. et al. Interspecific potato somatic hybrids between Solanum tuberosum and S. cardiophyllum, potential sources of late blight resistance breeding. Plant Cell Tiss Organ Cult 123, 579–589 (2015). https://doi.org/10.1007/s11240-015-0862-8

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  • DOI: https://doi.org/10.1007/s11240-015-0862-8

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