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Production and characterization of somatic hybrids between Solanum tuberosum L. and S. pinnatisectum Dun.

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Abstract

Interspecific somatic hybrids between the dihaploid Solanum tuberosum and the wild species S. pinnatisectum Dun. were produced via protoplast fusion. Protoplast isolation, electrofusion, culture of post-fusion products and regeneration of calli/shoots were undertaken following optimized protocols. Regenerants were characterized for hybridity, ploidy and resistance to Phytophthora infestans (Mont.) de Bery, causal fungal pathogen of late blight disease. From a total of 126 regenerated macrocalli, 12 somatic hybrids were confirmed by possessing species-specific diagnostic bands of their corresponding parents as revealed by RAPD, SSRs and cytoplasmic-DNA analyses. Tetraploid status of the 12 hybrids was determined using flow cytometry analysis. Intermediate phenotypes for leaf, flower, and tuber characteristics and high male fertility were observed in field-grown hybrid plants. Hybrids were highly resistant to foliage late blight based on field assessment for two seasons. In contrast, moderate level of resistance to foliage blight was observed in hybrids based on the detached leaf assay under laboratory conditions. Overall, somatic hybrids with moderate levels of resistance to foliage blight were identified, and these will be useful for in situ hybridization in potato breeding efforts.

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Abbreviations

AUDPC:

Area under the disease progress curve

CRBC:

Chicken red blood cell

CTAB:

Cetyltrimethylammonium bromide

DAP:

Days after planting

nDNA:

Nuclear DNA

EBN:

Endosperm balance number

FC:

Flow cytometry

HEPES:

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

MS:

Murashige and Skoog

MAS:

Marker assisted selection

PDS:

Protoplast digestion solution

RAPD:

Random amplified polymorphic DNA

RAUDPC:

Relative area under the disease progress curve

RCBD:

Randomized complete block design

SSR:

Simple sequence repeat

TPS:

True potato seed

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Acknowledgments

Authors are grateful to Dr. PS Ahuja, Director, Institute of Himalayan Bioresource and Technology (IHBT), Palampur, Himachal Pradesh, India for the facility provided for flow cytometry analysis. Authors thank to Mr. Sheeshram Thakur for the in vitro multiplication and field management. 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 also acknowledged. Useful comments and suggestions on the manuscript from the editor, associate editor and two anonymous reviewers are gratefully acknowledged.

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Author notes

  1. D. Sarkar

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

Authors and Affiliations

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

    D. Sarkar, Jagesh K. Tiwari, Sushruti Sharma,  Poonam, Sanjeev Sharma, J. Gopal, B. P. Singh, S. K. Pandey & D. Pattanayak

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

    S. K. Luthra

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  1. D. Sarkar
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Correspondence to Jagesh K. Tiwari.

Appendix

Appendix

See Tables 4, 5, 6 and Fig. 4.

Table 4 Number of protoplast fusion attempted, regenerated macrocalli/shoots/plantlets and identified somatic hybrids of S. tuberosum and S. pinnatisectum
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Table 5 Species-specific diagnostic DNA bands (bp) of the parents in the somatic hybrids S. tuberosum C-13 (+) S. pinnatisectum generated by 15 RAPD decamer primers
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Table 6 Species-specific diagnostic DNA bands (bp) of the parents in the somatic hybrids S. tuberosum C-13 (+) S. pinnatisectum generated by seven SSR markers
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Fig. 4
figure 4

Protoplast fusion between dihaploid S. tuberosum and diploid S. pinnatisectum. A heterokaryon (arrow) formed after electrofusion (left side) and undergoing post-fusion rounding up (right side)

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Sarkar, D., Tiwari, J.K., Sharma, S. et al. Production and characterization of somatic hybrids between Solanum tuberosum L. and S. pinnatisectum Dun.. Plant Cell Tiss Organ Cult 107, 427–440 (2011). https://doi.org/10.1007/s11240-011-9993-8

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