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Generation of interspecific hybrids between Trifolium vesiculosum and T. alexandrinum using embryo rescue

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Abstract

Interspecific hybrids were developed between Trifolium alexandrinum cultivar Wardan × Trifolium vesiculosum and T. alexandrinum cultivar BL1 × T. vesiculosum through embryo rescue, as the crosses failed to set seed under natural conditions. Trifolium vesiculosum was used as a donor/male parent in this study as it is reported to possess tolerance to stem rot and high forage yield. Fertilization in crossed florets of the crosses was manifested from the recovery of swollen ovaries (< 7.80%) and confirmed from the presence of one degenerated ovule in most (> 93.00%) of the swollen ovaries. The hybrid embryos at various developmental stages (heart, torpedo and cotyledonary) were rescued at a frequency of 2.56% from Wardan × T. vesiculosum and 6.12% from BL1 × T. vesiculosum. Differentiation occurred only in the cotyledonary stage embryos, resulting in 17 putative interspecific hybrid plantlets. The assessment of plantlet hybridity through SSR markers (for the alleles inherited from the donor parent), micromorphological leaf traits (leaf texture and stomata) and morphological characters (plant height, leaflet length and width) confirmed production of two interspecific hybrids designated as AV1 and BV3 representing both the crosses. AV1 displayed moderate resistance and BV3 was resistant to stem rot.

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Acknowledgements

The financial support from Department of Biotechnology, New Delhi (DBT Reference No. BT/PR8158/AGR/02/382/2006) on ‘Development of interspecific hybrids using embryo rescue in Trifolium’ in the network project on “Biotechnological approach towards forage crop improvement” is thankfully acknowledged.

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Authors and Affiliations

  1. School of Agricultural Biotechnology, Punjab Agricultural University, Ludhiana, 141 004, India

    Ajinder Kaur, Kamal Preet Kaur, Jagroop Gill Kahlon, Rajesh Sharma & Jagdeep Singh Sandhu

  2. Electron Microscopy and Nanoscience Laboratory, Punjab Agricultural University, Ludhiana, 141 004, India

    Anu Kalia

  3. Department of Plant Breeding and Genetics, Punjab Agricultural University, Ludhiana, 141 004, India

    Upasana Rani & Rahul Kapoor

  4. Crop Improvement Division, Indian Institute of Sugarcane Research, Lucknow, 226 002, India

    Devendra Malaviya

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  1. Ajinder Kaur
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Contributions

 AK attempted Trifolium alexandrinum cultivar Wardan x T. vesiculosum crosses, rescued embryos, recorded data on morphological traits of AV1 interspecific hybrid and drafted the manuscript. KPK attempted T. alexandrinum cultivar BL1 x T. vesiculosum crosses, rescued embryos, carried out SSR analysis of putative AV and BV interspecific hybrids and recorded data on morphological traits of BV3 interspecific hybrid. AK conducted SEM analysis. UR conducted preliminary screening of interspecific hybrids for disease reaction to stem rot. JGK captured embryo developmental stages, recorded pollen viability and seed formation of parents and interspecific hybrids under compound light microscope. RS clonally propagated putative hybrid plantlets and parents. DM provided seed material of Wardan, T. vesiculosum and imparted training on embryo culture. RK analyzed and interpreted data on morphological traits of interspecific hybrids. JSS planned, coordinated the experiments, interpreted results and drafted the manuscript. All authors have read and approved the final manuscript.

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Correspondence to Jagdeep Singh Sandhu.

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Kaur, A., Kaur, K.P., Kalia, A. et al. Generation of interspecific hybrids between Trifolium vesiculosum and T. alexandrinum using embryo rescue. Euphytica 213, 253 (2017). https://doi.org/10.1007/s10681-017-2042-x

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