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Asymmetric somatic hybridization between UV-irradiated Citrus unshiu and C. sinensis: regeneration and characterization of hybrid shoots

  • Genetic Transformation and Hybridization
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Abstract

In the present paper, attempts were made to explore the possibility of employing ultraviolet (UV) irradiation in citrus asymmetric fusion for transfer of limited amount of favorable traits from a desirable cultivar to a target one. Exposure of Satsuma mandarin (Citrus unshiu Marc.) embryogenic protoplasts to UV at an intensity of 300 μW cm−2 led to reduced viability, especially under long irradiation duration. The protoplasts could not grow during culture when they were irradiated for over 30 s. Terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) assay revealed extensive DNA fragmentation in the UV-irradiated protoplasts compared with those without UV treatment. Electrofusion between UV-irradiated protoplasts of Satsuma mandarin (donor) with those of Jincheng (C. sinensis Osbeck, recipient), a local cultivar of superior quality, gave rise to regeneration of several lines of shoots, which failed to root despite enormous endeavors. Ploidy analysis via flow cytometry and chromosome counting showed that four selected shoots were either diploid, triploid or tetraploid. Random amplified polymorphism DNA (RAPD) and amplified fragment length polymorphism (AFLP) confirmed the shoots, irrespective of their ploidy level, as putative somatic hybrids. Cleaved amplified polymorphism sequences (CAPS) demonstrated that the shoots predominantly got their cytoplasmic components, in terms of chloroplast (cp) and mitochondrion DNA, from Jincheng, along with possible recombination of cpDNA in some shoot lines. The current data indicated that UV-based asymmetric fusion could also be employed in citrus somatic hybridization with the intention of creating novel germplasms, which may provide an alternative approach for cultivar improvement.

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Abbreviations

AFLP :

Amplified fragment length polymorphism

CAPS :

Cleaved amplified polymorphism sequence

DAPI :

4′, 6-Diamidino-2-phenylindole

EIM :

Embryoid-induction medium

FCM :

Flow cytometry

FDA :

Fluorescein diacetate

H33258 :

Hoechst 33258

MT :

Murashige and Tucker

RAPD :

Random amplified polymorphism DNA

RFLP :

Restriction fragment length polymorphism

rpm:

Revolutions per minute

SSR :

Simple sequence repeat

TUNEL :

Terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling

UV :

Ultraviolet

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Acknowledgments

This research was partially supported by Natural Science Foundation of China, IRT 0548, the Ministry of Education, China, and Hubei Provincial Natural Science Foundation. The authors are grateful to Dr. JH Wu (HortResearch, New Zealand) for his critical reading of the paper.

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Authors and Affiliations

  1. National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

    Xiao-Yong Xu, Zhi-Yong Hu, Jun-Feng Li, Ji-Hong Liu & Xiu-Xin Deng

  2. College of Horticulture and Plant Protection, Yangzhou University, Yangzhou, 225009, People’s Republic of China

    Xiao-Yong Xu

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  1. Xiao-Yong Xu
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  2. Zhi-Yong Hu
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  3. Jun-Feng Li
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Correspondence to Ji-Hong Liu.

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Communicated by A. Atanassov.

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Xu, XY., Hu, ZY., Li, JF. et al. Asymmetric somatic hybridization between UV-irradiated Citrus unshiu and C. sinensis: regeneration and characterization of hybrid shoots. Plant Cell Rep 26, 1263–1273 (2007). https://doi.org/10.1007/s00299-007-0350-7

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  • DOI: https://doi.org/10.1007/s00299-007-0350-7

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