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Production and molecular characterization of diploid and tetraploid somatic cybrid plants between male sterile Satsuma mandarin and seedy sweet orange cultivars

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Abstract

Seedlessness, an important economic trait for fresh fruit, is among the prior goal for all citrus breeding programs. Symmetric somatic hybridization provides a new strategy for citrus seedless breeding by creating cybrids transferring mitochondrial DNA (mtDNA) controlled cytoplasmic male sterility (CMS) from the callus parent Satsuma mandarin (C. unshiu Marc.) to seedy cultivars. In this study, protoplast fusion was adopted to transfer CMS from C. unshiu Marc. cv. Guoqing No. 1 (G1) to three seedy sweet oranges (C. sinensis L. Osb.), i.e. ‘Early gold’, ‘Taoye’ and ‘Hongjiang’. Flow cytometry analysis showed that 12 of 13 regenerated plants from G1 + ‘Early gold’, 9 of 12 from G1 + ‘Taoye’ and both two plants from G1 + ‘Hongjiang’ were diploids, while the remaining regenerated plants were tetraploids. Molecular analysis using 23 simple sequence repeat (SSR) markers previously proven to map to the citrus genome showed that the nuclear DNA from all recovered diploid and tetraploid plants derived from their corresponding leaf parent, while cleaved amplified polymorphic sequence analysis showed that the mtDNA of all regenerated plants derived from the callus parent, indicating that the regenerated 2X and 4X plants from all these three combinations are authentic cybrids. Furthermore, the Chloroplast SSR analysis revealed that somatic cybrid plants from the three combinations possessed either of their parental chloroplast type in most cases. These results demonstrated that mtDNA of G1 Satsuma mandarin was successfully introduced into the three seedy sweet orange cultivars for potential seedlessness via symmetric fusion.

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Abbreviations

CAPS:

Cleaved amplified polymorphic sequence

CMS:

Cytoplasmic male sterility

cp-SSR:

Chloroplast simple sequence repeat

DAPI:

4′-6-Diamidino-2-phenylindole

G1:

C. unshiu Marc. cv. Guoqing No. 1

mtDNA:

Mitochondrial DNA

SSR:

Simple sequence repeat

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Acknowledgments

This research was financially supported by the Ministry of Science & Technology of China (nos. 2011CB100606, 2011AA100205), the National NSF of China (nos. 31125024, 31221062), and the Fundamental Research Funds for the Central Universities (No. 2013PY045).

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Authors and Affiliations

  1. Key Laboratory of Horticultural Plant Biology (Ministry of Education), Huazhong Agricultural University, Wuhan, 430070, China

    Shi-Xin Xiao, Manosh Kumar Biswas, Meng-Ya Li, Xiu-Xin Deng, Qiang Xu & Wen-Wu Guo

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  1. Shi-Xin Xiao
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  2. Manosh Kumar Biswas
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  3. Meng-Ya Li
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  4. Xiu-Xin Deng
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Correspondence to Wen-Wu Guo.

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Xiao, SX., Biswas, M.K., Li, MY. et al. Production and molecular characterization of diploid and tetraploid somatic cybrid plants between male sterile Satsuma mandarin and seedy sweet orange cultivars. Plant Cell Tiss Organ Cult 116, 81–88 (2014). https://doi.org/10.1007/s11240-013-0384-1

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  • DOI: https://doi.org/10.1007/s11240-013-0384-1

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