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Genetic analysis and identification of a candidate gene associated with in vitro regeneration ability of cucumber

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Abstract

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Candidate genes associated with in vitro regeneration were identified in cucumber.

Abstract

The ability to regenerate shoots or whole plants from differentiated plant tissues is essential for plant transformation. In cucumber (Cucumis sativus L.), regeneration ability varies considerably across accessions, but the genetic mechanism has not yet been demonstrated. In the present study, 148 recombinant inbred lines and a core collection were examined to identify candidate genes involved in cucumber regeneration. Four QTL for cotyledon regeneration that explained 9.7–16.6% of the phenotypic variation in regeneration were identified on cucumber chromosomes 1, 3, and 6. The loci Fcrms1.1 and Fcrms+1.1 were consistently detected in the same genetic interval on two regeneration media. A genome-wide association study revealed 18 SNPs (− log(p) > 5) significantly associated with cotyledon regeneration. Three candidate genes in this region were identified. RT-PCR analyses revealed that Csa1G642540 was significantly more highly expressed in genotypes with high cotyledon regeneration rates than in those with low regeneration. The Csa1G642540 CDS driven by its native promoter was transformed into cucumber line 9110Gt; molecular analyses showed that the T-DNA had integrated into the genomes of 8.6% of regenerated plantlets. The seeds from T0 plants expressing Csa1G642540 were tested for regeneration from cotyledon explants, and the segregate ratio in regeneration frequency is 3:1. The AT3G44110.1, the homologue gene of Csa1G642540 in Arabidopsis, has been reported as PM H+-ATPase activity regulation, integrating flowering signals and enlarging meristem function. These results demonstrate that Csa1G642540 might play an important role in regeneration in cucumber and could serve as a selectable marker for regeneration from cotyledons.

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Acknowledgements

This work was funded by Grants from the National Natural Science Foundation of China (NSFC: 31201626).

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Authors and Affiliations

  1. Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Key Laboratory of Biology and Genetic Improvement of Horticultural Crops of Ministry of Agriculture, Beijing, 100081, China

    Ye Wang, Yongshuo Ma, Han Miao, Shengping Zhang, Sanwen Huang, Zhonghua Zhang & Xingfang Gu

  2. Agricultural Genome Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen, 518124, China

    Qian Zhou, Guangtao Zhu & Sanwen Huang

  3. College of Horticulture, Northwest Agriculture and Forestry University, Yangling, 712100, Shanxi, China

    Shenhao Wang

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  1. Ye Wang
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  2. Qian Zhou
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  3. Guangtao Zhu
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  4. Shenhao Wang
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  5. Yongshuo Ma
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  6. Han Miao
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  9. Zhonghua Zhang
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  10. Xingfang Gu
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Contributions

YW performed the experiments and collected the data. YW, QZ, and GZ performed the bioinformatics analyses. SW, YM, HM, and SZ provided experimental assistance. SH and ZZ provided advice regarding research design. XG provided critical comments on the manuscript. All authors approved the final version of the manuscript.

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Correspondence to Zhonghua Zhang or Xingfang Gu.

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This work complies with the current laws of China.

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Communicated by Heiko C. Becker.

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Wang, Y., Zhou, Q., Zhu, G. et al. Genetic analysis and identification of a candidate gene associated with in vitro regeneration ability of cucumber. Theor Appl Genet 131, 2663–2675 (2018). https://doi.org/10.1007/s00122-018-3182-7

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  • DOI: https://doi.org/10.1007/s00122-018-3182-7

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