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In vitro induction of tetraploids in Vitis × Muscadinia hybrids

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Abstract

A protocol for in vitro induction of tetraploids of the grape rootstock 101-14 Millardet et de Grasset (Vitis riparia × V. rupestris) (101-14 Mgt) (2n = 2x = 38) × Muscadinia rotundifolia cv. Trayshed (2n = 2x = 40) hybrids was established. Different explant materials were exposed to the antimitotic chemical agents colchicine and oryzalin, at three concentrations, for 24, 48 and 72 h. Treated in vitro shoot tips, anthers and pre-embryogenic cultures were regenerated, and their ploidy levels were confirmed by chromosome counts and flow cytometry. It is generally considered that anatomical characteristics of leaves, including leaf stomatal size and number of chloroplasts in guard cells, correlate with ploidy levels of grape plants. Chromosome counts and flow cytometry are labor intensive and difficult, while the number of chloroplasts in stomatal guard cells and stomatal size are easily measured and can be stable and reliable markers. Analysis of stomata revealed clear differences between regenerated diploid and tetraploid plants, and these measurements were used as a preliminary screen, followed by chromosome counts and flow cytometry for confirmation. This study presents the results obtained by various combinations of explants, antimitotic agents, exposure time, concentration and ploidy analysis to evaluate in vitro chromosome doubling. These results provide an easy and effective method for production of tetraploid, potentially fertile Vitis × Muscadinia hybrids for utilization in rootstock breeding programs.

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Acknowledgments

The authors are grateful to Dr. Changmou Xu for his help with the statistical analysis. This work was supported by the China Scholarship Council (CSC) scholarship in conjunction with the assistance of the Department of Viticulture and Enology at University of California, Davis. Research funding for supplies was received from the California Grape Rootstock Improvement Commission.

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Authors and Affiliations

  1. College of Horticulture, Northwest A&F University, Yangling, 712100, Shaanxi, China

    Xiaoqing Xie & Yuejin Wang

  2. Department of Viticulture and Enology, University of California, Davis, 2152 RMI North, 595 Hilgard Lane, Davis, CA, 95616-3014, USA

    Xiaoqing Xie, Cecilia B. Agüero & M. Andrew Walker

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  1. Xiaoqing Xie
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  2. Cecilia B. Agüero
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  3. Yuejin Wang
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  4. M. Andrew Walker
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Correspondence to M. Andrew Walker.

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Xie, X., Agüero, C.B., Wang, Y. et al. In vitro induction of tetraploids in Vitis × Muscadinia hybrids. Plant Cell Tiss Organ Cult 122, 675–683 (2015). https://doi.org/10.1007/s11240-015-0801-8

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  • DOI: https://doi.org/10.1007/s11240-015-0801-8

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