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Marker-Aided Selection of Polyploid Poplars

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Abstract

Polyploid breeding is an important means for creating elite varieties for the development of poplar plantations. However, polyploid poplars are rare in natural stands. In this study, we established an analytical toolkit to perform marker-aided selection of polyploid poplars. This toolkit contains 12 SSR primer pairs with sites located in the exonic DNA regions and resulting amplified microsatellites in the intronic/intergenic regions. Highly conserved primer pairs were selected by testing in eight species from four poplar sections. The amplified loci’s variability was examined using trees from a germplasm collection of Populus deltoides. Subsequently, copy numbers amplified by the highly variable primers were experimentally determined using progeny of a full-sib diploid pedigree. Based on the above tests, a subset of primers were finally selected and used for marker-aided selection of polyploid poplars from a set of natural Populus tomentosa stands. The reliability of the established analytical toolkit was further verified using a flow cytometer. We established a fast and reliable technique to screen polyploid poplars from natural stands.

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Acknowledgments

Funding for this work was provided by the Key Forestry Public Welfare Project (201304102), the 973 project (2012CB114505), and the Natural Science Foundation of China (31125008). It was also funded by the Doctorate Fellowship Foundation (CXZZ11_0508) and the PCSIRT program of Jiangsu province and the Chinese Educational Department.

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Authors and Affiliations

  1. The Key Laboratory of Forest Genetics and Biotechnology, Nanjing Forestry University, 159#, Longpan Road, Nanjing, 210037, China

    Fanming Kong, Jingjing Liu, Yingnan Chen, Zhibing Wan & Tongming Yin

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  1. Fanming Kong
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  2. Jingjing Liu
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  3. Yingnan Chen
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  4. Zhibing Wan
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  5. Tongming Yin
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Corresponding author

Correspondence to Tongming Yin.

Additional information

Fanming Kong, Jingjing Liu, and Yingnan Chen contributed equally to this work.

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Supplemental Table 1

Alleles generated by 12 SSR primer pairs in 16 P. tomentosa trees. Note: Digital numbers in this table refer to the allele’s size. Loci with only one allele are homozygous, and those with more than one allele are heterozygous. Shadow labeled cells indicate samples that have three alleles at the corresponding loci. (DOC 114 kb)

Supplemental Table 2

Marker segregation in a F1 pedigree (P. deltoides × P. cathayana). Note: Digital numbers in column of “peaks” refer to the size of amplicons by different primers, and “Fail” means unsuccessful amplification. Fully informative markers are those primers amplified heterozygous locus in both parents. The primers in gray shadow generate fully informative markers but peaks are not neat. The primers in yellow shadow generate fully informative markers with neat peaks and they are used in marker-aided selection for polyploidy poplars. These primers were subsequently renamed as Ploidp-01 toPloidp-10. (XLS 89 kb)

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Kong, F., Liu, J., Chen, Y. et al. Marker-Aided Selection of Polyploid Poplars. Bioenerg. Res. 6, 984–990 (2013). https://doi.org/10.1007/s12155-013-9331-6

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