Chromosome identification for the carnivorous plant Genlisea margaretae

Abstract

Genlisea margaretae, subgenus Genlisea, section Recurvatae (184 Mbp/1C), belongs to a plant genus with a 25-fold genome size difference and an extreme genome plasticity. Its 19 chromosome pairs could be distinguished individually by an approach combining optimized probe pooling and consecutive rounds of multicolor fluorescence in situ hybridization (mcFISH) with bacterial artificial chromosomes (BACs) selected for repeat-free inserts. Fifty-one BACs were assigned to 18 chromosome pairs. They provide a tool for future assignment of genomic sequence contigs to distinct chromosomes as well as for identification of homeologous chromosome regions in other species of the carnivorous Lentibulariaceae family, and potentially of chromosome rearrangements, in cases where more than one BAC per chromosome pair was identified.

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Acknowledgments

We thank Ines Walde and Kristin Langanke for technical assistance; Tomáš Beseda, Jan Vrána, and Jana Dostálová for assistance with flow sorting and BAC library construction; and Giang T.H. Vu and Hieu X. Cao for helpful discussions. This work was supported by a grant of the Deutsche Forschungsgemeinschaft to IS and JF (SCHU 951/16-1), by the European Social Fund (CZ.1.07/2.3.00/20.0189) to IS, by the Ministry of Education, Youth and Sports of the Czech Republic (grant LO1204 from the National Program of Sustainability I) to HŠ and JD, and by a PhD scholarship of the Vietnam Ministry of Education and Training to TDT.

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Correspondence to Jörg Fuchs.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Supplemental Fig. 1
figure6

Phylogenetic relationship of Genlisea species which belong to the subgenus Tayloria (in blue) or the three sections Africanae (in green), Recurvatae (in red), and Genlisea (in violet) of the subgenus Genlisea. Subject of this study, G. margaretae, is labeled in red. The tree was modified based on Vu et al. (2015). (JPG 534 kb)

Supplemental Fig. 2
figure7figure7

The first series of successive mcFISH assigning BACs of pools PP_01 to PP_04 on nine chromosome pairs of G. margaretae. (A) BAC pools PP_01, PP_02 and PP_04 subsequently hybridized to the same metaphase plate. Note that FISH signals of BAC clone B03 of pool PP_02 are missing in this cell but identifiable in another cell (e.g., in panel C). Due to the degradation of cytological preparations after probe stripping, the chromosomal localization of pool PP_03 in comparison with pool PP_01 (B) or with pools PP_02 and PP_04 (C) was investigated in different metaphases. (D) Nine chromosome pairs of G. margaretae are hallmarked by either four BACs that identify a single chromosome each or by five groups of BACs each of which located on a different chromosome (connected by blue lines) when the results of sequential mcFISH from three metaphase cells are combined, as indicated in the right panel of A–C. Underlined BACs were used to prepare chromosome pools CP_01 and CP_02 for the second successive mcFISH experiment. Bars represent 5 μm. (JPG 600 kb)

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Supplemental Fig. 3
figure9

The second successive mcFISH experiment assigning repeat-free BACs of pools PP_05 to PP_08 and CP_01 and CP_02 on 15 chromosome pairs of G. margaretae. BAC B10 of pool PP_06 yielded no signal and thus was excluded. The co-localizations of BACs after six hybridizations are indicated by the blue lines connecting corresponding probes. Bar represents 5 μm. (JPG 480 kb)

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Tran, T.D., Šimková, H., Schmidt, R. et al. Chromosome identification for the carnivorous plant Genlisea margaretae . Chromosoma 126, 389–397 (2017). https://doi.org/10.1007/s00412-016-0599-0

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Keywords

  • Genlisea
  • Multicolor fluorescence in situ hybridization (mcFISH)
  • Reprobing
  • BACs
  • Karyotyping