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Chromosomal view of Lippia alba, a tropical polyploid complex under genome stabilization process

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Abstract

Lippia alba is a phenotypically variable tropical shrub thought to comprise a young autopolyploid complex. Chromosome numbers in L. alba include 2n = 30, 38, 45, 60, and 90. High levels of chemical and phenotypic variation associated with economic and medicinal importance were reported. However, the genetic background including chromosome composition remains under-explored. Furthermore, the occurrence of at least four ploidal levels in L. alba and the lack of data for polyploid plants in tropical areas also merit further study of L. alba. Here we assessed the chromosome composition using two new satellite repeats (CL98 and CL66) applied as FISH probes to mitotic chromosomes, and we proposed to calculate the degree of homozygosis for CL66 satDNA (named as index h) and to associate it to meiotic instability. The CL98 mapping showed few variations in both number of signals and position. However, the levels of structural homozygosity for a satellite repeat CL66 were very variable. The numbers of CL66-bearing-chromosomes were under-represented in tetraploids relative to diploids implying that CL66 arrays have been lost in tetraploid lineages as a result of increased meiotic instability. High percentage of irregularities was observed in meiotic cells, especially in polyploids. L. alba complex comprised a mixture of homomorphic and heteromorphic chromosomes. Overall, the polyploid complex presents features typical of both young and older stable polyploids. It seems that L. alba genome is still in the process of stabilization.

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Funding

This work was supported by CNPq (432412/2016-6 and 313740/2017-8), FAPEMIG, and CAPES.

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Authors and Affiliations

  1. Department of Biology, Federal University of Juiz de Fora, Juiz de Fora, MG, 36036-900, Brazil

    Aryane Campos Reis, Saulo Marçal de Sousa, Victória Rabelo Campos, Laís Stehling de Queiroz Nascimento, Sebastião Pacheco Júnior, Ana Luiza Franco & Lyderson Facio Viccini

  2. Royal Botanic Gardens, Kew, Richmond, TW9 3AE, UK

    Michael Chester

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  1. Aryane Campos Reis
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Supplementary information

Fig. S1

Circular graph layouts of CL98 (A) and CL66 (B) clusters. (PNG 104 kb)

High resolution image (TIF 1838 kb)

Fig. S2

representative meiocytes in different meiosis stages. On the left, DAPI fluorochrome staining and on the right, CL66 probing stained with texas red fluorochrome. a) haploid microspore with 15 chromosomes and five CL66 sites (2x, BGEN-29); b) cytomixis process between meiocytes with unequal distribution of CL 66 sites (4x, BGEN-92); c) prophase I with pairing errs, multivalents (4x, BGEN-03); d) metaphase I with chromosome losses (3x, BGEN-01); e) telophase II with lost chromosomes and unequal segregation of CL 66 sites (4x, BGEN-61); f) triad with unbalanced CL 66 sites (3x, BGEN-59); g-h) polyad with unequal distribution of chromosomes and CL 66 sites at the final of the division (6x, BGEN-42; 4x BGEN-92, respectively). Bar = 5 μm. (PNG 5855 kb)

High resolution image (TIF 70866 kb)

Fig. S3

Representative karyograms of Lippia alba (2n = 2x = 30) showing ten CL66-bearing chromosomes (red signals) and the proportion of homomorphic pairs (h-index) for each accession assessed. (PNG 511 kb)

High resolution image (TIF 989 kb)

Fig. S4

Representative karyograms of Lippia alba (2n = 2x = 30) showing 11 CL66-bearing chromosomes (red signals) and the proportion of homomorphic pairs (h-index) for each accession assessed. (PNG 326 kb)

High resolution image (TIF 701 kb)

Fig. S5

Representative karyograms of Lippia alba (2n = 2x = 30) showing 12 CL66-bearing chromosomes (red signals) and the proportion of homomorphic pairs (h-index) for each accession assessed. (PNG 287 kb)

High resolution image (TIF 642 kb)

Fig. S6

Representatives karyograms of Lippia alba (2n = 2x = 30) showing 13, 14 and15 CL66-bearing chromosomes (red signals) and the proportion of homomorphic pairs (h-index) for each accession assessed. (PNG 427 kb)

High resolution image (TIF 820 kb)

Fig. S7

Representative karyograms of Lippia alba (2n = 2x = 30) showing 16 and18 CL66-bearing chromosomes (red signals) and the proportion of homomorphic pairs (h-index) for each accession assessed. (PNG 394 kb)

High resolution image (TIF 800 kb)

Fig. S8

Representative karyograms of Lippia alba (2n = 3x = 45) showing 16, and 21CL66-bearing chromosomes (red signals) and the proportion of homomorphic pairs (h-index) for each accession assessed. (PNG 566 kb)

High resolution image (TIF 1005 kb)

Fig. S9

Representative karyograms of Lippia alba (2n = 3x = 45) showing 23 CL66-bearing chromosomes (red signals) and the proportion of homomorphic pairs (h-index) for each accession assessed. (PNG 403 kb)

High resolution image (TIF 797 kb)

Fig. S10

Representative karyograms of Lippia alba (2n = 4x = 60) showing ten CL66-bearing chromosomes (red signals) and the proportion of homomorphic pairs (h-index) for each accession assessed. (PNG 281 kb)

High resolution image (TIF 632 kb)

Fig. S11

Representative karyograms of Lippia alba (2n = 4x = 60) showing 18 CL66-bearing chromosomes (red signals) and the proportion of homomorphic pairs (h-index) for each accession assessed. (PNG 484 kb)

High resolution image (TIF 869 kb)

Fig. S12

Representative karyograms of Lippia alba (2n = 4x = 60) showing 19 and 20 CL66-bearing chromosomes (red signals) and the proportion of homomorphic pairs (h-index) for each accession assessed. (PNG 451 kb)

High resolution image (TIF 815 kb)

Fig. S13

Representative karyograms of Lippia alba (2n = 38) showing 13 and 18 CL66-bearing chromosomes and L. alba hexaploid (2n = 6x = 90) showing 20 CL66-bearing chromosomes (red signals). The proportion of homomorphic pairs (h-index) was calculated for the hexaploid accession. (PNG 429 kb)

High resolution image (TIF 813 kb)

Fig. S14

Variation of interstitial-CL66 site among Lippia alba accessions. Differences in number of sites, location and signal size/intensity. (PNG 470 kb)

High resolution image (TIF 696 kb)

Table S1

Collection information for 73 voucher specimens collected in five Brazilian geographic regions. Each voucher number corresponds to specimens in the Universidade Federal de Juiz de Fora, Herbarium CESJ, Brazil (DOCX 67 kb)

Table S2

Meiotic behaviour in 60 accessions of Lippia alba. (DOCX 100 kb)

Table S3

Pollen viability in 60 diploid accessions of Lippia alba. Fluorescein diacetate (FDA) assay. (DOCX 51 kb)

Table S4

Data of Lippia alba seedlings emerged from diploid, triploid and tetraploid mother plants. (DOCX 21 kb)

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Reis, A.C., Chester, M., de Sousa, S.M. et al. Chromosomal view of Lippia alba, a tropical polyploid complex under genome stabilization process. Protoplasma 259, 33–46 (2022). https://doi.org/10.1007/s00709-021-01636-y

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