Abstract
Tissue culture in Passiflora has emerged as a strategy to propagate species with agronomic relevance, which is the main focus of most in vitro studies. Different morphogenic responses have been obtained under the same environmental in vitro conditions, mainly for species of the subgenus Passiflora with distinct 2n chromosome numbers. The aims of this study were to verify and compare the in vitro responses in Passiflora species with distinct 2n chromosome numbers, ploidy levels and nuclear 2C values. Under the same in vitro conditions, only friable calli occurred from mature zygotic embryo explants of Passiflora coriacea (2n = 2x = 12 chromosomes, 2C = 1.00 pg), Passiflora lindeniana (2n = 4x = 24, 2C = 2.42 pg) and Passiflora contracta (2n = 8x = 48, 2C = 4.78 pg). In contrast, plantlets were regenerated from Passiflora foetida (2n = 20, 2C = 1.04 pg) and Passiflora miniata (2n = 18, 2C = 3.40 pg) via indirect organogenesis and indirect somatic embryogenesis, respectively. By now, from mature zygotic embryo explants, de novo shoot organogenesis and somatic embryos have been recovered for Passiflora species with 2n = 18 chromosomes and relative high nuclear 2C value (more than 2C = 2.93 pg—Passiflora cincinata), and only de novo shoot organogenesis for P. foetida with 2n = 20 chromosomes and relative low 2C value (2C = 1.04 pg). Therefore, in a taxonomic and evolutive context, this study showed that the in vitro morphogenic pathways pretty varied between the Passiflora species with distinct karyotype features.
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Abbreviations
- IZE:
-
Immature zygotic embryos
- IO:
-
Indirect organogenesis
- ISE:
-
Indirect somatic embryogenesis
- MZE:
-
Mature zygotic embryos
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Acknowledgements
We would like to thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brasília—DF, Brazil, grants 443801/2014-2 and 448671/2014-0), Fundação de Amparo à Pesquisa do Espírito Santo (FAPES, Vitória—ES, Brazil, grants 65942604/2014 and 0438/2015) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brasília—DF, Brazil) for financial support. We also thank M. S. Joaquim Gasparini dos Santos for images of in vitro tissue culture.
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The authors CTL, DATF, MMP-F and WRC conceived, designed and conducted the tissue culture experiments. The authors ATV, CRC and WRC conducted the flow cytometry analysis. The authors ATV and WRC contributed with chromosome number determination and karyotype characterization. CTL, WRC and AF conceived, designed and conducted the statistical analyses. All authors equally contributed to the manuscript editing and revision and approved the final manuscript version for submission.
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Communicated by Sergei Krasnyanski.
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11240_2018_1536_MOESM1_ESM.tif
Response surface showing the influence of time (in days) (x – time) and 2,4-D concentration (in μM) (y) on the explant responsive rate (z – rate) of the P. coriacea (a), P. lindeniana (b) and P. contracta (c). As in the Fig. 2, the graphics show that, for three species, the explant responsive rate (friable calli formation rate) increased in relation to exposure time and increasing 2,4-D concentration. The highest explant responsive rates were observed in 2,4-D concentrations up to 72.48 μM: 97% in M8 (72.48 μM 2,4-D, arrow) for P. coriacea, 73% in M7 (36.24 μM 2,4-D, arrow) for P. lindeniana, and 46% in M6 (27.18 μM 2,4-D, arrow) for P. contracta. Note that the lowest friable calli formation rate was observed for P. contracta, which possesses 2n = 48. Fitted quadratic models were significant (P < 0.05) by regression analysis for: a) P. coriacea, z = 0.5619 + 21.0x + 38.7544y + 3271.8428y2; b) P. lindeniana, z = 0.2209 + 21.0x + 47.8857y + 4191.9973y2; and c) P. contracta, z = 0.1342 + 21.0x + 38.7544y + 3271.8428y2. Supplementary material 1 (TIF 69938 KB)
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Leite, C.T., Ferreira, D.A.T., Vieira, A.T. et al. In vitro responses in Passiflora species with different chromosome numbers, ploidy levels and nuclear 2C values: revisiting and providing new insights. Plant Cell Tiss Organ Cult 136, 549–560 (2019). https://doi.org/10.1007/s11240-018-01536-9
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DOI: https://doi.org/10.1007/s11240-018-01536-9