Abstract
Tillandsia (Bromeliaceae) species have high endemism, and due to their strong ornamental potential, predatory extraction is threatening the extinction or drastic population reduction of many of them. In light of this scenario, it is necessary to find strategies for the conservation of these endangered species. The objective of this study was to evaluate two seed preservation strategies (freezing at − 5 °C and cryopreservation at − 196 °C) for 20 Tillandsia species occurring in the state of Bahia. We initially evaluated the morphometry, thousand-seed weight, and water content, followed by tests of germination and desiccation. After selecting the best result of the germination test (Germitest paper and incubation at 30 °C) and desiccation (3 h on silica gel), we established conservation tests utilizing two temperatures (freezing at − 5 °C and liquid nitrogen at − 196 °C), with storage times of 1, 7, 30, 180 and 450 days. Analysis of variance indicated that the 20 species had different behaviors when submitted to the two temperatures and different storage times. After 450 days there was a reduction in the germination percentage and germination speed index (GSI) of all the species studied when the seeds were preserved in the freezer. The storage in liquid nitrogen was efficient for the preservation of Tillandsia seeds when dried to a moisture content of approximately 7%. Our results support the establishment of a cryobank for Tillandsia to conserve these endemic species.
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Abbreviations
- HURB:
-
Herbario do Recôncavo da Bahia
- GSI:
-
Germination speed index
- MS:
-
Murashige and Skoog (1962) culture medium
References
Araújo S, Pagano A, Dondi D, Lazzaroni S, Pinela E, Macovei A, Balestrazzi A (2019) Metabolic signatures of germination triggered by kinetin in Medicago truncatula. Sci Rep 9:10466. https://doi.org/10.1038/s41598-019-46866-6
Barfuss MHJ, Till W, Leme EMC, Pinzón JP, Manzanares JM, Halbritter H, Samuel R, Brown JK (2016) Taxonomic revision of Bromeliaceae subfam. Tillandsioideae based on a multi-locus DNA sequence phylogeny and morphology. Phytotaxa 279(1):1–97. https://doi.org/10.11646/phytotaxa.279.1.1
Bastos MJSM, Bastos LP, Souza EH, Soares TL, Morais DV, Souza FVD, Costa MAPC (2017) Floral and reproductive biology of Alcantarea nahoumii (Bromeliaceae), a vulnerable endemic species of the Atlantic Forest. Acta Bot Bras 31(4):665–676. https://doi.org/10.1590/0102-33062017abb0102
Benzing DH (2000) Bromeliaceae: profile of an adaptive radiation. University Press, Cambrigde
Brasil. Ministério da Agricultura, Pecuária E Abastecimento (2009) Regras para análise de sementes. Ministério da agricultura, pecuária e abastecimento. Secretaria de Defesa Agropecuária Mapa/ACS, Brasília
Castillo RF, Trujillo‐Argueta S, Rivera‐García R, Gómez‐Ocampo Z, Mondragón‐Chaparro D (2013) Possible combined effects of climate change, deforestation, and harvesting on the epiphyte: a multidisciplinary approach. Ecol Evol 3(11):3935–3946. https://doi.org/10.1002/ece3.765
Castro RD, Bradford KJ, Hilhorst HWM (2004) Desenvolvimento de sementes e conteúdo de água. In: Ferreira AG, Borghetti F (eds) Germinação: do básico ao aplicado. Artmed, Porto Alegre, pp 51–67
Coelho MFB, Vieira SN, Chig LA, Santos LW, Albuquerque MCF (2011) Superação da dormência em sementes de Bromelia balansae (Bromeliaceae). Hortic Bras 29(4):472–476. https://doi.org/10.1590/S0102-05362011000400005
Dar JA, Kareem M, Zargar SM, Wani AA, Rasool S, Bhat KA (2020) Strategies for conservation of genetic resources. In: Salgotra R, Zargar S (eds) Rediscovery of genetic and genomic resources for future food security. Springer, Singapore, pp 315–334. https://doi.org/10.1007/978-981-15-0156-2_12
Dematê MESP (2005) Information on Brazilian ornamental species of the genus Tillandsia L. (Bromeliaceae). Acta Hort 683:293–200. https://doi.org/10.17660/ActaHortic.2005.683.35
Dinato NB, Santos IRI, Vigna BBZ, Paula AF, Fávero AP (2020) Pollen cryopreservation for plant breeding and genetic resources conservation. CryoLetters 41(3):115–127
Estrella-Parra E, Flores-Cruz M, Blancas-Flores G, Koch SD, Alarcón-Aguilar FJ (2019) The Tillandsia genus: history, uses, chemistry, and biological activity. Bol Latinoam Caribe Plantas Med Aromat 18(3):239–264
Fernandez LV, Beltramo J, Caldiz DO (1989) Germinacion y longevidad de semillas de Tillandsia recurvata. Revista de la Facultad de Agronomia 65(1–2):81–85
Ferrari EAP, Colombo RC, Faria RT, Takane RJ (2016) Cryopreservation of seeds of Encholirium spectabile Martius ex Schultes f. by the vitrification method. Rev Ciênc Agron 47(1):127–177. https://doi.org/10.5935/1806-6690.20160020
Figueiredo JC, David AMSS, Silva CD, Amaro HTR, Alves DD (2017) Maturação de sementes de pimenta em função de épocas de colheita dos frutos. Sci Agr 18(3):1–7. https://doi.org/10.5380/rsa.v18i3.51324
Flora do Brasil 2020 (cont.updated) Flora do Brasil 2020. Jardim Botânico do Rio de Janeiro. http://floradobrasil.jbrj.gov.br/reflora/floradobrasil/FB6361. Accessed 12 june 2020
Flores-Palacios A, Valencia-Díaz S (2007) Local illegal trade reveals unknown diversity and involves a high species richness of wild vascular epiphytes. Biol Conserv 136(3):372–387. https://doi.org/10.1016/j.biocon.2006.12.017
Forzza RC, Costa AF, Leme EMC, Versieux LM, Wanderley MGL, Louzada RB, Monteiro RF, Judice DM, Fernandez EP, Borges RAX, Penedo TSA, Monteiro NP, Moraes MA (2013) Bromeliaceae. In: Martinelli G, Moraes MA (eds) Livro vermelho da flora do Brasil. Andrea Jakobsson & amp, Rio de Janeiro, pp 315–397
Generoso AL, Carvalho VS, Walter R, Campbell G, Araújo LS, Santana JGS, Cunha M (2019) Mature-embryo culture in the cryopreservation of passion fruit (Passiflora edulis Sims) seeds. Sci Hortic 256:108638. https://doi.org/10.1016/j.scienta.2019.108638
Gosling PG (2003) Viability Testing. In: Smith RD, Dickie JB, Linington SH, Pritchard HW, Probert RJ (eds) Seed conservation: turning science into practice. Royal Botanic Gardens, Kew, pp 445–481
Gouda EJ, Butcher D, Gouda K (2021) Encyclopaedia of bromeliads. http://bromeliad.nl/bromNames/. Accessed 17 March 2021
Haeckel IB (2008) The “Arco Floral”: ethnobotany of Tillandsia and Dasylirion spp. in a Mexican religious adornment. Econ Bot 62:90–95. https://doi.org/10.1007/s12231-008-9009-8
Hietz P, Ausserer J, Schindler G (2002) Growth, maturation and survival of epiphytic bromeliads in a Mexican humid montane forest. J Trop Ecol 18(2):177–191. https://doi.org/10.1017/S0266467402002122
Hornung-Leoni CT (2011) Avances sobre usos etnobotánicos de las Bromeliaceae en Latinoamérica. Bol Latinoam Caribe Plant Med Aromat 10:297–314
Jenderek MM, Reed BM (2017) Cryopreserved storage of clonal germplasm in the USDA national plant germplasm system. Vitro Cell Dev Biol-Plant 53:299–308. https://doi.org/10.1007/s11627-017-9828-3
Khanna S, Jenkins H, Bucalo K, Determann RO, Cruse-Sanders JM, Pullman GS (2014) Effects of seed cryopreservation, stratification and scarification on germination for five rare species of pitcher plants. CryoLetters 35(1):29–39
Li DZ, Pritchard HW (2009) The science and economics of ex situ plant conservation. Trends Plant Sci 14(11):614–621. https://doi.org/10.1016/j.tplants.2009.09.005
Machado LC, Oliveira VC, Paraventi MD, Cardoso RNR, Martins DS, Ambrósio CE (2016) Maintenance of Brazilian biodiversity by germplasm bank. Pesq Vet Bras 36(1):62–66. https://doi.org/10.1590/S0100-736X2016000100010
Magalhães RI, Mariath JEA (2012) Seed morphoanatomy and its systematic relevance to Tillandsioideae (Bromeliaceae). Plant Syst Evol 298:1881–1895. https://doi.org/10.1007/s00606-012-0688-3
Malhotra N, Panatu S, Singh B, Negi N, Singh D, Singh M, Chandora R (2019) Genetic resources: collection, conservation, characterization and maintenance. In: Singh M (ed) Lentils: potential resources for enhancing genetic gains. Elsevier, pp 21–41. https://doi.org/10.1016/B978-0-12-813522-8.00003-0
Martins CC, Nakagawa J, Bovi MLA (2000) Influência do peso das sementes de palmito-vermelho (Euterpe espiritosantensis Fernandes) na porcentagem e na velocidade de germinação. Rev Bras Sem 22(1):47–53. https://doi.org/10.17801/0101-3122/rbs.v22n1p47-53
Masetto TE, Faria JMR (2019) In situ DNA fragmentation during the re-establishment of desiccation tolerance in germinated seeds of Cedrela fissilis Vell. J Seed Sci 41(2):244–249. https://doi.org/10.1590/2317-1545v42n2207417
Mello JIO, Barbedo CJ, Salatino A, Figueiredo-Ribeiro RCL (2010) Reserve carbohydrates and lipids from the seeds of four tropical tree species with different sensitivity to desiccation. Braz Arch Biol Technol 53(4):889–899. https://doi.org/10.1590/S1516-89132010000400019
Mercier H, Guerreiro Filho O (1990) Propagação sexuada de algumas bromélias nativas da Mata Atlântica: efeito da luz e da temperatura na germinação. Hoehnea 17(1):19–26
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Nara AK, Webber AC (2002) Biologia floral e polinização de Aechmea beeriana (Bromeliaceae) em vegetação de baixio na Amazônia Central. Acta Amaz 32(4):571–588. https://doi.org/10.1590/1809-43922002324588
Negrelle RRB, Mitchell D, Anacleto A (2012) Bromeliad ornamental species: conservation issues and challenges related to commercialization. Acta Sci Biol Sci 34(1):91–100. https://doi.org/10.4025/actascibiolsci.v34i1.731
NOM-059-SEMARNAT (2019) Lista de especies en riesgo de la Norma Oficial Mexicana. Protección ambiental-Especies nativas de México de flora y fauna silvestres-Categorías de riesgo y especificaciones para su inclusión, exclusión o cambio-Lista de especies en riesgo. Diario Oficial de la Federación 14/11/2019
Panis B, Lambardi M (2006) Status of cryopreservation technologies in plants (crops and forest trees). The Role of Biotechnol 5(7):43–54
Pegg DE (2010) The relevance of ice crystal formation for the cryopreservation of tissues and organs. Cryobiol 60(3):36–44. https://doi.org/10.1016/j.cryobiol.2020.01.005
Pence VC (1991) Cryopreservation of immature embryos of Theobroma cacao. Plant Cell Rep 10:144–147. https://doi.org/10.1007/BF00232046
Pereira TS (1988) Bromelioideae (Bromeliaceae): morfologia do desenvolvimento pós-seminal de algumas espécies. Arq Jardim Bot Rio de Janeiro 29:115–154
Pereira AR, Pereira TS, Andrade ACS (2008) Morfologia de sementes e do desenvolvimento pós-seminal de espécies de Bromeliaceae. Acta Bot Bras 22(4):1150–1162. https://doi.org/10.1590/S0102-33062008000400026
Pereira CC, Francine L, Panobianco M (2010) Germinação e armazenamento de sementes de Nidularium innocentii (Lem.). Rev Bras Sem 32(2):36–41. https://doi.org/10.1590/S0101-31222010000200004
Pinheiro F, Borghetti F (2003) Light and temperature requirements for germination of seeds of Aechmea nudicaulis (l.) Griesebach and Streptocalyx floribundus (Martius ex Shultes f.) Mez (Bromeliaceae). Acta Bot Bras 17(1):27–35. https://doi.org/10.1590/S0102-33062003000100003
Pritchard HW (2007) Cryopreservation of desiccation-tolerant seeds. In: Day JG, Stacey GN (eds) Cryopreservation and freeze-drying protocols methods in molecular biologyTM. Humana Press, pp 185–201. https://doi.org/10.1007/978-1-59745-362-2_13
Pritchard HW, Nadarajan J (2008) Cryopreservation of orthodox (desiccation tolerant) seeds. In: Reed BM (ed) Plant cryopreservation: a practical guide. Springer, New York, pp 485–501. https://doi.org/10.1007/978-0-387-72276-4_19
Pritchard HW, Poyner ALC, Seaton PT (1999) Interspecific variation in orchid seed longevity in relation to ultra-dry storage and cryopreservation. Lindleyana 14:92–101
R Development Core Team (2019) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna
Rao VR, Parthasarathy V (2019) Horticultural genetic resources conservation: priorities, challenges and way forward. In: Rajasekharan PE, Rao VR (eds) Conservation and utilization of horticultural genetic resource. Springer, pp 27–47. https://doi.org/10.1007/978-981-13-3669-0_2
Rodrigues LR, Dornelles ALC, Wittmann MTS (1999) Poliembrionia e número de sementes por fruto de quatro cultivares de tangerineira. Cienc Rural 29(3):469–474. https://doi.org/10.1590/S0103-84781999000300015
Rodrigues ARP, Forzza RC, Andrade ACS (2014) Physiological characteristics underpinning successful cryopreservation of endemic and endangered species of Bromeliaceae from the Brazilian Atlantic Forest. Bot J Linn Soc 176(4):567–578. https://doi.org/10.1111/boj.12219
Rueden CT, Schindelin J, Hiner MC, Dezonia BR, Walter AE, Arena ET, Eliceiri K (2017) W. Image J2: imageJ for the next generation of scientific image data. BMC Bioinformatics 18(529):1–26. https://doi.org/10.1186/s12859-017-1934-z
Salomão AN (2002) Tropical seed species’ responses to liquid nitrogen exposure. Braz J Plant Physiol 14(2):133–138. https://doi.org/10.1590/S1677-04202002000200008
Scrok GJ, Varassin IG (2011) Reproductive biology and pollination of Aechmea distichantha Lem. (Bromeliaceae). Acta Bot Bras 25(3):571–576. https://doi.org/10.1590/S0102-33062011000300009
Silva LMM, Rodrigues TJD, Aguiar IB (2002) Efeito da luz e da temperatura na germinação de sementes de aroeira (Myracrodruon urundeuva Allemão). Rev Árvore 26(6):691–697
Silva KS, Mendonça V, Medeiros LF, Freitas PSC, Góis GB (2010) Influência do tamanho da semente na germinação e vigor de mudas de jaqueira (Artocarpus heterophyllus Lam.). Rev Verde Agroecologia Desenvolv Sustent 5(4):217–221
Silva RL, Ferreira CF, Ledo CAS, Souza EH, Silva PH, Costa MAPC, Souza FVD (2016) Viability and genetic stability of pineapple germplasm after ten years of in vitro conservation. Plant Cell Cult Microprop 127(1):123–133
Silva SSS, Souza EH, Souza FVD, Max DAS, Rossi ML, Costa MAPC (2021) Post-seminal development and cryopreservation of endemic or endangered bromeliads. An Acad Bras Ciênc
Smith LB, Downs RJ (1977) Tillandsioideae (Bromeliaceae). Fl Neotrop Monogr 14(2):663–1492
Socolowski F, Vieira DCM, Takaki M (2011) Seed mass of Tecoma stans L. Juss. ex Kunth (Bignoniaceae): effects on emergence and seedling development under full sun and shade. Biota Neotrop 11(2):171–178. https://doi.org/10.1590/S1676-06032011000200017
Souza EH, Souza FVD, Rossi ML, Brancalleão N, Ledo CAS, Martinelli AP (2015) Viability, storage and ultrastructure analysis of Aechmea bicolor (Bromeliaceae) pollen grains, an endemic species to the Atlantic forest. Euphytica 204:13–28. https://doi.org/10.1007/s10681-014-1273-3
Souza FVD, Kaya E, Vieira LJ, Souza EH, Amorim VBO, Skogerboe D, Matsumoto T, Alves AAC, Ledo CAS, Jenderek MM (2016) Droplet-vitrification and morphohistological studies of cryopreserved shoot tips of cultivated and wild pineapple genotypes. Plant Cell Tissue Org Cult 126:351–360. https://doi.org/10.1007/s11240-015-0899-8
Souza EH, Versieux LM, Souza FVD, Rossi ML, Costa MAPC, Martinelli AP (2017) Interspecific and intergeneric hybridization in Bromeliaceae and their relationships to breeding systems. Sci Hortic 221(1):53–61. https://doi.org/10.1016/j.scienta.2017.04.027
Stanwood PC (1985) Cryopreservation of seed germplasm for genetic conservation. In: Kartha K (ed) Cryopreservation of plant cells and organs. CRC Press, Boca Raton, pp 199–225
Tarré E, Pires BBM, Guimarães APM, Carneiro LA, Forzza RC, Mansur E (2007) Germinability after desiccation, storage and cryopreservation of seeds from endemic Encholirium Mart. ex Schult. and Schult. f. and Dyckia Schult. and Schult. f. species (Bromeliaceae). Acta Bot Bras 21(4):777–783. https://doi.org/10.1590/S0102-33062007000400003
Toledo-Aceves T, García-Franco JG, López-Bezerra F (2014) Bromeliad rain: An opportunity for cloud forest management. Forest Ecol Manag 329:129–136. https://doi.org/10.1016/j.foreco.2014.06.022
Vasak V (1969) The viability of bromeliad seeds. Bromeliad Soc Bull 19:102–104
Vettorazzi RG, Carvalho VS, Teixeira MC, Campostrini E, Cunha M, Matos EM, Viccini LF (2019) Cryopreservation of immature and mature seeds of Brazilian orchids of the genus Cattleya. Sci Hortic 256:108603. https://doi.org/10.1016/j.scienta.2019.108603
Vieira DCM, Socolowski F, Takaki M (2007) Germinação de sementes de Dyckia tuberosa (Vell.) Beer (Bromeliaceae) sob diferentes temperaturas em luz e escuro. Braz J Bot 30(2):183–188. https://doi.org/10.1590/S0100-84042007000200003
Villela FA, Peres WB (2004) Coleta, beneficiamento e armazenamento. In: Borghetti F, Ferreira AG (eds) Germinação: do básico ao aplicado. Artmed, Porto Alegre, pp 265–281
Vivian R, Silva AA, Gimenes Junior M, Fagan EB, Ruiz ST, Labonia V (2008) Dormência em sementes de plantas daninhas como mecanismo de sobrevivência: breve revisão. Planta Daninha 26(3):695–706. https://doi.org/10.1590/S0100-83582008000300026
Voronkova NM, Kholina AB, Koldaeva MN, Nakonechnaya OV, Nechaev VA (2018) Morphophysiological dormancy, germination, and cryopreservation in Aristolochia contorta seeds. Plant Ecol Evol 151(1):77–86. https://doi.org/10.5091/plecevo.2018.1351
Walters C, Wheeler L, Stanwood PC (2004) Longevity of cryogenically stored seeds. Cryobiology 48(3):229–244. https://doi.org/10.1016/j.cryobiol.2004.01.007
Wang M, Lambardi M, Engelmann F, Pathirana R, Panis B, Volk GM, Wang QC (2020) Advances in cryopreservation of in vitro-derived propagules: technologies and explant sources. Plant Cell Tiss Organ Cult. https://doi.org/10.1007/s11240-020-01770-0
Wester S, Zotz G (2011) Seed comas of bromeliads promote germination and early seedling growth by wick-like water uptake. J Trop Ecol 27:115–119. https://doi.org/10.1017/S0266467410000593
Zotz G (2013) A longer story than expected: seeds of several species (Tillandsioideae) remain viable for up to two years. J Bromeliad Soc 63(1):83–86
Acknowledgments
This paper is part of RSO MSc dissertation and this study was financed in part by the Fundação de Amparo a Pesquisa do Estado da Bahia (BOL0177/2018) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES)—Finance Code 001. The authors would like to thank the support from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (PROCAD—88887.124186/2014-00). EHS acknowledges the support of CAPES/PNPD/UFRB (88882.315208/2019-01), FVDS currently holds a productivity grant from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq—304269/2018-2).
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RSO, ILS and SOS: data curation, formal analysis, investigation, methodology, visualization, writing—original draft. FVDS and LYSA: conceptualization, funding acquisition, project administration, resources, supervision, writing—review and editing. EHS: conceptualization, formal analysis, funding acquisition, methodology, project administration, resources, supervision, writing—original draft, writing—review and editing.
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de Oliveira, R.S., Souza, F.V.D., dos Santos, I.L. et al. Cryopreservation and low-temperature storage of seeds of Tillandsia species (Bromeliaceae) with ornamental potential. 3 Biotech 11, 186 (2021). https://doi.org/10.1007/s13205-021-02730-x
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DOI: https://doi.org/10.1007/s13205-021-02730-x