Morpho-physiological studies on in vitro germination of Berberis empetrifolia Lam. (Berberidaceae)

Tatiana Carvalho de  Castro; Danúsia da Silva   Santos; Claudia  Simões-Gurgel; Enrique Montiel-Eulefi; Marco Paredes Honorato; Norma Albarello

doi:10.32712/2446-4775.2023.1545

Morpho-physiological studies on in vitro germination of Berberis empetrifolia Lam. (Berberidaceae)

Vol. 17 No. 4 (2023)
490-501
08/02/2023
20/12/2023
https://doi.org/10.32712/2446-4775.2023.1545

Tatiana Carvalho de Castro
Danúsia da Silva Santos
Claudia Simões-Gurgel
Enrique Montiel-Eulefi
Marco Paredes Honorato
Norma Albarello

Keywords

Dormancy

Gibberellic acid

Medicinal plant

Post-seminal development

Pre-germinative treatments

Abstract

Berberis empetrifolia is an endemic species from Chile that produces some bioactive compounds, highlighting the alkaloid berberine, which demonstrated antitumor properties. Taking into account the constant plant supply demand required for medicinal use, the establishment of in vitro propagation protocols allows plant multiplication, without affecting natural populations. Moreover, the in vitro germination is an efficient strategy to obtain axenic plants. The aim of this study was to establish the in vitro germination and to characterize seeds morphology and seedling development of B. empetrifolia. After decontamination of seeds, pre-germinative treatments were applied. Seed viability ranged from 75-90% and the embedding curve revealed permeable integument. The germination was characterized as epigeal phanerocotylar type. Seeds not submitted to pre-germinative treatments reached 10-30% germination rate (GR) and showed asynchronous germination. The most efficient pre-germinative treatment was the storage of seeds at 3˚C for 12 days followed by overnight soaking in GA₃solution associated to seed coat removal. This treatment allowed the seed dormancy to be overcome, reaching GR above 95% and resulting in synchronous germination. The in vitro germination protocol established for B.empetrifolia was efficient to produce phenotypically normal axenic seedlings, allowing their use as source of plant material for biotechnological, phytochemical and pharmacological studies.

References

Khan I, Najeebullah S, Ali M, Shinwari ZK. Phytopharmacological and ethnomedicinal uses of the Genus Berberis (Berberidaceae): a review. Trop J Pharmac Res. 2016; 15(9): 2047-2057. [https://doi.org/10.4314/tjpr.v15i9.33].
Srivastava S, Srivastava M, Misra A, Pandey G, Rawat AKS. A review on biological and chemical diversity in Berberis (Berberidaceae). EXCLI J. 2015; 14: 247-267. [http://dx.doi.org/10.17179/excli2014-399] [https://pubmed.ncbi.nlm.nih.gov/26535033/].
Muñoz MO, Montes M, Wilkomirsky T. Plantas medicinales de uso en Chile: química y farmacología. 2ª edición. Santiago, Chile: Universidad de Chile. Vicerrectoría de Asuntos Académicos:Universitaria. 2004. 330p.
Silva F. Flora agropecuaria de Aysén. Ministerio de Agricultura de Chile. Servicio Agrícola y Ganadero Región de Aysén. 2010. 520p.
Singh R, Kumari P, Kumar S. Nanotechnology for enhanced bioactivity of bioactive phytomolecules. Nutr Deliv. 2017; 1(1): 413-456. [https://doi.org/10.1016/b978-0-12-804304-2.00011-1].
Castro TC, Simões-Gurgel C, Ribeiro IG, Coelho MGP, Albarello N. Morphological aspects of fruits, seeds, seedlings and in vivo and in vitro germination of species of the genus Cleome. J Seed Sci. 2014; 36(3): 326-335. [https://doi.org/10.1590/2317-1545v36n31013].
Baskin JM, Baskin CC. A Classification System for Seed Dormancy. Seed Sci Res. 14(1): 1-16. 2004. [https://doi.org/10.1079/SSR2003150].
Celedón-Neghme C, San Martin LA, Victoriano PF, Cavieres LA. Legitimate seed dispersal by lizards in an alpine habitat: The case of Berberis empetrifolia (Berberidaceae) dispersed by Liolaemus belii (Tropiduridae). Acta Oecologica. 33(1): 265-271. 2008. [https://doi.org/10.1016/j.actao.2007.11.006].
Silva ML, Chagas EA, Villaça R, Smiderle OJ, Moura EA, Chagas PC. Osmopriming duration in Araçá-boi seeds germination. Rev Agro@amb. 2016; 10(1): 17-21. [https://doi.org/10.18227/1982-8470ragro.v10i1.2808].
Barroso GM, Amorim MP, Peixoto AL, Ichaso CLF. Frutos e sementes. Morfologia aplicada à sistemática de dicotiledonôneas. Viçosa, MG: UFV. 2004. 444p.
Brasil. Brasília: Ministério da Agricultura, Pecuária e Abastecimento. Regras para Análise de Sementes. SDA/MAPA/ACS. 2009. 399p.
Lloyd G, Mccown B. Commercially-feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture. Proceedings of the International Plant Propagators’ Society.1981; 30: 421-427.
Labouriau LGA. Germinação das Sementes. Washington: Secretaria-Geral da Organização dos Estados Americanos. 1983. 174p.
Pandey A, Brijwal L, Tamta S. In vitro propagation and phytochemical assessment of Berberis chitria: An important medicinal shrub of Kumaun Himalaya, India. J Medic Pl Res. 2013; 7(15): 930-937. [https://doi.org/10.5897/JMPR13.4435].
Ribeiro MNO, Pasqual M, Villa F, Pio LAS, Hilhorst HWM. In vitro seed germination and seedling development of Annona crassiflora Mart. Sci Agricol. 2009;66(3): 410-413. [https://doi.org/10.1590/S0103-90162009000300017].
Li H, Zhang D. In vitro seed germination of Kalmia latifolia L. hybrids: a means for improving germination and speeding up breeding cycle. Hortscience. 2018; 53(4): 535–540. [https://doi.org/10.21273/HORTSCI12829-17].
Graeber K, Nakabayashi K, Miatton E, Leubner-Metzger G, Soppe WJJ. Molecular mechanisms of seed dormancy. Pl Cell Environment. 2012; 35(10): 1769-1786. [https://doi.org/10.1111/j.1365-3040.2012.02542.x].
Nikolaeva MG, Vorobèva NS. The role of abscisic acid and indolic compoundsin dormancy of the seeds of ash species. Soviet Pl Physiol (USA).1979; 26(12): 105-113. ISSN 0038-5719.
Baskin CC, Baskin JM. Seeds. Ecology, biogeography, and evolution of dormancy and germination. 1sted. San Diego: Academic Press. 2001. 666p.
Baskin CC, Baskin JM. Seeds: Ecology, biogeography, and evolution of dormancy and germination. 2nded. Kentucky, USA. 2014. 1601p.
Brijwal L, Pandey A, Tamta S. In vitro propagation of the endangered species Berberis aristata DC. via leaf-derived callus. In Vitro Cellular Develop Biol– Plant. 2015; 51(6):637-647. [https://doi.org/10.1007/s11627-015-9716-7].
Rhie YH, Lee SY, Kim KS. Seed dormancy and germination in Jeffersonia dubia (Berberidaceae) as affected by temperature and gibberellic acid. Plant Biology 2014;17: 327-334. [https://doi.org/10.1111/plb.12235].
Yamauchi Y, Ogawa M, Kuwahara A, Hanada A, Kamiya Y, Yamaguchi S. Activation of gibberellin biosynthesis and response pathways by low temperature during imbibition of Arabidopsis thaliana seeds. Plant Cell. 2004; 16(2): 367-378. [https://doi.org/10.1105/tpc.018143].
Bazin J, Batlla D, Dussert S, El-Maarouf-Bouteau H, Bailly C. Role of relative humidity, temperature, and water status in dormancy alleviation of sunflower seeds during dry after-ripening. J Experim Bot. 2011; 62(2): 627-640. [https://doi.org/10.1093/jxb/erq314].
Bewley JD, Black M. Seeds. Physiology, development and germination.2nd edition. New York, Plenum Press. 1994. 445p. [https://doi.org/10.1007/978-1-4899-1002-8].
Kim DH. Practical methods for rapid seed germination from seed coat-imposed dormancy of Prunus yedoensis. Scientia Horticulturae. 2019; 243: 451-456. [https://doi.org/10.1016/j.scienta.2018.08.039].
Tewari B, Tewari A. Seed germination in Prunus cerasoides influenced by natural seed desiccation and varying temperature in Central Himalayan region of Uttarakhand, India. Inter J Bioassays. 2016; 5(5): 4567-4572. [https://doi.org/10.21746/ijbio.2016.05.007].
Muralidhara BM, Reddy YTN, Srilatha V, Akshitha HJ. Effect of seed coat removal treatments on seed germination and seedling attributes in mango varieties. Inter J Fruit Sci. 2016; 16(1): 1-9. [https://doi.org/10.1080/15538362.2015.1021885].
Chen S, Kuo S, Chien C. Roles of gibberellins and abscisic acid in dormancy and germination of red bayberry (Myrica rubra) seeds. Tree Physiol. 2008; 28(9): 1431-1439. [https://doi.org/10.1093/treephys/28.9.1431].
Webster RE, Waterworth W.M, Stuppy W, West CE, Ennos R, Bray CM, Pritchard HW. Biomechanical, biochemical, and morphological mechanisms of heat shock-mediated germination in Carica papaya seed. J Experim Bot. 2016;67(22): 6373-6384. [https://doi.org/10.1093/jxb/erw402].

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Author(s)

Tatiana Carvalho de Castro
Rio de Janeiro State University (UERJ), Roberto Alcantara Gomes Institute of Biology (IBRAG), Plant Biotechnology Laboratory (Labplan), Rua São Francisco Xavier, 524, PHLC, Maracanã, CEP 20550-013, Rio de Janeiro, Brazil.
https://orcid.org/0000-0003-0554-6873
Danúsia da Silva Santos
Rio de Janeiro State University (UERJ), Roberto Alcantara Gomes Institute of Biology (IBRAG), Plant Biotechnology Laboratory (Labplan), Rua São Francisco Xavier - from 280 to 530 - even side, Maracanã, CEP 20550-013, Rio de Janeiro, RJ, Brazil.
https://orcid.org/0000-0003-4061-8086
Claudia Simões-Gurgel
Rio de Janeiro State University (UERJ), Roberto Alcantara Gomes Institute of Biology (IBRAG), Plant Biotechnology Laboratory (Labplan), Rua São Francisco Xavier - from 280 to 530 - even side, Maracanã, CEP 20550-013, Rio de Janeiro, RJ, Brazil.
https://orcid.org/0000-0001-8358-2568
Enrique Montiel-Eulefi
Universidade de La Frontera (UFRO), Faculty of Medicine, Department of Basic Sciences, Claro Solar, 180, Temuco, Chile.
https://orcid.org/0000-0002-0982-444X
Marco Paredes Honorato
Universidade de La Frontera (UFRO), Faculty of Medicine, Department of Basic Sciences, Claro Solar, 180, Temuco, Chile.
https://orcid.org/0000-0003-3538-6954
Norma Albarello
Rio de Janeiro State University (UERJ), Roberto Alcântara Gomes Institute of Biology (IBRAG), Plant Biotechnology Laboratory (Labplan), Rua São Francisco Xavier, 524, PHLC, Maracanã, CEP 20550-013, Rio de Janeiro, Brazil.
https://orcid.org/0000-0001-5803-2070

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How to Cite

Morpho-physiological studies on in vitro germination of Berberis empetrifolia Lam. (Berberidaceae). Rev Fitos [Internet]. 2023 Dec. 20 [cited 2026 Mar. 3];17(4):490-501. Available from: https://revistafitos.far.fiocruz.br/index.php/revista-fitos/article/view/1545

This work is licensed under a Creative Commons Attribution 4.0 International License.

[1] Khan I, Najeebullah S, Ali M, Shinwari ZK. Phytopharmacological and ethnomedicinal uses of the Genus Berberis (Berberidaceae): a review. Trop J Pharmac Res. 2016; 15(9): 2047-2057. [https://doi.org/10.4314/tjpr.v15i9.33].

[2] Srivastava S, Srivastava M, Misra A, Pandey G, Rawat AKS. A review on biological and chemical diversity in Berberis (Berberidaceae). EXCLI J. 2015; 14: 247-267. [http://dx.doi.org/10.17179/excli2014-399] [https://pubmed.ncbi.nlm.nih.gov/26535033/].

[3] Muñoz MO, Montes M, Wilkomirsky T. Plantas medicinales de uso en Chile: química y farmacología. 2ª edición. Santiago, Chile: Universidad de Chile. Vicerrectoría de Asuntos Académicos:Universitaria. 2004. 330p.

[4] Silva F. Flora agropecuaria de Aysén. Ministerio de Agricultura de Chile. Servicio Agrícola y Ganadero Región de Aysén. 2010. 520p.

[5] Singh R, Kumari P, Kumar S. Nanotechnology for enhanced bioactivity of bioactive phytomolecules. Nutr Deliv. 2017; 1(1): 413-456. [https://doi.org/10.1016/b978-0-12-804304-2.00011-1].

[6] Castro TC, Simões-Gurgel C, Ribeiro IG, Coelho MGP, Albarello N. Morphological aspects of fruits, seeds, seedlings and in vivo and in vitro germination of species of the genus Cleome. J Seed Sci. 2014; 36(3): 326-335. [https://doi.org/10.1590/2317-1545v36n31013].

[7] Baskin JM, Baskin CC. A Classification System for Seed Dormancy. Seed Sci Res. 14(1): 1-16. 2004. [https://doi.org/10.1079/SSR2003150].

[8] Celedón-Neghme C, San Martin LA, Victoriano PF, Cavieres LA. Legitimate seed dispersal by lizards in an alpine habitat: The case of Berberis empetrifolia (Berberidaceae) dispersed by Liolaemus belii (Tropiduridae). Acta Oecologica. 33(1): 265-271. 2008. [https://doi.org/10.1016/j.actao.2007.11.006].

[9] Silva ML, Chagas EA, Villaça R, Smiderle OJ, Moura EA, Chagas PC. Osmopriming duration in Araçá-boi seeds germination. Rev Agro@amb. 2016; 10(1): 17-21. [https://doi.org/10.18227/1982-8470ragro.v10i1.2808].

[10] Barroso GM, Amorim MP, Peixoto AL, Ichaso CLF. Frutos e sementes. Morfologia aplicada à sistemática de dicotiledonôneas. Viçosa, MG: UFV. 2004. 444p.

[11] Brasil. Brasília: Ministério da Agricultura, Pecuária e Abastecimento. Regras para Análise de Sementes. SDA/MAPA/ACS. 2009. 399p.

[12] Lloyd G, Mccown B. Commercially-feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture. Proceedings of the International Plant Propagators’ Society.1981; 30: 421-427.

[13] Labouriau LGA. Germinação das Sementes. Washington: Secretaria-Geral da Organização dos Estados Americanos. 1983. 174p.

[14] Pandey A, Brijwal L, Tamta S. In vitro propagation and phytochemical assessment of Berberis chitria: An important medicinal shrub of Kumaun Himalaya, India. J Medic Pl Res. 2013; 7(15): 930-937. [https://doi.org/10.5897/JMPR13.4435].

[15] Ribeiro MNO, Pasqual M, Villa F, Pio LAS, Hilhorst HWM. In vitro seed germination and seedling development of Annona crassiflora Mart. Sci Agricol. 2009;66(3): 410-413. [https://doi.org/10.1590/S0103-90162009000300017].

[16] Li H, Zhang D. In vitro seed germination of Kalmia latifolia L. hybrids: a means for improving germination and speeding up breeding cycle. Hortscience. 2018; 53(4): 535–540. [https://doi.org/10.21273/HORTSCI12829-17].

[17] Graeber K, Nakabayashi K, Miatton E, Leubner-Metzger G, Soppe WJJ. Molecular mechanisms of seed dormancy. Pl Cell Environment. 2012; 35(10): 1769-1786. [https://doi.org/10.1111/j.1365-3040.2012.02542.x].

[18] Nikolaeva MG, Vorobèva NS. The role of abscisic acid and indolic compoundsin dormancy of the seeds of ash species. Soviet Pl Physiol (USA).1979; 26(12): 105-113. ISSN 0038-5719.

[19] Baskin CC, Baskin JM. Seeds. Ecology, biogeography, and evolution of dormancy and germination. 1sted. San Diego: Academic Press. 2001. 666p.

[20] Baskin CC, Baskin JM. Seeds: Ecology, biogeography, and evolution of dormancy and germination. 2nded. Kentucky, USA. 2014. 1601p.

[21] Brijwal L, Pandey A, Tamta S. In vitro propagation of the endangered species Berberis aristata DC. via leaf-derived callus. In Vitro Cellular Develop Biol– Plant. 2015; 51(6):637-647. [https://doi.org/10.1007/s11627-015-9716-7].

[22] Rhie YH, Lee SY, Kim KS. Seed dormancy and germination in Jeffersonia dubia (Berberidaceae) as affected by temperature and gibberellic acid. Plant Biology 2014;17: 327-334. [https://doi.org/10.1111/plb.12235].

[23] Yamauchi Y, Ogawa M, Kuwahara A, Hanada A, Kamiya Y, Yamaguchi S. Activation of gibberellin biosynthesis and response pathways by low temperature during imbibition of Arabidopsis thaliana seeds. Plant Cell. 2004; 16(2): 367-378. [https://doi.org/10.1105/tpc.018143].

[24] Bazin J, Batlla D, Dussert S, El-Maarouf-Bouteau H, Bailly C. Role of relative humidity, temperature, and water status in dormancy alleviation of sunflower seeds during dry after-ripening. J Experim Bot. 2011; 62(2): 627-640. [https://doi.org/10.1093/jxb/erq314].

[25] Bewley JD, Black M. Seeds. Physiology, development and germination.2nd edition. New York, Plenum Press. 1994. 445p. [https://doi.org/10.1007/978-1-4899-1002-8].

[26] Kim DH. Practical methods for rapid seed germination from seed coat-imposed dormancy of Prunus yedoensis. Scientia Horticulturae. 2019; 243: 451-456. [https://doi.org/10.1016/j.scienta.2018.08.039].

[27] Tewari B, Tewari A. Seed germination in Prunus cerasoides influenced by natural seed desiccation and varying temperature in Central Himalayan region of Uttarakhand, India. Inter J Bioassays. 2016; 5(5): 4567-4572. [https://doi.org/10.21746/ijbio.2016.05.007].

[28] Muralidhara BM, Reddy YTN, Srilatha V, Akshitha HJ. Effect of seed coat removal treatments on seed germination and seedling attributes in mango varieties. Inter J Fruit Sci. 2016; 16(1): 1-9. [https://doi.org/10.1080/15538362.2015.1021885].

[29] Chen S, Kuo S, Chien C. Roles of gibberellins and abscisic acid in dormancy and germination of red bayberry (Myrica rubra) seeds. Tree Physiol. 2008; 28(9): 1431-1439. [https://doi.org/10.1093/treephys/28.9.1431].

[30] Webster RE, Waterworth W.M, Stuppy W, West CE, Ennos R, Bray CM, Pritchard HW. Biomechanical, biochemical, and morphological mechanisms of heat shock-mediated germination in Carica papaya seed. J Experim Bot. 2016;67(22): 6373-6384. [https://doi.org/10.1093/jxb/erw402].