Biologia plantarum 64: 224-233, 2020 | DOI: 10.32615/bp.2020.006

Induction of somatic embryogenesis and evaluation of genetic stability in regenerated plants of Magnolia dealbata

A. CHÁVEZ-CORTAZAR1*, M. MATA-ROSAS1, K. OYAMA2, M.S. SAMAIN3, M. QUESADA2
1 Instituto de Ecología, A.C., Xalapa, Veracruz 91073, Mexico
2 Escuela Nacional de Estudios Superiores Unidad Morelia, Universidad Nacional Autónoma de México, Morelia, Michoacán 58190, Mexico
3 Instituto de Ecología, A.C., Centro Regional del Bajío, Pátzcuaro, Michoacán 61600, Mexico

The utility of plant tissue culture for the mass propagation of trees is well known, but continuous in vitro multiplication of plant material may increase the possibility of somaclonal variation; therefore, it is essential to evaluate the genetic integrity of regenerants from species-specific in vitro protocols prior to mass production and implementation. The objectives of this study were: 1) to determine the effect of 2,4-dichlorophenoxyacetic acid (2,4-D) concentration over two cycles of secondary somatic embryogenesis in Magnolia dealbata; and 2) to verify the genetic stability of the regenerants obtained. The embryogenic response was not significantly affected by the concentration of 2,4-D but did vary across cycles of induction. The addition of 4.52 μM 2,4-D induced the highest total number of embryos (100.5), the mean number of somatic embryos (25.1) and somatic embryos per explant (80.6). In both 2,4-D concentration (2.26 or 4.52 μM), genetic integrity between the donor and the propagated clones was 0.90, and the low genetic instability (≤ 0.10 in both PGR treatments) might be due to effect of cyclic somatic embryogenesis or the different response of the explants at stress in in vitro culture conditions. However, it is necessary to examine more cell lines and somatic embryogenesis cycles.

Keywords: 2,4-dichlorophenoxyacetic acid, in vitro culture, simple sequence repeats, somaclonal variation.

Received: August 5, 2019; Revised: January 12, 2020; Accepted: January 14, 2020; Published online: March 11, 2020Show citation

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CHÁVEZ-CORTAZAR, A., MATA-ROSAS, M., OYAMA, K., SAMAIN, M.S., & QUESADA, M. (2020). Induction of somatic embryogenesis and evaluation of genetic stability in regenerated plants of Magnolia dealbata. Biologia plantarum, 64, 224-233. doi: 10.32615/bp.2020.006.
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    References

    1. Alonso-Castro, A.J., Domínguez, F., García-Regalado, A., González-Sánchez, M.A.C., García-Carrancá, A.: Magnolia dealbata seeds extract exert cytotoxic and chemopreventive effects on MDA-MB231 breast cancer cells. - Pharm Biol. 52: 621-627, 2014. Go to original source...
    2. Bhattacharyya, P., Kumar, V., Staden, J.V.: Assessment of genetic stability amongst micropropagated Ansellia africana, a vulnerable medicinal orchid species of Africa using SCoT markers. - South Afr. J. Bot. 108: 294-302, 2017.
    3. Bobadilla, L.R., Cenci, A., Georget, F., Bertrand, B., Camayo, G., Dechamp, E., Herrera, J.C., Santoni, S., Lashermes, P., Simpson, J., Etienne, H.: High genetic and epigenetic stability in Coffea arabica plants derived from embryogenic suspensions and secondary embryogenesis as revealed by AFLP, MSAP and the phenotypic variation rate. - PLoS ONE 8: e56372, 2013. Go to original source...
    4. Bradaï, F., Pliego-Alfaro, F., Sánchez-Romero, C.: Somaclonal variation in olive (Olea europaea L.) plants regenerated via somatic embryogenesis: influence of genotype and culture age on phenotypic stability. - Sci. Hort. 213: 208-215, 2016. Go to original source...
    5. Burg, K., Helmersson, A., Bozhkov, P., Von Arnold, S.: Developmental and genetic variation in nuclear microsatellite stability during somatic embryogenesis in pine. - J. exp. Bot. 58: 687-698, 2007.
    6. Carloni, E., Ribotta, A., López, C.E., Griffa, S., Quirog, M., Tommasino, E., Grunberg, K.: Somatic embryogenesis from in vitro anther culture of apomictic buffel grass genotypes and analysis of regenerated plants using flow cytometry .- Plant Cell Tissue Organ Cult. 117: 311-322, 2014. Go to original source...
    7. Carra, A., Sajeva, M., Abbate, L., Siragusa, M., Pathirana, R., Carimi, F.: Factors affecting somatic embryogenesis in eight Italian grapevine cultivars and the genetic stability of embryo-derived regenerants as assessed by molecular markers. - Sci. Hort. 204: 123-127, 2016. Go to original source...
    8. Chapuis, M.-P., Estoup, A.: Microsatellite null alleles and estimation of population differentiation. - Mol. Biol. Evol. 24: 621-631, 2007. Go to original source...
    9. Clarindo, W.R., Carvalho, C.R., Mendonça, M.A.C.: Ploidy instability in long-term in vitro cultures of Coffea arabica L. monitored by flow cytometry. - Plant Growth Regul. 68: 533-538, 2012. Go to original source...
    10. Clarindo, W.R., De Carvalho, C.R., Araújo, F.S., De Abreu, I.S., Otoni, W.C.: Recovering polyploid papaya in vitro regenerants as screened by flow cytometry. - Plant Cell Tissue Organ Cult. 92: 207-214, 2008. Go to original source...
    11. Corral-Aguirre, J., Sánchez-Velásquez, L.R.: Seed ecology and germination treatments in Magnolia dealbata: an endangered species. - Flora 201: 227-232, 2006. Go to original source...
    12. DeVerno, L.L., Park, Y.S., Bonga, J.M., Barrett, J.D.: Somaclonal variation in cryopreserved embryogenic clones of white, spruce [Picea glauca (Moench) Voss.]. - Plant Cell Rep. 18: 948-953, 1999. Go to original source...
    13. Dey, T., Saha, S., Ghosh, P.D.: Somaclonal variation among somatic embryo derived plants - evaluation of agronomically important somaclones and detection of genetic changes by RAPD in Cymbopogon winterianus. - South Afr. J. Bot. 96: 112-121, 2015. Go to original source...
    14. Domínguez, F., Chávez, M., Garduño-Ramírez, M.L., Chávez-Ávila, V.M., Mata, M., Cruz-Sosa, F.: Honokiol and magnolol production by in vitro micropropagated plants of Magnolia dealbata, an endangered endemic mexican species. - Nat. Prod. Commun. 5: 235-240, 2010. Go to original source...
    15. Domínguez, F., Chávez, M., Garduño-Ramírez, M.L., Chávez-Ávila, V.M., Mata, M., Cruz-Sosa, F.: Production of honokiol and magnolol in suspension cultures of Magnolia dealbata Zucc. - Nat. Prod. Commun. 4: 939-943, 2009. Go to original source...
    16. Doyle, J.J., Doyle, J.L.: A rapid DNA isolation procedure for small quantities of fresh leaf material. - Phytochem. Bull. 19: 11-15, 1987.
    17. Endemann, M., Hristoforoglu, K., Stauber, T., Wilhelm, E.: Assessment of age-related polyploidy in Quercus robur L. somatic embryos and regenerated plants using DNA flow cytometry. - Biol. Plant. 44: 339-345, 2001. Go to original source...
    18. Flores-Estévez, N., Vasquez-Morales, S.G., Cano-Medina, T., Sánchez-Velásquez, L.R., Noa-Carrazana, J.C., Díaz-Fleischer, F.: Insecticidal activity of raw ethanolic extracts from Magnolia dealbata Zucc on a tephritid pest. - J. Environ. Sci. Health, Part B 48: 582-586, 2013. Go to original source...
    19. Fourré, J.-L., Berger, P., Niquet, L., André, P.: Somatic embryogenesis and somaclonal variation in Norway spruce: morphogenetic, cytogenetic and molecular approaches. - Theor. appl. Genet. 94: 159-169, 1997. Go to original source...
    20. Gaj, M.D.: Direct somatic embryogenesis as a rapid and efficient system for in vitro regeneration of Arabidopsis thaliana. - Plant Cell Tissue Organ Cult. 64: 39-46, 2001. Go to original source...
    21. Gaj, M.D.: Factors influencing somatic embryogenesis induction and plant regeneration with particular reference to Arabidopsis thaliana (L.) Heynh. - Plant Growth Regul. 43: 27-47, 2004. Go to original source...
    22. Giridhar, P., Kumar, V., Ravishankar, G.A.: Somatic embryogenesis, organogenesis, and regeneration from leaf callus culture of Decalepis hamiltonii Wight & Arn., an endangered shrub. - In Vitro cell. dev. Biol. Plant 40: 567-571, 2004. Go to original source...
    23. Gutierrez, L., Vovides, A.P.: An in situ study of Magnolia dealbata Zucc. in Veracruz state: and endangered endemic tree of Mexico. - Biodivers. Conserv. 6: 89-97, 1997. Go to original source...
    24. Harvengt, L., Trontin, J.F., Reymond, I., Canlet, F., Paques, M.: Molecular evidence of true-to-type propagation of a 3-year-old Norway spruce through somatic embryogenesis. - Planta 213: 828-832, 2001. Go to original source...
    25. Henao, R.A.M., De la Hoz, V.T., Ospina, O.T.M. Atehortúa, G.L., Urrea, T.A.I.: Evaluation of the potential of regeneration of different Colombian and commercial genotypes of cocoa (Theobroma cacao L.) via somatic embryogenesis. - Sci. Hort. 229: 148-156, 2018. Go to original source...
    26. Kumari, S., Yadav, K., Singh, N.: Evaluation of genetic fidelity among micropropagated plants of Salvadora persica L. using DNA-based markers. - Meta Gene 14: 129-133, 2017. Go to original source...
    27. Li, X., Wang, X., Luan, C., Yang, J., Cheng, S., Dai, Z., Mei, P., Huang, C.: Somatic embryogenesis from mature zygotic embryos of Distylium chinense (Fr.) Diels and assessment of genetic fidelity of regenerated plants by SRAP markers. - Plant Growth Regul. 74: 11-21, 2014. Go to original source...
    28. Lloyd, G., McCown, B.: Commercially-feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture. - Int. Plant Propag. Soc. Proc. 30: 421-427, 1980.
    29. Lopes, T., Capelo, A., Brito, G., Loureiro, J., Santos, C.: Genetic variability analyses of the somatic embryogenesis induction process in Olea spp. using nuclear microsatellites. - Trees - Struct. Funct. 23: 29-36, 2009. Go to original source...
    30. Lopes, T., Pinto, G., Loureiro, J., Costa, A., Santos, C.: Determination of genetic stability in long-term somatic embryogenic cultures and derived plantlets of cork oak using microsatellite markers. - Tree Physiol. 26: 1145-1152, 2006. Go to original source...
    31. López, C.M.R., Wetten, A.C., Wilkinson, M.J.: Detection and quantification of in vitro-culture induced chimerism using simple sequence repeat (SSR) analysis in Theobroma cacao (L.). - Theor. appl. Genet. 110: 157-166, 2004.
    32. Mamedes-Rodrigues, T.C., Batista, D.S., Vieira, N.M., Matos, E.M., Fernandes, D., Nunes-Nesi, A., Cruz, C.D., Viccini, L.F., Nogueira, F.T.S., Otoni, W.C.: Regenerative potential, metabolic profile, and genetic stability of Brachypodium distachyon embryogenic calli as affected by successive subcultures. - Protoplasma 255: 655-667, 2018. Go to original source...
    33. Martínez, A.L., Domínguez, F., Orozco, S. Chávez, M., Salgado, H., González, M., González-Trujano, M.E.: Neuropharmacological effects of an ethanol extract of the Magnolia dealbata Zucc. leaves in mice. - J. Ethnopharmacol. 106: 250-255, 2006. Go to original source...
    34. Marum, L., Rocheta, M., Maroco, J., Oliveira, M.M., Miguel, C.: Analysis of genetic stability at SSR loci during somatic embryogenesis in maritime pine (Pinus pinaster). - Plant Cell Rep. 28: 673-682, 2009. Go to original source...
    35. Mata-Rosas, M., Jiménez-Rodríguez, Á., Chávez-Avila, V.M.: Somatic embryogenesis and organogenesis in Magnolia dealbata Zucc. (Magnoliaceae), an endangered, endemic mexican species. - HortScience 41: 1325-1329, 2006. Go to original source...
    36. Mehta, R., Sharma, V., Sood, A., Sharma, M., Kumar, S.R.: Induction of somatic embryogenesis and analysis of genetic fidelity of in vitro-derived plantlets of Bambusa nutans Wall., using AFLP markers. - Eur. J. Forest Res. 130: 729-736, 2011. Go to original source...
    37. Molina, D.M., Aponte, M.E., Cortina, H., Moreno, G.: The effect of genotype and explant age on somatic embryogenesis of coffee. - Plant Cell Tissue Organ Cult. 71: 117-123, 2002. Go to original source...
    38. Morais-Lino, L.S., Santos-Serejo, J.A., Amorim, E.P., Ferreira, de S.??, J.R., Pasqual, M., De Oliveira, S.S.: Somatic embryogenesis, cell suspension, and genetic stability of banana cultivars. - In vitro cell. dev. Biol. Plant 52: 99-106, 2016. Go to original source...
    39. Müller, E., Brown, P.T.H., Hartke, S., Lörz, H.: DNA variation in tissue-culture-derived rice plants. - Theor. appl. Genet. 80: 673-679, 1990. Go to original source...
    40. Naz, R., Anis, M., Alatar, A.A.: ISSR marker-based detection of genomic stability in Cassia occidentalis L. plantlets derived from somatic embryogenesis. - Eng. Life Sci. 16: 17-24, 2016. Go to original source...
    41. Nehra, N.S., Kartha, K.K., Stushnoff, C., Giles, K.L.: The influence of plant growth regulator concentrations and callus age on somaclonal variation in callus culture regenerants of strawberry. - Plant Cell Tissue Organ Cult. 29: 257-268, 1992. Go to original source...
    42. Niazian, M., Sadat, N.S.A., Galuszka, P., Tohidfar, M., Mohammad, S., Mortazavian, M.: Genetic stability of regenerated plants via indirect somatic embryogenesis and indirect shoot regeneration of Carum copticum L. - Ind. Crop. Prod. 97: 330-337, 2017. Go to original source...
    43. Pavlović, S., Vinterhalter, B., Zdravković-Koraé, S., Vinterhalter, D., Zdravković, J., Cvikić, D., Mitić, N.: Recurrent somatic embryogenesis and plant regeneration from immature zygotic embryos of cabbage (Brassica oleracea var . capitata ) and cauliflower (Brassica oleracea var . botrytis ). - Plant Cell Tissue Organ Cult. 113: 397-406, 2013. Go to original source...
    44. Prado, M.J., Rodriguez, E., Rey, L., González, M.V., Santos, C., Rey, M.: Detection of somaclonal variants in somatic embryogenesis-regenerated plants of Vitis vinifera by flow cytometry and microsatellite markers. - Plant Cell Tissue Organ Cult. 103: 49-59, 2010. Go to original source...
    45. Priyono, Florin, B., Rigoreau, M., Ducos, J.P., Sumirat, U., Mawardi, S., Lambot, C., Broun, P., Pétiard, V., Wahyudi, T., Crouzillat, D.: Somatic embryogenesis and vegetative cutting capacity are under distinct genetic control in Coffea canephora Pierre. - Plant Cell Rep. 29: 343-357, 2010. Go to original source...
    46. Raemakers, C.J.J.M., Jacobsen, E., Visser, R.G.F.: Secondary somatic embryogenesis and applications in plant breeding. - Euphytica 81: 93-107, 1995. Go to original source...
    47. Rai, M.K., Phulwaria, M., Harish, Gupta, A.K., Shekhawat, N.S., Jaiswal, U.: Genetic homogeneity of guava plants derived from somatic embryogenesis using SSR and ISSR markers. - Plant Cell Tissue Organ Cult. 111: 259-264, 2012. Go to original source...
    48. Raji, M.R., Lotfi, M., Tohidfar, M., Zahedi, B., Carra, A., Abbate, L., Carimi, F.: Somatic embryogenesis of muskmelon (Cucumis melo L.) and genetic stability assessment of regenerants using flow cytometry and ISSR markers. - Protoplasma 255: 873-883, 2018. Go to original source...
    49. Raymond, M., Rousset, F.: GENEPOP (version 1.2): population genetics software for exact test and ecumenicism. - J. Hered. 86: 248-249, 1995. Go to original source...
    50. Remakanthan, A., Menon, T.G., Soniya, E.V.: Somatic embryogenesis in banana (Musa acuminata AAA cv. Grand Naine): Effect of explant and culture conditions. - In vitro cell. dev. Biol. Plant 50: 127-136, 2014. Go to original source...
    51. Rivers, M., Beech, E., Murphy, L., Oldfield, S. (ed.): The Red List of Magnoliaceae - Revised and Extended. - BGCI, Richmond, 2016.
    52. Sánchez-Velásquez, L.R., Pineda-López, M. del R.: Comparative demographic analysis in contrasting environments of Magnolia dealbata: An endangered species from Mexico. - Popul. Ecol. 52: 203-210, 2010. Go to original source...
    53. Sedov, K.A., Fomenkov, A.A., Solov'yova, A.I., Nosov, A.V., Dolgikh, Y.I.: The level of genetic variability of cells in prolonged suspension culture of Arabidopsis thaliana. - Biol. Bull. 41: 493-499, 2014. Go to original source...
    54. Selkoe, K.A., Toonen,R.J.: Microsatellites for ecologists: a practical guide to using and evaluating microsatellite markers. - Ecol. Lett. 9: 615-629, 2006. Go to original source...
    55. Shahzad, A., Parveen, S., Sharma, S., Shaheen, A., Saeed, T., Yadav, V., Akhtar, R., Ahmad, Z., Upadhyay, A.: Plant tissue culture: applications in plant improvement and conservation. - In: Abdin, Z.M., Kiran, U., Kamauddin, Ali, A. (ed.): Plant Biotechnology: Principles and Applications. Pp 3-72. Springer, Singapure 2017. Go to original source...
    56. Sharma, S.K., Bryan, G.J., Winfield, M.O., Millam, S.: Stability of potato (Solanum tuberosum L.) plants regenerated via somatic embryos, axillary bud proliferated shoots, microtubers and true potato seeds: a comparative phenotypic, cytogenetic and molecular assessment. - Planta 226: 1449-1458, 2007. Go to original source...
    57. Soares, D.M.M., Sattler, M.C., Ferreira, M.F. da S., Praça-Fontes, M.M.: Assessment of genetic stability in three generations of in vitro propagated Jatropha curcas L. plantlets using ISSR markers .- Trop. Plant Biol. 9: 229-238, 2016. Go to original source...
    58. Tremblay, L., Levasseur, C., Tremblay, F.M.: Frequency of somaclonal variation in plants of black spruce (Picea mariana, Pinaceae) and white spruce (P. glauca, Pinaceae) derived from somatic embryogenesis and identification of some factors involved in genetic instability. - Amer. J. Bot. 86: 1373-1381, 1999. Go to original source...
    59. Triqui, Z.E.A., Guédira, A., Chlyah, A., Chlyah, H., Souvannavong, V., Haïcour, R., Sihachakr, D.: Effect of genotype, gelling agent, and auxin on the induction of somatic embryogenesis in sweet potato (Ipomoea batatas Lam.). - Compt. rend. Biol. 331: 198-205, 2008. Go to original source...
    60. Valladares, S., Sánchez, C., Martínez, M.T., Ballester, A., Vieitez, A.: Plant regeneration through somatic embryogenesis from tissues of mature oak trees: true-to-type conformity of plantlets by RAPD analysis. - Plant Cell Rep. 25: 879-886, 2006. Go to original source...
    61. Van Oosterhout, C., Hutchinson, W.F., Wills, D.P.M., Shipley, P.: MICRO-CHECKER: software for identifying and correcting genotyping errors in microsatellite data.- Mol. Ecol. Notes 4: 535-538, 2004. Go to original source...
    62. Vasic, D., Alibert, G., Skoric, D.: Protocols for efficient repetitive and secondary somatic embryogenesis in Helianthus maximiliani (Schrader). - Plant Cell Rep. 20: 121-125, 2001. Go to original source...
    63. Velazco-Macías, C.G., Foroughbakhch-Pournavab, R., Alanís-Flores, G.J., Alvarado-Vázquez, M.A.: Magnolia dealbata in Nuevo León, México. - Rev. Mex. Biodivers. 79: 459-463, 2008.
    64. Veltjen, E., Asselman, P., Hernández, R.M., Palmarola, B.A., Testé, L.E., González, T.L.R., Goetghebeur, P., Larridon, I., Samain, M.-S.: Genetic patterns in neotropical magnolias (Magnoliaceae) using de novo developed microsatellite markers. -Heredity 122: 485-500, 2019. Go to original source...
    65. Vinoth, A., Ravindhran, R.: Efficient plant regeneration of watermelon (Citrullus lanatus Thunb.) via somatic embryogenesis and assessment of genetic fidelity using ISSR markers. - In Vitro cell. dev. Biol. Plant 52: 107-115, 2016. Go to original source...
    66. Wilhelm, E., Hristoforoglu, K., Fluch, S., Burg, K.: Detection of microsatellite instability during somatic embryogenesis of oak (Quercus robur L.). - Plant Cell Rep. 23: 790-795, 2005. Go to original source...
    67. Yang, X.M., An, L.Z., Xiong, Y.C., Zhang, J.P., Li, Y., Xu, S.J.: Somatic embryogenesis from immature zygotic embryos and monitoring the genetic fidelity of regenerated plants in grapevine. - Biol. Plant. 52: 209-214, 2008. Go to original source...
    68. Zoghlami, N., Mliki, A., Ghorbel, A.: Evaluation of genetic diversity among Tunisian grapevines by RAPD markers. - Vitis 40: 31-37, 2001.

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