Effect of different sources and concentrations of carbohydrates on the morphological and physiological characteristics of Vanilla planifolia Jacks. RH350
Palavras-chave:
Sources of carbon; micropropagation; tissue culture; in vitro, photosynthetic pigments; orchids.Resumo
Veracruz is the leading vanilla-producing state in Mexico; however, its plantations face significant challenges from pathogens like Fusarium oxysporum f. sp. vanillae (Fov). Therefore, it is crucial to develop an effective protocol to ensure the availability of in vitro plants of the RH350 genotype, which exhibits 50% resistance to Fov. This genotype was developed through in vitro selection against Fov filtrates using the pathogenic strain Fov H₃. This study evaluated the effects of two concentrations (15 and 30 g L⻹) and five different carbon sources (maltose, fructose, sucrose, galactose, and glucose) on the morphological (number of shoots, leaves, and roots, shoot, leaf, and root length) and physiological (photosynthetic pigment content) characteristics of in vitro plants. This study was conducted to meet the growing demand for high-quality RH350 plants. The results revealed significant differences among the carbohydrates and their respective concentrations; however, the 30 g L⻹ concentration of fructose yielded the greatest number of shoots and leaves. A similar concentration of maltose positively impacted shoot length, while 15 g L⻹ of this carbohydrate yielded the best results in terms of chlorophyll content. These findings emphasize the importance of both the concentration and the type of carbon source in micropropagating plants such as V. planifolia, RH350.
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Referências
Abdelsalam, A., Chowdhury, K., & El Bakry, A. (2018). Efficient adventitious morphogenesis from in vitro cultures of the medicinal plant Cymbopogon schoenanthus. Plant Tissue Culture and Biotechnology, 28(2), 147-160. https://doi.org/10.3329/ptcb.v28i2.3967
Akyüz, B. (2025). Effect of different carbon sources and concentrations on in vitro propagation of chestnut. Plant Cell, Tissue and Organ Culture (PCTOC), 160(2), 25. https://doi.org/10.1007/s11240-024-02960-w
Bernal, L., Coello, P., & Martínez-Barajas, J. E. (2022). Regulación de la degradación del almidón en las hojas. Revista Fitotecnia Mexicana, 45(4), 503-507. https://doi.org/10.35196/rfm.2022.4.503
Blanc, G., Lardet, L., Martin, A., Jacob, J. L., & Carron, M. P. (2002). Blanc, G., Lardet, L., Martin, A., Jacob, J. L., & Carron, M. P. (2002). Differential carbohydrate metabolism conducts morphogenesis in embryogenic callus of Hevea brasiliensis (Mull. Arg.). Journal of Experimental Botany, 53(373), 1453-1462. https://doi.org/10.1093/jexbot/53.373.1453
Carrión-Pereira, A. V., Takamori, L. M., Yaguinuma, D. H., de Oliveira, A. M., & Ribas, A. F. (2019). Maltose in culture media improves the in vitro regeneration of Urochloa brizantha cv. ‘Marandu’ plants. Biotecnología Vegetal, 19(3), 205–213.
Jhon, N. B., Kawamitsu, Y., Yonemori, S., Hayashi, M., & Murayama, S. (2000). Effect of various carbon sources and their combinations on in vitro growth and photosynthesis of banana plantlets. Plant Production Science, 3(4), 392-397. https://doi.org/10.1626/pps.3.392
Chen, Y., Wang, H., Hu, W., Wang, S., Snider, J. L., & Zhou, Z. (2017). Co-occurring elevated temperature and waterlogging stresses disrupt cellulose synthesis by altering the expression and activity of carbohydrate balance-associated enzymes during fiber development in cotton. Environmental and Experimental Botany, 135, 106-117. https://doi.org/10.1016/j.envexpbot.2016.12.012
Cuenca, B., & Vieitez, A. M. (2000). Influence of carbon source on shoot multiplication and adventitious bud regeneration in in vitro beech cultures. Plant Growth Regulation, 32(1), 1-12. https://doi.org/10.1023/A:1006329510280
Flores-Hernández, A. B., Pérez-García, C. D., & López-Martínez, E. F. (2017). Beneficial effects of sucrose as carbon source in plant tissue culture. Plant Cell, Tissue and Organ Culture, 128(2), 123–134. https://doi.org/10.1007/s11240-016-1150-1
Fowler, M. W. (1982). Substrate utilization by plant-cell cultures. Journal of Chemical Technology and Biotechnology, 32(1), 338–346. https://doi.org/10.1002/jctb.5030320139
Gabryszewska, E. (2010). The effects of glucose and growth regulators on in vitro organogenesis of Paeonia lactiflora. Journal of Fruit and Ornamental Plant Research, 18(2), 309–320.
García, J. L., Troncoso, J., Sarmiento, R., & Troncoso, A. (2002). Influence of carbon source and concentration on the in vitro development of olive zygotic embryos and explants raised from them. Plant Cell, Tissue and Organ Culture, 69(1), 95–100. https://doi.org/10.1023/A:1015086104389
Hernández-Bernal, A. F., Gregorio-Jorge, J., & León, P. (2022). El papel de los azúcares como moléculas de señalización en las plantas. TIP Revista Especializada en Ciencias Químico-Biológicas, 25, 1–11. https://doi.org/10.22201/fesz.23958723e.2022.519
Joyia, F. A., & Khan, M. S. (2013). Scutellum-derived callus-based efficient and reproducible regeneration system for elite varieties of indica rice in Pakistan. International Journal of Agriculture and Biology, 15(1), 27–33.
Juturu, V. N., Mekala, G. K., & Kirti, P. B. (2015). Current status of tissue culture and genetic transformation research in cotton (Gossypium spp.). Plant Cell, Tissue and Organ Culture, 120(3), 813–839. https://doi.org/10.1007/s11240-014-0640-z
Kaur, R., & Kapoor, M. (2016). Plant regeneration through somatic embryogenesis in sugarcane. Sugar Tech, 18(1), 93–99. https://doi.org/10.1007/s12355-015-0380-3
Kumar, G. P., Subiramani, S., Govindarajan, S., Sadasivam, V., Manickam, V., Mogilicherla, K., Kumar, S., Thiruppathi, K., & Narayanasamy, J. (2015). Evaluation of different carbon sources for high frequency callus culture with reduced phenolic secretion in cotton (Gossypium hirsutumL.) cv. Svpr-2. Biotechnology Reports, 7, 72–80. https://doi.org/10.1016/j.btre.2015.05.005
Marino, G., Bertazza, G., Magnanini, E., & Altan, A. D. (1993). Comparative effects of sorbitol and sucrose as main carbon energy sources in micropropagation of apricot. Plant Cell, Tissue and Organ Culture, 34(3), 235–244. https://doi.org/10.1007/BF00029712
Martins, J. P. R., Rodrigues, L. C. A., Conde, L. T., Gontijo, A. B. P. L., & Falqueto, A. R. (2019). Anatomical and physiological changes of in vitro-propagated Vriesea imperialis (Bromeliaceae) in the function of sucrose and ventilated containers. Plant Biosystems, 154(1), 87–99. https://doi.org/10.1080/11263504.2019.1635223
Muller, B., Pantin, F., Génard, M., Turc, O., Freixes, S., Piques, M., & Gibon, Y. (2011). Water deficits uncouple growth from photosynthesis, increase C content, and modify the relationships between C and growth in sink organs. Journal of Experimental Botany, 62(6), 1715–1729. https://doi.org/10.1093/jxb/erq438
Murashige, T., & Skoog, F. A. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15(3), 473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Nowak, B., MiczyÅ„ski, K., & Hudy, L. (2004). Sugar uptake and utilization during adventitious bud differentiation on in vitro leaf explants of ‘Wegierka ZwykÅ‚a’ plum (Prunus domestica). Plant Cell, Tissue and Organ Culture, 76(3), 255–260. https://doi.org/10.1023/B:TICU.0000009247.94819.02
Ondo-Ovono, P., Claire-Kevers, C., & Dommes, J. (2009). Effects of reducing sugar concentration on in vitro tuber formation and sprouting in yam (Dioscorea cayenensis-D. rotundata complex). Plant Cell, Tissue and Organ Culture, 99(1), 55–59. https://doi.org/10.1007/s11240-009-9575-1
Pereira, A. V. C., Takamori, L. M., Yaguinuma, D. H., Oliveira, A. M., & Ribas, A. F. (2019). Maltose in culture media improves the in vitro regeneration of Urochloa brizantha cv.‘Marandu’plants. Biotecnología Vegetal, 19, 205-213.
Pérez-Pérez, J., García-Rodríguez, L., Veitía, N., Bermúdez-Caraballoso, I., Collado, R., & Roque, B. (2019). Influencia de maltosa y sacarosa en la maduración de embriones somáticos de Glycine max cv. ‘Incasoy-27’. Biotecnología Vegetal, 19(1), 35–42.
Pérez-Martínez, A. B., & Castañeda-Garzón, S. L. (2016). Propagación in vitro de orquídeas nativas como contribución para la conservación in vitro. Biotecnología Vegetal, 16(3), 143–151.
Porras, R. J., Thompson, W. A., & Kiederman, P. (1989). Determination of accurate extraction and simultaneous equations for assaying chlorophylls a and b extracted with different solvents: Verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochimica et Biophysica Acta (BBA) - Bioenergetics, 975(3), 384–394. https://doi.org/10.1016/S0005-2728(89)80347-0
Pullman, G. S., & Bucalo, K. (2014). Pine somatic embryogenesis: Analyses of seed tissue and medium to improve protocol development. New Forests, 45(3), 353–377. https://doi.org/10.1007/s11056-014-9407-y
Ramírez-Mosqueda, M. A., Iglesias-Andreu, L. G., Noa-Carrazana, J. C., & Armas-Silva, A. A. (2018). In vitro propagation of Vanilla pompona Schiede. Cuadernos de Biodiversidad, 54, 9–14. https://doi.org/10.14198/cdbio.2018.54.02
Ramos-Castellá, A., Iglesias-Andreu, L. G., Bello-Bello, J., & Lee-Espinosa, H. (2014). Improved propagation of vanilla (Vanilla planifolia Jacks. ex-Andrews) using a temporary immersion system. In Vitro Cellular & Developmental Biology - Plant, 50(5), 576–587. https://doi.org/10.1007/s11627-014-9602-8
Ren, J. P., Wang, X. G., & Yin, J. (2010). Dicamba and sugar effects on callus induction and plant regeneration from mature embryo culture of wheat. Agricultural Sciences in China, 9(1), 31–37. https://doi.org/10.1016/S1671-2927(09)60064-0
Sami, F., Yusuf, M., Faizan, M., Faraz, A., & Hayat, S. (2016). Role of sugars under abiotic stress. Plant Physiology and Biochemistry, 109, 54-61. https://doi.org/10.1016/j.plaphy.2016.09.005
Silos-Espino, H., Escalera-García, R., Moncada-González, D., Valera-Montero, L. L., Flores-Benítez, S., Ortiz-Morales, M., Guzmán-Maldonado, H. S., Escalante-García, N., & Olvera-González, E. (2023). Interaction of culture medium and artificial light type on pigmentation of micro-propagated Opuntia plants. Horticulturae, 9(12). https://doi.org/10.3390/horticulturae9121348
Teixeira da Silva, J. A. (2004). The effect of carbon source on in vitro organogenesis of Chrysanthemum thin cell layers. Bragantia, 63(2), 165–177. https://doi.org/10.1590/S0006-87052004000200002
Velázquez-Rosas, N., Sinaca Colin, S., Vázquez-Domínguez, G., Velasco-Murguía, A., Silva Rivera, E., Ruíz-Guerra, B., Friedrich, F. L., Cortés Galindo, R., Armenta-Montero, S., & Martínez-Mota, R. (2025). Importance of traditional vanilla cultivation in the conservation of plant diversity in tropical forests in northern Veracruz, Mexico. Sustainability, 17(6). https://doi.org/10.3390/su17062598
Zahara, M., Datta, A., Boonkorkaew, P., & Mishra, A. (2017). The effects of different media, sucrose concentrations and natural additives on plantlet growth of Phalaenopsis hybrid ‘Pink’. Brazilian Archives of Biology and Technology, 60, e17160149. https://doi.org/10.1590/1678-4324-2017160149
Zhu, T., Li, L., Feng, L., Mo, H., & Ren, M. (2020). Target of rapamycin regulates genome methylation reprogramming to control plant growth in Arabidopsis. Frontiers in Genetics, 11. https://doi.org/10.3389/fgene.2020.00186
Akyüz, B. (2025). Effect of different carbon sources and concentrations on in vitro propagation of chestnut. Plant Cell, Tissue and Organ Culture (PCTOC), 160(2), 25. https://doi.org/10.1007/s11240-024-02960-w
Bernal, L., Coello, P., & Martínez-Barajas, J. E. (2022). Regulación de la degradación del almidón en las hojas. Revista Fitotecnia Mexicana, 45(4), 503-507. https://doi.org/10.35196/rfm.2022.4.503
Blanc, G., Lardet, L., Martin, A., Jacob, J. L., & Carron, M. P. (2002). Blanc, G., Lardet, L., Martin, A., Jacob, J. L., & Carron, M. P. (2002). Differential carbohydrate metabolism conducts morphogenesis in embryogenic callus of Hevea brasiliensis (Mull. Arg.). Journal of Experimental Botany, 53(373), 1453-1462. https://doi.org/10.1093/jexbot/53.373.1453
Carrión-Pereira, A. V., Takamori, L. M., Yaguinuma, D. H., de Oliveira, A. M., & Ribas, A. F. (2019). Maltose in culture media improves the in vitro regeneration of Urochloa brizantha cv. ‘Marandu’ plants. Biotecnología Vegetal, 19(3), 205–213.
Jhon, N. B., Kawamitsu, Y., Yonemori, S., Hayashi, M., & Murayama, S. (2000). Effect of various carbon sources and their combinations on in vitro growth and photosynthesis of banana plantlets. Plant Production Science, 3(4), 392-397. https://doi.org/10.1626/pps.3.392
Chen, Y., Wang, H., Hu, W., Wang, S., Snider, J. L., & Zhou, Z. (2017). Co-occurring elevated temperature and waterlogging stresses disrupt cellulose synthesis by altering the expression and activity of carbohydrate balance-associated enzymes during fiber development in cotton. Environmental and Experimental Botany, 135, 106-117. https://doi.org/10.1016/j.envexpbot.2016.12.012
Cuenca, B., & Vieitez, A. M. (2000). Influence of carbon source on shoot multiplication and adventitious bud regeneration in in vitro beech cultures. Plant Growth Regulation, 32(1), 1-12. https://doi.org/10.1023/A:1006329510280
Flores-Hernández, A. B., Pérez-García, C. D., & López-Martínez, E. F. (2017). Beneficial effects of sucrose as carbon source in plant tissue culture. Plant Cell, Tissue and Organ Culture, 128(2), 123–134. https://doi.org/10.1007/s11240-016-1150-1
Fowler, M. W. (1982). Substrate utilization by plant-cell cultures. Journal of Chemical Technology and Biotechnology, 32(1), 338–346. https://doi.org/10.1002/jctb.5030320139
Gabryszewska, E. (2010). The effects of glucose and growth regulators on in vitro organogenesis of Paeonia lactiflora. Journal of Fruit and Ornamental Plant Research, 18(2), 309–320.
García, J. L., Troncoso, J., Sarmiento, R., & Troncoso, A. (2002). Influence of carbon source and concentration on the in vitro development of olive zygotic embryos and explants raised from them. Plant Cell, Tissue and Organ Culture, 69(1), 95–100. https://doi.org/10.1023/A:1015086104389
Hernández-Bernal, A. F., Gregorio-Jorge, J., & León, P. (2022). El papel de los azúcares como moléculas de señalización en las plantas. TIP Revista Especializada en Ciencias Químico-Biológicas, 25, 1–11. https://doi.org/10.22201/fesz.23958723e.2022.519
Joyia, F. A., & Khan, M. S. (2013). Scutellum-derived callus-based efficient and reproducible regeneration system for elite varieties of indica rice in Pakistan. International Journal of Agriculture and Biology, 15(1), 27–33.
Juturu, V. N., Mekala, G. K., & Kirti, P. B. (2015). Current status of tissue culture and genetic transformation research in cotton (Gossypium spp.). Plant Cell, Tissue and Organ Culture, 120(3), 813–839. https://doi.org/10.1007/s11240-014-0640-z
Kaur, R., & Kapoor, M. (2016). Plant regeneration through somatic embryogenesis in sugarcane. Sugar Tech, 18(1), 93–99. https://doi.org/10.1007/s12355-015-0380-3
Kumar, G. P., Subiramani, S., Govindarajan, S., Sadasivam, V., Manickam, V., Mogilicherla, K., Kumar, S., Thiruppathi, K., & Narayanasamy, J. (2015). Evaluation of different carbon sources for high frequency callus culture with reduced phenolic secretion in cotton (Gossypium hirsutumL.) cv. Svpr-2. Biotechnology Reports, 7, 72–80. https://doi.org/10.1016/j.btre.2015.05.005
Marino, G., Bertazza, G., Magnanini, E., & Altan, A. D. (1993). Comparative effects of sorbitol and sucrose as main carbon energy sources in micropropagation of apricot. Plant Cell, Tissue and Organ Culture, 34(3), 235–244. https://doi.org/10.1007/BF00029712
Martins, J. P. R., Rodrigues, L. C. A., Conde, L. T., Gontijo, A. B. P. L., & Falqueto, A. R. (2019). Anatomical and physiological changes of in vitro-propagated Vriesea imperialis (Bromeliaceae) in the function of sucrose and ventilated containers. Plant Biosystems, 154(1), 87–99. https://doi.org/10.1080/11263504.2019.1635223
Muller, B., Pantin, F., Génard, M., Turc, O., Freixes, S., Piques, M., & Gibon, Y. (2011). Water deficits uncouple growth from photosynthesis, increase C content, and modify the relationships between C and growth in sink organs. Journal of Experimental Botany, 62(6), 1715–1729. https://doi.org/10.1093/jxb/erq438
Murashige, T., & Skoog, F. A. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15(3), 473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Nowak, B., MiczyÅ„ski, K., & Hudy, L. (2004). Sugar uptake and utilization during adventitious bud differentiation on in vitro leaf explants of ‘Wegierka ZwykÅ‚a’ plum (Prunus domestica). Plant Cell, Tissue and Organ Culture, 76(3), 255–260. https://doi.org/10.1023/B:TICU.0000009247.94819.02
Ondo-Ovono, P., Claire-Kevers, C., & Dommes, J. (2009). Effects of reducing sugar concentration on in vitro tuber formation and sprouting in yam (Dioscorea cayenensis-D. rotundata complex). Plant Cell, Tissue and Organ Culture, 99(1), 55–59. https://doi.org/10.1007/s11240-009-9575-1
Pereira, A. V. C., Takamori, L. M., Yaguinuma, D. H., Oliveira, A. M., & Ribas, A. F. (2019). Maltose in culture media improves the in vitro regeneration of Urochloa brizantha cv.‘Marandu’plants. Biotecnología Vegetal, 19, 205-213.
Pérez-Pérez, J., García-Rodríguez, L., Veitía, N., Bermúdez-Caraballoso, I., Collado, R., & Roque, B. (2019). Influencia de maltosa y sacarosa en la maduración de embriones somáticos de Glycine max cv. ‘Incasoy-27’. Biotecnología Vegetal, 19(1), 35–42.
Pérez-Martínez, A. B., & Castañeda-Garzón, S. L. (2016). Propagación in vitro de orquídeas nativas como contribución para la conservación in vitro. Biotecnología Vegetal, 16(3), 143–151.
Porras, R. J., Thompson, W. A., & Kiederman, P. (1989). Determination of accurate extraction and simultaneous equations for assaying chlorophylls a and b extracted with different solvents: Verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochimica et Biophysica Acta (BBA) - Bioenergetics, 975(3), 384–394. https://doi.org/10.1016/S0005-2728(89)80347-0
Pullman, G. S., & Bucalo, K. (2014). Pine somatic embryogenesis: Analyses of seed tissue and medium to improve protocol development. New Forests, 45(3), 353–377. https://doi.org/10.1007/s11056-014-9407-y
Ramírez-Mosqueda, M. A., Iglesias-Andreu, L. G., Noa-Carrazana, J. C., & Armas-Silva, A. A. (2018). In vitro propagation of Vanilla pompona Schiede. Cuadernos de Biodiversidad, 54, 9–14. https://doi.org/10.14198/cdbio.2018.54.02
Ramos-Castellá, A., Iglesias-Andreu, L. G., Bello-Bello, J., & Lee-Espinosa, H. (2014). Improved propagation of vanilla (Vanilla planifolia Jacks. ex-Andrews) using a temporary immersion system. In Vitro Cellular & Developmental Biology - Plant, 50(5), 576–587. https://doi.org/10.1007/s11627-014-9602-8
Ren, J. P., Wang, X. G., & Yin, J. (2010). Dicamba and sugar effects on callus induction and plant regeneration from mature embryo culture of wheat. Agricultural Sciences in China, 9(1), 31–37. https://doi.org/10.1016/S1671-2927(09)60064-0
Sami, F., Yusuf, M., Faizan, M., Faraz, A., & Hayat, S. (2016). Role of sugars under abiotic stress. Plant Physiology and Biochemistry, 109, 54-61. https://doi.org/10.1016/j.plaphy.2016.09.005
Silos-Espino, H., Escalera-García, R., Moncada-González, D., Valera-Montero, L. L., Flores-Benítez, S., Ortiz-Morales, M., Guzmán-Maldonado, H. S., Escalante-García, N., & Olvera-González, E. (2023). Interaction of culture medium and artificial light type on pigmentation of micro-propagated Opuntia plants. Horticulturae, 9(12). https://doi.org/10.3390/horticulturae9121348
Teixeira da Silva, J. A. (2004). The effect of carbon source on in vitro organogenesis of Chrysanthemum thin cell layers. Bragantia, 63(2), 165–177. https://doi.org/10.1590/S0006-87052004000200002
Velázquez-Rosas, N., Sinaca Colin, S., Vázquez-Domínguez, G., Velasco-Murguía, A., Silva Rivera, E., Ruíz-Guerra, B., Friedrich, F. L., Cortés Galindo, R., Armenta-Montero, S., & Martínez-Mota, R. (2025). Importance of traditional vanilla cultivation in the conservation of plant diversity in tropical forests in northern Veracruz, Mexico. Sustainability, 17(6). https://doi.org/10.3390/su17062598
Zahara, M., Datta, A., Boonkorkaew, P., & Mishra, A. (2017). The effects of different media, sucrose concentrations and natural additives on plantlet growth of Phalaenopsis hybrid ‘Pink’. Brazilian Archives of Biology and Technology, 60, e17160149. https://doi.org/10.1590/1678-4324-2017160149
Zhu, T., Li, L., Feng, L., Mo, H., & Ren, M. (2020). Target of rapamycin regulates genome methylation reprogramming to control plant growth in Arabidopsis. Frontiers in Genetics, 11. https://doi.org/10.3389/fgene.2020.00186
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2026-04-13
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Biotecnologia
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Effect of different sources and concentrations of carbohydrates on the morphological and physiological characteristics of Vanilla planifolia Jacks. RH350. (2026). Acta Scientiarum. Biological Sciences, 48(1), e77900. https://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/77900
