Effect of 2,4-D and BA on the establishment of cell suspension from nodes and internodes of Capsicum annuum cv. Etna

MAURICIO REGINALDO ALVES DOS SANTOS; CAROLINA  SOUZA

doi:10.31686/ijier.vol7.iss5.1460

Authors

MAURICIO REGINALDO ALVES DOS SANTOS

EMBRAPA RONDONIA
CAROLINA AUGUSTO DE SOUZA

EMBRAPA RONDONIA

DOI:

https://doi.org/10.31686/ijier.vol7.iss5.1460

Keywords:

Callogenesis, growth curve, secondary metabolites

Abstract

In vitro cell suspension cultivation systems have been largely reported as safe and standardized methods for production of secondary metabolites with medicinal and agricultural interest. Capsicum annuum is one of the most widely grown vegetable in the world and its biological activities have been demonstrated against insects, fungi, bacteria and other groups of organisms. The determination of procedures for the dedifferentiation of cells into callus cells and the subsequent study of the callus growth pattern are necessary for the establishment of cell suspensions and also to subsidize studies regarding the bioactivity of its secondary metabolites. The objective of this study was to establish a protocol for dedifferentiation of leaf cells of the cultivar C. annuum cv. Etna and to determine the growth pattern of the calluses with a focus on the deceleration phase, when the callus cells must be subcultured into a liquid medium in order to establish cell suspension cultivations aiming at the production of secondary metabolites. treatment that resulted in the highest %CI, ACCC and callus weight was the combination of 4.52 µM 2,4-D + 0.44 µM BA. The calluses produced were friable and whitish and their growth pattern followed a sigmoid shape. The deceleration phase started on the 23rd day of cultivation. Callus induction in leaf explants of C. annuum cv. Etna can be achieved in MS medium supplemented with 4.52 µM 2,4-D + 0.44 µM BA, which results in high cellular proliferation; in order to start a cell suspension culture, callus cells on the 23rd day of culture should be used.

References

Coste, A., L. Vlase, A. Halmagyi, C. Deliu and G.Coldea. 2011. Effects of plant growth regulators and elicitors on production of secondary metabolites in shoot cultures of Hypericum hirsutum and Hypericum maculatum. Plant Cell, Tissue and Organ Culture, 106(2): 279–288.
Far, M.M.M.E. and H.A.A. Taie, 2009. Antioxidant activities, total anthocyanins, fhenolics and flavonoids contents of some sweetpotato genotypes under stress of different concentrations of sucrose and sorbitol. Australian Journal of Basic and Applied Sciences, 3(4): 3609-3616.
Feitosa, L.S., A.S. Costa, M.F. Arrigoni-Blank, R. Dibax, M.P. Botânico and A.F. Blank, 2013. Indução e análise histológica de calos em explantes foliares de Jatropha curcas L.(Euphorbiaceae). Bioscience Journal, 29(2): 370-377.
Gershenzon J. and Engelberth J. E. Metabólitos secundários e Defesa Vegetal. In: Taiz, L. and Zeiger, E. 2013. Fisiologia Vegetal. 5. ed. Porto Alegre: Artmed.
Gomes, F.P. 2009. Curso de Estatística Experimental. Fealq, Piracicaba.
Kang, S.-M., H.-Y. Jung, Y.-M. Kang, J.-Y. Min, C.S. Karigar, J.-K. Yang, S.-W. Kim, Y.-R. Ha, S.-H. Lee and M.-S. Choi. 2005. Biotransformation and impact of ferulic acid on phenylpropanoid and capsaicin levels in Capsicum annuum L. cv. P1482 cell suspension cultures. Journal of Agricultural and Food Chemistry, 53(9): 3449-3453.
Kintzios, S., J.B. Drossopoulos, E. Shortsianitis and D. Peppes. 2000. Induction of somatic embryogenesis from young, fully expanded leaves of chilli pepper (Capsicum annuum L.): effect of leaf position, illumination and explant pretreatment with high cytokinin concentrations. Scientia Horticulturae, 85(1-2): 137-144.
Kittipongpatana, N., P. Maneerat, P. Pattanakitkosol and O.S. Kittipongpatana. 2007. Effect of some factors on the growth of Capsicum annuum L. cell suspension culture and biotransformation of hydroquinone to arbutin. CMU Journal of Natural Sciences, 6(2): 207-218.
Luz, F.J.F. Caracterizações morfológica e molecular de acessos de pimenta (Capsicum chinense Jaqc.). 2007. 70 f. Tesis – Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, São Paulo.
Mendonça, E.G., V.C. Stein, F.P. Balieiro, C.D.F. Lima, B.R. Santos and L.V. Paiva. 2013. Genetic transformation of Eucaliptus camaldulensis by agrobalistic method. Revista Árvore, 37(3): 419-429.
Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologiae Plantarum, 15(3): 473-497.
Murthy, H.N., E.-J. Lee and K.-Y. Paek. 2014. Production of secondary metabolites from cell and organ cultures: strategies and approaches for biomass improvement and metabolite accumulation. Plant Cell, Tissue and Organ Culture, 118(1): 1-16.
Parsaeimehr, A. and B. Mousavi, 2009. Producing friable callus for suspension culture in Glycyrrhiza glabra. Advances in Environmental Biology, 3(2): 125-128.
Peixoto, C.P., T. V. Cruz and M.F.S.P. Peixoto, 2011. Análise quantitativa do crescimento de plantas: Conceitos e Prática. Enciclopédia Biosfera, 7(13):51-76.
Sanatombi, K. and Sharma, G. J. Capsaicin content and pungency of different Capsicum spp. cultivars. Notulae Botanicae Horti Agrobotanici, v. 36, n. 2, p. 89-90, 2008.
Santos, M.R.A., M.G.R. Ferreira, V. Sarubo, 2010. Determination of callus growth curve in Conilon coffee. Revista Caatinga, 23(1): 133-136.
Santos, M.R.A.; C.A. Souza, M.C.M. Guimarães, C.V. Smozinski, G.M.O. Magalhães, W.O. Nogueira and E.S. Paz. 2015. Growth pattern of friable callus from Piper Carniconnectivum leaf explants. Australian Journal of Basic and Applied Sciences, 9(27): 226-231.
Santos, M.R.A. and Smozinski, C.V. 2017. Proliferation of dedifferentiated cells from internodes of bell pepper (Capsicum annuum cv. Yolo Wonder). Australian Journal of Basic and Applied Sciences, 11(12): 20-24.
Santos, M.R.A. and Souza, C.A. 2016. Dedifferentiation of leaf cells and growth pattern of calluses of Capsicum annuum cv. Etna. Australian Journal of Basic and Applied Sciences, v. 10, n. 12, p. 362-368.
Souza, J.M.M., M.L. Tomaz, S.C.C. Arruda, C.G.B. Demétrio, W.N. Venables, A.P. Martinelli, 2011. Callus sieving is effective in improving synchronization and frequency of somatic embryogenesis in Citrus sinensis. Biologia Plantarum, 55(4): 703-307.
Souza, J.M.M.; Berkov, S. and Santos, A.S. 2014. Improvement of friable callus production of Boerhaavia paniculata Rich and the investigation of its lipid profile by GC/MS. Anais da Academia Brasileira de Ciências, 86(3): 1015-1027.
Umamaheswari, A. and V. Lalitha. 2007. In vitro effect of various growth hormones in Capsicum annuum L. on the callus induction and production of capsaicin. Journal of Plant Sciences, 2(5): 545-551.
Vanisree, M., C.-Y. Lee, S.-F. Lo, S.M. Nalawade, C.Y. Lin, H.-S. Tsay. 2004. Studies on the production of some important secondary metabolites from medicinal plants by plant tissue cultures. Botanical Bulletin of Academia Sinica, 45(1): 1-22.