Growth of Kaffir Lime (Citrus hystrix DC) Cell Line Derived from Seed Explant After Yeast Elicitation Using Pure and Technical Grade Yeast

https://doi.org/10.22146/jtbb.68650

Dewi Yuliana Rizqi(1), Frisca Damayanti(2), Ghea Putri Cristy(3), Alisa Julia Nurulita(4), Aries Bagus Sasongko(5), Endang Semiarti(6), Woro Anindito Sri Tunjung(7*)

(1) Postgraduate Student Faculty of Biology, Universitas Gadjah Mada, Jl. Teknika Selatan, Sekip Utara, Yogyakarta, 55281, Indonesia
(2) Postgraduate Student Faculty of Biology, Universitas Gadjah Mada, Jl. Teknika Selatan, Sekip Utara, Yogyakarta, 55281, Indonesia; Research Center for Plant Conservation and Botanic Gardens, National Research and Innovation Agency, Jl. Ir. H. Juanda No. 13, 16122, West Java, Indonesia
(3) Undergraduate Student Faculty of Biology, Universitas Gadjah Mada, Jl. Teknika Selatan, Sekip Utara, Yogyakarta, 55281, Indonesia; Division of Biotechnology, Generasi Biologi (Genbinesia) Foundation, Jl. Swadaya Barat no. 4, Gresik Regency 61171, East Java, Indonesia
(4) Undergraduate Student Faculty of Biology, Universitas Gadjah Mada, Jl. Teknika Selatan, Sekip Utara, Yogyakarta, 55281, Indonesia
(5) Faculty of Biology, Universitas Gadjah Mada, Jl. Teknika Selatan, Sekip Utara, Yogyakarta, 55281, Indonesia
(6) Faculty of Biology, Universitas Gadjah Mada, Jl. Teknika Selatan, Sekip Utara, Yogyakarta, 55281, Indonesia
(7) Faculty of Biology, Universitas Gadjah Mada, Jl. Teknika Selatan, Sekip Utara, Yogyakarta, 55281, Indonesia
(*) Corresponding Author

Abstract


The addition of elicitors in kaffir lime (Citrus hystrix DC.) culture is one of  strategies for obtaining and increasing the production of secondary metabolites.  Saccharomyces cerevisiae is one of the elicitors that can be used to increase secondary metabolites such as terpenoids. However, in its use, the pure cultures of S. cerevisiae are expensive. Therefore, the first objective of this study was to analyze the ability of technical grade (commercial baker’s yeast) to be used as an elicitor and measure the growth of kaffir lime cell line after being elicited by pure and technical grade (commercial baker’s yeast). The second objective is to determine the best time to subculture kaffir lime cell line after elicitation. We observed the morphology and measured the growth curve of pure and technical grade yeast until the 4th subculture generation. Furthermore, we used both grades of yeast for elicitation. Kaffir lime cell suspension was treated with 10 ppm pure grade or 5 ppm and 10 ppm technical grade yeast for 4 days. After elicitation, kaffir lime cell lines were subcultured and their growth was analyzed. The result showed that the morphology and growth curve of technical grade until 4th subculture generations was similar to the pure grade. On the other hand, after elicitation using pure and technical grade yeast and being subcultured, the growth of the elicitated kaffir lime cell line had the same pattern as the control group, but the cell density of the control group was higher than the elicitated group. The initial stationary phase of kaffir lime cell line was on the 17th day which is the best time to subculture. The subculturing process is important to maintain the viability of the kaffir lime cell line.

 


Keywords


cell line; elicitation; kaffir lime; S. cerevisiae; technical grade yeast

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References

Aleu, J. et al., 1999. Biotransformation of the fungistatic sesquiterpenoid ginsenol by Botrytis cinerea. Journal of the Chemical Society - Perkin Transactions 1, (6), pp.727–730. doi: 10.1039/A808044E.

Arano-Varela, H. et al., 2020. Verbascoside production in long-term Buddleja cordata Kunth cell suspension cultures. 3 Biotech, 10(6), pp.1–9. doi: 10.1007/s13205-020-02222-4.

Bourgaud, F. et al., 2001. Production of Plant Secondary Metabolites: A Historical Perspective. Plant Science, 161, pp. 839-851.

Chen, H. & Chen, F., 2000. Effect of yeast elicitor on the secondary metabolism of Ti-transformed Salvia miltiorrhiza cell suspension cultures. Plant Cell Reports, 19(7), pp.710–717. doi: 10.1007/s002999900166.

Chodisetti, B. et al., 2013. Improved gymnemic acid production in the suspension cultures of Gymnema sylvestre through biotic elicitation. Plant Biotechnology Reports, 7(4), pp.519–525. doi: 10.1007/s11816-013-0290-3.

Damayanti, F. et al., 2020. Variation of 2,4-dichlorophenoxyacetic acid (2,4-D) concentration on kaffir lime callus growth as raw material for cell suspension. AIP Conference Proceedings, 2260. doi: 10.1063/5.0016420.

Dwimahyani, I., 2007. Metode Suspensi Sel Untuk Membentuk Spot Hijau Pada Kultur In-Vitro Galur Mutan Tanaman Jarak Pagar (Jatropha curcas L). Jurnal Ilmiah Aplikasi Isotop dan Radiasi, 3(2), pp.55–79. doi: 10.17146/jair.2007.3.2.559.

Fajarina, S. et al., 2021. Evaluation of Anticancer Bioactive Compounds and Cytotoxicity of Kaffir Lime (Citrus hystrix Dc.) Callus Extract Post Preservation. Indonesian Journal of Pharmacy, 32(2), pp.179–192. doi: 10.22146/ijp.1062.

Gueven, A., 2003. Plant Tissue Cultures in Production of Secondary Metabolites. Scientific Works of UFT, pp.553–556.

Habibah, N.A. et al., 2017. Flavonoid production, growth, and differentiation of Stelechocarpus burahol (Bl.) hook. f. and th. cell suspension culture. Pakistan Journal of Biological Sciences, 20(4), pp.197–203. doi: 10.3923/pjbs.2017.197.203.

Jamil, S.Z.M.R., et al., 2018. Metabolite profiles of callus and cell suspension cultures of mangosteen. 3 Biotech, 8(8), pp.1–14. doi: 10.1007/s13205-018-1336-6.

Khanpour-Ardestani, N., Sharifi, M. & Behmanesh, M., 2015. Establishment of callus and cell suspension culture of Scrophularia striata Boiss.: an in vitro approach for acteoside production. Cytotechnology, 67(3), pp.475–485. doi: 10.1007/s10616-014-9705-4

Klis, F.M. et al., 2002. Dynamics of cell wall structure in Saccharomyces cerevisiae. FEMS Microbiology Reviews, 26(3), pp.239–256. doi: 10.1111/j.1574-6976.2002.tb00613.x.

Leksonowati, A., Witjaksono & Ratnadewi, D., 2017. Induction of Callus Culture and Cell Suspension Culture of Aquilaria malaccensis Lam. Berita Biologi, 16(1), pp.1–11. doi: 10.14203/beritabiologi.v16i1.2687

Montes de Oca, R. et al., 2016. Yeast : Description and Structure Chapter 2 Yeast : Description and Structure. Yeast Additive and Animal Production, (February), pp.3–13.

Moscatiello, R., Baldan, B. & Navazio, L., 2013. Plant Cell Suspension. Plant Mineral Nutrients, 953(July), pp.195–207. doi: 10.1007/978-1-62703-152-3_5

Murthy, H.N., Lee, E.J. & Paek, K.Y., 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), pp.1–16. doi: 10.1007/s11240-014-0467-7

Namdeo, A.G., 2007. PHCOG REV .: Review Article Plant Cell Elicitation for Production of Secondary Metabolites : A Review. Review Literature And Arts Of The Americas, 1(1), pp.69–79.

Pasitvilaiturm, M. & Pankasemsuk, T., 2012. Endosperm culture of Jatropha curcas L. Chiang Mai University Journal of Natural Sciences, 11(1 SPECIAL ISSUE), pp.97–102.

Pereira, P.S. et al., 2007. Enhanced triterpene production in Tabernaemontana catharinensis cell suspension cultures in response to biotic elicitors. Quimica Nova, 30(8), pp.1849–1852. doi: 10.1590/S0100-40422007000800010.

Ramirez-Estrada, K. et al., 2016. Elicitation, an effective strategy for the biotechnological production of bioactive high-added value compounds in plant cell factories. Molecules, 21(2). doi: 10.3390/molecules21020182

Ratnasari, J., Siregar, A.H. & R. E., R., 2001. Pengaruh Pemberian Elisitor Ekstrak Khamir Saccharomyces Terhadap Kandungan Ajmalisin dalam Kultur Agregat Sel Catharanthus roseus ( L .) G . Don. Berita Biologi, 5(April), pp.349–355. doi: 10.14203/beritabiologi.v5i4.1118.

dos Santos, A.L.W. et al., 2010. Biochemical and morphological changes during the growth kinetics of Araucaria angustifolia suspension cultures. Brazilian Archives of Biology and Technology, 53(3), pp.497–504. doi: 10.1590/S1516-89132010000300001.

Sierra, M.I. et al., 1992. Stability of alkaloid production in cell suspension cultures of Tabernaemontana divaricata during long-term subculture. Plant Cell, Tissue and Organ Culture, 28(1), pp.59–68. doi: 10.1007/BF00039916.

Sitinjak, R.R., Siregar, A.H. & Rizkita, R., 2000. Pengaruh Pemberian Ekstrak Saccharomyces cerevisiae Hansen terhadap Kandungan Gosipol pada Kultur Kalus Gossypium hirsutum L. Berita Biologi, 5(2), pp.131–136. doi: 10.14203/beritabiologi.v5i2.1133.

Sparapano, L. & Bruno, G., 2004. Cupressus callus and cell suspension cultures: Effect of seiridins on their growth and sensitivity. In Vitro Cellular and Developmental Biology - Plant, 40(6), pp.617–625. doi: 10.1079/IVP2004581.

Sukmara, E., Sukamto, L.A. & Bintang, M., 2014. Induksi dan Karakter Pertumbuhan Kalus Triploid dari Endosperma Avokad (Persea americana Mill.). Current Biochemistry, 1(1), pp.20–28.

Thorat, A.S. et al., 2017. Plant regeneration from cell suspension culture in Saccharum officinarum L. and ascertaining of genetic fidelity through RAPD and ISSR markers. 3 Biotech, 7(1), pp.1–12. doi: 10.1007/s13205-016-0579-3.

Trejo-Tapia, G. et al., 2008. Effect of screening and subculture on the production of betaxanthins in Beta vulgaris L. var. “Dark Detroit” callus culture. Innovative Food Science and Emerging Technologies, 9(1), pp.32–36. doi: 10.1016/j.ifset.2007.04.009.

Tunjung, W.A.S. et al., 2015. Anti-Cancer Effect of Kaffir Lime (Citrus hystrix DC) Leaf Extract in Cervical Cancer and Neuroblastoma Cell Lines. Procedia Chemistry, 14, pp.465-468. doi: 10.1016/j.proche.2015.03.062.



DOI: https://doi.org/10.22146/jtbb.68650

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