Multiple plant regeneration from matricular substance released from explants of Citrus jambhiri

POOJA MANCHANDA; ANU KALIA; GURUPKAR S SIDHU; H S RATTANPAL; KULJOT KAUR; SIMRANJEET KAUR

doi:10.56093/ijas.v90i1.98546

Authors

POOJA MANCHANDA Assistant Professor, Punjab Agricultural University, Ludhiana 141 001, India
ANU KALIA Assistant Professor, Punjab Agricultural University, Ludhiana 141 001, India
GURUPKAR S SIDHU Assistant Horticulturist, Punjab Agricultural University, Ludhiana 141 001, India
H S RATTANPAL Senior Horticulturist, Department of Fruit Science, Punjab Agricultural University, Ludhiana, India
KULJOT KAUR Research Scholars, Punjab Agricultural University, Ludhiana 141 001, India
SIMRANJEET KAUR Research Scholars, Punjab Agricultural University, Ludhiana 141 001, India

https://doi.org/10.56093/ijas.v90i1.98546

Keywords:

Cotyledons, Epicotyl, FT-IR Spectroscopy, LC-MS, SEM

Abstract

Comparative performance of epicotyl-, hypocotyl-, and cotyledonary (Cot) explants for direct organogenesis was evaluated with best response on BAP (3.5 mg/l) + NAA (0.5 mg/l) supplemented MS medium for Cot explants. This differential behaviour can be attributed to diversity and concentration of phytohormone(s), signal phenolics and other compounds. This study identifies the spatio-specific variabilities of these compounds leading to emergence of multiple shoots from explant’s injured regions indicated by formation of foamy-white exudate, ‘matrix’. Topographically, matrix appeared amorphous containing granulation. Further, FT-IR spectra indicated possible presence of woundinduced sugars and phytohormones. Likewise, LC-MS study revealed presence of diverse sugars, phytohormone (PH)/ PH-like and signal compounds. This study illustrates possible interactive roles played by spatial co-localization of these compounds at wounded cut ends of explants for multiple in vitro shoot organogenesis. This study put-forth a novel concept of localized occurrence of multiple primary and secondary metabolites at cut/wounded ends of citrus explants which led to emergence of multiple shoots. These results are promising and could serve the basis for further investigations on various other citrus species for diversity and quantity of various metabolites during in vitro cultivation.

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References

Brito A, Ramirez J E, Areche C, Sepúlveda B and Simirgiotis M J. 2014. HPLC-UV-MS profiles of phenolic compounds and antioxidant activity of fruits from three citrus species consumed in Northern Chile. Molecules 19: 17400–21. DOI: https://doi.org/10.3390/molecules191117400

Griffth A. 2007. SPSS for Dummies. Wiley Publishing, Inc. Indiana Pdis, Indiana. 363p.

Hart D S, Keightley A, Sappinton D, Nguyen P T M, Chritton C, Seckinger G R and Torres K C. 2016. Stability of adenine-based cytokinins in aqueous solution. In vitro Cellular and Developmental Biology-Plant 52: 1–9. DOI: https://doi.org/10.1007/s11627-015-9734-5

Horai H, Arita M, Kanaya S, Nihei Y, Ikeda T, Suwa K, Ojima Y, Tanaka K, Tanaka S, Aoshima K, Oda Y, Kakazu Y, Kusano M, Tohge T, Matsuda F, Sawada Y, Hirai MY, Nakanishi H, Ikeda K, Akimoto N, Maoka T, Takahashi H, Ara T, Sakurai N, Suzuki H, Shibata D, Neumann S, Iida T, Tanaka K, Funatsu K, Matsuura F, Soga T, Taguchi R, Saito K and Nishioka T. 2010. MassBank: a public repository for sharing mass spectral data for life sciences. Journal of Mass Spectrometry 45: 703–14. DOI: https://doi.org/10.1002/jms.1777

Hu W, Fagundez S, Katin-Grazzini L, Li Y, Li W, Chen Y, Wang X, Deng Z, Xie S, McAvoy R J and Li Y. 2017. Endogenous auxin and its manipulation influence in vitro shoot organogenesis of citrus epicotyl explants. Horticulture Research 4 b 17071; doi:10.1038/hortres.2017.71. DOI: https://doi.org/10.1038/hortres.2017.71

Ikeuchi M, Iwase A, Rymen B, Lambolez A, Kojima M, Takebayashi Y, Heyman J, Watanabe S, Seo M, Veylder L D, Sakakibara H and Sugimoto K. 2017. Wounding triggers callus formation via dynamic hormonal and transcriptional changes. Plant Physiology 175: 1158–74. DOI: https://doi.org/10.1104/pp.17.01035

Manchanda P and Gosal S S. 2012. Effect of activated charcoal, carbon sources and gelling agents on direct somatic embryogenesis and regeneration in sugarcane via leaf roll segments. Sugar Tech 14: 168–73. DOI: https://doi.org/10.1007/s12355-012-0143-3

Murashige T and Skoog F. 1962. A revised medium for rapid growth and Bio-assays with tobacco tissue cultures. Physiologia Plantarum 15: 473–97. DOI: https://doi.org/10.1111/j.1399-3054.1962.tb08052.x

Ng T L M, Karim R, Tan Y S, The H F, Danial A D, Ho L S, Khalid N, Appleton D R and Harikrishna J A. 2016. Amino acid and secondary metabolite production in embryogenic and non-embryogenic callus of fingerroot ginger (Boesenbergia rotunda). PLoS ONE 11(6): e0156714. doi:10.1371/journal. pone.0156714. DOI: https://doi.org/10.1371/journal.pone.0156714

Rani G, Singh B, Sharma S, Rehan L, Zaidi A A, Nagpal A and Virk G S. 2004. Micropropagation of Kinnow (Citrus nobilis × Citrus deliciosa) through nodal segments. Journal of Indian Botanical Society 83: 26–9.

Rattanpal H S, Kaur G and Gupta M. 2011. In vitro plant regeneration in rough lemon (Citrus jambhiri Lush) by direct organogenesis. African Journal of Biotechnology 10: 13724–28. DOI: https://doi.org/10.5897/AJB09.1264

Saini H K, Gill M S and Gill M I S. 2010. Direct shoot organogenesis and plant regeneration in rough lemon (Citrus jambhiri Lush.). Indian Journal of Biotechnology 9: 419–23.

Savita, Singh B, Virk G S and Nagpal A K. 2011. An efficient plant regeneration protocol from callus cultures of Citrus jambhiri Lush. Physiology and Molecular Biology- Plants 17: 161–9. DOI: https://doi.org/10.1007/s12298-011-0055-9

Sharma C, Kumari T, Pant G, Bajpai V, Srivastava M, Mitra K, Kumar B and Arya K R. 2015. Plantlet formation via somatic embryogenesis and LC-ESI-Q-TOF MS determination of secondary metabolites in Buteamono sperma (Lam.) Kuntze. Acta Physiologia Plantarum 37: 239. DOI 10.1007/s11738-015-1973-x DOI: https://doi.org/10.1007/s11738-015-1973-x

Schafer M, Brutting C, Baldwin I T and Kallenbach M. 2016. High-throughput quantification of more than 100 primary- and secondary-metabolites, and phytohormones by a single solid-phase extraction based sample preparation with analysis by UHPLC–HESI–MS/MS. Plant Methods 12: 30 DOI 10.1186/ s13007-016-0130-x. DOI: https://doi.org/10.1186/s13007-016-0130-x

Stfaan P, Werbrouck O and Debergh P A C.1994. Applied aspects of plant regeneration in plant cell culture: A practical approach. Oxford Unversity Press, Oxford, pp 127–45.

Tamas I A and Davies P J. 2016. Dynamics and control of phloem loading of indole-3-acetic acid in seedling cotyledons of Ricinus communis. Journal of Experimental Botany 67: 4755–65. DOI: https://doi.org/10.1093/jxb/erw255

Usman M, Muhammad S and Fatima B. 2005. In vitro multiple shoot induction from nodal explants of citrus cultivars. Journal of Central European Agriculture 6: 435–42.

Vatasescu-Balcan R A, Predoi D, Ungureanu F and Costache M. 2008. Study of iron oxide nanoparticles coated with dextrin obtained by co-precipitation. Journal of Optoelectronics and Advanced Materials 10: 693–6.