Influencing in vitro clonal propagation of Chonemorpha fragrans (moon) Alston by culture media strength, plant growth regulators, carbon source and photo periodic incubation

Abstract

Chonemorpha fragrans is an endangered medicinal woody climber, regarded among alternative plant sources of camptothecin. Camptothecin is a monoterpene indole anti-cancer alkaloid with annual trade value of over three billion U.S. dollars in the recent, and is used in the production of its analog drugs approved for the chemotherapy of cancer of varied types. Effects of plant growth regulators, culture media strength and photoperiodic duration on the micropropagation efficiency of C. fragrans from nodal segment explants were studied on Murashige and Skoog (MS) medium amended with Thidiazuron (TDZ), Benzylaminopurine (BAP) or Kinetin (Kin). Thidiazuron was more efficient over BAP and Kin when half basal MS medium was used over full or quarter strength. Results of carbon source experiment showed sucrose as the most effective over glucose, fructose, and maltose in the clonal production. Studies on the photoperiodic incubation duration showed 12 h as the best light period and sub or supra-optimal resulted in the production of abnormal and albino micro shoots. Experimental results on the evaluation of physiological, biochemical parameters showed the role of pigment molecules and antioxidant systems in the production of albino micro shoots.

Keywords

Micropropagation / Micro shoot / Plantlets / Chonemorpha fragrans / Plant physiology / Albino shoot / Basal callus / PGRs (plant growth regulators) / Antioxidants

References

[1] Abou Rayya MS, Kassim NE, Ali EAM. Effect of different cytokinins concentrations and carbon sources on shoot proliferation of bitter almond nodal cuttings. J Am Sci, 2011, 6(9): 135-139.

[2] Afreen F. Kozai T, Afreen F, Zobayed SMA. Physiological and anatomical characteristics of in vitro photoautotrophic plants. Photoautotrophic (sugar-free medium) micropropagation as a new micropropagation and transplant production system, 2005, Dordrecht: Springer 61 90

[3] Alexieva V, Sergiev I, Mapelli S, Karanov E. The effect of drought and ultraviolet radiation on growth and stress markers in pea and wheat. Plant Cell Environ, 2001, 24(12): 1337-1344.

[4] Ali M, Isah T, Mujib A, Dipti T. Shahzad A, Sharma S, Siddiqui SA. Climber plants: medicinal importance and conservation strategies. Biotechnological strategies for the conservation of medicinal and ornamental climbers, 2016, Cham: Springer International Publishing 101 138

[5] Alikina O, Chernobrovkina M, Dolgov S, Miroshnichenko D. Tissue culture efficiency of wheat species with different genomic formulas. Crop Breed Appl Biotechnol, 2016, 16(4): 307-314.

[6] Arnon DI. Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol, 1949 24 1 1

[7] Babiker AGT, Parker C, Suttle JC. Induction of Striga seed germination by TDZ. Weed Res, 1992, 32: 243-248.

[8] Baskakov YA, Shapovalov AA, Zhirmunskaya NM, Ovsyannikova TV, Nauk DA. Interrelationship of growth-regulating activity and phytotoxicity of synthetic cytokinins. Dokl Akad Nauk SSSR, 1981, 267: 1514-1517.

[9] Bates LS, Waldren RP, Teare ID. Rapid determination of free proline for water-stress studies. Plant Soil, 1973, 39(1): 205-207.

[10] Becana M, Moran JF, Iturbe-Ormaetxe I. Iron-dependent oxygen free radical generation in plants subjected to environmental stress: toxicity and antioxidant protection. Plant Soil, 1986, 201: 137-147.

[11] Bhadrawale D, Mishra JP, Mishra Y. An improvised in vitro vegetative propagation technique for Bambusa tulda: influence of season, sterilization and hormones. J For Res, 2017, 4: 1-6.

[12] Bode S, Quentmeier CC, Liao P-N, Hafi N, Barros T, Wilk L, Bittner F, Walla PJ, Fleming GR. On the regulation of photosynthesis by excitonic interactions between carotenoids and chlorophylls. Proc Natl Acad Sci USA, 2009, 106: 12311-12316.

[13] Cakmak I, Marschner H. Magnesium deficiency and high light intensity enhance activities of superoxide dismutase, ascorbate peroxidase, and glutathione reductase in bean leaves. Plant Physiol, 1992, 98(4): 1222-1227.

[14] Cappelletti R, Sabbadini S, Mezzetti B. The use of TDZ for the efficient in vitro regeneration and organogenesis of strawberry and blueberry cultivars. Sci Hortic, 2016, 207: 117-124.

[15] Cardoso JC, Ono EO. In vitro growth of Brassocattleya orchid hybrid in different concentrations of KNO3, NH4NO3, and benzyl-aminopurine. Hortic Bras, 2011, 29(3): 359-363.

[16] Chinnusamy V, Jagendorf A, Zhu JK. Understanding and improving salt tolerance in plants. Crop Sci, 2005, 45(2): 437-448.

[17] Christensen B, Sriskandarajah S, Serek M, Muller R. In vitro culture of Hibiscus rosa-sinensis L.: influence of iron, calcium and BAP on the establishment and multiplication. Plant Cell Tissue Organ Cult, 2008, 93(2): 151-161.

[18] Duarte-Ake F, Castillo-Castro E, Pool FB, Espadas F, Santamaría JM, Robert ML, De-la-Peña C. Physiological differences and changes in global DNA methylation levels in Agave angustifolia Haw. albino variant somaclones during the micropropagation process. Plant Cell Rep, 2016, 35(12): 2489-2502.

[19] Fadel D, Kintzios S, Economou AS, Moschopoulou G, Constantinidou HA. Effect of different strength of medium on organogenesis, phenolic accumulation and antioxidant activity of Spearmint (Mentha spicata l.). Open Hortic J, 2010, 3: 31-35.

[20] Flors V, Paradis M, Garcia-Andrade J, Cerezo M, Gonzalez-Bosch C, García-Agustín P. A tolerant behavior in salt-sensitive tomato plants can be mimicked by chemical stimuli. Plant Signal Behav, 2007, 2(1): 50-57.

[21] Gibson SI. Plant sugar-response pathways. Part of a complex regulatory web. Plant Physiol, 2000, 124: 1532-1539.

[22] Gnasekaran P, Rathinam X, Sinniah UR, Subramaniam S. A study on the use of organic additives on the protocorm-like bodies (PLBs) growth of Phalaenopsis violaceae orchid. J Phytol, 2010, 2: 29-33.

[23] Goyal D, Bhadauria S. In vitro propagation of Ceropegia bulbosa using nodal segments. Indian J Biotechnol, 2006, 5: 565-567.

[24] Graner EM, Oberschelp GPJ, Brondani GE, Bataginpiotto KD, de Almeida CV, de Almeida M. TDZ pulsing evaluation on the in vitro morphogenesis of peach palm. Physiol Mol Biol Plants, 2013, 19(2): 283-288.

[25] Grieve CM, Grattan SR. Rapid assay for determination of water soluble quaternary ammonium compounds. Plant Soil, 1983, 70(2): 303-307.

[26] Guo B, Abbasi BH, Zeb A, Xu LL, Wei YH. Thidiazuron: a multi-dimensional plant growth regulator. Afr J Biotechnol, 2011, 10(45): 8984-9000.

[27] Heath RL, Packer L. Photoperoxidation in isolated chloroplasts: I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys, 1968, 125(1): 189-198.

[28] Hemeda HM, Klein BP. Effects of naturally occurring antioxidants on peroxidase activity of vegetable extracts. J Food Sci, 1990, 55(1): 184-185.

[29] Hoang NN, Kitaya Y, Morishita T, Endo R, Shibuya T. A comparative study on growth and morphology of wasabi plantlets under the influence of the micro-environment in shoot and root zones during photoautotrophic and photomixotrophic micropropagation. Plant Cell Tissue Organ Cult, 2017, 130: 255-263.

[30] Hollander-Czytko H, Grabowski J, Sandorf I, Weckermann K, Weiler EW. Tocopherol content and activities of tyrosine aminotransferase and cystine lyase in Arabidopsis under stress conditions. J Plant Physiol, 2005, 162(7): 767-770.

[31] Horton P, Ruban AV, Walters RG. Regulation of light harvesting in green plants. Ann Rev Plant Biol, 1996, 47(1): 655-684.

[32] Huetteman CA, Preece JE. Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tissue Organ Cult, 1993, 33(2): 105-119.

[33] Isah T. Adjustments to in vitro culture conditions and associated anomalies in plants. Acta Biol Cracov ser Bot, 2015, 57(2): 9-28.

[34] Isah T. Anticancer alkaloids from trees: development into drugs. Pharmacogn Rev, 2016, 10(20): 90-99.

[35] Jain N, Babbar SB. Effect of carbon source on the shoot proliferation potential of epicotyl explants of Syzygium cuminii. Biol Plant, 2003, 47(1): 133-136.

[36] Jin HC, Sun Y, Yang QC, Chao YH, Kang JM, Jin H, Li Y, Margaret G. Screening of genes induced by salt stress from Alfalfa. Mol Biol Rep, 2010, 37(2): 745-753.

[37] Jung KH, Hur J, Ryu CH, Choi Y, Chung YY, Miyao A, Hirochika H, An GH. Characterization of a rice chlorophyll-deficient mutant using the T-DNA gene-trap system. Plant Cell Physiol, 2003, 44(5): 463-472.

[38] Kai G, Wu C, Gen L, Zhang L, Cui L, Ni X. Biosynthesis and biotechnological production of anti-cancer drug Camptothecin. Phytochem Rev, 2015, 14(3): 525-539.

[39] Kaul K, Sabharwal PS. Effects of sucrose and kinetin on growth and chlorophyll synthesis in tobacco tissue cultures. Plant Physiol, 1971, 47(5): 691-695.

[40] Khan S, Karnat NM, Shankar D. India’s foundation for revitalization of local health traditions, pioneering in situ conservation strategies for medicinal plants and local cultures. Herbalgram, 2005, 68: 34-45.

[41] Kishore K, Singh M. Effect of bacosides A alcoholic extract of Bacopa monnieri Linn. (Brahmi) on experimental amnesia in mice. Indian J Exp Biol, 2005, 43: 640-645.

[42] Koch KE. Carbohydrate-modulated gene expression in plants. Ann Rev Plant Biol, 1996, 47(1): 509-540.

[43] Kulkarni AV, Malpathak NP. Micropropagation of Chonemorpha fragrans (moon) Alston: an endangered medicinal plant. Indian Drugs Bombay, 2006 43 12 1001

[44] Kulkarni AV, Patwardhan AA, Lele U, Malpathak NP. Production of camptothecin in the cultures of Chonemorpha grandiflora. Pharmacogn Res, 2010, 2(5): 296-299.

[45] Kumari M, Clarke HJ, Small I, Siddique KH. Albinism in plants: a major bottleneck in wide hybridization, androgenesis and doubled haploid culture. Crit Rev Plant Sci, 2009, 28(6): 393-409.

[46] Lalita MP, Murthy HN, Dandin VS, Koli SP, Manohar SH, Payamalle SD. Micropropagation of yellow mangosteen: a valuable endemic tree of India. J For Res, 2016, 27(1): 161-165.

[47] Larkin PJ, Scowcroft WR. Somaclonal variation—a novel source of variability from cell cultures for plant improvement. Theor Appl Genet, 1981, 60(4): 197-214.

[48] Lin CH, Lee LY, Tseng MJ. The effect of stratification and TDZ treatment on germination and protein synthesis of Pyrus serotina Rehd cv. Niauli. Ann Bot, 1994, 73: 515-523.

[49] Lipavska H, Konradova H. Somatic embryogenesis in conifers: the role of carbohydrate metabolism. In Vitro Cell Dev Biol Plant, 2004, 40(1): 23-30.

[50] Liu NT, Jane WN, Tsay HS, Wu H, Chang WC, Lin CS. Chloroplast genome aberration in micropropagation-derived albino Bambusa edulis mutants, ab1 and ab2. Plant Cell Tissue Organ Cult, 2007, 88(2): 147-156.

[51] Lokhande VH, Nikam TD, Penna S. Biochemical, physiological and growth changes in response to salinity in callus cultures of Sesuvium portulacastrum L. Plant Cell Tissue Organ Cult, 2010, 102(1): 17-25.

[52] Lokhande VH, Nikam TD, Patade VY, Ahire ML, Suprasanna P. Effects of optimal and supra-optimal salinity stress on antioxidative defence, osmolytes and in vitro growth responses in Sesuvium portulacastrum L. Plant Cell Tissue Organ Cult, 2011, 104(1): 41-49.

[53] Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem, 1951, 193(1): 265-275.

[54] Madke SS, Cherian KJ, Badere RS. A modified Murashige and Skoog media for efficient multiple shoot induction in Gmelina arborea Roxb. J For Res, 2014, 25(3): 557-564.

[55] Makara AM, Rubaihayo PR, Magambo MJS. Carry-over effect of Thidiazuron on banana in vitro proliferation at different culture cycles and light incubation conditions. Afr J Biotechnol, 2010, 9(21): 3079-3085.

[56] Mallik S, Nayak M, Sahu BB, Panigrahi AK, Shaw BP. Response of antioxidant enzymes to high NaCl concentration in different salt-tolerant plants. Biol Plant, 2011, 55(1): 191-195.

[57] Mohanty M, Bhola N, Mohanty S. Albino seedlings in Pongamia pinnata (L.) pierre. Indian For, 2005, 131: 127-128.

[58] Mohiuddin AKM, Karim NH, Sultana S, Ferdous Z. Recovery of green plantlets from albino shoot primordia derived from anther culture of Indica Rice (Oryza sativa L.). Trop Life Sci Res, 2011 22 1 1

[59] Mok DWS, Mok MC. Cytokinin metabolism and action. Ann Rev Plant Physiol Plant Mol Biol, 2001, 52: 89-118.

[60] Moyo M, Finnie JF, van Staden J. Recalcitrant effects associated with the development of basal callus-like tissue on caulogenesis and rhizogenesis in Sclerocarya birrea. Plant Growth Regul, 2011, 63(2): 187-195.

[61] Moyo M, Koetle MJ, van Staden J. Photoperiod and plant growth regulator combinations influence growth and physiological responses in Pelargonium sidoides. In Vitro Cell Dev Biol Plant, 2014, 50: 487-492.

[62] Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant, 1962, 15(3): 473-497.

[63] Murthy S, Saxena PK (1997) Morpho-physiological role of Thidiazuron in plants. Thesis submitted to the University of Guelph, pp 154–167. https://www.collectionscanada.gc.ca/obj/s4/f2/dsk2/ftp03/NQ27462.pdf. Accessed 24 Jan 2017

[64] Nakano Y, Asada K. Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol, 1981, 22(5): 867-880.

[65] Nambiar N, Tee CS, Maziah M. Effects of organic additives and different carbohydrates sources on proliferation of protocorm-like bodies in Dendrobium Alya Pink. Plant Omics J, 2012, 5: 10-18.

[66] Park YG, Kim SJ, Kang YM, Jung HY, Prasad DT, Kim SW, Chung YG, Choi MS. Production of ginkgolides and bilobalide from optimized the Ginkgo biloba cell culture. Biotechnol Bioprocess Eng, 2004, 9(1): 41-46.

[67] Patil JG, Ahire ML, Nikam TD. Influence of plant growth regulators on in vitro seed germination and seedling development of Digitalis purpurea L. Asian Australas J Plant Sci Biotechnol, 2012, 6: 12-18.

[68] Paul DK (1972) A handbook of nursery practice for Pinus caribaea var. hondurensis and other conifers in West Malaysia. Working Paper Number 19, FO: SF/MAL 12, UNDP/FAO, Kuala Lumpur, Malaysia

[69] Perveen S, Varshney A, Anis M, Aref IM. Influence of cytokinins, basal media and pH on adventitious shoot regeneration from excised root cultures of Albizia lebbeck. J For Res, 2011, 22(1): 47-52.

[70] Peter RH, Evert RF, Eichhorn SE. Biology of plants, 2005 7 New York: W.H. Freeman and Company Publishers 504 508

[71] Pierik RLM. In vitro culture of higher plants, 1987, Dordrecht: Martinus Nijhoff

[72] Pinero MC, Houdusse F, Garcia-Mina JM, Garnica M, del Amor FM. Regulation of hormonal responses of sweet pepper as affected by salinity and elevated CO2 concentration. Physiol Plant, 2014, 151(4): 375-389.

[73] Pogson B, Rissler H, Frank H. Wydrzynski T, Satoh K. The roles of carotenoids in photosystem II of higher plants. Photosystem II: The water/plastoquinone oxidoreductase in photosynthesis, 2006, Dordrecht: Springer 516 536

[74] Prathanturarug S, Pheakkoet R, Jenjittikul T, Chuakul W, Saralamp P. In vitro propagation of Stemona hutanguriana W. Chuakul, an endangered medicinal plant. Physiol Mol Biol Plants, 2012, 18(3): 281-286.

[75] Rahman MH, Alsadon AA. Photoautotrophic and photomixotrophic micropropagation of three potato cultivars. J Bio-Sci, 2007, 15: 111-116.

[76] Rastogi RP, Mehrotra BN. Compendium of Indian medicinal plants, 1993, New Delhi: CDRI, Publication Information Directorate 178

[77] Rezali NI, Sidik NJ, Saleh A, Osman NI, Adam NAM. The effects of different strength of MS media in solid and liquid media on in vitro growth of Typhonium flagelliforme. Asian Pac J Trop Biomed, 2017, 7(2): 151-156.

[78] Richet N, Afif D, Tozo K, Maillard P, Huber F, Priault P, Banvoy J, Gross P, Dizengremel P, Lapierre C, Perré P, Cabané M. Elevated CO₂ and/or ozone modify lignification in the wood of poplars (Populus tremula × alba). J Exp Bot, 2012, 63(11): 4291-4301.

[79] Rinaldi LMR. Germination of seeds of olive (Olea europaea L.) and ethylene production: effects of harvesting time and Thidiazuron treatment. J Hortic Sci Biotechnol, 2000, 75(6): 727-732.

[80] Rodriguez M, Costa TH, Festucci-Buselli RA, Silva LC, Otoni WC. Effects of flask sealing and growth regulators on in vitro propagation of neem (Azadirachta indica A. Juss.). In Vitro Cell Dev Biol Plant, 2012, 48(1): 67-72.

[81] Roh KS, Choi BY. Sucrose regulates growth and activation of rubisco in tobacco leaves in vitro. Biotechnol Bioprocess Eng, 2004, 9(3): 229-235.

[82] Sankar TYD (2010) In vitro culture of Camptotheca acuminate (Decaisne) in temporary immersion system (TIS): growth, development and production of secondary metabolites. Ph.D. thesis, University Hamburg, Germany

[83] Scandalios JG, Guan L, Polidoros AN. Scandalios JG. Catalases in plants: gene structure, properties, regulation, and expression. Oxidative stress and the molecular biology of antioxidant defenses, 1997, New York: Cold Spring Harbor Laboratory Press 343 406

[84] Schnapp SR, Preece JE. In vitro growth reduction of tomato and carnation microplants. Plant Cell Tissue Organ Cult, 1986, 6(1): 3-8.

[85] Sergio L, De Paola A, Cantore V, Pieralice M, Cascarano NA, Bianco VV, Di Venere D. Effect of salt stress on growth parameters, enzymatic antioxidant system, and lipid peroxidation in wild chicory (Cichorium intybus L.). Acta Physiol Plant, 2012, 34(6): 2349-2358.

[86] Shigeoka S, Ishikawa T, Tamoi M, Miyagawa Y, Takeda T, Yabuta Y, Yoshimura K. Regulation and function of ascorbate peroxidase isoenzymes. J Exp Bot, 2002, 53(372): 1305-1319.

[87] Srilatha T, Anithadevi U, Ugandhar T. Efficient plantlet regeneration from nodal explant culture of blackgram (Vigna mungo L.) Hepper. Biosci Discov, 2014, 5(2): 131-138.

[88] Sudipta KM, Kumara Swamy M, Anuradha M. Influence of various carbon sources and organic additives on in vitro growth and morphogenesis of Leptadenia reticulate (Wight & Arn), a valuable medicine plant of India. Int J Pharm Sci Rev Res, 2013, 21: 174-179.

[89] Us-Camas R, Castillo-Castro E, Aguilar-Espinosa M, Limones-Briones V, Rivera-Madrid R, Robert-Díaz ML, De-la-Peña C. Assessment of molecular and epigenetic changes in the albinism of Agave angustifolia Haw. Plant Sci, 2017, 263: 156-167.

[90] Wang Y, Yao RL. Plantlet regeneration of adult Pinus massoniana Lamb. trees using explants collected in March and thidiazuron in culture medium. J For Res, 2017, 28(6): 1169-1175.

[91] Watanabe S, Kojima K, Ide Y, Sasaki S. Effects of saline and osmotic stress on proline and sugar accumulation in Populus euphratica in vitro. Plant Cell Tissue Organ Cult, 2000, 63(3): 199-206.

[92] Wellburn AR. The spectral determination of chlorophylls a and b, as well as total carotenoids, using various solvents with spectrophotometers of different resolution. J Plant Physiol, 1994, 144(3): 307-313.

[93] Wu CH, Dewir YS, Hahn EJ, Paek KY. Optimization of culturing conditions for the production of biomass and phenolic from adventitious roots of Echinacea angustifolia. J Plant Biol, 2006, 49: 193-199.

[94] Yaacob JS, Mahmad N, Mat Taha R, Mohamed N, Mad Yussof AI, Saleh A. Optimization of culture conditions (sucrose, pH, and photoperiod) for in vitro regeneration and early detection of somaclonal variation in ginger lime (Citrus assamensis). Sci World J, 2014, 3: 262710.

[95] Yaseen M, Ahmad T, Sablok G, Standardi A, Hafiz IA. Role of carbon sources for in vitro plant growth and development. Mol Biol Rep, 2013, 40(4): 2837-2849.

[96] Yildiz M, Onde S, Ozgen M. Sucrose effects on phenolic concentration and plant regeneration from sugarbeet leaf and petiole explants. J Sugar Beet Res, 2007, 44(1/2): 1-15.

[97] Yoon YJ, Mobin M, Hahn EJ, Paek KY. Impact of in vitro CO2 enrichment and sugar deprivation on acclamatory responses of Phalaenopsis plantlets to ex vitro conditions. Environ Exp Bot, 2009, 65(2): 183-188.