Abstract
In the present study, influence of nitrate concentration on in vitro rooting and its ex vitro hardened tubers were investigated in D. hamiltonii. In vitro multiple shoots established on the MS medium were used as explants for this study and concentration of indole-3-butyric acid (1.23 µM) required for root initiation was determined. The effect of different nitrate concentrations in MS medium for in vitro rooting was investigated and positive (¼ and ½ strength) influence was observed on 14 days inoculation. The in vitro rooted plants (IVP) showed 80% survival upon hardening and exhibited similar growth pattern to seedling plants. The 1-year grew ex vitro hardened plants were evaluated for their tuber quality with reference to their yield and flavor metabolite 2-hydroxy-4-methoxy benzaldehyde (2H4MB) content. The IVP grown from ¼ nitrate strength media produced 155 ± 4.85 g FW of tuber biomass which is observed to be higher than full nitrate strength IVP (112 ± 2.52 g) which are grown under similar conditions in the greenhouse. The flavor metabolite content, total phenolics, total flavonoids, and antioxidant potential of these IVP tubers were evaluated. Upregulation of flavor biosynthetic pathway genes DhPAL, DhC4H and DhCoMT were observed in tubers of potted plants that developed from low nitrate strength culture media. In this study, superiority of tissue cultured plants was evident, wherein in vitro plants developed in low nitrate strength medium and acclimatized could produce a higher yield in tuber biomass and maintain relative content of flavor compounds in this endangered plant.
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References
Ahmad Z, Shahzad A, Sharma S (2018) Enhanced multiplication and improved ex vitro acclimatization of Decalepis arayalpathra. Biol Plant 62:1–10. https://doi.org/10.1007/s10535-017-0746-3
Bensaddek L, Gillet F, Saucedo JEN, Fliniaux M-A (2001) The effect of nitrate and ammonium concentrations on growth and alkaloid accumulation of Atropa belladonna hairy roots. J Biotechnol 85:35–40. https://doi.org/10.1016/S0168-1656(00)00372-2
Ch M, Reddy R, Murthy KSR (2013) A review on Decalepis hamiltonii Wight Arn. J Med Plant Res 7:3014–3029. https://doi.org/10.5897/JMPR2013.5099
Chashmi NA, Sharifi M, Karimi F, Rahnama H (2010) Differential production of tropane alkaloids in hairy roots and in vitro cultured two accessions of Atropa belladonna L. under nitrate treatments. Z Naturforsch 65:373–379. https://doi.org/10.1515/znc-2010-5-609
Chen JJ, Liao YJ (1993) Nitrogen-induced changes in the growth and metabolism of cultured potato tubers. J AM Soc Hortic Sci 118:831–834. https://doi.org/10.21273/JASHS.118.6.831
Collazo P, Montoliu L, Puigdomènech P, Rigau J (1992) Structure and expression of the lignin O-methyltransferase gene from Zea mays L. Plant Mol Biol 20:857–867. https://doi.org/10.1007/BF00027157
Damodaran S, Strader LC (2019) Indole 3-butyric acid metabolism and transport in Arabidopsis thaliana. Front in Plant Sci 10:851. https://doi.org/10.3389/fpls.2019.00851
Davis JM, Loescher WH, Hammond MX, Thornton RE (1986) Response of potatoes to nitrogen form and to change in nitrogen form at tuber initiation. J AM Soc Hortic Sci 111:70–72
Ezzine M, Ghorbel MH (2006) Physiological and biochemical responses resulting from nitrite accumulation in tomato (Lycopersicon esculentum Mill. cv. Ibiza F1). J Plant Physiol 163:1032–1039. https://doi.org/10.1016/j.jplph.2005.07.013
Francini A, Giro A, Ferrante A (2019) Chapter 11 - Biochemical and molecular regulation of phenylpropanoids pathway under abiotic stresses. In: Khan MIR, Reddy PS, Ferrante A, Khan NA (eds) Plant Signaling Molecules. Woodhead Publishing, pp 183–192
Frick EM, Strader CL (2018) Roles for IBA-derived auxin in plant development. J Exp Bot 69(2):169–177. https://doi.org/10.1093/jxb/erx298
Fritz C, Palacios-Rojas N, Feil R, Stitt M (2006) Regulation of secondary metabolism by the carbon–nitrogen status in tobacco: nitrate inhibits large sectors of phenylpropanoid metabolism. Plant J 46:533–548
Gallage NJ, Hansen EH, Kannangara R, Olsen CE, Motawia MS, Jorgensen K, Holme I, Hebelstrup K, Grisoni M, Moller BL (2014) Vanillin formation from ferulic acid in Vanilla planifolia is catalysed by a single enzyme. Nat Commun 5:1–14
Gangaprasad A, William Decruse S, Seeni S, Nair GM (2005) Micropropagation and ecorestoration of Decalepis arayalpathra (Joseph & Chandra.) Venter—An endemic and endangered ethnomedicinal plant of Western Ghats. Ind J of Biotechnol 4:265–270
GhoochaniKhorasani A, Roohollahi I, Golkar P (2022) Effect of different concentrations of nitrogen and 2, 4-D on callus and plantlet production of Zamioculcas zamiifolia Engl under in vitro condition. J Hortic Sci 35:55–62
Giridhar P (2007) Micro propagation of vanilla: a foremost orchid spice crop of humid tropics. J Ind Bot Soc 86:51–53
Giridhar P, Reddy BO, Ravishankar GA (2001) Silver nitrate influences in vitro shoot multiplication and root formation in Vanilla planifolia Andr. Curr Sci 81:1166–1170
Giridhar P, Gururaj HB, Ravishankar GA (2005a) In vitro shoot multiplication through shoot tip cultures of Decalepis hamiltonii Wight & Arn., a threatened plant endemic to Southern India. In Vitro Cell Dev Biol-Plant 41:77–80. https://doi.org/10.1079/IVP2004600
Giridhar P, Rajasekaran T, Ravishankar GA (2005b) Improvement of growth and root specific flavour compound 2-hydroxy-4-methoxy benzaldehyde of micropropagated plants of Decalepis hamiltonii Wight & Arn., under triacontanol treatment. Sci Hortic 106:228–236. https://doi.org/10.1016/j.scienta.2005.02.024
Giridhar P, Rajasekaran T, Ravishankar GA (2005c) Production of a root-specific flavour compound, 2-hydroxy-4-methoxy benzaldehyde by normal root cultures of Decalepis hamiltonii Wight and Arn (Asclepiadaceae). J Sci Food Agric 85:61–64
Huang D, Ou B, Prior RL (2005) The chemistry behind antioxidant capacity assays. J Agric Food Chem 53:1841–1856
Inoue K, Sewalt VJ, Ballance GM et al (1998) Developmental expression and substrate specificities of alfalfa caffeic acid 3-O-methyltransferase and caffeoyl coenzyme A 3-O-methyltransferase in relation to lignification. Plant Physiol 117:761–770
Izquierdo AM, del Torres MPN, Jiménez GS, Sosa FC (2011) Changes in biomass allocation and phenolic compounds accumulation due to the effect of light and nitrate supply in Cecropia peltata plants. Acta Physiol Plant 33:2135
Kamireddy K, Matam P, Priyanka PS, Parvatam G (2017) Biochemical characterization of a key step involved in 2H4MB production in Decalepis hamiltonii. J Plant Physiol 214:74–80. https://doi.org/10.1016/j.jplph.2017.04.006
Kamireddy K, Sonbarse PP, Mishra Shashank K, Agrawal L, Chauhan PS, Lata C, Parvatam G (2021) Proteomic approach to identify the differentially abundant proteins during flavour development in tuberous roots of Decalepis hamiltonii Wight & Arn. 3 Biotech 11:173. https://doi.org/10.1007/s13205-021-02714-x
Kiran K, Sonbarse PP, Veeresh L, Shetty NP, Parvatam G (2018) Expression profile of phenylpropanoid pathway genes in Decalepis hamiltonii tuberous roots during flavour development. 3 Biotech 8:365. https://doi.org/10.1007/s13205-018-1388-7
Kolachevskaya OO, Lomin SN, Arkhipov DV, Romanov GA (2019) Auxins in potato: molecular aspects and emerging roles in tuber formation and stress resistance. Plant Cell Rep 38:681–698. https://doi.org/10.1007/s00299-019-02395-0
Kumar V, Kim SH, Priatama RA et al (2020) NH4+ Suppresses NO3–-dependent lateral root growth and alters gene expression and gravity response in OsAMT1 RNAi mutants of rice (Oryza sativa). J Plant Biol 63:391–407. https://doi.org/10.1007/s12374-020-09263-5
Lea US, Slimestad R, Smedvig P, Lillo C (2007) Nitrogen deficiency enhances expression of specific MYB and bHLH transcription factors and accumulation of end products in the flavonoid pathway. Planta 225:1245–1253. https://doi.org/10.1007/s00425-006-0414-x
Lee JH, Lee JM, Lee S-H, Kim SM, Cha SW (2015) Comparison of artemisinin content and antioxidant activity from various organs of Artemisia species. Hortic Environ Biotechnol 56:697–703. https://doi.org/10.1007/s13580-015-0143-9
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Matam P, Parvatam G (2017a) Putrescine and polyamine inhibitors in culture medium alter in vitro rooting response of Decalepis hamiltonii Wight & Arn. Plant Cell Tiss Organ Cult 128:273–282. https://doi.org/10.1007/s11240-016-1108-0
Matam P, Parvatam G (2017b) Arbuscular mycorrhizal fungi promote enhanced growth, tuberous roots yield and root specific flavour 2-hydroxy-4-methoxybenzaldehyde content of Decalepis hamiltonii Wight & Arn. Acta Sci Pol Hortorum Cultus 16:3–10
McAlister FM, Jenkins CLD, Watson JM (1998) Sequence and expression of a stem-abundant caffeic acid O-methyltransferase cDNA from perennial ryegrass (Lolium perenne). Functional Plant Biol 25:225–235. https://doi.org/10.1071/pp97127
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Murthy KNC, Rajasekaran T, Giridhar P, Ravishankar GA (2006) Antioxidant property of Decalepis hamiltonii Wight & Arn. Indian J Exp Biol 44(10):832–837
Obul Reddy B, Giridhar P, Ravishankar GA (2002) The effect of triacontanol on micropropagation of Capsicum frutescens and Decalepis hamiltonii W & A. Plant Cell Tissue Organ Cult 71:253–258. https://doi.org/10.1023/A:1020342127386
Olsen KM, Lea US, Slimestad R, Verheul M, Lillo C (2008) Differential expression of four Arabidopsis PAL genes; PAL1 and PAL2 have functional specialization in abiotic environmental-triggered flavonoid synthesis. J Plant Physiol 165:1491–1499. https://doi.org/10.1016/j.jplph.2007.11.005
Pichersky E, Gang DR (2000) Genetics and biochemistry of secondary metabolites in plants: an evolutionary perspective. Trends Plant Sci 5:439–445. https://doi.org/10.1016/S1360-1385(00)01741-6
Pradeep M, Kiran K, Giridhar P (2016) A Biotechnological perspective towards improvement of decalepis hamiltonii: potential applications of its tubers and bioactive compounds of nutraceuticals for value Addition. In: Shahzad A, Sharma S, Siddiqui SA (eds) Biotechnological strategies for the conservation of medicinal and ornamental climbers. Springer International Publishing, Cham, pp 217–238
Pradeep M, Shetty NP, Giridhar P (2019) HPLC and ESI-MS analysis of vanillin analogue 2-hydroxy-4-methoxy benzaldehyde in swallow root – the influence of habitat heterogeneity on antioxidant potential. Acta Sci Pol Hortorum Cultus 18:21–28. https://doi.org/10.24326/.2019.2.3
Reddy BO, Giridhar P, Ravishankar GA (2001) In vitro rooting of Decalepis hamiltonii Wight & Arn., an endangered shrub, by auxins and root-promoting agents. Curr Sci 81:1479–1482
Rodrigues V, Kumar A, Gokul S, Verma RS, Rahman L, Sundaresan V (2020) Micropropagation, encapsulation, and conservation of Decalepis salicifolia, a vanillin isomer containing medicinal and aromatic plant. In Vitro Cell Dev Biol Plant 56(4):526–537
Roycewicz P, Malamy JE (2012) Dissecting the effects of nitrate, sucrose and osmotic potential on Arabidopsis root and shoot system growth in laboratory assays. Philos Trans R Soc Lond B, Biol Sci 367:1489–1500. https://doi.org/10.1098/rstb.2011.0230
Sadasivam S, Manickam A (2008) Biochemical Methods, 3rd. New Age International Publishers, New Delhi, India
Schmitz U, Lorz H (1990) Nutrient uptake in suspension cultures of gramineae. II. Suspension cultures of rice (Oryza sativa L.). Plant Sci 66:95–111. https://doi.org/10.1016/0168-9452(90)90174-M
Seginer I (2003) A dynamic model for nitrogen-stressed lettuce. Ann Bot 91:623–635. https://doi.org/10.1093/aob/mcg069
Seginer I, Buwalda F, Straten G (1999) Lettuce growth limited by nitrate supply. Acta Hortic. https://doi.org/10.17660/1999.507.16
Sharma S, Shahzad A (2012) Encapsulation technology for short-term storage and conservation of a woody climber, Decalepis hamiltonii Wight and Arn. Plant Cell Tiss Organ Cult 111:191–198. https://doi.org/10.1007/s11240-012-0183-0
Shekhawat MS, Kannan N, Manokari M, Ravindran CP (2015) In vitro regeneration of shoots and ex vitro rooting of an important medicinal plant Passiflora foetida L. through nodal segment cultures. J Genet Eng Biotechnol 13:209–214. https://doi.org/10.1016/j.jgeb.2015.08.002
Sirju-Charran G, Wickham LD (1988) The development of alternative storage sink sites in sweet potato Ipomoea batatas. Ann Bot 61:99–102. https://doi.org/10.1093/oxfordjournals.aob.a087533
Soare JR, Dinis TCP, Cunha AP, Almeida L (1997) Antioxidant activities of some extracts of Thymus zygis. Free Radic Res 26:469–478. https://doi.org/10.3109/10715769709084484
Srivastava A, Harish SR, Shivanandappa T (2006) Antioxidant activity of the roots of Decalepis hamiltonii (Wight & Arn.). LWT Food Sci Technol 39:1059–1065. https://doi.org/10.1016/j.lwt.2005.07.005
Srivastava A, Jagan Mohan Rao L, Shivanandappa T (2007) Isolation of ellagic acid from the aqueous extract of the roots of Decalepis hamiltonii: antioxidant activity and cytoprotective effect. Food Chem 103:224–233. https://doi.org/10.1016/j.foodchem.2006.08.010
The Wealth of India (1990) A dictionary of indian raw materials & industrial products. Publications & Information Directorate, Council of Scientific & Industrial Research, vol 1, p 161
Umashankar K, Giridhar P (2022) Biomass production and bioactive secondary metabolites of bioreactor grown callus suspension cultures of Swallow root (Decalepis hamiltonii Wight & Arn. J Emerg Technol Innov Res 9(3):g153–g158
Umashankar K, Pradeep M, Giridhar P (2022) In vitro elicitation supports the enrichment of 2H4MB production in callus suspension cultures of D hamiltonii Wight & Arn. Romanian Biotechnol Lett 27(1):3302–3308. https://doi.org/10.25083/rbl/27.1/3302-3308
Ved D, Saha D, Ravikumar K, Haridasan K (2015) Decalepis hamiltonii. The IUCN Red list of threatened species of threatened species 2015: e.T50126587A50131330. https://doi.org/10.2305/IUCN.UK.2015- 2.RLTS.T50126587A50131330.en. Accessed 13 Nov 2020
Wojtania A, Skrzypek E, Marasek-Ciolakowska A (2019) Soluble sugar, starch and phenolic status during rooting of easy- and difficult-to-root magnolia cultivars. Plant Cell Tiss Organ Cult 136:499–510. https://doi.org/10.1007/s11240-018-01532-z
Wu X, Wu J, Luo Y, Bragg J, Anderson O, Vogel J, Gu YQ (2013) Phylogenetic, molecular, and biochemical characterization of caffeic acid o-methyltransferase gene family in Brachypodium distachyon. Int J Plant Genomics 2013:423189
Zhang J, Xu L, Wang F, Deng M, Yi K (2012) Modulating the root elongation by phosphate/nitrogen starvation in an OsGLU3 dependant way in rice. Plant Signal Behav 7:1144–1145
Acknowledgements
The authors acknowledge the Science & Engineering Research Board (SERB), Council for Scientific and Industrial Research (CSIR) Government of India, New Delhi for the research Grants EMR_2016_001049 and 12 FYP-Network Program-BSC0106, respectively.
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The study was supported by SERB, CSIR Government of India, New Delhi for the research Grant EMR_2016_001049 and 12 FYP-Network Program-BSC0106, respectively.
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KU: investigation, analysis, and compiling the data. MP: conceptualization, investigation, analysis, manuscript writing. PG: funding acquisition, overall supervision of research work, interpretation, writing and editing.
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Koppada, U., Matam, P. & Parvatam, G. In vitro-derived plants grown from low nitrate medium produced quality tubers in Decalepis hamiltonii Wight & Arn. 3 Biotech 12, 200 (2022). https://doi.org/10.1007/s13205-022-03256-6
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DOI: https://doi.org/10.1007/s13205-022-03256-6