Response surface methodology-based optimization of hairy roots cultures for in vitro AM production
DOI:
https://doi.org/10.14719/pst.5116Keywords:
Arbuscular mycorrhizal fungi (AMF), Hairy root production, Response surface methodology (RSM), Carrot root organ culture, Rhizobium rhizogenesAbstract
Arbuscular mycorrhizal fungi can do wonders in promoting the crop growth as well maintaining soil health. In vitro root organ culture technology is an exciting avenue for AM biofertilizer production. This study aims to optimize the key influencing factors for enhancing the hairy root production used for in vitro AMF culturing using response surface methodology, a statistical and mathematical tool used for designing optimization studies. Study uses Rhizobium rhizogenes MTCC 2364 for transformation in carrot explant. Factors considered for optimization are concentration of gelling agent (phytagel), carbon source (sucrose) and pH of the Modified Strullu and Romand medium (MSR). Design Expert software uses second order polynomial regression equation for predicting the outcome of each experiment. Totally, 17 experiments were run following Box-Behnken design and average hairy root length and average side branch emergence were taken as response. ANOVA analysis reveals that the concentration of phytagel and sucrose had a strong influence on root length, while the phytagel and pH of the medium had a strong effect on side branch emergence. Overall, taking into account the both responses, concentration of phytagel had a significant impact on hairy root production. The maximum average hairy root length obtained was 1.12 cm and number of side branches emerged were 6.78 per day. Based on these results, the optimal parameters were MSR medium with 3gL-1 phytagel, 11gL-1 sucrose and a pH of 4 for boosting hairy root development. This study is a cost-effective approach and minimizes the time taken for establishing the hairy root technology.
Downloads
References
Nations U. Peace, dignity and equality on a healthy planet [Internet]. Available from: https://www.un.org/en/global-issues/population.
Sreethu S, Chhabra V, Kaur G, Ali B. Biofertilizers as a greener alternative for increasing soil fertility and improving food security under climate change condition. Commun Soil Sci Plant Anal. 2024;55(2):261-85. https://doi.org/10.1080/00103624.2023.2265945
Mahdi SS, Hassan G, Samoon S, Rather H, Dar SA, Zehra B. Bio-fertilizers in organic agriculture. J Phytol. 2010;2:42-54.
Nosheen S, Ajmal I, Song Y. Microbes as biofertilizers, a potential approach for sustainable crop production. Sustain. 2021;13(4):1868. https://doi.org/10.3390/su13041868
Subhesh saurabh jha, Songachan LS. The usage of arbuscular mycorrhizal fungi (AMF) as a biofertilizer. Res Sq. [Preprint]. [cited 2023 Feb 08]. Available from: https://doi.org/10.21203/rs.3.rs-2559546/v1
Mitra PK, Adhikary R, Mandal V. Current status of mycorrhizal biofertilizer in crop improvement and its future prospects. In: Microbial Symbionts and Plant Health: Trends and Applications for Changing Climate. Springer Sci Rev; 2023.465-85. https://doi.org/10.1007/978-981-99-0030-5_17
Rai S, Shukla N. Biofertilizer: An alternative of synthetic fertilizers. Plant Archives. 2020;20(2):1374-79.
Ghorui M, Chowdhary S, Prakash B, Krishnan K, Djearamane S, Manjunathan J, et al. A review: In vitro cultivation of arbuscular mycorrhizal fungus for commercialisation. Oxid Commun. 2023;46(3).
St-Arnaud M, Hamel C, Vimard B, Caron M, Fortin JA. Enhanced hyphal growth and spore production of the arbuscular mycorrhizal fungus Glomus intraradices in an in vitro system in the absence of host roots. Mycol Res. 1996;100(3):328-32. https://doi.org/10.1016/S0953-7562(96)80164-X
Diop T. Ecophysiologie des champignons mycorhiziens à vésicules et arbuscules associés à acacia albida dans les zones sahéliennes et soudano-guinéenne du Sénégal. Applied Biological and Fundamental Sciences, Doctoral [dissertation]. Angers (France): University of Angers;1995.
Subramaniam S, Karunanandham K, Raja AS, Shukla SK, Uthandi S. EnZolv delignification of cotton spinning mill waste and optimization of process parameters using response surface methodology (RSM). Biotechnol Biofuels Bioprod. 2024;17(1):37. https://doi.org/10.1186/s13068-024-02473-w
Box GE, Behnken DW. Some new three level designs for the study of quantitative variables. Technometrics. 1960;2(4):455-75. https://doi.org/10.1080/ 00401706.1960.10489912
Bidondo LF, Pergola M, Silvani V, Colombo R, Bompadre J, Godeas A. Continuous and long-term monoxenic culture of the arbuscular mycorrhizal fungus Gigaspora decipiens in root organ culture. Fungal Biol. 2012;116(6):729-35. https://doi.org/10.1016/j.funbio.2012.04.007
Declerck S, Strullu DG, Plenchette C. In vitro mass-production of the arbuscular mycorrhizal fungus, Glomus versiforme, associated with Ri T-DNA transformed carrot roots. Mycol Res. 1996;100(10):1237-42. https://doi.org/10.1016/S0953-7562(96)80186-9
Devikrishna S, Kumutha K, Santhanakrishnan P, Priya LS. Standardization of process for hairy root production using Agrobacterium rhizogenes for monoxenic culture of arbuscular mycorrhizal fungi. Indian J Agric Sci. 2010;80(11):993-97.
Bécard G, Fortin JA. Early events of vesicular–arbuscular mycorrhiza formation on Ri T-DNA transformed roots. New Phytol. 1988;108(2):211-18. https://doi.org/10.1111/j.1469-8137.1988.tb03698.x
Bécard G, Piché Y. Establishment of vesicular-arbuscular mycorrhiza in root organ culture: review and proposed methodology. Methods in Microbiology. 1992;24:89-108. https://doi.org/10.1016/S0580-9517(08)70089-8
Declerck S, Strullu DG, Plenchette C. Monoxenic culture of the intraradical forms of Glomus sp. isolated from a tropical ecosystem: a proposed methodology for germplasm collection. Mycol. 1998;90(4):579-85. https://doi.org/10.1080/00275514.1998.12026946
Elsen A, Declerck S, De Waele D. Use of root organ cultures to investigate the interaction between Glomus intraradices and Pratylenchus coffeae. AEM. 2003;69(7):4308-11. https://doi.org/10.1128/AEM.69.7.4308-4311.2003
Srinivasan M, Kumar K, Kumutha K, Marimuthu P. Establishing monoxenic culture of arbuscular mycorrhizal fungus Glomus intraradices through root organ culture. J Apl Nat Sci. 2014;6(1):290-93. https://doi.org/10.31018/jans.v6i1.417
Radha V, Marimuthu RP, Kumutha K. Sporulation of arbuscular mycorrhizal fungus, Glomus intraradices, through root organ culture. Appli Biol Res. 2015;17(1):38-47. http://dx.doi.org/10.5958/0974-4517.2015.00006.3
Ridgway HJ, Kandula J, Stewart A. Optimising production of carrot hairy roots. N Z Plant Prot. 2004;57:77-80. https://doi.org/10.30843/nzpp.2004.57.6893
Klimaszewska K, Bernier-Cardou M, Cyr DR, Sutton BC. Influence of gelling agents on culture medium gel strength, water availability, tissue water potential and maturation response in embryogenic cultures of Pinus strobus L. In Vitro Cellular and Developmental Biology-Plant. 2000;36:279-86. https://doi.org/10.1007/s11627-000-0051-1
Zhou Y, Yan J, Xu BY, Wang BC. The study on mechanical properties of Phytagel medium. In: IOP Conf Ser Earth Environ Sci; 2019.346(1):012089. https://doi.org/10.1088/1755-1315/346/1/012089
Sandle T. Assessment of culture media in pharmaceutical microbiology. Am Pharm Review. 2014;17(4). https://doi.org/10.4155/pbp.13.64
George EF, Sherrington PD. Plant propagation by tissue culture: A hand book and directory of commercial laboratories. England: Exgetics Ltd.;1984.
George EF, Hall MA, Klerk GJ. The components of plant tissue culture media II: organic additions, osmotic and pH effects and support systems. In: Plant Propagation by Tissue Culture. The Netherlands, Dordrecht. Springer; 2008. p.115-73. https://doi.org/10.1007/978-1-4020-5005-3_4
Hunault G. Interactions acides organiques, pH ammonium et nitrate sur la croissance des tissus d'asperge cultivés in vitro. Botanique, Paris. Ann Sci Naturelles. 1985;13:63-75.
Smith DL, Krikorian AD. pH control of carrot somatic embryogenesis. In: Progress in Plant Cellular and Molecular Biology: Proceedings of the VIIth International Congress on Plant Tissue and Cell Culture, Amsterdam, The Netherlands, Dordrecht. Springer; 1990. p. 449-53. https://doi.org/10.1007/978-94-009-2103-0_69
Leifert C, Waites WM. Bacterial growth in plant tissue culture media. J Appl Bacteriol. 1992;72(6):460-66. https://doi.org/10.1111/j.1365-2672.1992.tb01859.x
Praveen N, Murthy HN. Synthesis of withanolide A depends on carbon source and medium pH in hairy root cultures of Withania somnifera. Ind Crops Prod. 2012;35(1):241-43. https://doi.org/10.1016/j.indcrop.2011.07.009
Praveen N, Chung IM. Effect of media strength and pH on the growth of hairy roots and production of gymnemic acid from Gymnema Sylvestre. MJS. 2019;18(4):31. https://doi.org/10.12723/mjs.51.3
Sivakumar G, Yu KW, Hahn EJ, Paek KY. Optimization of organic nutrients for ginseng hairy roots production in large-scale bioreactors. Curr Sci. 2005;89(4)641-49.
Merkli A, Christen P, Kapetanidis I. Production of diosgenin by hairy root cultures of Trigonella foenum-graecum L. Plant Cell Rep. 1997;16:632-36. https://doi.org/10.1007/BF01275505
Ahmad T, Abbasi NA, Hafiz IA, Ali A. Comparison of sucrose and sorbitol as main carbon energy sources in microprogation of peach rootstock GF-677. Pak J Bot. 2007;39(4):1269-75.
Wang Y, Weathers PJ. Sugars proportionately affect artemisinin production. Plant Cell Rep. 2007;26:1073-81. https://doi.org/10.1007/s00299-006-0295-2
Verma PC, Singh H, Negi AS, Saxena G, Rahman LU, Banerjee S. Yield enhancement strategies for the production of picroliv from hairy root culture of Picrorhiza kurroa Royle ex Benth. Plant Signaling Behav. 2015;10(5):e1023976. https://doi.org/10.1080/15592324.2015.1023976
Ashoka Babu VL, Banu S, Azamthulla M. Comparative anti-inflammatory potential of betalains of hairy root culture of Beta vulgaris and Beta vulgaris. WJPPS. 2022;11(2)1166-75. https://doi.org/10.20959/wjpps20222-21123
Yeo HJ, Park CH, Kim JK, Sathasivam R, Jeong JC, Kim CY, Park SU. Effects of chilling treatment on baicalin, baicalein and wogonin biosynthesis in Scutellaria baicalensis plantlets. Plants. 2022;11(21):2958. https://doi.org/10.3390/plants11212958
Yeo HJ, Kwon MJ, Han SY, Jeong JC, Kim CY, Park SU, Park CH. Effects of carbohydrates on rosmarinic acid production and in vitro antimicrobial activities in hairy root cultures of Agastache rugosa. Plants. 2023;12(4):797. https://doi.org/10.3390/plants12040797
Xu H, Park JH, Kim YK, Park NI, Lee SY, Park SU. Optimization of growth and pyranocoumarins production in hairy root culture of Angelica gigas Nakai. J Med Plants Res. 2009;3(11):978-81.
Ahn JC, Chong WS, Kim YS, Hwang B. Optimization of the sucrose and ion concentrations for saikosaponin production in hairy root culture of Bupleurum falcatum. BBE. 2006;11:121-26. https://doi.org/10.1007/BF02931895
Kusakari K, Yokoyama M, Inomata S. Enhanced production of saikosaponins by root culture of Bupleurum falcatum L. using two-step control of sugar concentration. Plant Cell Rep. 2000;19:1115-20. https://doi.org/10.1007/s002990000240
Massah M, Rabiei M. Effect of acetosyringone, sucrose and nutrients on transgenic hairy root induction in Chenopodium quinoa using different Rhizobium rhizogenes strains. https://doi.org/10.21203/rs.3.rs-2833983/v1
Downloads
Published
Versions
- 15-11-2024 (2)
- 09-11-2024 (1)
How to Cite
Issue
Section
License
Copyright (c) 2024 Priya PV, K Kumutha, R Subhashini, R Renuka, T Sivakumar, P Kannan
This work is licensed under a Creative Commons Attribution 4.0 International License.
Copyright and Licence details of published articles
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
Open Access Policy
Plant Science Today is an open access journal. There is no registration required to read any article. All published articles are distributed under the terms of the Creative Commons Attribution License (CC Attribution 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited (https://creativecommons.org/licenses/by/4.0/). Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
