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Induction of in vitro micro rhizomes and assessment of yield, quality, and clonal fidelity in ex vitro established plants of ginger (Zingiber officinale Rosc.)

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Abstract

In vitro micro rhizome technology is a highly effective approach in combating seed-borne diseases and ensuring the production of healthy and high-quality planting material in ginger (Zingiber officinale Rosc.). To gauge the efficiency of micro rhizome production and their viability ex vitro, an experiment was conducted on several ginger varieties viz., IISR Varada, IISR Mahima, IISR Rejatha, and Karthika, at ICAR- Indian Institute of Spices Research, Kozhikode, Kerala, India. This experiment adhered to established protocols and standardized procedures. All four varieties exhibited varying rates of micro rhizome production after 180 days of culture. Among these, IISR Varada demonstrated the highest mean weight of cultured plant mass (96.0 ± 4.41 g), followed by Karthika (91.4 ± 5.72 g), IISR Rejatha (78.45 ± 5.59 g), and IISR Mahima (72.4 ± 3.56 g). IISR Rejatha exhibited the maximum number of micro rhizomes per bottle (11.35 ± 0.81) compared to IISR Mahima (10.8 ± 0.54), Karthika (9.8 ± 0.58), and IISR Varada (9.0 ± 0.63). The highest total weight of micro rhizome and mean weight of a single micro rhizome per bottle were recorded in IISR Varada (32.6 ± 1.92 g and 3.9 ± 0.29 g, respectively), followed by Karthika (27.1 ± 1.19 g and 2.9 ± 0.16 g, respectively), IISR Rejatha (27.0 ± 1.79 g and 2.5 ± 0.18 g, respectively) and IISR Mahima (24.5 ± 1.10 g and 2.4 ± 0.16 g, respectively). Besides, IISR Varada, followed by Karthika, emerged as the most promising varieties for micro rhizome production in terms of their multiplication rate. The evaluation extended to the first and second-generation progenies of micro rhizomes from IISR Varada. Results indicated the successful establishment of first-generation micro rhizomes in grow bags and second-generation micro rhizomes in the field, employing both direct planting and transplanting methods. Assessment of quality parameters revealed that the second-generation (V2) transplanted plants of micro rhizomes of IISR Varada exhibited the highest essential oil content 0.78%. The total phenolic content was highest in second-generation (V2) rhizomes directly planted in soil (23 mg GAE/g), whereas the first-generation micro rhizomes raised in grow bags registered the highest total flavonoid content (TFC) of 1.39 mg QE/g. Moreover, the genetic fidelity test conducted on the first and second generations (V1 and V2, respectively) of micro rhizome-derived plants, using molecular markers, exhibited a monomorphic banding pattern similar to that of the mother plant, confirming their genetic stability.

Key message

In vitro micro rhizomes of ginger have the potential to combat pathogens transmitted through seed materials, while simultaneously preserving clonal fidelity and ensuring high quality standards.

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References

  • Abbas M, Aly U, Taha H, Gaber ES (2014) In vitro production of microrhizomes in ginger (ZingiberOfficinaleRosc). J Microbiol Biotechnol Food Sci 04(02):142–148. https://doi.org/10.15414/jmbfs.2014.4.2.142-148

    Article  CAS  Google Scholar 

  • Akshitha HJ, Prasath D, Umesha K, Venkataravanappa V (2022) Molecular characterization of ginger genotypes using RAPD and SSR markers. J Horticult Sci 17(1):95–102. https://doi.org/10.24154/jhs.v17i1.1052

    Article  Google Scholar 

  • Akshitha HJ (2018) Molecular and chemo profiling of ginger (Zingiber Officinale Rosc) genotypes. Ph.D. thesis, UHS, Bagalkot

  • American Spice Trade Association (1997) ASTA 21.0 method. Official analytical methods of the American Spice Trade Association (ASTA)

  • Archana CP, Geetha SP, Balachandran I (2013b) Microrhizome and minirhizome production in three high yielding cultivars of ginger (zingiberofficinalerosc.). Int J Curr Microbiol App Sci 2(10):477–84

    Google Scholar 

  • Archana CP, Geetha SP, Balachandran I (2013c) In vitro microrhizome induction in three high yielding cultivars of Zingiberofficinale rosc. and their phytopathological analysis. Int J Adv Biotechnol Res 4(3):296–300

    CAS  Google Scholar 

  • Archana TS, Mesta RK, Basavarajappa MP, Kiran Kumar KC (2023) Promoting resilience in ginger: Elicitor-driven strategies to combat the rhizome rot disease. J Phytopathol 171(11–12):627–641. https://doi.org/10.1111/jph.13223

    Article  CAS  Google Scholar 

  • Archana Cheethaparambil, Geetha S Pillai, Indira Balachandran (2013a) Comparative studies on in vitro micro rhizome induction in three varieties of Curcuma longa (turmeric) - the role of two stress hormones. Int J Sci Res Manag: 230–237. https://bipublication.com/files/IJABR-V4I3-2013-02.pdf

  • Askari N, Visser RG, De Klerk GJ (2018) Growth of lily bulblets in vitro, a review. Int J Horticult Sci Technol 5(2):133–143. https://doi.org/10.22059/ijhst.2018.268870.263

    Article  CAS  Google Scholar 

  • Babu KN, Samsudeen K, Divakaran M, Pillai GS, Sumathi V, Praveen K, Ravindran PN, Peter KV (2016) Protocols for in vitro propagation, conservation, synthetic seed production, embryo rescue, microrhizome production, molecular profiling, and genetic transformation in ginger (Zingiberofficinale Roscoe.). Methods Mol Biol 28:403–426. https://doi.org/10.1007/978-1-4939-3332-7_28

    Article  CAS  Google Scholar 

  • Babu KN, Samsudeen K, Minoo D, Geetha S. Pillai and Ravindran PN (2005) Tissue culture and biotechnology of ginger, pp. 181- 210. In PN Ravindran and K Nirmal Babu (eds). Ginger –The genus Zingiber, CRC Press, Boca Raton, USA. ISBN: 9780429120770

  • Chang C, Yang M, Wen H, Chern J (2002) Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J Food Drug Anal 10:178–182. https://doi.org/10.38212/2224-6614.2748

    Article  CAS  Google Scholar 

  • Cyriac A, Paul R, Prasath D, Deepesh PV, Parthasarathy VA (2016) Transferability of ginger, turmeric and large cardamom SSR primers to small cardamom (Elettaria cardamomum Maton). J Trop Agric 53(2):107–115. https://jtropag.kau.in/index.php/ojs2/article/view/340

  • Dev H, Sharma V (2022) Genetic variability in ginger (Zingiberofficinale Rosc.). Int J Bio-resource Stress Manag 13(7):709–717. https://doi.org/10.23910/1.2022.2941a

    Article  Google Scholar 

  • Doyle J (1991) DNA protocols for plants. In: Molecular techniques in taxonomy. Springer, Berlin Heidelberg, pp 283–293. https://doi.org/10.1007/978-3-642-83962-7_18

  • Dusabumuremyi JC, Duhoranimana E, Chen X (2022) Health benefits of turmeric and ginger. Handb Nutraceuticals Nat Prod: Biological, Medicinal, and Nutritional Properties and Applications 2:259–268. https://doi.org/10.1002/9781119746843.ch24

    Article  CAS  Google Scholar 

  • Estouka I, Alhagdow M, Bughrara S (2021) Simple micropropagtion method of ginger (Zingiberofficinale Rosc.). J Genet Genom Plant Breed 5(4):106–114

    CAS  Google Scholar 

  • George NM, Raghav SB, Prasath D (2022) Direct in vitro regeneration of medicinally important Indian and exotic red-colored ginger (Zingiberofficinale Rosc.) and genetic fidelity assessment using ISSR and SSR markers. In Vitro Cell Dev Biol-Plant 58(4):551–558. https://doi.org/10.1007/s11627-022-10268-7

    Article  CAS  Google Scholar 

  • Gezahegn G, Feyisa T, Rezene Y (2024) Induction of micro-rhizomes for in vitro ginger (Zingiberofficinale Rosco) disease-free planting materials regeneration. Biotechnol Rep 41:e00820. https://doi.org/10.1016/j.btre.2023.e00820

    Article  CAS  Google Scholar 

  • Guji MJ, Yetayew HT, Kidanu ED (2019) Yield loss of ginger (Zingiberofficinale) due to bacterial wilt (Ralstoniasolanacearum) in different wilt management systems in Ethiopia. Agric Food Sec 8:1–11. https://doi.org/10.1186/s40066-018-0245-6

    Article  Google Scholar 

  • Hemanthakumar AS, Preetha TS (2023) Macro-and micropropagation of plants for income generation. In: Conservation and Sustainable Utilization of Bioresources. Springer Nature, Singapore, pp 409–450. https://doi.org/10.1007/978-981-19-5841-0_17

  • Higgins S, Rose K, Wilson R, Francis RD, Watson CT, Riley CK (2020) In vitro propagation of ZingiberOfficinale Rosc. Biotech Res 6(1):1–6

    Google Scholar 

  • Hussain MJ, Abbas Y, Nazli N, Fatima S, Drouet S, Hano C, Abbasi BH (2022) Root cultures, a boon for the production of valuable compounds: a comparative review. Plants 11(3):439. https://doi.org/10.3390/plants11030439

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Islam MA, Kloppstech K, Jacobsen HJ (2004) Efficient Procedure for in vitro microrhizome induction in Curcuma longa L. (Zingiberaceae) - A medicinal plant of tropical Asia. Plant Tissue Cult 14(2):123–134

    Google Scholar 

  • Kandiannan K, Aravind S, Aarthi S, Akshitha HJ, Babu KN (2020) Innovative techniques in quality planting material production of spices. Int J Innov Hortic 9(2):104–119. https://doi.org/10.5958/2582-2527.2020.00014.7

    Article  Google Scholar 

  • Kavyashree R (2009) An efficient in vitro protocol for clonal multiplication of ginger var Varada. Int J Biotechnol 8:328–331

    Google Scholar 

  • Lo-Apirukkul S, Jenjittikul T, Saralamp P, Prathanturarug S (2012) Micropropagation of a Thai medicinal plant for women’s health, Curcuma comosa Roxb., via shoot and microrhizome inductions. J Nat Med 66:265–270. https://doi.org/10.1007/s11418-011-0577-z

    Article  CAS  PubMed  Google Scholar 

  • Macalalad AE, Christopher Jay T. Robidillo, Eufrocinio C Marfori (2016) Influence of different cytokinins on the growth, [6]-gingerol production and antioxidant activity of in vitro multiple shoot culture of ginger (Zingiber officinale Roscoe). Res J Med Plants 10:194–200. https://scialert.net/abstract/?doi=rjmp.2016.194.200

  • Mehaboob VM, Faizal K, Shamsudheen KM, Raja P, Thiagu G, Shajahan A (2019) Direct organogenesis and microrhizome production in ginger (Zingiberofficinale Rosc.). J Pharmacogn Phytochem 8(3):2880–2883

    CAS  Google Scholar 

  • Miri SM (2020) Micropropagation, callus induction and regeneration of ginger (Zingiberofficinale Rosc.). Open Agricult 5(1):75–84. https://doi.org/10.1515/opag-2020-0008

    Article  Google Scholar 

  • Moghaddam M, Mehdizadeh L (2017) Chemistry of essential oils and factors influencing their constituents. In: Soft chemistry and food fermentation. Academic Press, pp 379–419. https://doi.org/10.1016/B978-0-12-811412-4.00013-8

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Plant Physiol 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x

    Article  CAS  Google Scholar 

  • Nayak S, Kaur T, Mohanty S, Ghosh G, Choudhury R, Acharya L, Subudhi E (2011) In vitro and ex vitro evaluation of long-term micropropagated turmeric as analyzed through cytophotometry, phytoconstituents, biochemical and molecular markers. Plant Growth Regul 64:91–98. https://doi.org/10.1007/s10725-010-9541-2

    Article  CAS  Google Scholar 

  • Nazir R, Gupta S, Dey A, Kumar V, Yousuf M, Hussain S, Pandey DK (2021) In vitro propagation and assessment of genetic fidelity in Dioscoreadeltoidea, a potent diosgenin yielding endangered plant. S Afr J Bot 140:349–355. https://doi.org/10.1016/j.sajb.2020.07.018

    Article  CAS  Google Scholar 

  • Prasath D, Kandiannan K, Srinivasan V, Leela NK, Anandaraj M (2018) Comparison of conventional and transplant production systems on yield and quality of ginger (Zingiberofficinale). Indian J Agric Sci 88(4):615–20

    Article  CAS  Google Scholar 

  • Ravi Y, Gangadharappa PM, Shantappa T, Prashant A (2022) Performance evaluation of ginger genotypes on their yield and nutrient uptake under the coastal Karnataka, India. Int J Bio-res Stress Manag 13(Jun 6):630–637. https://doi.org/10.23910/1.2022.2780a

    Article  CAS  Google Scholar 

  • Rout GR, Palai SK, Samantary S, Das P (2001) Effect of growth regulator and culture conditions on shoot multiplication and rhizome formation in ginger (Zingiberofficianale Rosc.) in vitro. In Vitro Cell Dev Biol-Plant 37(6):814–819. https://doi.org/10.1007/s11627-001-0135-6

    Article  CAS  Google Scholar 

  • Sakamura F, Suga T (1989) Zingiber officinale Roscoe (Ginger): In Vitro propagation and the production of volatile constituents. Biotechnol Agric For 524–538. https://doi.org/10.1007/978-3-642-73617-9_29

  • Shaaban A, Elnfishy N, Aween Z, Abdelah E, Abughni E (2023) In vitro micropropagation of ginger plant (Zingiberofficinale). Sci J Fac Sci-Sirte University 3(2):154–161. https://doi.org/10.37375/sjfssu.v3i2.1659

    Article  Google Scholar 

  • Sharifi-Rad M, Varoni EM, Salehi B, Sharifi-Rad J, Matthews KR, Ayatollahi SA, Kobarfard F, Ibarahim SA, Mnayer D, Zakaria ZA (2017) Plants of the genus Zingiber as a source of bioactive phytochemicals: From tradition to pharmacy. Molecules 22(12):2145. https://doi.org/10.3390/molecules22122145

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shylaja MR, Parvathy KI, Lekha L (2017) Micro rhizome technology for large scale production of disease free seed rhizomes of ginger (ZingiberOfficinale Rosc.). Indian J Arecanut Spices Med Plants 19(2):6–9

    Google Scholar 

  • Shylaja MR, Sabitha KR, Kumar PKS, Narayanankutty C, Narayanankutty MC, Mathew D (2018) Production technology for In Vitro induced micro-rhizomes of ginger in high-tech poly-house. Indian J Arecanut Spices Med Plants 20(3):3–7. https://doi.org/10.2139/ssrn.3577809

    Article  Google Scholar 

  • Singleton V, Rossi J (1965) Colorimetry of total phenolic compounds with phosphomolybdic-phosphotungstic acid reagents. Am J Enol Vitic 16:144–158. https://doi.org/10.5344/ajev.1965.16.3.144

    Article  CAS  Google Scholar 

  • Swarnathilaka DBR, Kottearachchi NS, Weerakkody WJSK (2016) Factors affecting on induction of microrhizomes in ginger (ZingiberOfficinale Rosc.), cultivar local from Sri Lanka. British Biotechnol J 12(2):1–7. https://doi.org/10.9734/BBJ/2016/2390

    Article  Google Scholar 

  • Tewelde S, Patharajan S, Teka Z, Sbhatu DB (2020) Assessing the efficacy of broad-spectrum antibiotics in controlling bacterial contamination in the in vitro micropropagation of ginger (Zingiber officinale Rosc). Sci World J 2020. https://doi.org/10.1155/2020/6431301

  • Uma S, Karthic R, Kalpana S, Backiyarani S (2023) Evaluation of temporary immersion bioreactors for in vitro micropropagation of banana (Musa spp.) and genetic fidelity assessment using flow cytometry and simple-sequence repeat markers. S Afr J Bot 157:553–565. https://doi.org/10.1016/j.sajb.2023.04.006

    Article  CAS  Google Scholar 

  • Wardani RS, Schellack N, Govender T, Dhulap AN, Utami P, Malve V, Wong YC (2023) Treatment of the common cold with herbs used in Ayurveda and Jamu: monograph review and the science of ginger, liquorice, turmeric and peppermint. Drugs Context 12. https://doi.org/10.7573/dic.2023-2-12

  • Yan D, Wang Q, Li Y, Guo M, Guo X, Ouyang C, Cao A (2022) Efficacy and economics evaluation of seed rhizome treatment combined with preplant soil fumigation on ginger soilborne disease, plant growth, and yield promotion. J Sci Food Agric 102(5):1894–1902. https://doi.org/10.1002/jsfa.11526

    Article  CAS  PubMed  Google Scholar 

  • Zahid NA, Jaafar HZ, Hakiman M (2021a) Alterations in microrhizome induction, shoot multiplication and rooting of ginger (Zingiberofficinale Roscoe) var. Bentong with regards to sucrose and plant growth regulators application. Agronomy 11(2):320. https://doi.org/10.3390/agronomy11020320

    Article  CAS  Google Scholar 

  • Zahid NA, Jaafar HZ, Hakiman M (2021b) Micropropagation of ginger (Zingiberofficinale Roscoe) ‘Bentong’and evaluation of its secondary metabolites and antioxidant activities compared with the conventionally propagated plant. Plants 10(4):630. https://doi.org/10.3390/plants10040630

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhao X, Yu S, Wang Y, Jiang D, Zhang Y, Hu L, Zhu Y, Qie J, Yin J, Liu Y, Cai X (2023) Field performance of disease-free plants of ginger produced by tissue culture and agronomic, cytological, and molecular characterization of the morphological variants. Agronomy 13(1):74. https://doi.org/10.3390/agronomy13010074

    Article  CAS  Google Scholar 

  • Zheng Y, Liu Y, Ma M, Xu K (2008) Increasing in vitro microrhizome production of ginger (Zingiberofficinale Roscoe). Acta Physiol Plant 30:513–519. https://doi.org/10.1007/s11738-008-0149-3

    Article  CAS  Google Scholar 

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Acknowledgements

The authors are thankful to the Mission for Integrated Development in Horticulture (MIDH), Government of India for funding through DASD, Kozhikode and Director and Heads of ICAR-IISR, Kozhikode for facilities and guidance.

Funding

Partial financial support was received from Mission for Integrated Development in Horticulture (MIDH), Government of India.

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Authors and Affiliations

  1. ICAR-Indian Institute of Spices Research, Kozhikode, Kerala, India

    Sharon Aravind, Nisthar E, K. C. Chaithanya, R. Sivaranjani, K. Kandiannan, V. Srinivasan, S. Mukesh Sankar & K. Nirmal Babu

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  1. Sharon Aravind
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  2. Nisthar E
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  3. K. C. Chaithanya
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  4. R. Sivaranjani
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  7. S. Mukesh Sankar
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  8. K. Nirmal Babu
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Contributions

Sharon Aravind conceptualized the project, conducted investigation, developed the methodology, wrote the original draft, and reviewed the work. Nisthar E prepared cultures, induced micro rhizomes, and collected data. Chaithanya K.C analyzed genetic fidelity. R Sivaranjani studied the biochemical aspects of micro rhizomes and analyzed the data. K. Kandiannan performed formal analysis, curated data, and reviewed and revised the manuscript. V. Srinivasan administered the project, analyzed data, and reviewed and revised the manuscript. Mukesh Sankar conducted formal data analysis. K. Nirmal Babu contributed to the methodology, provided resources, and supervised the project.

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Correspondence to Sharon Aravind.

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Aravind, S., E, N., Chaithanya, K.C. et al. Induction of in vitro micro rhizomes and assessment of yield, quality, and clonal fidelity in ex vitro established plants of ginger (Zingiber officinale Rosc.). Plant Cell Tiss Organ Cult 157, 28 (2024). https://doi.org/10.1007/s11240-024-02751-3

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