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Development of an Effective Sterilization Protocol for Plant Tissue Culture Studies in Superfruit Aronia [Aronia melanocarpa (Michaux) Elliot]

Year 2024, Volume: 7 Issue: 6, 679 - 685, 15.11.2024
https://doi.org/10.47115/bsagriculture.1548432

Abstract

Effective sterilization protocols are crucial for a successful tissue culture study in Aronia. These protocols directly influence contamination rates, shoot health, and root development. In this context, the study aims is to develop an effective sterilization protocol for plant tissue culture studies in Aronia [Aronia melanocarpa (Michaux) Elliot], commonly known as the "superfruit." In the study, the Nero Aronia variety shoot tips were used as material. The sterilized shoot tips were transferred to the respective plant tissue culture media in a randomized parcels trial pattern with three replicates, each containing three explants per replicate. Various concentrations and combinations of sterilizing agents, such as sodium hypochlorite (NaOCl), hydrogen peroxide (H2O2), mercuric chloride (HgCl2), and ethanol (C2H5OH), were evaluated to determine their effectiveness in maintaining tissue health and reducing contamination. Twelve protocols were developed, incorporating different concentrations of these chemicals. The data were subjected to statistical analysis using the SAS software package (LSD0.05-Proc GLM). Analysis of variance (ANOVA) was performed to evaluate differences among sterilization treatments for each parameter. The results showed that the combination of 5% NaOCl and 3% H2O2 (10 min each) provided the lowest average contamination rate 0.0%, the highest average number uncontaminated explants 9.0 pieces, shoot length 3.0 cm and root length 2.5 cm demonstrating the sterilization efficiency of this combination. On the other hand, protocols containing HgCl2, especially at higher concentrations, resulted in impaired root development. High ethanol concentrations also contributed to effective sterilization, with the combination of 7% NaOCl (10 min) and 80% ethanol (5 min) yielding a low contamination rate (22.0%) and preserving tissue health. This study emphasizes balancing sterilization protocols between effective contamination control and tissue viability. The findings are expected to benefit the improvement and development of tissue culture techniques for Aronia and similar species, providing a basis for further research on effective sterilization practices, currently limited in Aronia tissue culture.

References

  • Agrawal A, Gowthami R, Chander S, Srivastava, V, Shankar M, Deepak DA. 2024. Biotechnological tools for conservation of plant genetic diversity. Sustainable Utilization and Conservation of Plant Genetic Diversity, Springer Nature, Singapore, pp: 711-752.
  • Ahmadpoor F, Zare N, Asghar R, Sheikhzadeh P. 2022. Sterilization protocols and the effect of plant growth regulators on callus induction and secondary metabolites production in in vitro cultures Melia azedarach L. AMB Express, 12: 1-12.
  • Al Ghasheem N, Stănică F, Petıcılă AG, Venat O. 2018. In vitro effect of various sterilization techniques on peach (Prunus persica (L.) Batsch) explants. Scient Papers, 227: 227-234.
  • Almokar HM M, Pırlak L. 2018. Propagation of Aronia (Aronia melanocarpa) with tissue culture. Selcuk J Agri Food Sci, 32(3): 549-558.
  • Angel LPL, Abu Bakar D, Sundram S. 2021. A non-invasive tissue culture system for performing artificial pathogenicity assays of Ganoderma boninense. Australasian Plant Pathol, 2021: 1-4.
  • Anuruddi HIGK, Nakandalage N, Fonseka DK. 2023. New insights for the production of medicinal plant materials: ex vitro and in vitro propagation. In Biosynthesis of Bioactive Compounds in Medicinal and Aromatic Plants: Manipulat Convent Biotechnol Approac, 2023: 181-212.
  • Bidabadi SS, Jain SM. 2020. Cellular, molecular, and physiological aspects of in vitro plant regeneration. Plants, 9(6): 702.
  • Cargnelutti D, Tabaldi LA, Spanevello RM, de Oliveira Jucoski G, Battisti V, Redin M, Schetinger MRC. 2006. Mercury toxicity induces oxidative stress in growing cucumber seedlings. Chemosphere, 65(6): 999-1006.
  • Cassells AC. 2012. Contamination and its impact in tissue culture. Plant Cell Tissue Organ Cult, 100(2): 87-90.
  • da Silva JA T, Winarto B, Dobránszki J, Cardoso JC, Zeng S. 2016. Tissue disinfection for preparation of culture. Folia Horticult, 28(1): 57-75.
  • Dagne H, Palanivel H, Yeshitila A, Benor S, Abera S, Abdi A. 2023. Advanced modeling and optimizing for surface sterilization process of grape vine (Vitis vinifera) root stock 3309C through response surface, artificial neural network, and genetic algorithm techniques. Heliyon, 9(8): e18628
  • El-Sherif NA. 2019. Impact of plant tissue culture on agricultural sustainability. Sustain Agri Environ Egypt: Part II: Soil-Water-Plant Nexus, 2019: 93-107.
  • Fidancı A. 2015. Türkiye için yeni bir minör meyve: Aronia bitkisi ve yetiştirme teknikleri. Bahçe, I(Özel Sayı): 1177-1180.
  • Fielder H, Beale T, Jeger MJ, Oliver G, Parnell S, Szyniszewska AM, Cunniffe NJ. 2024. A synoptic review of plant disease epidemics and outbreaks published in 2022. Phytopathology, 114(8): 1717-1732.
  • George EF, Sherrington PD. 1984. Plant propagation by tissue culture. Plant Cell Rep, 3(2): 85-91.
  • Hannan JM. 2013. Aronia berries profile. Iowa State University Extension and Outreach, Commercial Horticulture Field Specialist, October 2013, Ames, US.
  • Jurikova T, Mlcek J, Skrovankova S, Sumczynski D, Sochor J, Hlavacova I, Snopek L, Orsavova J. 2017. Fruits of black chokeberry Aronia melanocarpa in the prevention of chronic diseases. Molecules, 22: 944: 1-23.
  • Kampf G, Kampf G. 2018. Ethanol antiseptic stewardship: Biocide resistance and clinical implications, 2018: 9-35.
  • Karaokur OF, Kaya F, Yavuz E, Yenipinar, A. 2019. Comparison of commonly used statistics package programs. BSJ Eng Sci, 2(1): 26-32.
  • Kaya C, Sariyer T, Sahin E. 2023. Determination of effective surface sterilization protocol in in vitro tissue culture for Giant Snowdrop (Galanthus elwesii Hook) bulbs. Int J Agri Environ Food Sci, 7(2): 345-348. https://doi.org/10.31015 /jaefs.2023.2.12
  • Kaya Ç, Sarıyer T. 2024. Comparative biplot analysis of micropropagation of viking aronia cultivar in different plant tissue culture media. Anadolu Ege Tar Araş Enst Derg, 34(1): 90-96. https://doi.org/10.18615/anadolu.1494423
  • Knudson M. 2009. Plant Guide for black chokeberry (Photinia melanocarpa (Michx.) USDA-Natural Resources Conservation Service. URL= http://plant-materials.nrcs.usda.gov- (accessed date: June 10, 2024).
  • Kulling SE, Rawel HM. 2008. Chokeberry (Aronia melanocarpa) - a review on the characteristic components and potential health effects. Plant Medic, 74: 1625-1634.
  • Leifert C, Cassells AC. 2001. Microbial hazards in plant tissue and cell cultures. In Vitro Cell Developmental Biol-Plant, 37: 133-138.
  • Lema-Rumińska J, Sadowska K, Tymoszuk A, Andrzejewska J. 2023. Scutellarin and other metabolites as well as morphological and molecular characterization of the Scutellaria barbata lines from in vitro and in vivo cultivation. Indust Crops Prod, 195: 116464.
  • Mannino G, Gentile C, Ertani A, Serio G, Bertea C. M. 2021. Anthocyanins: Biosynthesis, distribution, ecological role, and use of biostimulants to increase their content in plant foods—A review. Agriculture, 11(3): 212.
  • Mekonnen T, Diro M, Sharma M. 2013. An alternative safer and cost effective surface sterilization method for sugarcane (Saccharum officinarum L.) explants. African J Biotechnol, 12(44): 6282-6286.
  • Misra AN, Misra M. 2012. Sterilisation techniques in plant tissue culture. Fakir Mohan University, Balasore, India.
  • Murashige T, Skoog F. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plantarum, 15(3): 473-497.
  • Oszmiański J, Wojdyło A. 2005. Aronia melanocarpa Phenolics and Their Antioxidant Activity. Eur Food Res Technol, 221(6): 809-813.
  • Park S. 2021. Plant tissue culture: techniques and experiments. Academic Press, London, UK, pp: 227.
  • Rai AC, Kumar A, Modi A, Singh M. 2022. Advances in plant tissue culture: current developments and future trends. Academic Press, London, UK, pp: 407.
  • Rout GR, Jain SM. 2020. Advances in tissue culture techniques for ornamental plant propagation. Achieving Sustainable Cultivation of Ornamental Plants. Taylor & Francis, New York, US, pp: 149-188.
  • Şen A. 2012. Oxidative stress studies in plant tissue culture. Antioxidant Enzyme, 3(3): 59-88.
  • Şengül E. 2012. In vitro propagation of black mulberry (Morus nigra L.). MSc Thesis, Bursa Uludag University, Bursa, Türkiye, pp: 60.
  • Sikora J, Broncel M, Mikiciuk-Olasik E. 2014. Aronia melanocarpa Elliot reduces the activity of angiotensin i‐converting enzyme—in vitro and ex vivo studies. Oxidat Medic Cell Longevity, 2014(1): 739721.
  • Singh J, Bajpai R, Gangwar RK. 2023. Biotechnology in environmental remediation. John Wiley & Sons, New York, US, pp: 1-8.
  • Soumare A, Diédhiou AG, Arora NK, Tawfeeq Al-Ani LK, Ngom M, Fall S, Sy MO. 2021. Potential role and utilization of plant growth promoting microbes in plant tissue culture. Front Microbiol, 12: 649878.
  • Soylu A, U Erturk. 1999. Researchs on micropropagation of chestnut. Acta Hort, 494: 247-253
  • Vinh BVT, Tung HT, Bien LT, Nhut DT. 2024. Silver nanoparticles as a sterilant in plant micropropagation. In: Metal Nanopart Plant Cell Tissue Organ Cult, Springer Nature, Singapore, pp: 105-122.
Year 2024, Volume: 7 Issue: 6, 679 - 685, 15.11.2024
https://doi.org/10.47115/bsagriculture.1548432

Abstract

References

  • Agrawal A, Gowthami R, Chander S, Srivastava, V, Shankar M, Deepak DA. 2024. Biotechnological tools for conservation of plant genetic diversity. Sustainable Utilization and Conservation of Plant Genetic Diversity, Springer Nature, Singapore, pp: 711-752.
  • Ahmadpoor F, Zare N, Asghar R, Sheikhzadeh P. 2022. Sterilization protocols and the effect of plant growth regulators on callus induction and secondary metabolites production in in vitro cultures Melia azedarach L. AMB Express, 12: 1-12.
  • Al Ghasheem N, Stănică F, Petıcılă AG, Venat O. 2018. In vitro effect of various sterilization techniques on peach (Prunus persica (L.) Batsch) explants. Scient Papers, 227: 227-234.
  • Almokar HM M, Pırlak L. 2018. Propagation of Aronia (Aronia melanocarpa) with tissue culture. Selcuk J Agri Food Sci, 32(3): 549-558.
  • Angel LPL, Abu Bakar D, Sundram S. 2021. A non-invasive tissue culture system for performing artificial pathogenicity assays of Ganoderma boninense. Australasian Plant Pathol, 2021: 1-4.
  • Anuruddi HIGK, Nakandalage N, Fonseka DK. 2023. New insights for the production of medicinal plant materials: ex vitro and in vitro propagation. In Biosynthesis of Bioactive Compounds in Medicinal and Aromatic Plants: Manipulat Convent Biotechnol Approac, 2023: 181-212.
  • Bidabadi SS, Jain SM. 2020. Cellular, molecular, and physiological aspects of in vitro plant regeneration. Plants, 9(6): 702.
  • Cargnelutti D, Tabaldi LA, Spanevello RM, de Oliveira Jucoski G, Battisti V, Redin M, Schetinger MRC. 2006. Mercury toxicity induces oxidative stress in growing cucumber seedlings. Chemosphere, 65(6): 999-1006.
  • Cassells AC. 2012. Contamination and its impact in tissue culture. Plant Cell Tissue Organ Cult, 100(2): 87-90.
  • da Silva JA T, Winarto B, Dobránszki J, Cardoso JC, Zeng S. 2016. Tissue disinfection for preparation of culture. Folia Horticult, 28(1): 57-75.
  • Dagne H, Palanivel H, Yeshitila A, Benor S, Abera S, Abdi A. 2023. Advanced modeling and optimizing for surface sterilization process of grape vine (Vitis vinifera) root stock 3309C through response surface, artificial neural network, and genetic algorithm techniques. Heliyon, 9(8): e18628
  • El-Sherif NA. 2019. Impact of plant tissue culture on agricultural sustainability. Sustain Agri Environ Egypt: Part II: Soil-Water-Plant Nexus, 2019: 93-107.
  • Fidancı A. 2015. Türkiye için yeni bir minör meyve: Aronia bitkisi ve yetiştirme teknikleri. Bahçe, I(Özel Sayı): 1177-1180.
  • Fielder H, Beale T, Jeger MJ, Oliver G, Parnell S, Szyniszewska AM, Cunniffe NJ. 2024. A synoptic review of plant disease epidemics and outbreaks published in 2022. Phytopathology, 114(8): 1717-1732.
  • George EF, Sherrington PD. 1984. Plant propagation by tissue culture. Plant Cell Rep, 3(2): 85-91.
  • Hannan JM. 2013. Aronia berries profile. Iowa State University Extension and Outreach, Commercial Horticulture Field Specialist, October 2013, Ames, US.
  • Jurikova T, Mlcek J, Skrovankova S, Sumczynski D, Sochor J, Hlavacova I, Snopek L, Orsavova J. 2017. Fruits of black chokeberry Aronia melanocarpa in the prevention of chronic diseases. Molecules, 22: 944: 1-23.
  • Kampf G, Kampf G. 2018. Ethanol antiseptic stewardship: Biocide resistance and clinical implications, 2018: 9-35.
  • Karaokur OF, Kaya F, Yavuz E, Yenipinar, A. 2019. Comparison of commonly used statistics package programs. BSJ Eng Sci, 2(1): 26-32.
  • Kaya C, Sariyer T, Sahin E. 2023. Determination of effective surface sterilization protocol in in vitro tissue culture for Giant Snowdrop (Galanthus elwesii Hook) bulbs. Int J Agri Environ Food Sci, 7(2): 345-348. https://doi.org/10.31015 /jaefs.2023.2.12
  • Kaya Ç, Sarıyer T. 2024. Comparative biplot analysis of micropropagation of viking aronia cultivar in different plant tissue culture media. Anadolu Ege Tar Araş Enst Derg, 34(1): 90-96. https://doi.org/10.18615/anadolu.1494423
  • Knudson M. 2009. Plant Guide for black chokeberry (Photinia melanocarpa (Michx.) USDA-Natural Resources Conservation Service. URL= http://plant-materials.nrcs.usda.gov- (accessed date: June 10, 2024).
  • Kulling SE, Rawel HM. 2008. Chokeberry (Aronia melanocarpa) - a review on the characteristic components and potential health effects. Plant Medic, 74: 1625-1634.
  • Leifert C, Cassells AC. 2001. Microbial hazards in plant tissue and cell cultures. In Vitro Cell Developmental Biol-Plant, 37: 133-138.
  • Lema-Rumińska J, Sadowska K, Tymoszuk A, Andrzejewska J. 2023. Scutellarin and other metabolites as well as morphological and molecular characterization of the Scutellaria barbata lines from in vitro and in vivo cultivation. Indust Crops Prod, 195: 116464.
  • Mannino G, Gentile C, Ertani A, Serio G, Bertea C. M. 2021. Anthocyanins: Biosynthesis, distribution, ecological role, and use of biostimulants to increase their content in plant foods—A review. Agriculture, 11(3): 212.
  • Mekonnen T, Diro M, Sharma M. 2013. An alternative safer and cost effective surface sterilization method for sugarcane (Saccharum officinarum L.) explants. African J Biotechnol, 12(44): 6282-6286.
  • Misra AN, Misra M. 2012. Sterilisation techniques in plant tissue culture. Fakir Mohan University, Balasore, India.
  • Murashige T, Skoog F. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plantarum, 15(3): 473-497.
  • Oszmiański J, Wojdyło A. 2005. Aronia melanocarpa Phenolics and Their Antioxidant Activity. Eur Food Res Technol, 221(6): 809-813.
  • Park S. 2021. Plant tissue culture: techniques and experiments. Academic Press, London, UK, pp: 227.
  • Rai AC, Kumar A, Modi A, Singh M. 2022. Advances in plant tissue culture: current developments and future trends. Academic Press, London, UK, pp: 407.
  • Rout GR, Jain SM. 2020. Advances in tissue culture techniques for ornamental plant propagation. Achieving Sustainable Cultivation of Ornamental Plants. Taylor & Francis, New York, US, pp: 149-188.
  • Şen A. 2012. Oxidative stress studies in plant tissue culture. Antioxidant Enzyme, 3(3): 59-88.
  • Şengül E. 2012. In vitro propagation of black mulberry (Morus nigra L.). MSc Thesis, Bursa Uludag University, Bursa, Türkiye, pp: 60.
  • Sikora J, Broncel M, Mikiciuk-Olasik E. 2014. Aronia melanocarpa Elliot reduces the activity of angiotensin i‐converting enzyme—in vitro and ex vivo studies. Oxidat Medic Cell Longevity, 2014(1): 739721.
  • Singh J, Bajpai R, Gangwar RK. 2023. Biotechnology in environmental remediation. John Wiley & Sons, New York, US, pp: 1-8.
  • Soumare A, Diédhiou AG, Arora NK, Tawfeeq Al-Ani LK, Ngom M, Fall S, Sy MO. 2021. Potential role and utilization of plant growth promoting microbes in plant tissue culture. Front Microbiol, 12: 649878.
  • Soylu A, U Erturk. 1999. Researchs on micropropagation of chestnut. Acta Hort, 494: 247-253
  • Vinh BVT, Tung HT, Bien LT, Nhut DT. 2024. Silver nanoparticles as a sterilant in plant micropropagation. In: Metal Nanopart Plant Cell Tissue Organ Cult, Springer Nature, Singapore, pp: 105-122.
There are 40 citations in total.

Details

Primary Language English
Subjects Agricultural Engineering (Other)
Journal Section Research Articles
Authors

Çağlar Kaya 0000-0002-7054-3081

Şehnaz Özatay 0000-0003-0268-105X

Publication Date November 15, 2024
Submission Date September 11, 2024
Acceptance Date October 15, 2024
Published in Issue Year 2024 Volume: 7 Issue: 6

Cite

APA Kaya, Ç., & Özatay, Ş. (2024). Development of an Effective Sterilization Protocol for Plant Tissue Culture Studies in Superfruit Aronia [Aronia melanocarpa (Michaux) Elliot]. Black Sea Journal of Agriculture, 7(6), 679-685. https://doi.org/10.47115/bsagriculture.1548432

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