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Effects of different surface sterilization protocols on fungal load and germination of black henbane seeds

Year 2024, Volume: 37 Issue: 3, 115 - 120, 06.12.2024
https://doi.org/10.29136/mediterranean.1530967

Abstract

Black henbane (Hyoscyamus niger L.) is an important medicinal plant rich in tropane alkaloids with significant pharmacological effects. To extract these valuable metabolites, seeds are mostly used as the initial material in tissue culture techniques. Due to the intricate surface structure of the seeds, coupled with cultivation of the plant under unsuitable ecological conditions, a high risk of contamination during in vitro culture arises. As such, it is important to determine the most suitable sterilization method for successful germination of black henbane seeds under in vitro conditions. In this study, the effects of 10 different sterilization protocols with ethyl alcohol (EtOH), sodium hypochlorite (NaOCl), copper sulfate (CuSO4), hydrogen peroxide (H2O2), mercury chloride (HgCl2), and silver nitrate (AgNO3), on reduction of the fungal contamination of black henbane seeds was determined by the agar test method. Additionally, the germination rates, as well as shoot lengths and fresh plant weights of the germinated seedlings, were investigated. As a result, it was found that Alternaria, Fusarium, and Penicillium species were the most common fungi on black henbane seeds. Among the chemicals used in the sterilization protocols, AgNO3 was found as the most effective one, completely inhibiting fungal growth. Sterilization protocols with AgNO3 also yielded the highest germination rates.

References

  • Abdi G, Salehi H, Khosh-Khui M (2008) Nanosilver: a novel nanomaterial for removal of bacterial contaminants in valerian (Valeriana officinalis L.) tissue culture. Acta Physiologiae Plantarum 30: 709-714.
  • Al Ghasheem N, Stanica F, Peticila AG, Venat O (2018) In vitro effect of various sterilization techniques on peach (Prunus persica (L.) Batsch) explants. Scientific Papers, Series B, Horticulture LXII: 227-234.
  • Aljibouri AMJ, Al-Samarraei KW, Abd AS, Mageed DM, Ali AJA (2012) Alkaloids production from callus of Hyoscyamus niger L. in vitro. Journal of Life Sciences 6: 874-882.
  • Amza J (2018) Seed borne fungi; food spoilage, the negative impact and their management: A review. Food Science and Quality Management 81: 70-79.
  • Barampuram S, Allen G, Krasnyanski S (2014) Effect of various sterilization procedures on the in vitro germination of cotton seeds. Plant Cell, Tissue and Organ Culture (PCTOC) 118: 179-185.
  • Battal A, Görmez G, Türker M (2019) The efficient germination method for marshmallows with medicinal importance: The nicking on seed coat. Bitlis Eren University Journal of Science 8: 843-851.
  • Blanco R, Aveling TAS (2018) Seed-borne Fusarium pathogens in agricultural crops. Acta Horticulturae 1204: 161-170.
  • Carisse O, Ouimet A, Toussaint V, Philion, V (2000) Evaluation of the effect of seed treatments, bactericides, and cultivars on bacterial leaf spot of lettuce caused by Xanthomonas campestris pv. vitians. Plant Disease 84: 295-299.
  • Danso KE, Azu E, Elegba W, Asumeng A, Amoatey HM, Klu GYP (2011) Effective decontamination and subsequent plantlet regeneration of sugarcane (Sacchrum officinarum L.) in vitro. International Journal of Integrative Biology 11: 90-96.
  • Ditommaso A, Nurse RE (2004) Impact of sodium hypochlorite concentration and exposure period on germination and radicle elongation of three annual weed species. Seed Science and Technology 32: 377-391.
  • Ellis MB (1971) Dematiaceous Hyphomycetes. CAB International, Wallingford, UK.
  • Ghorbanpour M, Hatami M, Khavazi K (2013) Role of plant growth promoting Rhizobacteria on antioxidant enzyme activities and tropane alkaloid production of Hyoscyamus niger under water deficit stress. Turkish Journal of Biology 37: 350-360.
  • Kane ME (2000) Culture indexing for bacterial and fungal contaminants. In: Trigiano RN, Gray DJ (Eds), Plant Tissue Culture Concepts and Laboratory Exercises. CRC Press, Florida, pp. 427-432.
  • Kaygusuz T, Coşkuntuna A (2022) Determination of seed-bornefungi in some medicinal and aromatic plants. Applied Ecology and Environmental Research 20: 2553-2564.
  • Kesho A, Abebe W (2020) Detection of seed borne fungi associated with some cereals and legume crops of seeds grown in main season at Holetta Agricultural Research Center. American Journal of Life Sciences 8: 91-95.
  • Martin I, Galvez L, Guasch L, Palmero D (2022) Fungal pathogens and seed storage in the drystate. Plants 11: 3167.
  • Misra AN, Misra M (2012) Sterilization techniques in plant tissue culture. In: Sharma HP, Dogra JVV, Misra AN (Eds), Plant Tissue Culture: Totipotency to Transgenic. Agrobios, India. pp. 31-41.
  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum 15: 473-497.
  • Nartop P (2019) Silver nanoparticles: ecofriendly surface sterilization of plant seeds in different shapes and sizes. The Journal of Animal & Plant Sciences 29: 453-460.
  • Oyebanji OB, Nweke O, Odebunmi O, Galadima NB, Idris MS, Nnodi UN, Afolabi AS, Ogbadu GH (2009) Simple, effective and economical explant-surface sterilization protocol for cowpea, rice and sorghum seeds. African Journal of Biotechnology 8: 5395-5399.
  • Pinto ACR, Demattê MESP, Creste S, Barbosa JC (2012) Seed and seedling surface-sterilization for in vitro culture of Tillandsia gardneri (Bromeliaceae). Acta Horticulturae 961: 383-389.
  • Rai M, Ingle AP, Pandit R, Paralikar P, Shende S, Gupta I, Biswas JK, da Silva SS (2018) Copper and copper nanoparticles: role in management of insect-pests and pathogenic microbes. Nanotechnology Reviews 7: 303-315.
  • Rajmohan K, Soni KB, Swapna A, Nazeem PA, Suku SS (2010) Use of copper sulphate for controlling systemic contamination in black pepper (Piper nigrum L.) cultures. Journal of Food, Agriculture and Environment 8: 569-571.
  • Ramandi A, Javan IY, Tazehabadi FM, Asl GI, Khosravanian R, Ebrahimzadeh MH (2019) Improvement in seed surface sterilization and in vitro seed germination of ornamental and medicinal plant Catharanthus roseus (L.). Chiang Mai Journal of Science 46: 1107-1112.
  • Rao GS, Narayana S, Bhadraiah B, Manoharachary C (2014) Biochemical changes due to fungal infestation in stored seeds of some vegetable crops. Indian Phytopathology 67: 159-163.
  • Rathod S (2012) Seed borne Alternaria species: A review. Current Botany 3: 21-23.
  • Samson RA, Hoekstra ES, Frisvad JC, Filtenborg O (1995) Introduction to Food-borne Fungi. Fourth Edition. Centraalbureau Voor Schimmelcultures, Baarn, Delft.
  • Sen MK, Jamal MAHM, Nasrin S (2013) Sterilization factors affect seed germination and proliferation of Achyranthes aspera cultured in vitro. Environmental and Experimental Biology 11: 119-123.
  • Shabana HA, Mahmoud T, Gairola S, Al Ketbi A, Aljasmi M, Al Sallani M (2021) Effect of storage conditions and sodium hypochlorite treatment on germination of Cucumis prophetarum (Cucurbitaceae) seeds from arid Arabian deserts. Research Square. doi: 10.21203/rs.3.rs-201765/v1.
  • Tort N (1997) Anason (Pimpinella anisum L.) ve pamuk (Gossypium hirsutum L.) tohumlarının in vitro ortamda sterilizasyonu üzerinde bir araştırma. Anadolu Ege Tarımsal Araştırma Enstitüsü Dergisi 7: 41-50.
  • Ugur R (2020) Development of in vitro sterilization protocol for DO-1 (Prunus domestica) rootstock. Applied Ecology and Environmental Research 18: 2339-2349.
  • Wahyono T, Hardani SNW, Sugoro I (2018) Low irradiation dose for sorghum seed sterilization: hydroponic fodder system and in vitro study. Buletin Peternakan 42: 215-221.
  • Watanabe T (2002) Pictorial Atlas of Soil and Seed Fungi, Morphologies of Cultured Fungi and Key to Species. Second Edition. CRC Press, Florida, pp. 486.

Effects of different surface sterilization protocols on fungal load and germination of black henbane seeds

Year 2024, Volume: 37 Issue: 3, 115 - 120, 06.12.2024
https://doi.org/10.29136/mediterranean.1530967

Abstract

Black henbane (Hyoscyamus niger L.) is an important medicinal plant rich in tropane alkaloids with significant pharmacological effects. To extract these valuable metabolites, seeds are mostly used as the initial material in tissue culture techniques. Due to the intricate surface structure of the seeds, coupled with cultivation of the plant under unsuitable ecological conditions, a high risk of contamination during in vitro culture arises. As such, it is important to determine the most suitable sterilization method for successful germination of black henbane seeds under in vitro conditions. In this study, the effects of 10 different sterilization protocols with ethyl alcohol (EtOH), sodium hypochlorite (NaOCl), copper sulfate (CuSO4), hydrogen peroxide (H2O2), mercury chloride (HgCl2), and silver nitrate (AgNO3), on reduction of the fungal contamination of black henbane seeds was determined by the agar test method. Additionally, the germination rates, as well as shoot lengths and fresh plant weights of the germinated seedlings, were investigated. As a result, it was found that Alternaria, Fusarium, and Penicillium species were the most common fungi on black henbane seeds. Among the chemicals used in the sterilization protocols, AgNO3 was found as the most effective one, completely inhibiting fungal growth. Sterilization protocols with AgNO3 also yielded the highest germination rates.

References

  • Abdi G, Salehi H, Khosh-Khui M (2008) Nanosilver: a novel nanomaterial for removal of bacterial contaminants in valerian (Valeriana officinalis L.) tissue culture. Acta Physiologiae Plantarum 30: 709-714.
  • Al Ghasheem N, Stanica F, Peticila AG, Venat O (2018) In vitro effect of various sterilization techniques on peach (Prunus persica (L.) Batsch) explants. Scientific Papers, Series B, Horticulture LXII: 227-234.
  • Aljibouri AMJ, Al-Samarraei KW, Abd AS, Mageed DM, Ali AJA (2012) Alkaloids production from callus of Hyoscyamus niger L. in vitro. Journal of Life Sciences 6: 874-882.
  • Amza J (2018) Seed borne fungi; food spoilage, the negative impact and their management: A review. Food Science and Quality Management 81: 70-79.
  • Barampuram S, Allen G, Krasnyanski S (2014) Effect of various sterilization procedures on the in vitro germination of cotton seeds. Plant Cell, Tissue and Organ Culture (PCTOC) 118: 179-185.
  • Battal A, Görmez G, Türker M (2019) The efficient germination method for marshmallows with medicinal importance: The nicking on seed coat. Bitlis Eren University Journal of Science 8: 843-851.
  • Blanco R, Aveling TAS (2018) Seed-borne Fusarium pathogens in agricultural crops. Acta Horticulturae 1204: 161-170.
  • Carisse O, Ouimet A, Toussaint V, Philion, V (2000) Evaluation of the effect of seed treatments, bactericides, and cultivars on bacterial leaf spot of lettuce caused by Xanthomonas campestris pv. vitians. Plant Disease 84: 295-299.
  • Danso KE, Azu E, Elegba W, Asumeng A, Amoatey HM, Klu GYP (2011) Effective decontamination and subsequent plantlet regeneration of sugarcane (Sacchrum officinarum L.) in vitro. International Journal of Integrative Biology 11: 90-96.
  • Ditommaso A, Nurse RE (2004) Impact of sodium hypochlorite concentration and exposure period on germination and radicle elongation of three annual weed species. Seed Science and Technology 32: 377-391.
  • Ellis MB (1971) Dematiaceous Hyphomycetes. CAB International, Wallingford, UK.
  • Ghorbanpour M, Hatami M, Khavazi K (2013) Role of plant growth promoting Rhizobacteria on antioxidant enzyme activities and tropane alkaloid production of Hyoscyamus niger under water deficit stress. Turkish Journal of Biology 37: 350-360.
  • Kane ME (2000) Culture indexing for bacterial and fungal contaminants. In: Trigiano RN, Gray DJ (Eds), Plant Tissue Culture Concepts and Laboratory Exercises. CRC Press, Florida, pp. 427-432.
  • Kaygusuz T, Coşkuntuna A (2022) Determination of seed-bornefungi in some medicinal and aromatic plants. Applied Ecology and Environmental Research 20: 2553-2564.
  • Kesho A, Abebe W (2020) Detection of seed borne fungi associated with some cereals and legume crops of seeds grown in main season at Holetta Agricultural Research Center. American Journal of Life Sciences 8: 91-95.
  • Martin I, Galvez L, Guasch L, Palmero D (2022) Fungal pathogens and seed storage in the drystate. Plants 11: 3167.
  • Misra AN, Misra M (2012) Sterilization techniques in plant tissue culture. In: Sharma HP, Dogra JVV, Misra AN (Eds), Plant Tissue Culture: Totipotency to Transgenic. Agrobios, India. pp. 31-41.
  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum 15: 473-497.
  • Nartop P (2019) Silver nanoparticles: ecofriendly surface sterilization of plant seeds in different shapes and sizes. The Journal of Animal & Plant Sciences 29: 453-460.
  • Oyebanji OB, Nweke O, Odebunmi O, Galadima NB, Idris MS, Nnodi UN, Afolabi AS, Ogbadu GH (2009) Simple, effective and economical explant-surface sterilization protocol for cowpea, rice and sorghum seeds. African Journal of Biotechnology 8: 5395-5399.
  • Pinto ACR, Demattê MESP, Creste S, Barbosa JC (2012) Seed and seedling surface-sterilization for in vitro culture of Tillandsia gardneri (Bromeliaceae). Acta Horticulturae 961: 383-389.
  • Rai M, Ingle AP, Pandit R, Paralikar P, Shende S, Gupta I, Biswas JK, da Silva SS (2018) Copper and copper nanoparticles: role in management of insect-pests and pathogenic microbes. Nanotechnology Reviews 7: 303-315.
  • Rajmohan K, Soni KB, Swapna A, Nazeem PA, Suku SS (2010) Use of copper sulphate for controlling systemic contamination in black pepper (Piper nigrum L.) cultures. Journal of Food, Agriculture and Environment 8: 569-571.
  • Ramandi A, Javan IY, Tazehabadi FM, Asl GI, Khosravanian R, Ebrahimzadeh MH (2019) Improvement in seed surface sterilization and in vitro seed germination of ornamental and medicinal plant Catharanthus roseus (L.). Chiang Mai Journal of Science 46: 1107-1112.
  • Rao GS, Narayana S, Bhadraiah B, Manoharachary C (2014) Biochemical changes due to fungal infestation in stored seeds of some vegetable crops. Indian Phytopathology 67: 159-163.
  • Rathod S (2012) Seed borne Alternaria species: A review. Current Botany 3: 21-23.
  • Samson RA, Hoekstra ES, Frisvad JC, Filtenborg O (1995) Introduction to Food-borne Fungi. Fourth Edition. Centraalbureau Voor Schimmelcultures, Baarn, Delft.
  • Sen MK, Jamal MAHM, Nasrin S (2013) Sterilization factors affect seed germination and proliferation of Achyranthes aspera cultured in vitro. Environmental and Experimental Biology 11: 119-123.
  • Shabana HA, Mahmoud T, Gairola S, Al Ketbi A, Aljasmi M, Al Sallani M (2021) Effect of storage conditions and sodium hypochlorite treatment on germination of Cucumis prophetarum (Cucurbitaceae) seeds from arid Arabian deserts. Research Square. doi: 10.21203/rs.3.rs-201765/v1.
  • Tort N (1997) Anason (Pimpinella anisum L.) ve pamuk (Gossypium hirsutum L.) tohumlarının in vitro ortamda sterilizasyonu üzerinde bir araştırma. Anadolu Ege Tarımsal Araştırma Enstitüsü Dergisi 7: 41-50.
  • Ugur R (2020) Development of in vitro sterilization protocol for DO-1 (Prunus domestica) rootstock. Applied Ecology and Environmental Research 18: 2339-2349.
  • Wahyono T, Hardani SNW, Sugoro I (2018) Low irradiation dose for sorghum seed sterilization: hydroponic fodder system and in vitro study. Buletin Peternakan 42: 215-221.
  • Watanabe T (2002) Pictorial Atlas of Soil and Seed Fungi, Morphologies of Cultured Fungi and Key to Species. Second Edition. CRC Press, Florida, pp. 486.
There are 33 citations in total.

Details

Primary Language English
Subjects Phytopathology, Plant Biotechnology in Agriculture
Journal Section Makaleler
Authors

Gursel Hatat Karaca 0000-0002-5159-2734

Hikmet Deveci 0000-0003-4393-8552

Ilknur Albayrak 0000-0003-3158-3440

Nilgun Gokturk Baydar 0000-0002-5482-350X

Publication Date December 6, 2024
Submission Date August 15, 2024
Acceptance Date October 10, 2024
Published in Issue Year 2024 Volume: 37 Issue: 3

Cite

APA Hatat Karaca, G., Deveci, H., Albayrak, I., Gokturk Baydar, N. (2024). Effects of different surface sterilization protocols on fungal load and germination of black henbane seeds. Mediterranean Agricultural Sciences, 37(3), 115-120. https://doi.org/10.29136/mediterranean.1530967
AMA Hatat Karaca G, Deveci H, Albayrak I, Gokturk Baydar N. Effects of different surface sterilization protocols on fungal load and germination of black henbane seeds. Mediterranean Agricultural Sciences. December 2024;37(3):115-120. doi:10.29136/mediterranean.1530967
Chicago Hatat Karaca, Gursel, Hikmet Deveci, Ilknur Albayrak, and Nilgun Gokturk Baydar. “Effects of Different Surface Sterilization Protocols on Fungal Load and Germination of Black Henbane Seeds”. Mediterranean Agricultural Sciences 37, no. 3 (December 2024): 115-20. https://doi.org/10.29136/mediterranean.1530967.
EndNote Hatat Karaca G, Deveci H, Albayrak I, Gokturk Baydar N (December 1, 2024) Effects of different surface sterilization protocols on fungal load and germination of black henbane seeds. Mediterranean Agricultural Sciences 37 3 115–120.
IEEE G. Hatat Karaca, H. Deveci, I. Albayrak, and N. Gokturk Baydar, “Effects of different surface sterilization protocols on fungal load and germination of black henbane seeds”, Mediterranean Agricultural Sciences, vol. 37, no. 3, pp. 115–120, 2024, doi: 10.29136/mediterranean.1530967.
ISNAD Hatat Karaca, Gursel et al. “Effects of Different Surface Sterilization Protocols on Fungal Load and Germination of Black Henbane Seeds”. Mediterranean Agricultural Sciences 37/3 (December 2024), 115-120. https://doi.org/10.29136/mediterranean.1530967.
JAMA Hatat Karaca G, Deveci H, Albayrak I, Gokturk Baydar N. Effects of different surface sterilization protocols on fungal load and germination of black henbane seeds. Mediterranean Agricultural Sciences. 2024;37:115–120.
MLA Hatat Karaca, Gursel et al. “Effects of Different Surface Sterilization Protocols on Fungal Load and Germination of Black Henbane Seeds”. Mediterranean Agricultural Sciences, vol. 37, no. 3, 2024, pp. 115-20, doi:10.29136/mediterranean.1530967.
Vancouver Hatat Karaca G, Deveci H, Albayrak I, Gokturk Baydar N. Effects of different surface sterilization protocols on fungal load and germination of black henbane seeds. Mediterranean Agricultural Sciences. 2024;37(3):115-20.

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