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Cited by 27Plant tissue culture and regenerationPlant Disease Management TechniquesAntimicrobial agents and applications

A Fumigation-Based Surface Sterilization Approach for Plant Tissue Culture

Open access
Sivanesan, Iyyakkannu, Muthu, Manikandan, Gopal, Judy, Tasneem, Shadma, Kim, Doo-Hwan, Oh, Jae-Wook·2021·International Journal of Environmental Research and Public Health
DOI 10.3390/ijerph18052282PMC PMC7967719
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Abstract

Plant tissue culture has led to breakthroughs in understanding and applying the fundamental knowledge towards harnessing more from plants. Microbial contamination is one of the serious problems limiting the successful extrapolation of plant tissue culture practices. Sources of in vitro contamination include culture containers, media, explants, equipment, the environment of the culture room and transfer area, and operating personnel. The successful initiation of in vitro culture mostly depends on surface sterilization of explants because this is the primary source. Usually, surface sterilization is done using chemicals, or toxic nanomaterials, this is the first time such an approach has been demonstrated. Numerous surface microflora attached to plant surfaces grow faster than the cultured explants and release phytotoxic substances into the culture media, hindering positive outcomes. In the current work, for the first time, the applicability of turmeric and benzoin resin-based fumigation of explants is demonstrated. The results showed that fumigation methods for surface sterilization were promising and could lead to fifty and even 100% contamination-free plant tissue culture. Nanoparticulate carbon was identified in the turmeric and benzoin smoke and coined the key player in the surface sterilization effect. These studies open a whole new avenue for the use of fumigation-based methods for riddance of microbial contamination.

AI snapshot

Written from the paper’s full text.

TL;DR

Turmeric and benzoin resin smoke were tested as fumigation-based surface sterilization methods for explants of three ornamental plants, with carbon nanoparticles identified in the smoke. Fumigation of explants or pretreatment with soot suspensions reduced contamination, in some cases achieving 100% inhibition, while media-only fumigation was largely ineffective.

Key findings

  • Direct explant fumigation (Method B) inhibited contamination by 65% (TurFE) and 80% (BRFE) in Explant 1; 80% (TurFE) and 20% (BRFE) in Explant 2; and 100% (TurFE) and 80% (BRFE) in Explant 3.
  • Soot-suspension pretreatment (Method C) left 16% contamination with TurSSTE and 31% with BRSSTE in Explant 1; gave 20% (TurSSTE) versus 50% (BRSSTE) inhibition in Explant 2; and gave 80% (TurSSTE) versus 100% (BRSSTE) inhibition in Explant 3.
  • Media-only fumigation (Method A) produced a nearly two-order magnitude decrease in surface contaminants for Explants 1 and 3 with turmeric-fumigated plates, but no fumigation effect for Explant 2; benzoin-fumigated plates showed no effect for all three explants.
  • Characterization found UV absorption maxima between 220 and 250 nm, FTIR bands consistent with carbon nanoparticles, and TEM carbon dots of 5–50 nm in turmeric and smaller than 10 nm in benzoin, with larger particles attributed to fly ash debris.

Methods

  • Explants were actively growing shoots of *Rhododendron yedoense* var. Poukhanense (Explant 1), *Hedera helix* (Explant 2), and *Rosa hybrida* ‘Red Sandra’ (Explant 3), collected in June from Konkuk University campus, Seoul, Korea; they were washed, defoliated, treated with mild detergent, rinsed with sterile distilled water, and blotted dry.
  • Fumigation used turmeric rhizome or benzoin resin: Method A fumigated culture media plates for 3 min at 15 cm; Method B directly fumigated explants for 10 sec at 30 cm; Method C immersed explants for 5 min in smoke soot dispersed in 25 mL of 20% ethanol. Controls were non-fumigated media plates and untreated explants.
  • Explants were cultured on Murashige and Skoog medium with 0.8% (w/v) plant agar and 3% (w/v) sucrose, pH 5.6, at 25 ± 2 °C under a 16 h photoperiod at 45 µmol m−2 s−1; the experiment was conducted in triplicate with 25 explants per treatment, with bacterial contamination assessed after 2 days and fungal contamination after 1 week. Fumigants were characterized by UV–Vis spectrophotometry, TEM, and FTIR.

Limitations

  • The paper states that the proposed mode of action—carbon nanodots functionalized with active ingredients from turmeric or benzoin smoke—is speculative and that more studies are required to validate it and identify the exact mechanism.
  • The paper states that Method B showed varying degrees of sterilization and that there is scope for extension and improvement to arrive at absolute results.
  • Evident from the text: only three ornamental explant types from one collection time and location were tested; no statistical analysis is reported despite triplicate experiments; and no explant viability, regeneration, or phytotoxicity outcomes are reported after fumigation or soot treatment.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.3390/ijerph18052282

Cites

43 of 43 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

15 of 72 cited sources checked · 9 supported · 0 flagged · accuracy not yet measured

  • Premisecitation

    Turmeric is well established for anti-inflammatory, antiseptic, antibacterial, antifungal, anticancer, and disinfectant properties.

    “Turmeric is well established for its powerful anti-inflammatory, antiseptic, antibacterial, antifungal, anticancer, and disinfectant properties and is used in several different ways, including burning and inhaling the smoke [36,37,38,39,40].”
    Quote in the paperSubstantial claim1 of 5 sources checked
    • cites [36] Govindarajan, 1980, Turmeric-chemistry, technology, and quality, Crit. Rev. Food Sci. Nutr. source
    • cites [37] Lai, 2004, Antimicrobial and chemopreventive properties of herbs and spices, Curr. Med. Chem. source
    • cites [38] Maheshwari, 2006, Multiple biological activities of curcumin: A short review, Life Sci. source
    • cites [39] Araujo, 2001, Biological activities of Curcuma longa L. Mem, Inst. Oswaldo Cruz source

      The Historical Background and Anti-protozoal activity sections describe anti-inflammatory and antibacterial effects, and the Anti-HIV section reports anti-tumor findings for curcumin. The passages do not establish the claimed antifungal, antiseptic, or disinfectant properties; the reported effects also do not establish all these properties as well established for turmeric generally. No page numbers are available in our copy. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites [40] Burt, 2004, Essential oils: Their antibacterial properties and potential applications in foods: A review, Int. J. Food Microbiol. source
  • Premisecitation

    Several studies have reported phytotoxicity of antimicrobial agents to cell, organ, and tissue cultures in plant species.

    “Several studies have reported the phytotoxicity of antimicrobial agents to cell, organ, and tissue cultures in plant species [5,6,7,8,10].”
    Quote in the paperSubstantial claim1 of 5 sources checked
    • cites [5] Leifert, 1992, Effect of combinations of antibiotics on micropropagated Clematis, Delphinium, Hosta, Iris and Photinia, Plant Cell Tissue Organ Cult. source
    • cites [6] Silva, 2003, The effect of antibiotics on the in vitro growth response of chrysanthemum and tobacco stem transverse thin cell layers (tTCLs), Sci. Hortic. source
    • cites [7] Qin, 2011, Response of in vitro strawberry to antibiotics, Plant Growth Regul. source
    • cites [8] Tambarussi, 2015, Influence of antibiotics on indirect organogenesis of Teak, Ann. For. Res. source
    • cites [10] Leifert, 2013, Microbial hazards in plant tissue and cell cultures, In Vitro Cell. Dev. Biol. Plant source
  • Interpretationcitation

    Both fumigation sources have a history of antimicrobial abilities.

    “Thus, both the fumigation sources we have employed have a history of antimicrobial abilities.”
    Quote in the paperSubstantial claim1 of 7 sources checked
    • cites [36] Govindarajan, 1980, Turmeric-chemistry, technology, and quality, Crit. Rev. Food Sci. Nutr. source
    • cites [37] Lai, 2004, Antimicrobial and chemopreventive properties of herbs and spices, Curr. Med. Chem. source
    • cites [38] Maheshwari, 2006, Multiple biological activities of curcumin: A short review, Life Sci. source
    • cites [39] Araujo, 2001, Biological activities of Curcuma longa L. Mem, Inst. Oswaldo Cruz source

      The anti-bacterial activity passage (page unavailable) reports that Curcuma oil inhibited the growth of two Staphylococcus species, supporting a history of antimicrobial activity for Curcuma longa. The paper does not establish antimicrobial activity for both fumigation sources, nor does it test them as fumigants. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [40] Burt, 2004, Essential oils: Their antibacterial properties and potential applications in foods: A review, Int. J. Food Microbiol. source
    • cites [41] Duke, J.A. (2008). Benzoin (Styrax benzoin Dryander.), Duke’s Handbook of Medicinal Plants of the Bible, Taylor & Francis. source
    • cites [42] Burdock, G.A. (2010). Benzoin Resin, Fenaroli’s Handbook of Flavor Ingredients, Taylor & Francis. [6th ed.].

      Not checked in the free run.

  • Premisecitation

    Chun et al. described and confirmed nanoparticulate carbon role in antimicrobial activity of smoke.

    “Chun et al. [30] described and confirmed the nanoparticulate carbon role in the antimicrobial activity of smoke in their recent article.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [30] Chun, 2016, The ethanopharmacological aspect of carbon nanodots in turmeric smoke, Sci. Rep. source
  • Premisecitation

    Role of carbon nanomaterials in turmeric smoke resulting in antimicrobial activity has been reported.

    “Further, the role of carbon nanomaterials in turmeric smoke resulting in antimicrobial activity has also been reported [30].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [30] Chun, 2016, The ethanopharmacological aspect of carbon nanodots in turmeric smoke, Sci. Rep. source
  • Classified as probably not a substantial claim. 16
  • May combine two claims. 9

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Sources of in vitro contamination include culture containers, media, explants, equipment, the environment of the culture room and transfer area, and operating personnel.

    “Sources of in vitro contamination include culture containers, media, explants, equipment, the environment of the culture room and transfer area, and operating personnel.”
    • Also stated in: § 1. Introduction · hedged
      “Several possible sources of in vitro tissue culture contamination are culture containers, media, explants, equipment, the environment of the culture room and the transfer area, and operators [4,5,6,7,8,9].”
    Quote in the paperNot a substantial claim2 of 6 sources checked
    • cites [4] Leifert, 1994, Ecology of microbial saprophytes and pathogens in tissue culture and field-grown plants: Reasons for contamination problems in vitro, Crit. Rev. Plant Sci. source
    • cites [5] Leifert, 1992, Effect of combinations of antibiotics on micropropagated Clematis, Delphinium, Hosta, Iris and Photinia, Plant Cell Tissue Organ Cult. source
    • cites [6] Silva, 2003, The effect of antibiotics on the in vitro growth response of chrysanthemum and tobacco stem transverse thin cell layers (tTCLs), Sci. Hortic. source
    • cites [7] Qin, 2011, Response of in vitro strawberry to antibiotics, Plant Growth Regul. source
    • cites [8] Tambarussi, 2015, Influence of antibiotics on indirect organogenesis of Teak, Ann. For. Res. source

      The Introduction and Discussion describe eliminating Agrobacterium from plant tissue cultures, and the Plant material and antibiotics section describes sterilizing teak seeds. Our copy does not identify the claimed sources of in vitro contamination; searches for contamination sources, culture-room conditions, and operating personnel found no such passage. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [9] Kim, 2017, Nanomaterials in plant tissue culture: The disclosed and undisclosed, RSC Adv. source

      Section 2.1 (page not provided) identifies explants and the laboratory environment as sources of microbial contaminants. It does not specifically identify culture containers, media, equipment, the transfer area, or operating personnel as contamination sources; the Introduction (page not provided) discusses vessels and media as factors in culture success, not as sources of contamination. (LLM, conf=0.86)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Ag NP treatments led to successful surface sterilization of cotyledons, leaves or seeds of Arabidopsis, G × N15 rootstock, potato, Rosa hybrida, and tomato.

    “Few reports have confirmed that Ag NP treatments led to successful surface sterilization of cotyledons, leaves or seeds of Arabidopsis, G × N15 (hybrid of almond × peach) rootstock, potato, Rosa hybrida, and tomato [16,17,18,19].”
    Quote in the paperNot a substantial claim1 of 4 sources checked
    • cites [16] Rostami, 2009, Nano-silver particles eliminate the in vitro contaminations of Olive ‘Mission’ explants, Asian J. Plant Sci. source
    • cites [17] Mahna, 2013, Plant in vitro culture goes nano: Nanosilver-mediated decontamination of ex vitro explants, J. Nanomed. Nanotechnol. source

      The Results and Discussion report nanosilver decontamination of Arabidopsis seeds, potato leaves, and tomato cotyledons. This paper does not report treatment of G × N15 rootstock or Rosa hybrida; those parts of the multi-citation sentence are not supported by this source. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [18] Arab, 2014, Effects of antimicrobial activity of silver nanoparticles on in vitro establishment of G × N15 (hybrid of almond × peach) rootstock, J. Genet. Eng. Biotechnol. source
    • cites [19] Shokri, 2014, The effects of different concentrations of nano-silver on elimination of bacterial contaminations and phenolic exudation of Rosae (Rosa hybrida L.) in vitro culture, Int. J. Farm. Alli. Sci. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Several researchers have reported adverse effects of NPs on survival and subsequent regeneration of explants while using cytotoxic NPs.

    “Several researchers have also reported adverse effects of NPs on the survival and subsequent regeneration of explants while using cytotoxic NPs [9].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [9] Kim, 2017, Nanomaterials in plant tissue culture: The disclosed and undisclosed, RSC Adv. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Our group has reported characterization and properties of soot-acquired carbon nanomaterials.

    “Our group has also reported the characterization and properties of soot-acquired carbon nanomaterials [30,31].”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [30] Chun, 2016, The ethanopharmacological aspect of carbon nanodots in turmeric smoke, Sci. Rep. source
    • cites [31] Anthonydhason, 2018, Nanocarbon Effect of Smoking Biofilms for Effective Control, J. Clust. Sci. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Nanoparticles have been applied for eliminating plant tissue culture contaminants in the recent decade.

    “In the recent decade, nanoparticles (NPs) have been applied for eliminating plant tissue culture contaminants [9].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [9] Kim, 2017, Nanomaterials in plant tissue culture: The disclosed and undisclosed, RSC Adv. source

  • May combine two claims.

    Findingown data

    Tur and BR smudges showed absorption bands identical to those expected of carbon nanoparticles in smoke smudges, confirming presence of nanoparticulate carbon in the smudges.

    “Thus, as observed from Figure 4B, the Tur and BR smudges showed absorption bands identical to those expected of carbon nanoparticles in smoke smudges, confirming the presence of nanoparticulate carbon in the smudges.”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [29] Song, 2013, Ferrocene-functionalized carbon nanoparticles, Nanoscale source
    • cites [30] Chun, 2016, The ethanopharmacological aspect of carbon nanodots in turmeric smoke, Sci. Rep. source

      The Results section (‘Characterization of the Turmeric smudges’; page unavailable) reports that turmeric smudge had absorption bands matching those expected of carbon nanoparticles, supporting the Tur portion of the claim. The paper does not report absorption bands for BR smudges, so it cannot support that portion. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [31] Anthonydhason, 2018, Nanocarbon Effect of Smoking Biofilms for Effective Control, J. Clust. Sci. source

  • Classified as probably not a substantial claim.

    Interpretationcitation

    All these reports narrowed down modulus operandi behind antimicrobial disinfection of carbon dots in smoke.

    “All these reports narrowed down the modulus operandi behind the antimicrobial disinfection of the carbon dots in the smoke.”
    Quote in the paperSubstantial claim1 of 3 sources checked
    • cites [30] Chun, 2016, The ethanopharmacological aspect of carbon nanodots in turmeric smoke, Sci. Rep. source
    • cites [31] Anthonydhason, 2018, Nanocarbon Effect of Smoking Biofilms for Effective Control, J. Clust. Sci. source
    • cites [44] Mohagheghzadeh, 2006, Medicinal smokes, J. Ethnopharmacol. source

  • May combine two claims.

    Findingown data

    Fumigation methods for surface sterilization were promising and could lead to fifty and even 100% contamination-free plant tissue culture.

    “The results showed that fumigation methods for surface sterilization were promising and could lead to fifty and even 100% contamination-free plant tissue culture.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In Explant 2, TurFE > BRSSTE > TurSSTE.

    “Summarizing the results, in the case of Explant 1, TurSSTE, followed by BRFE and BRSSTE were the most successful; explant 2: TurFE > BRSSTE > TurSSTE.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Turmeric and benzoin resin-based fumigation of explants is applicable and demonstrated for surface sterilization.

    “In the current work, for the first time, the applicability of turmeric and benzoin resin-based fumigation of explants is demonstrated.”
    • Also stated in: § 1. Introduction · boosted
      “However, for the first time, we have demonstrated a technique where biocompatible fumigants from plant sources have been used for surface sterilization of explants.”
    • Also stated in: § 4. Conclusions · boosted
      “The successful use of turmeric and benzoin resin-based fumigation for the inhibition of surface contaminants on plant explants has been demonstrated for the first time.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: No prior report has used fumigation for surface sterilization of plant explants.
    • Rests on an assumption: Turmeric and benzoin resin fumigants are biocompatible.

  • May combine two claims.

    Findingown data

    Carbon dots varying from 5 nm to 50 nm were well dispersed in Tur; in BR, particles smaller than 10 nm were observed.

    “Carbon dots varying from 5 nm to 50 nm were well dispersed in the case of Tur (Figure 5A) and in BR, as seen from Figure 5B, particles smaller than 10 nm were observed.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingreasoning

    Fumigation of explants and pretreatment of explants in carbon soot solution resulted in excellent surface sterilization.

    “As observed from the results, it was found that fumigation of the explants and pretreatment of the explants in carbon soot solution resulted in excellent surface sterilization.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisenone given

    Microbial contamination is one of the serious problems limiting the successful extrapolation of plant tissue culture practices.

    “Microbial contamination is one of the serious problems limiting the successful extrapolation of plant tissue culture practices.”
    • Also stated in: § 1. Introduction · plain
      “Microbial contamination is one other challenge facing plant tissue culture practices.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Dispersion of carbon nanoparticles in ethanol leads to their uniform distribution.

    “It is well established now that dispersion of carbon nanoparticles in ethanol lead to their uniform distribution [32].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [32] Liu, 2016, Carbon dots: Surface engineering and applications, J. Mater. Chem. B source

  • May combine two claims.

    Interpretationreasoning

    TurFE was the most effective surface decontamination method, followed by TurSSTE and BRSSTE.

    “Thus, it can be said that TurFE was the most effective surface decontamination method, followed by TurSSTE and BRSSTE.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Our group has reported use of fumigation for inhibition of biofilm on material surfaces.

    “We have reported the use of fumigation for inhibition of biofilm on material surfaces [31].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [31] Anthonydhason, 2018, Nanocarbon Effect of Smoking Biofilms for Effective Control, J. Clust. Sci. source

  • May combine two claims.

    Interpretationown data

    Method B was successful in surface sterilization of explants although varying degrees of sterilization were observed.

    “These results indicate that Method B was indeed successful in the surface sterilization of the explants although varying degrees of sterilization were observed.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Inclusion of Ag NPs, TiO2 NPs, Zn NPs or ZnO NPs in the culture medium reduces internal and external contamination in Araucaria excelsa, Bacopa monnieri, banana, barley, G × N15 rootstocks, olive, potato, Rosa hybrida, Vanilla planifolia, and tobacco.

    “Inclusion of Ag NPs, TiO2 NPs, Zn NPs or ZnO NPs in the culture medium is reported to reduce internal and external contamination in Araucaria excelsa, Bacopa monnieri, banana, barley, G × N15 (hybrid of almond × peach) rootstocks, olive, potato, Rosa hybrida, Vanilla planifolia, and tobacco [12,16,18,19,20,21,22,23,24,25,26,27].”
    Quote in the paperNot a substantial claim0 of 10 sources checked
    • cites [12] Sarmast, 2011, Nano silver treatment is effective in reducing bacterial contamination of Araucaria excelsa R. Br. var. glauca explants, Acta Biologica Hungarica source
    • cites [16] Rostami, 2009, Nano-silver particles eliminate the in vitro contaminations of Olive ‘Mission’ explants, Asian J. Plant Sci. source
    • cites [18] Arab, 2014, Effects of antimicrobial activity of silver nanoparticles on in vitro establishment of G × N15 (hybrid of almond × peach) rootstock, J. Genet. Eng. Biotechnol. source
    • cites [19] Shokri, 2014, The effects of different concentrations of nano-silver on elimination of bacterial contaminations and phenolic exudation of Rosae (Rosa hybrida L.) in vitro culture, Int. J. Farm. Alli. Sci. source
    • cites [20] Safavi, 2011, The study of nano silver (NS) antimicrobial activity and evaluation of using NS in tissue culture media, Int. Conf. Life Sci. Technol. IPCBEE
    • cites [21] Safavi, 2011, In vitro antibacterial activity of nanomaterial for using in tobacco plants tissue culture, World Acad. Sci. Eng. Technol. source
    • cites [22] Safavi, K. (2012, January 13–14). Evaluation of using nanomaterial in tissue culture media and biological activity. Proceedings of the 2nd International Conference on Ecological, Environmental and Biological Sciences (EEBS 2012), Bali, Indonesia.

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [23] Safavi, 2014, Effect of titanium dioxide nanoparticles in plant tissue culture media for enhance resistance to bacterial activity, Bull. Environ. Pharmacol. Life Sci.
    • cites [24] Mandeh, 2012, In vitro influences of TiO2 nanoparticles on Barley (Hordeum vulgare L.) tissue culture, Biol. Trace Elem. Res. source
    • cites [25] Helaly, 2014, Effect of nanoparticles on biological contamination of in vitro cultures and organogenic regeneration of banana, Aust. J. Crop Sci.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Mohagheghzadeh et al. summarized an exhaustive review of medicinal smoke highlighting antimicrobial activity of smoke.

    “Mohagheghzadeh et al. [44] summarized an exhaustive review of medicinal smoke, its sources, and uses, primarily highlighting the antimicrobial activity of smoke.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [44] Mohagheghzadeh, 2006, Medicinal smokes, J. Ethnopharmacol. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Antibiotics and antifungal compounds have been included in plant tissue culture media to destroy or prevent microbial growth.

    “Removing endophytic microbes is very challenging, and hence, antibiotics and antifungal compounds have been included in plant tissue culture media to destroy or prevent microbial growth [10].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [10] Leifert, 2013, Microbial hazards in plant tissue and cell cultures, In Vitro Cell. Dev. Biol. Plant source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Fumigation as a source of cleansing or purging air and atmosphere is a well-accepted fact.

    “Moreover, fumigation as a source of cleansing or purging the air and atmosphere is a well-accepted fact [43].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [43] Nautiyal, 2007, Medicinal smoke reduces airborne bacteria, J. Ethnopharmacol. source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Explants of Araucaria excelsa, Cynodon dactylon, Gerbera jamesonii, olive, Valeriana officinalis, and Vitis vinifera obtained from greenhouse- or field-grown plants were disinfected with ethanol or commercial bleach, and then silver nanoparticles reduced or eliminated the contaminants.

    “The explants of Araucaria excelsa, Cynodon dactylon, Gerbera jamesonii, olive, Valeriana officinalis, and Vitis vinifera obtained from greenhouse- or field-grown plants were disinfected with ethanol or commercial bleach, and then silver nanoparticles (AgNPs) eventually reduced or eliminated the contaminants [11,12,13,14,15].”
    Quote in the paperNot a substantial claim0 of 5 sources checked
    • cites [11] Abdi, 2008, Nano silver: A novel nanomaterial for removal of bacterial contaminants in valerian (Valeriana officinalis L.) tissue culture, Acta Physiol. Plant. source
    • cites [12] Sarmast, 2011, Nano silver treatment is effective in reducing bacterial contamination of Araucaria excelsa R. Br. var. glauca explants, Acta Biologica Hungarica source
    • cites [13] Fakhrfeshani, 2012, Disinfecting effects of nano silver fluids in Gerbera (Gerbera jamesonii) capitulum tissue culture, J. Biol. Environ. Sci.
    • cites [14] Gouran, A., Jirani, M., Mozafari, A.A., Saba, M.K., Ghaderi, N., and Zaheri, S. (2014, January 20). Effect of silver nanoparticles on grapevine leaf explants sterilization at in vitro conditions. Proceedings of the 2nd National Conference on Nanotechnology from Theory to Application, Isfahan, Iran.

      Not checked in the free run.

    • cites [15] Taghizadeh, 2014, The application of essential oils and silver nanoparticles for sterilization of sermuda grass explants in in vitro culture, J. Hortic. Sci. Technol. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Several chemicals have been used to disinfect explants before culturing.

    “Several chemicals have been used to disinfect the explants before culturing [4].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [4] Leifert, 1994, Ecology of microbial saprophytes and pathogens in tissue culture and field-grown plants: Reasons for contamination problems in vitro, Crit. Rev. Plant Sci. source

  • Classified as probably not a substantial claim.

    Premisecitation

    Judy et al. (2016) reported self-assembly of soot-derived carbon dots onto substrates exposed to them.

    “Judy et al., (2016) recently reported the self-assembly of the soot-derived carbon dots onto substrates exposed to them [45].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [45] Gopal, J., Muthu, M., and Chun, S. (2016). Autochthonous self-assembly of nature’s nanomaterials: Green, parsimonious and antibacterial carbon nanofilms on glass. Phys. Chem. Chem. Phys., 18670–18677. source
24 of 24

  • The paper relies on this without support.

    Successful initiation of in vitro culture mostly depends on surface sterilization of explants because this is the primary source.

    “The successful initiation of in vitro culture mostly depends on surface sterilization of explants because this is the primary source.”
    “The successful initiation of in vitro culture rests on effective surface sterilization of explants.”

  • The paper relies on this without support.

    A wide range of microbes attached to plant surfaces grow faster than cultured explants and release phytotoxic substances into culture media, interfering with culturing.

    “a wide range of microbes are attached to the surface of plants, and these grow faster than the cultured explants and release phytotoxic substances into the culture media, interfering with the culturing process.”

  • The paper relies on this without support.

    Surface sterilants are toxic to explants, and toxicity depends on dose and duration.

    “surface sterilants are also toxic to the explants, and the level of toxicity depends on the dose of disinfectants and duration of treatment.”

  • The paper relies on this without support.

    Some sterilants fail to eliminate contaminants in explants and profoundly affect morphogenesis due to phytotoxicity.

    “Some sterilants have even failed to eliminate contaminants in explants; rather, they profoundly affect morphogenesis due to their phytotoxicity.”

  • The paper relies on this without support.

    Removing endophytic microbes is very challenging.

    “Removing endophytic microbes is very challenging”

  • The paper relies on this without support.

    Effectiveness of NPs in eliminating microbial contaminants in plant tissue culture depends on their dimension, size, distribution, and type.

    “The effectiveness of NPs in the elimination of microbial contaminants in plant tissue culture depends on their dimension, size, distribution, and type.”

  • The paper relies on this without support.

    No prior report has used fumigation for surface sterilization of plant explants.

    “For the first time, we report the use of fumigation for surface sterilization of plant explants.”
    “The successful use of turmeric and benzoin resin-based fumigation for the inhibition of surface contaminants on plant explants has been demonstrated for the first time.”

  • The paper relies on this without support.

    Turmeric and benzoin resin fumigants are biocompatible.

    “biocompatible fumigants from plant sources have been used for surface sterilization of explants”
    “more cytofriendly nanomaterial such as carbon”
8 of 8

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Method A is described as effective despite reported no-effect results

    Minor

    The paper says all three methods led to efficient surface sterilization compared with the untreated control, but later reports that Method A's benzoin-resin plates had no fumigation effect for any of the three explants.

  • Song et al. reference year differs between text and reference list

    Minor

    The discussion cites Song et al. [29] as a 2011 report, while reference 29 lists its publication year as 2013.

2 of 2

Cites

Flagged first, then by citations

Research works printed but not linked: 3

  1. Gouran, A., Jirani, M., Mozafari, A.A., Saba, M.K., Ghaderi, N., and Zaheri, S. (2014, January 20). Effect of silver nanoparticles on grapevine leaf explants sterilization at in vitro conditions. Proceedings of the 2nd National Conference on Nanotechnology from Theory to Application, Isfahan, Iran.
  2. Safavi, K. (2012, January 13–14). Evaluation of using nanomaterial in tissue culture media and biological activity. Proceedings of the 2nd International Conference on Ecological, Environmental and Biological Sciences (EEBS 2012), Bali, Indonesia.
  3. Burdock, G.A. (2010). Benzoin Resin, Fenaroli’s Handbook of Flavor Ingredients, Taylor & Francis. [6th ed.].

Cited by

Sorted by citations

Related

Authors
Sivanesan, I., Muthu, M., Gopal, J., Tasneem, S., Kim, D.-H., & Oh, J.-W.
Affiliations
  • Konkuk University (KR)
  • Laboratory of Neo Natural Farming, Chunnampet, Tamil Nadu 603 401, India
  • Jazan University (SA)
Corresponding author
Oh, Jae-Wook
ORCID
Venue
International Journal of Environmental Research and Public Health
Publisher
MDPI AG
Published
2021-02-25
Volume
18
Issue
5
Pages
2282
ISSN
1660-4601 (ISSN-L)
DOI
10.3390/ijerph18052282
PMC
PMC7967719
OpenAlex
W3131701888
Semantic Scholar
cbbdef4c0af36ce0cd7a2430cf37eea9d76d7639
Cited by
27
References
42
Citations by year
2026: 5 · 2025: 11 · 2024: 3 · 2023: 3 · 2022: 3 · 2021: 2
FWCI
0.86
Type
journal article
Language
en
Open access
Licence
Locations
  • International Journal of Environmental Research and Public Health (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • Konkuk University
  • Ministry of Agriculture, Food and Rural Affairs
  • Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant tissue culture and regeneration
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Disease Management Techniques
  • Physical Sciences › Chemistry › Organic Chemistry › Antimicrobial agents and applications
Keywords
surface sterilization, fumigation, plant tissue culture, microbial contamination, explants, plant toxins
MeSH terms
Carbon, Culture Media, Fumigation, Plants, Sterilization

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-28.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-28.

Cited sources are checked by gpt-6-sol.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

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