Contamination In Tissue Culture Research Articles

Studies on In vitro Surface Sterilisation and Antioxidants on Pomegranate (Punica granatum L.) cv. Bhagwa

Pomegranate, regarded as the “Fruit of Paradise”, is one of the important fruit crops of tropical and subtropical regions. Pomegranate fruits are delicious and possess significant nutritional and medicinal benefits. Due to the increase in demand, the proliferation of pomegranate through tissue culture is essential to get high-quality planting material. However, microbial contamination in plant tissue culture is one of the bottlenecks for establishing aseptic cultures. Although multiple explant sterilisation methods have been developed for pomegranate, explant sterilisation methods can indeed vary from region to region due to the local environment and the mother plant. In the present study, 1-2 drops of tween 20 (20 min) + 500 mg/L carbendazim (30 min) + 500 mg/L streptomycin sulphate (10 min) + 100 mg/L citric acid (30 min) + 100 mg/L tebuconazole (folicur) (30 min) + 0.1% HgCl2 (1 min) + dipping explant after cutting the edges in sterile water (30 min) + dipping in 500 mg/L PVP (5 min) was found to be optimum in the prevention of microbial contamination with minimal cell death. Nodal segments-initiated callus at 54 days after culture, showing a callus induction percentage of 55.55% and a calli weight of 80 mg/explant.

The Efficiency of Fungicides on Fungal Growth Inhibition in Cultured Tissues of Banana (Musa acominata), and Their Effect on Tissues Traits

The goal of this study was to identify fungicides with a broad-spectrum effectiveness and their capacity to inhibit fungal contaminants in banana tissue cultures. Numerous fungi were isolated from tissues of banana cultures of the Grand 9 variety. Aspergillus flavus, Aspergillus niger, Cladosporium oxysporum, Penicillium digitatum, P. expansum, and Penicillium sp. were among the contaminated fungi isolated. These fungi were inhibited successfully using the fungicides Beltanol, Agrisave, and Zoxis. Hoever, Beltanol was outperformed compared with Agrisave and Zoxis fungicides, with an inhibition rate of 100% for concentrations of 0.25, 0.5, and 1 ml/l. Thus, the addition of Beltanol fungicide to the banana culture medium had a positive influence on the fresh and dry weight of the tissues, with the average fresh weight increasing to 2.93 g compared with 2.4 g in the control treatment. Also, the dry weight was increased from to 0.21 g while it was 0.17 g in the control treatment. Additionally, the results demonstrated the enzymatic effectiveness (cellulase enzyme) of isolated fungi without treatment, that showed high effectiveness in P. digitatum fungus with an activity rate reached 2.33 mm. In contrast, C. oxysporum and P. expansum fungi displayed moderate efficiency with activity rates that reached 1.5 and 1.41 mm respectively. On the other hand, A. flavus and A. niger fungi exhibited low efficacy with activity rates of 1.25 and 1.12 mm respectively. Furthermore, the results revealed that P. expansum fungus had a high enzymatic effectiveness index with a rate of 1.85 mm, while C. oxysporum, P. digitatum, and A. flavus fungi had moderate enzymatic effectiveness with rates of 1.46, 1.34, and 1.26, respectively. A. niger fungus had the lowest enzymatic effectiveness with a rate of 1.23. The enzymatic secretion of the isolated fungi was inhibited efficiently via addition of the fungicide Beltanol to these fungi.

Optimizing Surface Sterilization for Micropropagation of Gerbera jamesonii: A Study on Explant Survival and Contamination Control

In the commercial production of gerberas, micropropagation enables the large-scale production of plants in a limited space, as well as the obtaining of disease and pest-free uniform plants. This technique also ensures precision in production schedules and product quality, regarding plant homogeneity and vigour. Nevertheless, the occurrence of contamination in an in vitro culture is frequent, and might result in high damage. Plant materials have different kind of microbes i.e., exogenous and endogenous in nature which is the main reason for contamination in tissue culture. Pre-treatments to explants with fungicides and bactericides can be effectively used to eliminate the surface contamination. Surface sterilizing agents, their levels and duration of exposure are known to impact the in vitro culture establishment. In the present experiment, pre-treatment of explants was carried out with Teepol BleachTM solution (0.1%) and BavistinTM (0.1%) for 15 and 20 minutes respectively for effective control of fungal contaminations in all the cultures. For the control of bacterial contamination antibiotics were added in culture medium. Surface sterilization of explants with 0.1% mercuric chloride (HgCl2) for five minutes gave the maximum survival of the growing cultures.

The use of ZnO NPs and Ag NPs along with sterilizing agents for managing contamination in banana tissue culture

The use of ZnO NPs and Ag NPs along with sterilizing agents for managing contamination in banana tissue culture

Óleos essenciais de plantas condimentares e medicinais no controle de contaminantes da micropropagação de Myrciaria dubia

Abstract The main limitation of the micropropagation of camu-camu (Myrciaria dubia) is related to in vitro contamination. In order to overcome contamination, the effect of essential oils was studied as an alternative to conventional chemical treatments. This study aimed to analyze the action of essential oils from four condiment and medicinal plants (Oregano, Origanum vulgare L.; Garlic, Allium sativum L.; Citronella, Cymbopogon nardus L.;and Ginger, Zingiber officinale Rosc.) in reducing microbial contamination growth and on the survival rate of explants in the micropropagation of camu-camu. The antimicrobial activity of essential oils was analyzed on the in vitro contamination of tissue culture from camu-camu at concentrations of 0.5, 1, 2, 3 and 4 µL mL-1 in woody plant medium (WPM), emulsified with 1% Polysorbate 80 (Tween 80). The antibacterial activity of essential oils on strains isolated from camu-camu tissue culture was also analyzed using agar diffusion and broth microdilution methods. The use of essential oils allowed a reduction in the rate of in vitro contamination in tissue culture, it being observed that, from the concentration of 2 µL mL-1, there was no manifestation of fungal contaminants, with a significant reduction in the rate of bacterial contamination, with the exception of ginger essential oil that showed significant contamination in all analyzed concentrations. Inversely in relation to the reduction in microbial growth in vitro, there is an increase in explant oxidation as concentrations increase, with citronella and oregano oils showing phytotoxic potential from the lowest concentrations. Garlic essential oil showed better balance, with lower oxidation rates and greater control of microorganisms in tissue culture at concentrations of 0.5 and 1 µL mL-1. Oregano and citronella essential oils showed better antibacterial activity in the agar diffusion test.

Screening of potential probiotic lactic acid bacteria with antimicrobial properties and selection of superior bacteria for application as biocontrol using machine learning models

144 lactic acid bacteria (LAB) strains were screened to find those with the best antimicrobial activity and potential probiotic properties. Firstly, bacteria were tested for resistance to low pH and high bile salt, antimicrobial properties against the common agent of bacterial contamination in plant tissue cultures. Then, the selected strains were assayed for probiotic characteristics including autoaggregation, coaggregation, hydrophobicity, biofilm formation, and adhesion ability. Finally, ranking and selection of probiotic potential bacteria for harnessing as antibacterial agents in plant tissue cultures were performed using supervised machine learning models. The highest values of autoaggregation percentages were observed for Lactobacillus paracasei S23 (96.1%), while the highest coaggregation values with Staphylococcus aureus, Bacillus cereus, and Listeria monocytogenes were observed for strains Lactobacillus plantarum S57 (63.4%), L. plantarum S70 (57.4%), and Lactobacillus casei S81 (58.6%), respectively. Principal component analysis (PCA) and machine learning algorithms allowed to select and prioritizing of 15 out of 144 LAB strains, based on their probiotic potential properties or their antimicrobial activity, of which four strains including Lb. paracasei S23, Lb. plantarum S70, Lb. casei S81, and Lb. plantarum S57 were proposed as appropriate antimicrobial candidates for harnessing in plant tissue cultures.

Microbial Contaminants and Their Control Methods: A Review with Reference to Potato Tissue Culture

No cell or tissue culture problem is as universal as that of culture loss due to contamination, so that microbial hazards cause drastic economic losses in the plant tissue culture industries. A wide range of microorganisms (filamentous fungi, yeasts, bacteria, virus and viroid) and micro-arthropods (mites and thrips) have been identified as contaminants in plant tissue culture. With this paramount importance of in-vitro culture contaminants, various research work has been conducted from different corners of the world. This review paper compiles such important information to share experiences within a single amassed document. Different research and review articles, proceedings, protocol notes, scientific notes, conference papers, case reports as well as case studies, research communications and technical reports are included in this paper. Hence, this article aimed to review and provide insights about microbial contaminants and their management techniques and thereby to document the possible attempts made by different scholars to mitigate microbial contamination under plant tissue culture. Moreover, the paper skim through the current advancements made on culture loss due to microbes, their control methods and hostile effects on regeneration capability of culture plants. Keywords : Microbial contamination, bacteria, fungi, plant tissue culture, in vitro culture DOI: 10.7176/JNSR/12-10-01 Publication date: May 31 st 2021

Identification and Prevention of Microbial Contamination in Tissue Culture of Catharanthus roseus - An Important Medicinal Herb

The explants of two varities rosea and alba of Catharanthus roseus used for in vitro propagation and found to be more than 50% of the cultures became contaminated. The most common bacterial contaminants were Bacillus licheniformis, Micrococcus, Panibacillus and fungal contaminants were Fusarium, Alternaria, Cladosporium and Aspergillus. Combinations of different antibiotics (Penicillin, norfloxacin, tobramycin, gatifloxacin, ofloxacin) and fungicides (Bavastin, captan, fluconazole and trichoderma) were used to control the growth of the contaminants. Gatifloxacin and ofloxacin inhibited 100% growth of bacteria whereas, bavastin and captan appeared to be the most effective fungicides. Combination of gatifloxacin, ofloxacin with bavastin and captan inhibited the growth of contaminants at their minimum phytotoxic concentration (MIC). The observed minimum phytotoxic concentration (MPC) of ofloxacin, gatifloxacin, bavastin and captan was 15, 9, 6 and 5% at their respective MIC. More than 90% of the cultures responded for callus formation in the combination of gatifloxacin (4%) + bavastin (1%). While the combination of gatifloxacin and captan was highly toxic that reduces the growth of the culture.
 Plant Tissue Cult. & Biotech. 30(2): 297-305, 2020 (December)

Accuracy of Fuzzy Logic-based Contamination Grading System on Abaca Tissue Culture

This study aimed to support the coalition for abaca rehabilitation program of the different cooperating agencies of Southern Leyte with the use of intelligent systems in the grading of abaca tissue culture contamination. As the Southern Leyte State University Tissue Culture Laboratory has since used manual grading of contaminated specimen, the use of image acquisition technology can lessen the human effort involved in this activity. Fuzzy logic was used in the grading of the contamination. There were five inputs for the inference engine, namely red, green, blue, whitish and brownish. There were 62 rules identified covering the different combinations, and the triangular-shaped membership function was used for all the membership types of the inputs. Based on the result of the testing, it had an accuracy rate of 82.00% for the binary contamination in 50 sample specimens, and an over-all accuracy of 80.33% in multiclass contamination grading. The system could not match the precision of the human expert, but the model is comparable to that of the expected actual result, while minimizing human intervention in grading the contamination of tissue cultured abaca.

Screening of fungal contaminants in banana tissue cultures in Jkuat, Kenya

Tissue culture is prone to high costs of production arising from losses incurred from fungal contamination. The aim of the study was to characterise fungal contaminants and elucidate the exhibited mode of resistance to most preferred sterilants. Twenty nine fungal samples were collected at the different stages of tissue culture growth, using purposive sampling technique. Morphology results were confirmed by molecular characterization using fungal 18S rRNA sequences. Biochemical and antibiosis tests, identification of genes for capsulation and ATP binding Cassete (ABC) transporters, were performed to show the relationship between the fungi and sterilants resistance. Amylases and proteases were highly expressed by all isolates while xylanases and lipases were moderately expressed and esterases were lowly expressed. Only fourteen isolates had antagonistic activity for Candida albicans while nine of them had antagonistic activity for Pseudomonas aeruginosa. Three isolates were both antagonistic for Staphylococcus aureus and for Escherichia coli. Cunninghamella bainieri (10R) recorded a unique antibiosis and extra cellular enzymatic activity (p<0.05). All the isolates were positive for mdr1 gene and three isolates had CAP64 capsule genes. Aspergillus sp., Penincillium sp., Cladosporium sp., Cunninghamella sp. and Fusarium sp. were identified to be the major fungal contaminants of tissue culture banana cultures in JKUAT laboratories. Key words: Tissue culture, 18S rRNA, fungal contaminants, banana cultures.

Prevention of Fungal Contamination in Plant Tissue Culture Using Cyclic Lipopeptides Secreted by Bacillus amyloliquefaciens AB30a

Plant tissue culture has revolutionized the field of plant biotechnology. However, there are certain obstacles which overall restrain the output of the plant tissue culturing. One of them is contamination of the tissue culture stock which is a major problem limiting the output. Aegle marmelos (L.) is a medicinal plant whose genotype qualities are maintained through clonal propagation of nodal segment as an explant. It harbors plethora of fungi which curbs the successful in vitro propagation. Chemical fungicide like bavistin is used to prevent the contamination in tissue culture which raises the environmental concerns. Thus, use of microbially derived antifungals can help in preventing fungal growth with benefit of positively impacting the plant growth. Here, authors investigated the use of heat stable lipopeptides which are secondary metabolites derived from Bacillus amyloliquefaciens AB30a for prevention of contamination in tissue culturing of nodal explants of A. marmelos positively impacting its in-vitro propagation.&#x0D; Plant Tissue Cult. &amp; Biotech. 29(1): 111-119, 2019 (June)

Microbial contamination in tissue culture of Chlorophytum borivilianum, a rare medicinal herb: identification and prevention

When inflorescence axis was used as the explant for micropropagation of Chlorophytum borivilianum, 50% of tubes became contaminated at Stage-I of in vitro culture. Bacteria (Bacillus licheniformis, Micrococcus sp. and Panibacillus sp.) and fungi (Alternaria sp., Aspergillus sp., Cladosporium sphaerospermum and Fusarium sp.) were the most common contaminants.The effectiveness of synthetic antimicrobial compounds in inhibiting the growth of contaminants was compared. Five antibiotics, penicillin (PE), norfloxacin (NF), tobramycin (TB), gatifloxacin (GT) and ofloxacin (OF), at 1%–15%, and four fungicides, Bavastin (BV), Captan (CA), Fluconazole (FL) and Trichoderma biofungicide (TV), at 0.5–2%, were tested on the bacterial and fungal isolates respectively. GT (5%) and OF (8%) inhibited 100% growth of bacteria, but the former was more efficient. BV and CA at 1–2% appeared to be the most effective fungicides. MIC of antibiotics was higher (8–10%) than MIC of fungicides (1–2%). The two most effective antibiotics (GT, OF) and fungicides (BV, CA) were also evaluated in vitro for phytotoxicity. The observed MPCs for GT, OF, BV and CA were 4%, 9%, 1% and 1.5% respectively. They were then tested in combination at their respective MICs in amended MS media and culture survival was recorded at Stage-I. Ninety per cent of cultures survived with GT (4%) + BV(1%) combination, while the combination of GT and CA was highly toxic. Enhancement in culture survival by 40% resulted from synergistic or additive effects of GT and BV.

Negative hydrostatic pressure is an unnoticed but significant source of contamination in tissue culture©

Plants are characterized by a negative hydrostatic pressure, brought about by transpiration and by capillary activity of xylem vessels (Taiz and Zeiger, 2010). Because of this, a stem that is being cut sucks up what is nearby. Often this is air but it may also be liquid. The diameter of the xylem vessels is 50-100 μm, so when the liquid contains bacteria (that are typically 0.5-5.0 μm), they will enter deeply into the tissue (Askari et al., 2014; De Klerk et al., 2014). To our knowledge, this alleged source of contamination has never been examined.

Influence of media supplements on inhibition of oxidative browning and bacterial endophytes of Camellia sinensis var. sinensis.

Explant oxidative browning and necrosis of Camellia sinensis var. sinensis is a severe problem in tissue culture, often associated with the exuded phenolic compounds and microbial contamination from the explants. In this study, 2-aminoindane-2-phosphonic acid (AIP), an inhibitor of the polyphenol production-required enzyme phenylalanine ammonia lyase (PAL), and different antibiotics were tested to control tea explant necrosis and browning. These compounds were supplemented in the regular plant growth medium together with 6-benzylaminopurine and thidiazuron at different concentrations. Our data indicated that application of 2µM of AIP was able to effectively inhibit callus browning, significantly reduce EGC abundance, and greatly improve callus induction and growth. Moreover, the use of 150mg/L of timentin and 30mg/L gentamycin resulted in an effective elimination of the surface and endophytic microbes associated with explants of C. sinensis var. sinensis. Our study revealed that the inhibition of PAL using AIP combined with the two tested antibiotics could open up new doors to control oxidative tissue browning and endophyte contamination in tissue culture for tea genetic manipulation.

Molecular detection and characterization of sustainable intracellular contaminants in commercially used cell cultures for pre-clinical studies

Microbial contamination in cell and tissue culture is a constant problem, which can compromise development and applications of cell lines. An immediate consequence of cell culture contamination is loss of researcher time, money and effort spent developing cultures and setting up experiments. There are adverse effects detected on cultures suffering from undetected biological contamination. This hidden contamination can potentially achieve high densities altering the growth and characteristics of the cultures. The objective of this study was to assess the molecular detection and characterization of sustainable intracellular contaminants in commercially used cell cultures use for regulatory pre-clinical studies for developing new chemical entities. This study was prompted by a series of observations by multiple researchers that cell lines were harbouring visible black particulate contaminants, capable of intracellular mobility with secondary impacts on cell adherence and lyses. This was initially limited to human liver cells, HepG2, C3A and CaCO2 cell lines, but recently similar evidence has been found in a series of other cell lines as well. Giemsa staining did show many spore-like entities in the cytoplasm mainly of 0.5-1 μ diameter, rounded and transparent in colour. Electron microscope examination of C3A infected cell line revealed the presence of numerous intracellular bacteria located in vacuoles or free in the host cytoplasm. In addition, the interaction of this bacterium with epithelial cells was associated with the elongation of micro-villar extension that extruded from the host cell membranes and engulfed the bacteria. This internalization mechanism strongly resembles Salmonella- or Shigella-induced macro-pinocytosis. The strain was characterized using the 16S RNA sequence that amplified the gene from many genera. The closest phylogenetic relative was |HQ877772.1| Escherichia sp. A94 with 88% 16S ribosomal RNA gene sequence similarity. It was proposed that unidentified strain be assigned as type strain of species of the bacteria origin based on the 16S rRNA gene sequence search in Ribosomal Database project, small subunit rRNA and large subunit rRNA database together with the phylogenetic tree analysis.© 2017 International Formulae Group. All rights reserved.Keywords: Intracellular contaminants, cell cultures, bacteria culture, pre-clinical studies

Endogenous Production of Geosmin in Table Beet

The earthy flavor of table beet is due to an aromatic terpene derivative called geosmin. It has been hypothesized that geosmin presence in beet is due to geosmin-producing bacteria such as Streptomyces spp. that exist in the soil. However, recent findings suggest that beet may produce geosmin endogenously without microbial influence. The purpose of this study was to determine whether such endogenous production of geosmin occurred in beet by making use of an aseptic tissue culture (TC) environment to remove potential microbial influences on geosmin production. Four table beet accessions (‘Bull’s Blood’, ‘Touchstone Gold’, ‘W364B’, and ‘Pacemaker III’) were grown in three separate TC experiments and in the greenhouse and measured for geosmin concentration via gas chromatography–mass spectrometry (GC-MS). Sequencing of 16S ribosomal RNA was used to identify potential microbial contaminants in TC. Operational taxa units (OTUs) classification resulted in RNA sequences with homology to bacterial RNA of either chloroplast (98%) or mitochondria (2%) origin. Other OTUs identified were considered within the range of sequencing error. In 15 of the 16 TC–grown samples used for the 16S rRNA aseptic validation and in all the greenhouse-grown plant samples, geosmin was detected. Geosmin concentrations from bulked beet tissue of each accession were higher in the TC environment than the greenhouse environment. The lack of microbial detection in the TC environment and the subsequent identification of geosmin from beets grown in the aseptic environment is a strong indication that geosmin is produced endogenously by beets. This finding raises several interesting questions about the functional significance of this molecule for Beta vulgaris.

PENGARUH TEKNIK STERILISASI EXPLAN TERHADAP TINGKAT PEROLEHAN KULTUR JARINGAN AKSENIK RAMIN (Gonystylus bancanus)

Since 2005, ramin has been included in CITES Appendix II and according to IUCN red list, ramin is also regarded as vulnerable (VU A1cd). The Lack of information on explant sterilization techniques in ramin tissue culture is one reason why there has not been enough initiation to develop ramin micro propagation. Plant tissue culture contamination has economic impact due to its direct influence in losses during in vitro culture of plants. The aim of this study was to observe the influence of explants sterilisation technique on acquisition rate of ramin axenic tissue culture with the rate of contamination increase, acsenic culture acquisition and shoot elongation as parameters. Ramin seedlings as explants source were collected from Tumbang Nusa, Central Kalimantan. Fifty replicating explants of one nodule each in 3 techniques were used in this study based on in vitro incubation time: sterilization 1 (24 hours incubation), sterilization 2 (48 hours incubation) and sterilization 3 (72 hours incubation) with non metallic antimicrobial compounds namely detergent, ditiocarbamate with mankozeb active ingredient (compound A), a compound containing 5.25% NaOCl added sodium hypochlorite and hydrogen oxide (compound B), 70% alcohol (compound C) as explants surface sterilants. Murashige-Skoog was used as media for evaluation of sterilization technique and explants regeneration on one year incubation. The results of this study indicate that the lowest rate of contamination was found in sterilization 3 (28%) with the success number of axenic tissue culture at 46%. The average shoot elongation of the axenic culture was 5.91 cm after 12 months subcultured in every month.

Rapid re-identification of human samples using portable DNA sequencing.

DNA re-identification is used for a broad suite of applications, ranging from cell line authentication to forensics. However, current re-identification schemes suffer from high latency and limited access. Here, we describe a rapid, inexpensive, and portable strategy to robustly re-identify human DNA called 'MinION sketching'. MinION sketching requires as few as 3 min of sequencing and 60-300 random SNPs to re-identify a sample enabling near real-time applications of DNA re-identification. Our method capitalizes on the rapidly growing availability of genomic reference data for cell lines, tissues in biobanks, and individuals. This empowers the application of MinION sketching in research and clinical settings for periodic cell line and tissue authentication. Importantly, our method enables considerably faster and more robust cell line authentication relative to current practices and could help to minimize the amount of irreproducible research caused by mix-ups and contamination in human cell and tissue cultures.

A human monocytic NF-κB fluorescent reporter cell line for detection of microbial contaminants in biological samples.

Sensing of pathogens by innate immune cells is essential for the initiation of appropriate immune responses. Toll-like receptors (TLRs), which are highly sensitive for various structurally and evolutionary conserved molecules derived from microbes have a prominent role in this process. TLR engagement results in the activation of the transcription factor NF-κB, which induces the expression of cytokines and other inflammatory mediators. The exquisite sensitivity of TLR signalling can be exploited for the detection of bacteria and microbial contaminants in tissue cultures and in protein preparations. Here we describe a cellular reporter system for the detection of TLR ligands in biological samples. The well-characterized human monocytic THP-1 cell line was chosen as host for an NF-ᴋB-inducible enhanced green fluorescent protein reporter gene. We studied the sensitivity of the resultant reporter cells for a variety of microbial components and observed a strong reactivity towards TLR1/2 and TLR2/6 ligands. Mycoplasma lipoproteins are potent TLR2/6 agonists and we demonstrate that our reporter cells can be used as reliable and robust detection system for mycoplasma contaminations in cell cultures. In addition, a TLR4-sensitive subline of our reporters was engineered, and probed with recombinant proteins expressed in different host systems. Bacterially expressed but not mammalian expressed proteins induced strong reporter activity. We also tested proteins expressed in an E. coli strain engineered to lack TLR4 agonists. Such preparations also induced reporter activation in THP-1 cells highlighting the importance of testing recombinant protein preparations for microbial contaminations beyond endotoxins. Our results demonstrate the usefulness of monocytic reporter cells for high-throughput screening for microbial contaminations in diverse biological samples, including tissue culture supernatants and recombinant protein preparations. Fluorescent reporter assays can be measured on standard flow cytometers and in contrast to established detection methods, like luciferase-based systems or Limulus Amebocyte Lysate tests, they do not require costly reagents.

Volatile indicators of contamination in tissue cultures

Microbial contamination is the most important reason for losses in both scientific and commercial micropropagation systems. Latent and epiphytic bacteria can hitchhike imperceptibly during many subcultures. They reduce growth, multiplication and rooting or alter the response to in vitro plant growth regulators. In vitro plants, culture vessel, medium and possible endogenous bacteria release a set of volatile aromatic molecules (biogenic volatile organic compounds, BVOC). Because of its high throughput rate and extreme sensitivity, Selected Ion Flow Tube Mass Spectrometry (SIFT-MS) was chosen to distinguish between presumed clean and artificially Escherichia coli contaminated cultures of Ficus benjamina and Spathiphyllum wallisii. Contaminated cultures of F. benjamina and S. wallisii yielded a net BVOC fingerprint that was both different from each other and from the BVOC spectrum of E. coli. This suggests that they release volatiles that result from the specific interaction with E. coli. The availability of a true 100% bacteria free in vitro plant will be necessary to validate this technology for commercial use.