How do you justify the usage of antibiotics in PTC at the time of
1) explant sterilization, or 2) within the plant tissue culture media.
is it suggestable/ethical to use antibiotics in normal tissue culture experiments? (not for the transgenic selection etc)
if yes how?
if not why?
Since antibiotics have a negative effect on the growth and metabolism of plants, they are not recommended except in obligatory cases.
difficult to answer and make clear judgments. It depends on the procedures adopted. The same disposal procedures as for transgenic experiments should be used to avoid the dispersion of resistant microorganisms. Given these procedures and if other substances such as the plant preservative medium or other sterilizazion protocols are not enough, then it is possible to use them in experiments
To kill bacteria effectively antibiotics are essential otherwise by other means , it is very difficult to destroy bacteria.
Thank you Madhukar Baburao Deshmukh Sir for your reply.
When I asked a few researchers they were told, that antibiotics are not to be used in the medium. if we are using the antibiotics in the PTC Medium, the microbes will get a prolonged period of exposure/contact, so that if the concentration is not up to the mark there are chances of getting ANTIMICROBIAL RESISTANCE, which is unethical in their point of view. How do you comment on this view sir?
Isolate the contamination with loop, and check it under a light microscope. Sometimes it looks like bacteria by need eyes, but it might be yeast after a close look. Once you define the contamination type, apply an efficient chemicals. If it is bacteria, go through with its cell wall (- or +), then figure out the antibiotic and its concentration starting from 20 mg/L upto 100 mg/L. If it is not bacteria, go with an anti fungal agent depending on concentrations. Silver nitrate would be an effective chemical, but it reduces callus formations at high concentrations (>1 mg/L) in general. Good luck
Thank You Buhara Yucesan Sir
I will try to encounter it this way next time...!
Dear Reza, not all antibiotics cause damages to plants. Certainly, some aminoglycosides (Kanamicin, Streptomicin...) might have detrimental effects, depending of plant species and dosages. However, some betalactamics, like Cefotaxime (some authors have suggested that its metabolization has connections with citoquinins synthesis) and Carbenicillin, beside control bacteria, promote the in vitro growth of some species. Dear Bhadepuri, the usage of antibiotics in tissue culture shouldn't be considered as a routine, but under certain circumstances these offer huge benefits controlling endophytics.
1. Explant sterilisation does not need antibiotics. For this hypochlorite solution or alcohol is used depending on the nature of eggplant.
2. Antibiotics are not used in normal PTC medium as it inhibits the growth. However for selection purposes such as in transgenic experiments, obtaining somatic hybrids or resistant cell lines antibiotics have been used.
Thank You M. K. Razdan Sir for sharing your valuable experiences...
Badhepuri Mahesh Kumar
According to the latest reports, the incorporation of Phloroglucinol could eliminate the use of antibiotics in culture medium. PG would thus provide a potential application for use as an antimicrobial agent in vitro. Its ability to be autoclaved makes it an attractive alternative to antibiotics. It might also serve to “sterilize” tissue culture media without the need for autoclaving.
Dear M. Manokari it's an interesting (and amazing too) effect of PG. Could you please share the paper where do you read it? Thanks beforehand.
Thank you M. Manokari Madam
its quite interesting and new to know...
let me try this once
Dear Ricardo Julian Licea-Moreno
Please find the information in
Article Phloroglucinol in plant tissue culture
by Teixeira da Silva JA, Dobránszki J, Ross S (2013).
Dear M. Manokari I know very well this review. Regarding it, I've to make some comments. The inference of antimicrobial activity of PG is based on the paper of Miyake and Hiramitsu (2011). In this paper 4 antimicrobial substances were isolated from citrus peels. Among them, it's mentioned phloroglucinol 1-β-D-glucopyranoside; however, it's not PG (1, 3,5 trihydroxybenzene) but phlorin (3,5-dihydroxyphenyl beta-D-glucopyranoside), a more complex substance. Anyway, because of its phenolic nature (phenol is really caustic), it might acts as a antimicrobial. A last comment, some derivates from PG have antibacterial activity (see Casero et al 2015).
Dear Ricardo Julian Licea-Moreno
Thank you for the comments and sharing the references.
Thank you Ricardo Julian Licea-Moreno Sir for sharing valuable experience and knowledge...!
Badhepuri Mahesh Kumar I'd prefer to use as many as explants to find internal clean materials
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