![]() However, the formation of bacterial colonies on culture media and bacterial overgrowth of plant tissues are common and manifest as infection by a variety of pathogenic species or the non-fastidious proliferation of commensal endophytic bacteria that can be triggered by changes in environmental conditions or plant host physiology. In fact, several studies showed a beneficial effect of endophytic bacteria on the growth of in vitro cultures of tomato, grapevine, sweet cherry, apple, purple coneflower and tobacco. Some bacterial endophytes maintain habitually concealed lifestyles or, due to limited bacterial growth-supporting media conditions, remain latent over extended periods of in vitro tissue cultivation. Being protected inside explant or seed tissues, endophytic bacteria evade surface sterilization procedures used for cell culture initiation, and they are common in plant tissues grown in vitro. These results imply that the diversity of the plant-associated microbiome might represent a significant factor contributing to the efficient propagation of in vitro tissue culture.Įndophytes are a class of endosymbiotic microorganisms that internally inhabit plant tissues. Antibiotic treatment resulted in a decline in microbial diversity (the number of families was reduced 4.5-fold) and increased domination by the Mycobacteriaceae family. Microbiome analysis of the shoot samples using multivariable region-based 16S rRNA gene sequencing revealed a diverse microbial community in the control tobacco shoots, including 59 bacterial families however, it was largely dominated by Mycobacteriaceae. Moreover, the growth properties of shoots were only partially restored after transfer to a medium without the antibiotic. The study revealed that shoot cultivation on a medium supplemented with 250 mg L −1 timentin resulted in 29 ± 4% reduced biomass accumulation and a 1.2–1.6-fold higher level of oxidative stress injury compared to the control samples. Therefore, the aim of this study was to assess the effect of antibiotic treatment on the growth and stress level of tobacco ( Nicotiana tabacum L.) shoots in vitro as well as the composition of the plant-associated microbiome. Antibiotics are commonly used to decontaminate plant tissue culture or during genetic transformation however, the effect of antibiotic treatment on the diversity of indigenous microbial populations and the consequences on the performance of tissue culture is not completely understood.
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