Effects of Different β-Lactam Antibiotics on Indirect Tomato (Solanum lycopersicum L.) Shoot Organogenesis and Agrobacterium tumefaciens Growth Inhibition In Vitro

Abstract

A β-lactams that act by inhibiting the bacterial cell wall biosynthesis are one of the most common classes of antibiotics applied to suppress the growth of latent bacterial infection associated with the plant tissue culture, as well as in the Agrobacterium-mediated transformation techniques. Plant sensitivity to antibiotics usually is species-, genotype-, or even tissue-specific and mainly depends on concentrations, growth conditions, and culture system. In the presented article, we estimated a comparative effect of four β-lactam antibiotics (Claforan®, timentin, amoxicillin, and Amoxiclav®) at different concentrations in an agar-solidified Murashige and Skoog (MS) culture medium supplemented with 5 mg L−1 6-benzylaminopurine (6-BA) and 0.1 mg L−1 indole-3-acetic acid (IAA) on in vitro callus induction and shoot organogenesis from hypocotyl and cotyledon explants of two tomato cultivars (Rekordsmen, Moryana). The role of clavulanic acid in combination with amoxicillin (Amoxiclav®) in the shoot organogenesis frequency and number of shoots per explant has been demonstrated. Additionally, the growth inhibition of Agrobacterium tumefaciens AGL0 strain according to agar disk-diffusion assay was studied. As a result, both stimulatory (timentin, amoxicillin, and Amoxiclav®) and inhibitory (Claforan®) effects of β-lactam antibiotics on in vitro morphogenetic responses of tomato were noted. It was found that clavulanic acid, which is part of the commercial antibiotic Amoxiclav®, significantly increased the shoot regeneration frequency from cotyledon and hypocotyl explants of Rekordsmen tomato cultivar. Possible reasons for the stimulating effect of clavulanic acid on the induction of shoot organogenesis are discussed. According to agar disk-diffusion assay, the maximum diameter of growth inhibition zones (43.9 mm) was identified using 200 mg L−1 timentin. The in vitro antibacterial activity of tested β-lactam antibiotics was arranged in the following order: timentin > Claforan® > amoxicillin ≥ Amoxiclav®. Thus, to suppress the growth of internal and latent bacterial infection of tomato plant tissue culture, as well as for transformation of Moryana and Rekordsmen cultivars by A. tumefaciens strain AGL0, we recommend adding of 100–200 mg L−1 timentin or 400–800 mg L−1 Amoxiclav® to the shoot induction medium.

Ticarcillin degradation product thiophene acetic acid is a novel auxin analog that promotes organogenesis in tomato

“…These authors suggested that auxin-like degradation products of timentin could be playing a role in this increased regeneration. ( Varlamova et al., 2021 ) showed that timentin and Amoxicillin enhanced shoot organogenesis in tomato is concentration and genotype dependent. They suggested that the positive effects of these antibiotics on shoot organogenesis is probably due to the presence of clavulanic acid or its degradation products.…”

“…Previous studies have reported a positive effect of β-lactam ring-containing antibiotics such as timentin, carbenicillin, and penicillin G on shoot regeneration in species such as tomato, tobacco, and London plane tree. This effect was found to be dependent on the antibiotic concentration and plant genotype ( Nauerby et al., 1997 ; Ling et al., 1998 ; Costa et al., 2000 ; Li et al., 2007 ; Varlamova et al., 2021 ). In contrast, some studies have also reported no significant effect of these antibiotics on shoot regeneration in species such as tobacco and Siberian elm ( Cheng et al., 1998 ).…”

Aztreonam is a novel chemical inducer that promotes Agrobacteium transformation and lateral root development in soybean

“…Like beta-lactam antibiotics, cefotaxime, carbenicillin, penicillin G, and some other antibiotics were found effective during somatic embryogenesis, axillary bud formation, and shoot induction in dainthus and Solanum viarum ( Nakano and Mii, 1993 ; Mahadev et al, 2014 ). Amoxiclav was found to increase morphogenesis and shoot regeneration in maize, carrot, and tomato, respectively ( Danilova and Dolgikh, 2005 ; Grzebelus and Skop, 2014 ; Varlamova et al, 2021 ). However, if we utilized this during co-cultivation, its antibacterial activities against gram-negative bacteria might impede bacterial development.…”

Advancing Antibiotic Residue Analysis: LC-MS/MS Methodology for Ticarcillin Degradation Products in Tomato Leaves

“…Timentin is a formulation that provides stability to the β-lactam ring in the broad-spectrum semi-synthetic β-lactam antibiotic ticarcillin by combining it with clavulanic acid, a β-Lactamase competitive inhibitor [24,25]. Timentin has been demonstrated to have a favorable impact on shoot regeneration in several plant species, including London plane trees, tomato, and tobacco, along with a few other β-lactam ring-containing antibiotics like penicillin G and carbenicillin [23,[26][27][28]. Conversely, other research has also shown that these antibiotics have no appreciable impact on the regeneration of shoots in species like tobacco and Siberian elm [29,30].…”