In the face of climate change, progressive degradation of the environment, including agricultural land negatively affecting plant growth and development, endangers plant productivity. Seeking efficient and sustainable agricultural techniques to replace agricultural chemicals is one of the most important challenges nowadays. The use of plant growth-promoting microorganisms is among the most promising approaches; however, molecular mechanisms underneath plant–microbe interactions are still poorly understood. In this review, we summarized the knowledge on plant–microbe interactions, highlighting the role of microbial and plant proteins and metabolites in the formation of symbiotic relationships. This review covers rhizosphere and phyllosphere microbiomes, the role of root exudates in plant–microorganism interactions, the functioning of the plant’s immune system during the plant–microorganism interactions. We also emphasized the possible role of the stringent response and the evolutionarily conserved mechanism during the established interaction between plants and microorganisms. As a case study, we discussed fungi belonging to the genus Trichoderma. Our review aims to summarize the existing knowledge about plant–microorganism interactions and to highlight molecular pathways that need further investigation.

Among several mechanisms involved in the plant stress response, synthesis of guanosine tetra and pentaphosphates (alarmones), homologous to the bacterial stringent response, is of crucial importance. Plant alarmones affect, among others, photosynthetic activity, metabolite accumulation, and nutrient remobilization, and thus regulate plant growth and development. The plant RSH (RelA/SpoT homolog) genes, that encode synthetases and/or hydrolases of alarmones, have been characterized in a limited number of plant species, e.g., Arabidopsis thaliana, Oryza sativa, and Ipomoea nil. Here, we used dry-to-wet laboratory research approaches to characterize RSH family genes in the polyploid plant Brassica napus. There are 12 RSH genes in the genome of rapeseed that belong to four types of RSH genes: 6 RSH1, 2 RSH2, 3 RSH3, and 1 CRSH. BnRSH genes contain 13–24 introns in RSH1, 2–6 introns in RSH2, 1–6 introns in RSH3, and 2–3 introns in the CRSH genes. In the promoter regions of the RSH genes, we showed the presence of regulatory elements of the response to light, plant hormones, plant development, and abiotic and biotic stresses. The wet-lab analysis showed that expression of BnRSH genes is generally not significantly affected by salt stress, but that the presence of PGPR bacteria, mostly of Serratia sp., increased the expression of BnRSH significantly. The obtained results show that BnRSH genes are differently affected by biotic and abiotic factors, which indicates their different functions in plants.

The problem of handling zinc in the cell is of great importance because zinc is an indispensable micronutrient involved in most physiological processes in all living organisms. Moreover, our understanding of mechanisms governing the discrimination between micronutrients and toxic metals on the level of individual proteins to the whole-organism level is incomplete. Metallothioneins are able to bind heavy metal ions, and roles in zinc homeostasis have been proposed. Here, we have studied the in vitro and in vivo metal-binding abilities of Brassica napus type 4 metallothionein (BnMT4) and its expression in germinating seeds in response to metal treatment. Our studies on the regulation of MT4 expression by metals at early stages of ontogenic development have revealed for the first time that the mRNA levels of BnMT4 were elevated in response to cadmium and zinc. Given this unexpected metalloregulation, and the dramatic differences in protein folding as detected by 1H NMR spectroscopy, we suggest that the BnMT4 protein may not only have a role in zinc homeostasis in early ontogenesis, but also the potential to discriminate between zinc and cadmium, perhaps via differential recognition of Cd- and Zn-complexes by cellular components involved in protein turnover.

Germination is a crucial process in crop plant life cycle that determines future yield. Hydrogen peroxide plays a significant role in both dormancy release and seed ageing. The potential of the rapeseed metallothioneins (BnMT1‐4) as reactive oxygen species scavengers was investigated. In the presence of H2O2, bacterial cells expressing BnMT1‐4 had higher growth rate than control cells. However, such effect was not observed in an oxidative stress oversensitive yap‐1Δ yeast mutant strain expressing BnMT1‐4. Next, it was shown that H2O2, up to 100 and 10 mM, promotes rapeseed germination rate and seedling growth, respectively. Moreover, it was demonstrated that in seeds germinating in the presence of H2O2, the expression of seed‐specific BnMT4 decreases to a high extent in a dose‐ and time‐dependent manner. It was shown that the decline in BnMT4 mRNA level, accelerated by up to 10 mM H2O2 treatment, correlates positively with increased rapeseed germination and early growth, underscoring the plausible role of BnMT4 as a H2O2 scavenging protein that level has to be downregulated in conditions where the compound is needed. Finally, it was demonstrated that H2O2 could be used as a priming agent of rapeseeds.

Anthropogenic pollution with heavy metals is an on-going concern throughout the world, and methods to monitor release and impact of heavy metals are of high importance. With a view to probe its suitability as molecular biomarker of metal pollution, this study has determined a coding sequence for metallothionein of the African sharptooth catfish Clarias gariepinus. The gene product was recombinantly expressed in Escherichia coli in presence of Zn(II), Cd(II), or Cu, and characterised by Electrospray Ionisation Mass Spectrometry and elemental analysis. C. gariepinus MT displays typical features of fish MTs, including 20 conserved cysteines, and seven bound divalent cations (Zn(II) or Cd(II)) when saturated. Livers from wild C. gariepinus fish collected in all three seasons from four different sites on the Kafue River of Zambia were analysed for their metal contents and for MT expression levels by quantitative PCR. Significant correlations were found between Zn and Cu levels and MT expression in livers, with MT expression clearly highest at the most polluted site, Chililabombwe, which is situated in the Copperbelt region. Based on our findings, hepatic expression of MT from C. gariepinus may be further developed as a major molecular biomarker of heavy metal pollution resulting from mining activities in this region.

Osmotic stress frequently caused by drought is the most threatening environmental stress that remarkably reduces crop yield. Oat (Avena sativa L.) is sensitive to water deficiency during growth. Plant metallothioneins (pMTs) show tremendous promise in enhancing general stress tolerance in plants. This study aimed to verify whether pMTs and elements of the antioxidant defence system protect oat against osmotic stress.Coding and genomic regions of A. sativa L. MTs belonging to three different types were cloned. To evaluate the role of MTs in osmotic stress, the expression of genes encoding AsMT1-3 was checked by qRT-PCR in the roots and shoots of oat plants growing in a hydroponic culture in the presence of polyethylene glycol (PEG 6000) and mannitol. The expression of AsMT1-3 changed in response to osmotic stress; however, the changes depended on the type of MT and the treatment. The amount of AsMT3 transcript was about five-fold and nine-fold higher in shoots and roots, respectively, in the presence of mannitol. To further analyse the response of oat to osmotic stress, the level of phenolic compounds, soluble sugars, abscisic acid, and the activity of antioxidant enzymes were tested using spectrophotometric and chromatographic methods. During osmotic stress, the content of phenols, soluble sugars, abscisic acid, and the activity of catalase and peroxidase in shoots increased. In roots treated with PEG 6000, the amount of phenolic compounds was higher than that in roots treated with mannitol. The activity of superoxide dismutase was about 5-fold higher in roots than in shoots. MTs are involved in plant response to osmotic stress. In the future, insight provided in this study will lead to application in agriculture either by using MTs as molecular markers for stress-resistant crop cultivars or by a generation of genetically modified crops overexpressing MTs.

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