A large data array on plant gene expression accumulated thanks to comparative omic studies directs the efforts of researchers to the specific or fine effects of the target gene functions and, as a consequence, elaboration of relatively simple and concurrently effective approaches allowing for the insight into the physiological role of gene products. Numerous studies have convincingly demonstrated the efficacy of transient expression strategy for characterization of the plant gene functions. The review goals are (i) to consider the advantages and limitations of different plant systems and methods of transient expression used to find out the role of gene products; (ii) to summarize the current data on the use of the transient expression approaches for the insight into fine mechanisms underlying the gene function; and (iii) to outline the accomplishments in efficient transient expression of plant genes. In general, the review discusses the main and critical steps in each of the methods of transient gene expression in plants; areas of their application; main results obtained using plant objects; their contribution to our knowledge about the fine mechanisms of the plant gene functions underlying plant growth and development; and clarification of the mechanisms regulating complex metabolic pathways.

The complexities of translational strategies make this stage of implementing genetic information one of the most challenging to comprehend and, simultaneously, perhaps the most engaging. It is evident that this diverse range of strategies results not only from a long evolutionary history, but is also of paramount importance for refining gene expression and metabolic modulation. This notion is particularly accurate for organisms that predominantly exhibit biochemical and physiological reactions with a lack of behavioural ones. Plants are a group of organisms that exhibit such features. Addressing unfavourable environmental conditions plays a pivotal role in plant physiology. This is particularly evident with the changing conditions of global warming and the irrevocable loss or depletion of natural ecosystems. In conceptual terms, the plant response to abiotic stress comprises a set of elaborate and intricate strategies. This is influenced by a range of abiotic factors that cause stressful conditions, and molecular genetic mechanisms that fine-tune metabolic pathways allowing the plant organism to overcome non-standard and non-optimal conditions. This review aims to focus on the current state of the art in the field of translational regulation in plants under abiotic stress conditions. Different regulatory elements and patterns are being assessed chronologically. We deem it important to focus on significant high-performance techniques for studying the genetic information dynamics during the translation phase.

One of the unresolved questions in stress-response biology is how plants coordinate expression levels between the response and adaptation. In this work, we proposed a two-level analysis that examines both transcriptional and translational profiles of Solanum lycopersicum under conditions of short-term cold stress, hardening, and their combination. By combining polysome profiling and total transcriptome analysis, we revealed that expression under cold stress is not a simple linear process but a structurally distinct system with two coordinated regulation centres. Hardening triggers a strong transcriptional program focused on biogenesis, light signalling, and structural adaptations. In contrast, acute stress prompts selective translation of metabolic and defence proteins without prior transcriptional increase. Modular analysis (WGCNA) showed little overlap between transcriptional and translational networks, indicating functional differences between regulation levels. This work demonstrates that the cold response involves a strategic reallocation of resources between transcription and translation based on the type of signal. It bridges basic biology and applied breeding, providing targets promising for improving plant stress tolerance and advancing bioengineering of adaptive agriculture.

Background: Since the beginning of the use of reporter proteins for expression analysis, a variety of approaches have been developed and proposed; both qualitative and quantitative. The lack of simple methods for direct observation of gene expression in living organisms makes it necessary to continue to propose new methods. In this work, we consider a method for the quantitative analysis of the expression of thermostable lichenase from Clostridium thermocellum used as a sensitive reporter protein. Results: In this study, we report the design a high throughput fluorometric method for quantification of thermostable lichenase C. thermocellum using Congo red and further experimental verification of its relevance and efficiency in assessment of the functional role of regulatory sequences in the plant cell. Conclusions: The specific interaction between the dye Congo red and $$\beta$$β -d-glucans formed the background for designing a high-throughput fluorometric assay for quantification of C. thermocellum thermostable lichenase as a reporter protein for plants. This assay (i) makes it possible to precisely measure the amount of reporter protein in a plant sample; (ii) has shown a high sensitivity for quantification of thermostable lichenase; (iii) is more time- and cost-efficient as compared with the Somogyi–Nelson assay; and (iv) is to the least degree dependent on the presence of the tested buffer components as compared with the Somogyi–Nelson assay.

The intricacies of translational strategies make this stage of the implementation of genetic information one of the most difficult to understand and, at the same time, perhaps the most interesting. It also becomes clear that all this variety of strategies is not just a consequence of a long evolutionary history but is of paramount importance for fine-tuning gene expression and metabolic modulation. This is especially true for those organisms that demonstrate predominantly biochemical and physiological reactions with a lack of behavioural ones. Plants are just such a group of organisms. Overcoming unfavourable environmental conditions occupies a key place in plant physiology. This is especially true with the changing conditions of global warming and the irretrievable loss or depletion of natural ecosystems. Conceptually, the response of plants to abiotic stress is a set of complex and intricate strategies. This is dictated both by a variety of abiotic factors that provoke stressful conditions and by a variety of molecular genetic mechanisms that make it possible to fine-tune the metabolic pathways through which the plant organism overcomes non-standard and non-optimal conditions. In the presented review, we tried to focus on the state-of-the-art in the field of translational regulation in plants under abiotic stress conditions. Various regulatory elements and patterns are considered in relative chronological order. We also considered it necessary to pay attention to key high-performance methods for studying the dynamics of genetic information at the stage of translation.

One of the unresolved questions in stress-response biology remains the mechanism of coordination of expression levels in plants between the slow preparation phase and the rapid reactive response to stress exposure. In this work, we proposed a two-level analysis covering simultaneously transcriptional and translational profiles of Solanum lycopersicum under short-term cold stress, hardening and their combination. By combining polysome profiling and total transcriptome analysis, we revealed that expression under cold stress is not a linear chain but a structurally separated system with two coordinated centres of regulation. Hardening activates a robust transcriptional programme with a focus on biogenesis, light signalling and structural adaptations. In contrast, acute stress initiates selective translation of metabolic and defence proteins without prior enhancement of transcription. Modular analysis (WGCNA) showed low overlap between transcriptional and translational networks, indicating functional divergence between levels of regulation. In this work, we demonstrate that the cold response is a strategic reallocation of resources between expression levels depending on the type of signal. The work forms a bridge between basic biology and applied breeding, offering targets promising for targeted improvement of plant stress tolerance and further bioengineering of adaptive agrocultures.