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Complementary peptides represent a credible alternative to agrochemicals by activating translation of targeted proteins

Agriculture

Complementary peptides represent a credible alternative to agrochemicals by activating translation of targeted proteins

M. Ormancey, B. Guillotin, et al.

Discover how complementary peptides (CPEPs) could revolutionize agriculture by enhancing plant traits such as growth, pathogen resistance, and heat stress tolerance, all without genetic modification. This innovative research conducted by Mélanie Ormancey and colleagues shows the potential of CPEPs as a safe and effective alternative to traditional agrochemicals.

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Playback language: English
Introduction
Modern agriculture heavily relies on chemicals to boost crop yields, combating weeds and pathogens, and acting as growth regulators. However, challenges such as rising global populations, increasing temperatures, herbicide-resistant weeds, and concerns over chemical safety necessitate the development of novel, sustainable agricultural practices. This study investigates the potential of complementary peptides (CPEPs) as a safe and effective alternative to agrochemicals. CPEPs are short peptides designed to interact with specific mRNA sequences, thereby modulating protein expression. The researchers hypothesize that external application of synthetic CPEPs can enhance the abundance of targeted proteins, leading to improved plant traits and reduced weed growth. The study's significance lies in its potential to provide a more environmentally friendly and socially acceptable approach to crop improvement compared to conventional methods. This approach avoids the need for genetic manipulation, making it suitable for a wider range of plant species, including those challenging to transform genetically.
Literature Review
The paper references the significant increase in crop yields since 1945, largely attributed to the combined use of chemicals, agricultural mechanization, genetics, and crop management. It acknowledges the increasing challenges faced by modern agriculture, including rising global temperatures impacting crop yields, the depletion of mineral phosphorus-based fertilizers, the development of herbicide-resistant weeds, and societal concerns about the environmental and health impacts of agrochemicals. The limitations of existing alternatives such as CRISPR (difficult application in weed control) and small RNAs (poor penetration into plant cells) are also mentioned. The paper points to emerging research on short open reading frames (sORFs) producing small functional peptides, suggesting the possibility of naturally occurring CPEPs in plants.
Methodology
The study employed a variety of techniques. Initially, RNA immunoprecipitation (RIP) followed by PCR was used to validate the interaction between a designed cPEP and its target mRNA in Arabidopsis thaliana. Luciferase activity assays were conducted to assess the impact of CPEPs on protein expression levels. Quantitative PCR (qPCR) was used to measure mRNA abundance. Additional CPEPs targeting different sequences within the luciferase gene were designed and tested. The impact of peptide length, concentration, and treatment duration on protein expression were systematically examined. Proteomic analysis was performed to assess potential off-target effects of CPEPs. Fluorescence resonance energy transfer-fluorescence lifetime imaging microscopy (FRET-FLIM) was used to investigate the interaction between CPEPs and their target mRNAs in planta. The study further extended its scope to Medicago truncatula and investigated the impact of CPEPs on various plant traits, including root development, nodulation, and defense responses against pathogens. A range of experimental techniques were used across various plant species, including pathogenicity assays, measuring chlorophyll content, and seedling viability under heat stress, to assess the effect of CPEPs on different plant traits. In vitro transcription/translation assays were used to determine if CPEPs influence protein stability or translation. 5'P sequencing was used to study ribosome occupancy on target transcripts. Co-immunoprecipitation followed by mass spectrometry was employed to identify proteins interacting with the CPEPs. Polysome analysis was conducted to understand the mechanism of action of CPEPs. The study was not limited to model plants but was extended to plants of agronomic importance, including tomato and soybean, assessing plant defense, heat stress tolerance and growth promotion. Weed control potential was also evaluated using various weed species.
Key Findings
The core finding is that externally applied synthetic CPEPs specifically enhance the expression of their target proteins without altering mRNA levels. This effect was observed across various plant species and proteins, demonstrating the broad applicability of this approach. Specifically, CPEPs increased the activity of their targeted proteins, with optimal effects observed at a concentration of 50 µM and after 24 hours of treatment. Proteomic analysis did not reveal significant off-target effects. The research confirmed that the activity of CPEPs depends on the presence of their complementary RNA sequence on the target mRNA. The increased protein levels observed after cPEP treatment directly resulted from increased translation efficiency, not increased protein stability. 5'P sequencing revealed that CPEPs increase ribosome recruitment at translation initiation sites on target transcripts. Co-immunoprecipitation identified ribosomal protein RPL19 as a crucial factor involved in the cPEP's mechanism of action. The study demonstrated the ability to use CPEPs to modulate diverse plant phenotypes, including root development, nodulation, pathogen resistance, chlorophyll content, and heat stress tolerance. Notably, CPEPs exhibited additive effects when combined, leading to enhanced phenotypic changes. Importantly, the technology proved effective in plants of agronomic interest, improving plant defense in tomatoes, heat stress tolerance in soybeans, and showing promise in weed control using different weed species.
Discussion
The study provides compelling evidence for the use of CPEPs as a novel tool for modulating protein expression in plants. The finding that CPEPs enhance translation efficiency without impacting mRNA levels offers a new approach for precisely controlling protein levels. The success in manipulating diverse plant traits across various species showcases the wide applicability of CPEPs in agricultural applications. The observation of additive effects with cPEP combinations suggests the possibility of simultaneously targeting multiple pathways to achieve more robust phenotypic changes. This technology offers significant potential for developing sustainable agricultural practices by providing a safe and effective alternative to agrochemicals. The lack of observed off-target effects suggests high specificity, further strengthening its potential for practical applications.
Conclusion
This research demonstrates the potential of complementary peptides (CPEPs) as a novel and effective method to modulate plant traits. The ability to enhance protein expression by external application of synthetic peptides, without genetic modification, offers significant advantages over traditional methods, particularly in species resistant to genetic transformation. The successful application of CPEPs across various species and traits, including improvement of growth, stress resistance, and disease control, along with their potential for weed management, positions CPEPs as promising tools in sustainable agriculture. Future research should focus on identifying naturally occurring CPEPs (natCPEPs), exploring the full range of plant responses to CPEPs, and optimizing their formulation for field applications.
Limitations
While the study demonstrates significant progress, there are areas that require further investigation. The mechanism of CPEPs' interaction with ribosomes warrants further exploration to fully elucidate their mode of action. A thorough investigation into the long-term environmental impact of CPEPs and potential for degradation in soil is also crucial. Extensive field trials are needed to confirm the effectiveness and scalability of the technology under various environmental conditions.
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