-
1.
Recent Advances in Plant Nanoscience.
Zhang, Q, Ying, Y, Ping, J
Advanced science (Weinheim, Baden-Wurttemberg, Germany). 2022;(2):e2103414
Abstract
Plants have complex internal signaling pathways to quickly adjust to environmental changes and harvest energy from the environment. Facing the growing population, there is an urgent need for plant transformation and precise monitoring of plant growth to improve crop yields. Nanotechnology, an interdisciplinary research field, has recently been boosting plant yields and meeting global energy needs. In this context, a new field, "plant nanoscience," which describes the interaction between plants and nanotechnology, emerges as the times require. Nanosensors, nanofertilizers, nanopesticides, and nano-plant genetic engineering are of great help in increasing crop yields. Nanogenerators are helping to develop the potential of plants in the field of energy harvesting. Furthermore, the uptake and internalization of nanomaterials in plants and the possible effects are also worthy of attention. In this review, a forward-looking perspective on the plant nanoscience is presented and feasible solutions for future food shortages and energy crises are provided.
-
2.
Current progress and challenges in crop genetic transformation.
Anjanappa, RB, Gruissem, W
Journal of plant physiology. 2021;:153411
Abstract
Plant transformation remains the most sought-after technology for functional genomics and crop genetic improvement, especially for introducing specific new traits and to modify or recombine already existing traits. Along with many other agricultural technologies, the global production of genetically engineered crops has steadily grown since they were first introduced 25 years ago. Since the first transfer of DNA into plant cells using Agrobacterium tumefaciens, different transformation methods have enabled rapid advances in molecular breeding approaches to bring crop varieties with novel traits to the market that would be difficult or not possible to achieve with conventional breeding methods. Today, transformation to produce genetically engineered crops is the fastest and most widely adopted technology in agriculture. The rapidly increasing number of sequenced plant genomes and information from functional genomics data to understand gene function, together with novel gene cloning and tissue culture methods, is further accelerating crop improvement and trait development. These advances are welcome and needed to make crops more resilient to climate change and to secure their yield for feeding the increasing human population. Despite the success, transformation remains a bottleneck because many plant species and crop genotypes are recalcitrant to established tissue culture and regeneration conditions, or they show poor transformability. Improvements are possible using morphogenetic transcriptional regulators, but their broader applicability remains to be tested. Advances in genome editing techniques and direct, non-tissue culture-based transformation methods offer alternative approaches to enhance varietal development in other recalcitrant crops. Here, we review recent developments in plant transformation and regeneration, and discuss opportunities for new breeding technologies in agriculture.
-
3.
Emergent Damped Oscillation Induced by Nutrient-Modulating Growth Feedback.
Melendez-Alvarez, J, He, C, Zhang, R, Kuang, Y, Tian, XJ
ACS synthetic biology. 2021;(5):1227-1236
-
-
Free full text
-
Abstract
Growth feedback, the inherent coupling between the synthetic gene circuit and the host cell growth, could significantly change the circuit behaviors. Previously, a diverse array of emergent behaviors, such as growth bistability, enhanced ultrasensitivity, and topology-dependent memory loss, were reported to be induced by growth feedback. However, the influence of the growth feedback on the circuit functions remains underexplored. Here, we reported an unexpected damped oscillatory behavior of a self-activation gene circuit induced by nutrient-modulating growth feedback. Specifically, after dilution of the activated self-activation switch into the fresh medium with moderate nutrients, its gene expression first decreases as the cell grows and then shows a significant overshoot before it reaches the steady state, leading to damped oscillation dynamics. Fitting the data with a coarse-grained model suggests a nonmonotonic growth-rate regulation on gene production rate. The underlying mechanism of the oscillation was demonstrated by a molecular mathematical model, which includes the ribosome allocation toward gene production, cell growth, and cell maintenance. Interestingly, the model predicted a counterintuitive dependence of oscillation amplitude on the nutrition level, where the highest peak was found in the medium with moderate nutrients, but was not observed in rich nutrients. We experimentally verified this prediction by tuning the nutrient level in the culture medium. We did not observe significant oscillatory behavior for the toggle switch, suggesting that the emergence of damped oscillatory behavior depends on circuit network topology. Our results demonstrated a new nonlinear emergent behavior mediated by growth feedback, which depends on the ribosome allocation between gene circuit and cell growth.
-
4.
The Tryptophan-Induced tnaC Ribosome Stalling Sequence Exposes High Amino Acid Cross-Talk That Can Be Mitigated by Removal of NusB for Higher Orthogonality.
Sherman, MW, Sandeep, S, Contreras, LM
ACS synthetic biology. 2021;(5):1024-1038
Abstract
A growing number of engineered synthetic circuits have employed biological parts coupling transcription and translation in bacterial systems to control downstream gene expression. One such example, the leader sequence of the tryptophanase (tna) operon, is a transcription-translation system commonly employed as an l-tryptophan inducible circuit controlled by ribosome stalling. While induction of the tna operon has been well-characterized in response to l-tryptophan, cross-talk of this modular component with other metabolites in the cell, such as other naturally occurring amino acids, has been less explored. In this study, we investigated the impact of natural metabolites and E. coli host factors on induction of the tna leader sequence. To do so, we constructed and biochemically validated an experimental assay using the tna operon leader sequence to assess differential regulation of transcription elongation and translation in response to l-tryptophan. Operon induction was then assessed following addition of each of the 20 naturally occurring amino acids to discover that several additional amino acids (e.g., l-alanine, l-cysteine, l-glycine, l-methionine, and l-threonine) also induce expression of the tna leader sequence. Following characterization of dose-dependent induction by l-cysteine relative to l-tryptophan, the effect on induction by single gene knockouts of protein factors associated with transcription and/or translation were interrogated. Our results implicate the endogenous cellular protein, NusB, as an important factor associated with induction of the operon by the alternative amino acids. As such, removal of the nusB gene from strains intended for tryptophan-sensing utilizing the tna leader region reduces amino acid cross-talk, resulting in enhanced orthogonal control of this commonly used synthetic system.
-
5.
Genome-based engineering of ligninolytic enzymes in fungi.
Asemoloye, MD, Marchisio, MA, Gupta, VK, Pecoraro, L
Microbial cell factories. 2021;(1):20
Abstract
BACKGROUND Many fungi grow as saprobic organisms and obtain nutrients from a wide range of dead organic materials. Among saprobes, fungal species that grow on wood or in polluted environments have evolved prolific mechanisms for the production of degrading compounds, such as ligninolytic enzymes. These enzymes include arrays of intense redox-potential oxidoreductase, such as laccase, catalase, and peroxidases. The ability to produce ligninolytic enzymes makes a variety of fungal species suitable for application in many industries, including the production of biofuels and antibiotics, bioremediation, and biomedical application as biosensors. However, fungal ligninolytic enzymes are produced naturally in small quantities that may not meet the industrial or market demands. Over the last decade, combined synthetic biology and computational designs have yielded significant results in enhancing the synthesis of natural compounds in fungi. In this review, we gave insights into different protein engineering methods, including rational, semi-rational, and directed evolution approaches that have been employed to enhance the production of some important ligninolytic enzymes in fungi. We described the role of metabolic pathway engineering to optimize the synthesis of chemical compounds of interest in various fields. We highlighted synthetic biology novel techniques for biosynthetic gene cluster (BGC) activation in fungo and heterologous reconstruction of BGC in microbial cells. We also discussed in detail some recombinant ligninolytic enzymes that have been successfully enhanced and expressed in different heterologous hosts. Finally, we described recent advance in CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-Cas (CRISPR associated) protein systems as the most promising biotechnology for large-scale production of ligninolytic enzymes. SHORT CONCLUSION Aggregation, expression, and regulation of ligninolytic enzymes in fungi require very complex procedures with many interfering factors. Synthetic and computational biology strategies, as explained in this review, are powerful tools that can be combined to solve these puzzles. These integrated strategies can lead to the production of enzymes with special abilities, such as wide substrate specifications, thermo-stability, tolerance to long time storage, and stability in different substrate conditions, such as pH and nutrients.
-
6.
Multifloret spikelet improves rice yield.
Ren, D, Li, Y, He, G, Qian, Q
The New phytologist. 2020;(6):2301-2306
Abstract
The typical rice (Oryza sativa) spikelet contains a single fertile floret and produces only one grain; by contrast, Brachypodium distachyon spikelets contain multiple fertile florets and produce several grains. To increase yield, rice breeders have traditionally focused on panicle morphology (branch number and length, spikelet density), but have not considered the number of florets in each spikelet. Production of rice spikelets with more florets could further increase the number of grains per panicle. Here, we describe two novel approaches - altering meristem determinacy and restoring lateral floret formation - for breeding rice cultivars with a multifloret spikelet, thereby increasing the number of grains per panicle and potentially improving yield.
-
7.
Target-specific gene delivery in plant systems and their expression: Insights into recent developments.
Nandy, D, Maity, A, Mitra, AK
Journal of biosciences. 2020
Abstract
In order to improve crop plants in terms of their yield, drought resistance, pest resistance, nutritional value, etc., modern agriculture has relied upon plant genetic engineering. Since the advent of recombinant DNA technology, several tools have been used for genetic transformations in plants such as Agrobacterium tumefaciens, virus-mediated gene transfer, direct gene transfer systems such as electroporation, particle gun, microinjection and chemical methods. All these traditional methods lack specificity and the transgenes are integrated at random sites in the plant DNA. Recently novel techniques for gene targeting have evolved such as engineered nucleases such as Zinc Finger Nucleases, Transcription Activator like effector nucleases, Clustered regular interspaced short palindromic repeats. Other advances include improvement in tools for delivery of gene editing components which include carrier proteins, and carbon nanotubes. The present review focuses on the latest techniques for target specific gene delivery in plants, their expression and future directions in plant biotechnology.
-
8.
Engineering Translation Components Improve Incorporation of Exotic Amino Acids.
Katoh, T, Suga, H
International journal of molecular sciences. 2019;(3)
Abstract
Methods of genetic code manipulation, such as nonsense codon suppression and genetic code reprogramming, have enabled the incorporation of various nonproteinogenic amino acids into the peptide nascent chain. However, the incorporation efficiency of such amino acids largely varies depending on their structural characteristics. For instance, l-α-amino acids with artificial, bulky side chains are poorer substrates for ribosomal incorporation into the nascent peptide chain, mainly owing to the lower affinity of their aminoacyl-tRNA toward elongation factor-thermo unstable (EF-Tu). Phosphorylated Ser and Tyr are also poorer substrates for the same reason; engineering EF-Tu has turned out to be effective in improving their incorporation efficiencies. On the other hand, exotic amino acids such as d-amino acids and β-amino acids are even poorer substrates owing to their low affinity to EF-Tu and poor compatibility to the ribosome active site. Moreover, their consecutive incorporation is extremely difficult. To solve these problems, the engineering of ribosomes and tRNAs has been executed, leading to successful but limited improvement of their incorporation efficiency. In this review, we comprehensively summarize recent attempts to engineer the translation systems, resulting in a significant improvement of the incorporation of exotic amino acids.
-
9.
Cell-free gene-regulatory network engineering with synthetic transcription factors.
Swank, Z, Laohakunakorn, N, Maerkl, SJ
Proceedings of the National Academy of Sciences of the United States of America. 2019;(13):5892-5901
-
-
Free full text
-
Abstract
Gene-regulatory networks are ubiquitous in nature and critical for bottom-up engineering of synthetic networks. Transcriptional repression is a fundamental function that can be tuned at the level of DNA, protein, and cooperative protein-protein interactions, necessitating high-throughput experimental approaches for in-depth characterization. Here, we used a cell-free system in combination with a high-throughput microfluidic device to comprehensively study the different tuning mechanisms of a synthetic zinc-finger repressor library, whose affinity and cooperativity can be rationally engineered. The device is integrated into a comprehensive workflow that includes determination of transcription-factor binding-energy landscapes and mechanistic modeling, enabling us to generate a library of well-characterized synthetic transcription factors and corresponding promoters, which we then used to build gene-regulatory networks de novo. The well-characterized synthetic parts and insights gained should be useful for rationally engineering gene-regulatory networks and for studying the biophysics of transcriptional regulation.
-
10.
Report on the SCRA Nuts and Bolts Workshop II: case studies of citrus greening, Ultra-low Gossypol Cotton, and blight tolerant, low-acrylamide potato.
Hood, EE, Eversole, KA, Leach, L, Hogan, M, McHughen, A, Cordts, J, Rathore, K, Rood, T, Collinge, S, Irey, M
GM crops & food. 2019;(3):139-158
-
-
Free full text
-
Abstract
To be commercialized and grown in the US, genetically engineered (GE) crops typically go through an extensive food, feed, and environmental safety assessment process which, in certain instances, requires complex consultations with three different US regulatory agencies. Many small market, niche, and specialty crops have been genetically engineered using the modern tools of recombinant DNA but few have been commercialized due to real or perceived regulatory constraints. This workshop discussed the practical aspects of developing dossiers on GE specialty, niche, or small-market crops/products for submission to US regulatory agencies. This workshop focused on actual case studies, and provided an opportunity for public or private sector scientists and crop developers to spend time with regulatory officials to learn the specifics of compiling a dossier for regulatory approval. The objective of the workshop was to explain and demystify data requirements and regulatory dossier compilation by small companies, academics, and other developers.