Showing papers in "Frontiers in Plant Science in 2020"

Phytoremediation: A Promising Approach for Revegetation of Heavy Metal-Polluted Land

Abstract: Heavy metal accumulation in soil has been rapidly increased due to various natural processes and anthropogenic (industrial) activities. As heavy metals are non-biodegradable, they persist in the environment, have potential to enter the food chain through crop plants, and eventually may accumulate in the human body through biomagnification. Owing to their toxic nature, heavy metal contamination has posed a serious threat to human health and the ecosystem. Therefore, remediation of land contamination is of paramount importance. Phytoremediation is an eco-friendly approach that could be a successful mitigation measure to revegetate heavy metal-polluted soil in a cost-effective way. To improve the efficiency of phytoremediation, a better understanding of the mechanisms underlying heavy metal accumulation and tolerance in plant is indispensable. In this review, we describe the mechanisms of how heavy metals are taken up, translocated, and detoxified in plants. We focus on the strategies applied to improve the efficiency of phytostabilization and phytoextraction, including the application of genetic engineering, microbe-assisted and chelate-assisted approaches.

Editorial: Biostimulants in Agriculture.

Abstract: Recently, the agricultural sector is facing concomitant challenges of rising the productivity to feed the growing global population and increasing the resources use efficiency, while reducing the environmental impact on the ecosystems and human health. In fact, fertilizers and pesticides play a crucial role in agriculture, representing a powerful tool for growers to increase yield and guarantee continuous productivity throughout the seasons under both optimal and suboptimal conditions. In the last three decades, several technological innovations have been proposed to enhance the sustainability of agricultural production systems, through a significant reduction of synthetic agrochemicals like pesticides and fertilizers. A promising and environmental-friendly innovation would be the use of natural plant biostimulants (PBs) that enhance flowering, plant growth, fruit set, crop productivity, and nutrient use efficiency (NUE), and are able also to improve the tolerance against a wide range of abiotic stressors (Colla and Rouphael, 2015). PBs were initially defined by excluding some functionalities like fertilizers or plant protection products. In 1997, in Grounds Maintenance web-journal, Zhang and Schmidt from the Department of Crop and Soil Environmental Sciences of the Virginia Polytechnic Institute and State University, defined PBs as “materials that, in minute quantities, promote plant growth”. By using the statement “minute quantities” for describing PBs, the authors implicitly wanted to discriminate biostimulants from nutrients and soil amendments, which also promote plant growth, but are clearly applied in larger quantities. The PBs mentioned in this web article were two important categories such as humic acids and seaweed extracts, and their action on plants was proposed to be essentially hormonal. In 2012, the European Commission has assigned an ad hoc study on plant biostimulants to evaluate the substances and materials involved, which was published by du Jardin (2012) as: “The Science of Plant Biostimulants A bibliographic Analysis”. Based on the scientific literature (250 scientific articles using the term ‘biostimulant' in their titles and/or abstracts), the following definition was proposed: “Plant biostimulants are substances and materials, with the exception of nutrients and pesticides, which, when applied to plant, seeds or growing substrates in specific formulations, have the capacity to modify physiological processes of plants in a way that provides potential benefits to growth, development and/or stress responses”. du Jardin (2012) concluded that PBs are very heterogeneous materials, and proposed in his study eight categories of substances that acts as biostimulants: humic substances, complex organic materials (obtained from agro-industrial and urban waste products, sewage sludge extracts, composts, and manure), beneficial chemical elements (Al, Co, Na, Se, and Si), inorganic salts including phosphite, seaweed extracts (brown, red, and green

Modulation of the Root Microbiome by Plant Molecules: The Basis for Targeted Disease Suppression and Plant Growth Promotion

Abstract: Plants host a mesmerizing diversity of microbes inside and around their roots, known as the microbiome. The microbiome is composed mostly of fungi, bacteria, oomycetes, and archaea that can be either pathogenic or beneficial for plant health and fitness. To grow healthy, plants need to surveil soil niches around the roots for the detection of pathogenic microbes, and in parallel maximize the services of beneficial microbes in nutrients uptake and growth promotion. Plants employ a palette of mechanisms to modulate their microbiome including structural modifications, the exudation of secondary metabolites and the coordinated action of different defence responses. Here, we review the current understanding on the composition and activity of the root microbiome and how different plant molecules can shape the structure of the root-associated microbial communities. Examples are given on interactions that occur in the rhizosphere between plants and soilborne fungi. We also present some well-established examples of microbiome harnessing to highlight how plants can maximize their fitness by selecting their microbiome. Understanding how plants manipulate their microbiome can aid in the design of next-generation microbial inoculants for targeted disease suppression and enhanced plant growth.

Manganese in Plants: From Acquisition to Subcellular Allocation.

Abstract: Manganese (Mn) is an important micronutrient for plant growth and development and sustains metabolic roles within different plant cell compartments. The metal is an essential cofactor for the oxygen-evolving complex (OEC) of the photosynthetic machinery, catalyzing the water-splitting reaction in photosystem II (PSII). Despite the importance of Mn for photosynthesis and other processes, the physiological relevance of Mn uptake and compartmentation in plants has been underrated. The subcellular Mn homeostasis to maintain compartmented Mn-dependent metabolic processes like glycosylation, ROS scavenging, and photosynthesis is mediated by a multitude of transport proteins from diverse gene families. However, Mn homeostasis may be disturbed under suboptimal or excessive Mn availability. Mn deficiency is a serious, widespread plant nutritional disorder in dry, well-aerated and calcareous soils, as well as in soils containing high amounts of organic matter, where bio-availability of Mn can decrease far below the level that is required for normal plant growth. By contrast, Mn toxicity occurs on poorly drained and acidic soils in which high amounts of Mn are rendered available. Consequently, plants have evolved mechanisms to tightly regulate Mn uptake, trafficking, and storage. This review provides a comprehensive overview, with a focus on recent advances, on the multiple functions of transporters involved in Mn homeostasis, as well as their regulatory mechanisms in the plant's response to different conditions of Mn availability.

A CNN-RNN Framework for Crop Yield Prediction.

Abstract: Crop yield prediction is extremely challenging due to its dependence on multiple factors such as crop genotype, environmental factors, management practices, and their interactions. This paper presents a deep learning framework using convolutional neural networks (CNNs) and recurrent neural networks (RNNs) for crop yield prediction based on environmental data and management practices. The proposed CNN-RNN model, along with other popular methods such as random forest (RF), deep fully connected neural networks (DFNN), and LASSO, was used to forecast corn and soybean yield across the entire Corn Belt (including 13 states) in the United States for years 2016, 2017, and 2018 using historical data. The new model achieved a root-mean-square-error (RMSE) 9% and 8% of their respective average yields, substantially outperforming all other methods that were tested. The CNN-RNN has three salient features that make it a potentially useful method for other crop yield prediction studies. (1) The CNN-RNN model was designed to capture the time dependencies of environmental factors and the genetic improvement of seeds over time without having their genotype information. (2) The model demonstrated the capability to generalize the yield prediction to untested environments without significant drop in the prediction accuracy. (3) Coupled with the backpropagation method, the model could reveal the extent to which weather conditions, accuracy of weather predictions, soil conditions, and management practices were able to explain the variation in the crop yields.

Recognition and Localization Methods for Vision-Based Fruit Picking Robots: A Review.

Abstract: The utilization of machine vision and its associated algorithms improves the efficiency, functionality, intelligence, and remote interactivity of harvesting robots in complex agricultural environments. Machine vision and its associated emerging technology promise huge potential in advanced agricultural applications. However, machine vision and its precise positioning still have many technical difficulties, making it difficult for most harvesting robots to achieve true commercial applications. This article reports the application and research progress of harvesting robots and vision technology in fruit picking. The potential applications of vision and quantitative methods of localization, target recognition, 3D reconstruction, and fault tolerance of complex agricultural environment are focused, and fault-tolerant technology designed for utilization with machine vision and robotic systems are also explored. The two main methods used in fruit recognition and localization are reviewed, including digital image processing technology and deep learning-based algorithms. The future challenges brought about by recognition and localization success rates are identified: target recognition in the presence of illumination changes and occlusion environments; target tracking in dynamic interference-laden environments, 3D target reconstruction, and fault tolerance of the vision system for agricultural robots. In the end, several open research problems specific to recognition and localization applications for fruit harvesting robots are mentioned, and the latest development and future development trends of machine vision are described.

Salicylic Acid Biosynthesis in Plants

Abstract: Salicylic acid (SA) is an important plant hormone that is best known for mediating host responses upon pathogen infection. Its role in plant defense activation is well established, but its biosynthesis in plants is not fully understood. SA is considered to be derived from two possible pathways; the ICS and PAL pathway, both starting from chorismate. The importance of both pathways for biosynthesis differs between plant species, rendering it hard to make generalizations about SA production that cover the entire plant kingdom. Yet, understanding SA biosynthesis is important to gain insight into how plant pathogen responses function and how pathogens can interfere with them. In this review, we have taken a closer look at how SA is synthesized and the importance of both biosynthesis pathways in different plant species.

Drought and Salinity Stress Responses and Microbe-Induced Tolerance in Plants.

Abstract: Drought and salinity are among the most important environmental factors that hampered agricultural productivity worldwide. Both stresses can induce several morphological, physiological, biochemical, and metabolic alterations through various mechanisms, eventually influencing plant growth, development, and productivity. The responses of plants to these stress conditions are highly complex and depend on other factors, such as the species and genotype, plant age and size, the rate of progression as well as the intensity and duration of the stresses. These factors have a strong effect on plant response and define whether mitigation processes related to acclimation will occur or not. In this review, we summarize how drought and salinity extensively affect plant growth in agriculture ecosystems. In particular, we focus on the morphological, physiological, biochemical, and metabolic responses of plants to these stresses. Moreover, we discuss mechanisms underlying plant-microbe interactions that confer abiotic stress tolerance.

Insights Into Oxidized Lipid Modification in Barley Roots as an Adaptation Mechanism to Salinity Stress

Abstract: Lipidomics is an emerging technology, which aims at the global characterization and quantification of lipids within biological matrices including biofluids, cells, whole organs and tissues. The changes in individual lipid molecular species in stress treated plant species and different cultivars can indicate the functions of genes affecting lipid metabolism or lipid signaling. Mass spectrometry-based lipid profiling has been used to track the changes of lipid levels and related metabolites in response to salinity stress. We have developed a comprehensive lipidomics platform for the identification and direct qualification and/or quantification of individual lipid species, including oxidized lipids, which enables a more systematic investigation of peroxidation of individual lipid species in barley roots under salinity stress. This new lipidomics approach has improved with an advantage of analyzing the composition of acyl chains at the molecular level, which facilitates to profile precisely the 18:3-containing diacyl-glycerophosphates and allowed individual comparison of lipids across varieties. Our findings revealed a general decrease in most of the galactolipids in plastid membranes, and an increase of glycerophospholipids and acylated steryl glycosides, which indicate that plastidial and extraplastidial membranes in barley roots ubiquitously tend to form a hexagonal II (HII) phase under salinity stress. In addition, salt-tolerant and salt-sensitive cultivars showed contrasting changes in the levels of oxidized membrane lipids. These results support the hypothesis that salt-induced oxidative damage to membrane lipids can be used as an indication of salt stress tolerance in barley.

Tomato Diseases and Pests Detection Based on Improved Yolo V3 Convolutional Neural Network.

Abstract: Tomato is affected by various diseases and pests during its growth process. If the control is not timely, it will lead to yield reduction or even crop failure. How to control the diseases and pests effectively and help the vegetable farmers to improve the yield of tomato is very important, and the most important thing is to accurately identify the diseases and insect pests. Compared with the traditional pattern recognition method, the diseases and pests recognition method based on deep learning can directly input the original image. Instead of the tedious steps such as image preprocessing, feature extraction and feature classification in the traditional method, the end-to-end structure is adopted to simplify the recognition process and solve the problem that the feature extractor designed manually is difficult to obtain the feature expression closest to the natural attribute of the object. Based on the application of deep learning object detection, not only can save time and effort, but also can achieve real-time judgment, greatly reduce the huge loss caused by diseases and pests, which has important research value and significance. Based on the latest research results of detection theory based on deep learning object detection and the characteristics of tomato diseases and pests images, this study will build the dataset of tomato diseases and pests under the real natural environment, optimize the feature layer of Yolo V3 model by using image pyramid to achieve multi-scale feature detection, improve the detection accuracy and speed of Yolo V3 model, and detect the location and category of diseases and pests of tomato accurately and quickly. Through the above research, the key technology of tomato pest image recognition in natural environment is broken through, which provides reference for intelligent recognition and engineering application of plant diseases and pests detection.

How Does Proline Treatment Promote Salt Stress Tolerance During Crop Plant Development

Abstract: Soil salinity is one of the major abiotic stresses restricting the use of land for agriculture because it limits the growth and development of most crop plants. Improving productivity under these physiologically stressful conditions is a major scientific challenge because salinity has different effects at different developmental stages in different crops. When supplied exogenously, proline has improved salt stress tolerance in various plant species. Under high-salt conditions, proline application enhances plant growth with increases in seed germination, biomass, photosynthesis, gas exchange, and grain yield. These positive effects are mainly driven by better nutrient acquisition, water uptake, and biological nitrogen fixation. Exogenous proline also alleviates salt stress by improving antioxidant activities and reducing Na+ and Cl- uptake and translocation while enhancing K+ assimilation by plants. However, which of these mechanisms operate at any one time varies according to the proline concentration, how it is applied, the plant species, and the specific stress conditions as well as the developmental stage. To position salt stress tolerance studies in the context of a crop plant growing in the field, here we discuss the beneficial effects of exogenous proline on plants exposed to salt stress through well-known and more recently described examples in more than twenty crop species in order to appreciate both the diversity and commonality of the responses. Proposed mechanisms by which exogenous proline mitigates the detrimental effects of salt stress during crop plant growth are thus highlighted and critically assessed.

Emerging Technologies in Algal Biotechnology: Toward the Establishment of a Sustainable, Algae-Based Bioeconomy.

Abstract: Mankind has recognized the value of land plants as renewable sources of food, medicine, and materials for millennia. Throughout human history, agricultural methods were continuously modified and improved to meet the changing needs of civilization. Today, our rapidly growing population requires further innovation to address the practical limitations and serious environmental concerns associated with current industrial and agricultural practices. Microalgae are a diverse group of unicellular photosynthetic organisms that are emerging as next-generation resources with the potential to address urgent industrial and agricultural demands. The extensive biological diversity of algae can be leveraged to produce a wealth of valuable bioproducts, either naturally or via genetic manipulation. Microalgae additionally possess a set of intrinsic advantages, such as low production costs, no requirement for arable land, and the capacity to grow rapidly in both large-scale outdoor systems and scalable, fully contained photobioreactors. Here, we review technical advancements, novel fields of application, and products in the field of algal biotechnology to illustrate how algae could present high-tech, low-cost, and environmentally friendly solutions to many current and future needs of our society. We discuss how emerging technologies such as synthetic biology, high-throughput phenomics, and the application of internet of things (IoT) automation to algal manufacturing technology can advance the understanding of algal biology and, ultimately, drive the establishment of an algal-based bioeconomy.

The Response of Salinity Stress-Induced A. tricolor to Growth, Anatomy, Physiology, Non-Enzymatic and Enzymatic Antioxidants.

Abstract: An investigation was carried out to elucidate growth, anatomical, physiological, and major ROS detoxification pathways involved in the tolerance of A. tricolor under salinity stress. Both VA14 and VA3 varieties exhibited the reduction in relative water content (RWC), photosynthetic pigments, growth, increased electrolyte leakage (EL), and leaf anatomy adaptation under salinity stress, whereas VA14 was well adapted and performed better compared to VA3. Higher ROS accumulation was demonstrated in the sensitive variety (VA3) in comparison to the tolerant variety (VA14). Salinity stress changed the cellular antioxidant pool by increasing total carotenoids, ascorbate, proline, total polyphenol content (TPC), total flavonoid content (TFC), and total antioxidant capacity (TAC) in both varieties. Although a higher increment was demonstrated in the tolerant variety, the proline increment was much more pronounced in the sensitive variety. Non-enzymatic antioxidant, ascorbate, carotenoids, TPC, TFC, TAC, and antioxidant enzymes SOD and APX were noted to be a major H2O2 detoxifier in the tolerant A. tricolor variety, where there is a comparatively lower H2O2 load. It was complemented by GPOX and CAT activity at a comparatively higher H2O2 load (in the sensitive variety). SOD contributed to the dismutation of superoxide radical (SOR) both in the tolerant and sensitive varieties; however, it greatly contributed to the dismutation of SOR in the tolerant variety. The increase in SOD, ascorbate, and APX makes it predominantly evident that SOD and the AsA-GSH cycle had greatly contributed to quench reactive oxygen species (ROS) of the tolerant variety of A. tricolor.

Sensitivity and Responses of Chloroplasts to Heat Stress in Plants.

Abstract: Increased temperatures caused by global warming threaten agricultural production, as warmer conditions can inhibit plant growth and development or even destroy crops in extreme circumstances. Extensive research over the past several decades has revealed that chloroplasts, the photosynthetic organelles of plants, are highly sensitive to heat stress, which affects a variety of photosynthetic processes including chlorophyll biosynthesis, photochemical reactions, electron transport, and CO2 assimilation. Important mechanisms by which plant cells respond to heat stress to protect these photosynthetic organelles have been identified and analyzed. More recent studies have made it clear that chloroplasts play an important role in inducing the expression of nuclear heat-response genes during the heat stress response. In this review, we summarize these important advances in plant-based research and discuss how the sensitivity, responses, and signaling roles of chloroplasts contribute to plant heat sensitivity and tolerance.

C2H2 Zinc Finger Proteins: Master Regulators of Abiotic Stress Responses in Plants.

Abstract: Abiotic stresses such as drought and salinity are major environmental factors that limit crop yields. Unraveling the molecular mechanisms underlying abiotic stress resistance is crucial for improving crop performance and increasing productivity under adverse environmental conditions. Zinc finger proteins, comprising one of the largest transcription factor families, are known for their finger-like structure and their ability to bind Zn2+. Zinc finger proteins are categorized into nine subfamilies based on their conserved Cys and His motifs, including the Cys2/His2-type (C2H2), C3H, C3HC4, C2HC5, C4HC3, C2HC, C4, C6, and C8 subfamilies. Over the past two decades, much progress has been made in understanding the roles of C2H2 zinc finger proteins in plant growth, development, and stress signal transduction. In this review, we focus on recent progress in elucidating the structures, functions, and classifications of plant C2H2 zinc finger proteins and their roles in abiotic stress responses.

Effects of Potassium Levels on Plant Growth, Accumulation and Distribution of Carbon, and Nitrate Metabolism in Apple Dwarf Rootstock Seedlings.

Abstract: Nitrogen (N) is one of the most required mineral elements for plant growth, and potassium (K) plays a vital role in nitrogen metabolism, both elements are most widely used for fertilizer in agricultural production. However, the exact relationship between K and nitrogen use efficiency (NUE) remain unclear. Apple dwarf rootstock seedlings (M9T337) were used to study the impacts of different potassium levels on plant growth, nitrogen metabolism, and carbon assimilation in water culture experiments for two years. The results showed that both deficiency and excess in K inhibited the growth and root development of M9T337 seedlings. When the potassium supply concentration was 0 mM and 12 mM, the biomass of each organ, the root-shoot ratio, the root activity and NO3− ion flow rate decreased significantly, net photosynthetic rate (Pn) and photochemical efficiency (Fv/Fm) were lower. Meanwhile, seedlings treated with 6 mM K+ had higher nitrogen and carbon metabolizing enzyme activities and higher nitrate transporter gene expression levels (NRT1.1; NRT2.1). 13C and 15N labeling results showed that deficiency and excess K could not only reduce the 15N absorption and 13C assimilation accumulation of M9T337 seedlings but also reduce the 15N distribution ratio in leaves and the 13C distribution ratio in roots. These results suggest that appropriate K supply level (6 mM) was optimal as it enhanced photosynthetic products transport from the leaves to the roots and increased NUE by mediating photosynthesis, carbon and nitrogen metabolizing enzyme activities, nitrate assimilation gene activities, and nitrate transport.

Drought Stress Responses and Resistance in Plants: From Cellular Responses to Long-Distance Intercellular Communication.

Abstract: The drought stress responses of vascular plants are complex regulatory mechanisms because they include various physiological responses from signal perception under water deficit conditions to the acquisition of drought stress resistance at the whole-plant level. It is thought that plants first recognize water deficit conditions in roots and that several molecular signals then move from roots to shoots. Finally, a phytohormone, abscisic acid (ABA) is synthesized mainly in leaves. However, the detailed molecular mechanisms of stress sensors and the regulators that initiate ABA biosynthesis in response to drought stress conditions are still unclear. Another important issue is how plants adjust ABA propagation, stress-mediated gene expression and metabolite composition to acquire drought stress resistance in different tissues throughout the whole plant. In this review, we summarize recent advances in research on drought stress responses, focusing on long-distance signaling from roots to shoots, ABA synthesis and transport, and metabolic regulation in both cellular and whole-plant levels of Arabidopsis and crops. We also discuss coordinated mechanisms for acquiring drought stress adaptations and resistance via tissue-to-tissue communication and long-distance signaling.

Detection of Plant Viruses and Disease Management: Relevance of Genetic Diversity and Evolution.

Abstract: Plant viruses cause considerable economic losses and are a threat for sustainable agriculture. The frequent emergence of new viral diseases is mainly due to international trade, climate change, and the ability of viruses for rapid evolution. Disease control is based on two strategies: i) immunization (genetic resistance obtained by plant breeding, plant transformation, cross-protection, or others), and ii) prophylaxis to restrain virus dispersion (using quarantine, certification, removal of infected plants, control of natural vectors, or other procedures). Disease management relies strongly on a fast and accurate identification of the causal agent. For known viruses, diagnosis consists in assigning a virus infecting a plant sample to a group of viruses sharing common characteristics, which is usually referred to as species. However, the specificity of diagnosis can also reach higher taxonomic levels, as genus or family, or lower levels, as strain or variant. Diagnostic procedures must be optimized for accuracy by detecting the maximum number of members within the group (sensitivity as the true positive rate) and distinguishing them from outgroup viruses (specificity as the true negative rate). This requires information on the genetic relationships within-group and with members of other groups. The influence of the genetic diversity of virus populations in diagnosis and disease management is well documented, but information on how to integrate the genetic diversity in the detection methods is still scarce. Here we review the techniques used for plant virus diagnosis and disease control, including characteristics such as accuracy, detection level, multiplexing, quantification, portability, and designability. The effect of genetic diversity and evolution of plant viruses in the design and performance of some detection and disease control techniques are also discussed. High-throughput or next-generation sequencing provides broad-spectrum and accurate identification of viruses enabling multiplex detection, quantification, and the discovery of new viruses. Likely, this technique will be the future standard in diagnostics as its cost will be dropping and becoming more affordable.

Photosynthetic Regulation Under Salt Stress and Salt-Tolerance Mechanism of Sweet Sorghum.

Abstract: Sweet sorghum is a C4 crop with the characteristic of fast-growth and high-yields. It is a good source for food, feed, fiber, and fuel. On saline land, sweet sorghum can not only survive, but increase its sugar content. Therefore, it is regarded as a potential source for identifying salt-related genes. Here, we review the physiological and biochemical responses of sweet sorghum to salt stress, such as photosynthesis, sucrose synthesis, hormonal regulation, and ion homeostasis, as well as their potential salt-resistance mechanisms. The major advantages of salt-tolerant sweet sorghum include: 1) improving the Na+ exclusion ability to maintain ion homeostasis in roots under salt-stress conditions, which ensures a relatively low Na+ concentration in shoots; 2) maintaining a high sugar content in shoots under salt-stress conditions, by protecting the structures of photosystems, enhancing photosynthetic performance and sucrose synthetase activity, as well as inhibiting sucrose degradation. To study the regulatory mechanism of such genes will provide opportunities for increasing the salt tolerance of sweet sorghum by breeding and genetic engineering.

Reduced Enzymatic Browning in Potato Tubers by Specific Editing of a Polyphenol Oxidase Gene via Ribonucleoprotein Complexes Delivery of the CRISPR/Cas9 System

Abstract: Polyphenol Oxidases (PPOs) catalyze the conversion of phenolic substrates to quinones, leading to the formation of dark-colored precipitates in fruits and vegetables. This process, known as enzymatic browning, is the cause of undesirable changes in organoleptic properties and the loss of nutritional quality in plant-derived products. In potato (Solanum tubersoum L.), PPOs are encoded by a multi-gene family with different expression patterns. Here, we have studied the application of the CRISPR/Cas9 system to induce mutations in the StPPO2 gene in the tetraploid cultivar Desiree. We hypothesized that the specific editing of this target gene would result in a lower PPO activity in the tuber with the consequent reduction of the enzymatic browning. Ribonucleoprotein complexes (RNPs), formed by two sgRNAs and Cas9 nuclease, were transfected to potato protoplasts. Up to 68% of regenerated plants contained mutations in at least one allele of the target gene, while 24% of edited lines carried mutations in all four alleles. No off-target mutations were identified in other analyzed StPPO genes. Mutations induced in the four alleles of StPPO2 gene, led to lines with a reduction of up to 69% in tuber PPO activity and a reduction of 73% in enzymatic browning, compared to the control. Our results demonstrate that the CRISPR/Cas9 system can be applied to develop potato varieties with reduced enzymatic browning in tubers, by the specific editing of a single member of the StPPO gene family.

A Perspective on RNAi-Based Biopesticides.

Abstract: Sustainable agriculture relies on practices and technologies that combine effectiveness with a minimal environmental footprint. RNA interference (RNAi), a eukaryotic process in which transcript expression is reduced in a sequence-specific manner, can be co-opted for the control of plant pests and pathogens in a topical application system. Double-stranded RNA (dsRNA), the key trigger molecule of RNAi, has been shown to provide protection without the need for integration of dsRNA-expressing constructs as transgenes. Consequently, development of RNA-based biopesticides is gaining momentum as a narrow-spectrum alternative to chemical-based control measures, with pests and pathogens targeted with accuracy and specificity. Limitations for a commercially viable product to overcome include stable delivery of the topically applied dsRNA and extension of the duration of protection. In addition to the research focus on delivery of dsRNA, development of regulatory frameworks, risk identification, and establishing avoidance and mitigation strategies is key to widespread deployment of topical RNAi technologies. Once in place, these measures will provide the crop protection industry with the certainty necessary to expend resources on the development of innovative dsRNA-based products. Readily evident risks to human health appear minimal, with multiple barriers to uptake and a long history of consumption of dsRNA from plant material. Unintended impacts to the environment are expected to be most apparent in species closely related to the target. Holistic design practices, which incorporate bioinformatics-based dsRNA selection along with experimental testing, represent important techniques for elimination of adverse impacts.

Rational Improvement of Rice Yield and Cold Tolerance by Editing the Three Genes OsPIN5b, GS3, and OsMYB30 With the CRISPR-Cas9 System.

Abstract: Significant increases in rice yield and stress resistance are constant demands for breeders. However, high yield and high stress resistance are often antagonistic to each other. Here, we report several new rice mutants with high yield and excellent cold tolerance that were generated by simultaneously editing three genes, OsPIN5b (a panicle length gene), GS3 (a grain size gene) and OsMYB30 (a cold tolerance gene) with the CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-associated protein 9) system. We edited two target sites of each gene with high efficiency: 53% for OsPIN5b-site1, 42% for OsPIN5b-site2, 66% for GS3-site1, 63% for GS3-site2, 63% for OsMYB30-site1, and 58% for OsMYB30-site2. Consequently, the ospin5b mutants, the gs3 mutants, and the osmyb30 mutants exhibited increased panicle length, enlarged grain size and increased cold tolerance, respectively. Then nine transgenic lines of the ospin5b/gs3, six lines of ospin5b/osmyb30 and six lines of gs3/osmyb30 were also acquired, and their yield related traits and cold tolerance corresponded to the genes being edited. Additionally, we obtained eight ospin5b/gs3/osmyb30 triple mutants by editing all three genes simultaneously. Aside from the ospin5b/gs3/osmyb30-4 and ospin5b/gs3/osmyb30-25 mutants, the remaining six mutants had off-target events at the putative off-target site of OsMYB30-site1. The results also showed that the T2 generations of these two mutants exhibited higher yield and better cold tolerance compared with the wild type. Together, these results demonstrated that new and excellent rice varieties with improved yield and abiotic stress resistance can be generated through gene editing techniques and may be applied to rice breeding. Furthermore, our study proved that the comprehensive agronomic traits of rice can be improved with the CRISPR-Cas9 system.

Double-Stranded RNA Technology to Control Insect Pests: Current Status and Challenges

Abstract: Exploiting the RNA interference (RNAi) gene mechanism to silence essential genes in pest insects, leading to toxic effects, has surfaced as a promising new control strategy in the past decade. While the first commercial RNAi-based products are currently coming to market, the application against a wide range of insect species is still hindered by a number of challenges. In this review, we discuss the current status of these RNAi-based products and the different delivery strategies by which insects can be targeted by the RNAi-triggering double-stranded RNA (dsRNA) molecules. Furthermore, this review also addresses a number of physiological and cellular barriers, which can lead to decreased RNAi efficacy in insects. Finally, novel non-transgenic delivery technologies, such as polymer or liposomic nanoparticles, peptide-based delivery vehicles and viral-like particles, are also discussed, as these could overcome these barriers and lead to effective RNAi-based pest control.

A Deep-Learning-Based Real-Time Detector for Grape Leaf Diseases Using Improved Convolutional Neural Networks.

Abstract: Black rot, Black measles, Leaf blight and Mites of grape are four common grape leaf diseases that seriously affect grape yield. However, the existing research lacks a real-time detecting method for grape leaf diseases, which cannot guarantee the healthy growth of grape plants. In this article, a real-time detector for grape leaf diseases based on improved deep convolutional neural networks is proposed. This article first expands the grape leaf disease images through digital image processing technology, constructing the grape leaf disease dataset (GLDD). Based on GLDD and the Faster R-CNN detection algorithm, a deep-learning-based Faster DR-IACNN model with higher feature extraction capability is presented for detecting grape leaf diseases by introducing the Inception-v1 module, Inception-ResNet-v2 module and SE-blocks. The experimental results show that the detection model Faster DR-IACNN achieves a precision of 81.1% mAP on GLDD, and the detection speed reaches 15.01 FPS. This research indicates that the real-time detector Faster DR-IACNN based on deep learning provides a feasible solution for the diagnosis of grape leaf diseases and provides guidance for the detection of other plant diseases.

Genome Edited Crops Touch the Market: A View on the Global Development and Regulatory Environment.

Abstract: Products of genome editing as the most promising "New Plant Breeding Technology" (NPBT) have made the transition from the lab to the market in a short time. Globally, research activities employing genome editing are constantly expanding and more and more plants with market-oriented traits are being developed, and companies have already released the first genome edited crops to the market. Few countries, most of which are located in the Americas, have adapted legislations to these technologies or released guidelines supporting the use of genome editing. Other countries are debating the path to come either because there is no clarity on the legal classification or due consensus is hampered by a renewed GMO debate. In recent years (2017-2020), eight countries have introduced guidelines clarifying the legal status of genome edited products and many of those are actively committed to international harmonization of their policies. In this publication we give an overview on the current and potentially future international regulatory environment and an update on plants derived by genome editing with market-oriented traits.

Magnesium Fertilization Improves Crop Yield in Most Production Systems: A Meta-Analysis.

Abstract: Magnesium deficiency is a frequently occurring limiting factor for crop production due to low levels of exchangeable Mg (ex-Mg) in acidic soil, which negatively affects sustainability of agriculture development. How Mg fertilization affects crop yield and subsequent physiological outcomes in different crop species, as well as agronomic efficiencies of Mg fertilizers, under varying soil conditions remain particular interesting questions to be addressed. A meta-analysis was performed with 570 paired observations retrieved from 99 field research articles to compare effects of Mg fertilization on crop production and corresponding agronomic efficiencies in different production systems under varying soil conditions. The mean value of yield increase and agronomic efficiency derived from Mg application was 8.5% and 34.4 kg kg-1 respectively, when combining all yield measurements together, regardless of the crop type, soil condition, and other factors. Under severe Mg deficiency (ex-Mg < 60 mg kg-1), yield increased up to 9.4%, nearly two folds of yield gain (4.9%) in the soil containing more than 120 mg kg-1 ex-Mg. The effects of Mg fertilization on yield was 11.3% when soil pH was lower than 6.5. The agronomic efficiency of Mg fertilizers was negatively correlated with application levels of Mg, with 38.3 kg kg-1 at lower MgO levels (0 - 50 kg ha-1) and 32.6 kg kg-1 at higher MgO levels (50 - 100 kg ha-1). Clear interactions existed between soil ex-Mg, pH, and types and amount of Mg fertilizers in terms of crop yield increase. With Mg supplementation, Mg accumulation in the leaf tissues increased by 34.3% on average; and concentrations of sugar in edible organs were 5.5% higher compared to non-Mg supplemented treatments. Our analysis corroborated that Mg fertilization enhances crop performance by improving yield or resulting in favorable physiological outcomes, providing great potentials for integrated Mg management for higher crop yield and quality.

Hairy Root Cultures—A Versatile Tool With Multiple Applications

Abstract: Hairy roots derived from the infection of a plant by Rhizobium rhizogenes (previously referred to as Agrobacterium rhizogenes) bacteria, can be obtained from a wide variety of plants and allow the production of highly diverse molecules. Hairy roots are able to produce and secrete complex active glycoproteins from a large spectrum of organisms. They are also adequate to express plant natural biosynthesis pathways required to produce specialized metabolites and can benefit from the new genetic tools available to facilitate an optimized production of tailor-made molecules. This adaptability has positioned hairy root platforms as major biotechnological tools. Researchers and industries have contributed to their advancement, which represents new alternatives from classical systems to produce complex molecules. Now these expression systems are ready to be used by different industries like pharmaceutical, cosmetics, and food sectors due to the development of fully controlled large-scale bioreactors. This review aims to describe the evolution of hairy root generation and culture methods and to highlight the possibilities offered by hairy roots in terms of feasibility and perspectives.

The Evolution of Flavonoid Biosynthesis: A Bryophyte Perspective.

Abstract: The flavonoid pathway is one of the best characterized specialized metabolite pathways of plants. In angiosperms, the flavonoids have varied roles in assisting with tolerance to abiotic stress and are also key for signaling to pollinators and seed dispersal agents. The pathway is thought to be specific to land plants and to have arisen during the period of land colonization around 550-470 million years ago. In this review we consider current knowledge of the flavonoid pathway in the bryophytes, consisting of the liverworts, hornworts, and mosses. The pathway is less characterized for bryophytes than angiosperms, and the first genetic and molecular studies on bryophytes are finding both commonalities and significant differences in flavonoid biosynthesis and pathway regulation between angiosperms and bryophytes. This includes biosynthetic pathway branches specific to each plant group and the apparent complete absence of flavonoids from the hornworts.

Plant Unsaturated Fatty Acids: Biosynthesis and Regulation.

Abstract: In most plants, major unsaturated fatty acids (UFAs) are three C18 species, namely, oleic (18:1), linoleic (18:2), and α-linolenic (18:3) acids. These simple compounds play multiple crucial roles in planta and are also important economic traits of oil crops. The enzymatic steps of C18 UFA biosynthesis have been well established. However, the associated FA/lipid trafficking between the plastid and the endoplasmic reticulum remains largely unclear, as does the regulation of the expression and activities of the involved enzymes. In this review, we will revisit the biosynthesis of C18 UFAs with an emphasis on the trafficking, and present an overview of the key enzymes and their regulation. Of particular interest is the emerging regulatory network composed of transcriptional factors and upstream signaling pathways. The review thereby provides the promise of using physical, biochemical and/or genetic means to manipulate FA composition and increase oil yield in crop improvement.

Signaling Role of Glutamate in Plants.

Abstract: It is well known that glutamate (Glu), a neurotransmitter in human body, is a protein amino acid. It plays a very important role in plant growth and development. Nowadays, Glu has been found to emerge as signaling role. Under normal conditions, Glu takes part in seed germination, root architecture, pollen germination, and pollen tube growth. Under stress conditions, Glu participates in wound response, pathogen resistance, response and adaptation to abiotic stress (such as salt, cold, heat, and drought), and local stimulation (abiotic or biotic stress)-triggered long distance signaling transduction. In this review, in the light of the current opinion on Glu signaling in plants, the following knowledge was updated and discussed. 1) Glu metabolism; 2) signaling role of Glu in plant growth, development, and response and adaptation to environmental stress; as well as 3) the underlying research directions in the future. The purpose of this review was to look forward to inspiring the rapid development of Glu signaling research in plant biology, particularly in the field of stress biology of plants.