Experimental Study on the Effect of Wetting-Drying Cycles on Crack Development and Radon Retardation of the Covering Soil in Uranium Mill Tailing Impoundments.

The majority of uranium mill tailing impoundments in the southern part of China are located in humid subtropical regions where persistent rainfall and rapid evaporation of water after rain often occur. Under the prolonged influence of alternating wet and dry conditions, the covering soil layer of uranium mill tailing impoundments develops cracks, leading to the issue of degradation or even failure of the radon retardation effect. A beach surface of uranium mill tailing impoundments in the southern part of China is selected as the research object. Through use of a self-made simulation test device, a degradation experiment of uranium mill tailing covering soil models under wetting-drying cycles was conducted indoors. The experimental results indicate that with a constant amplitude of wetting-drying cycles, microcracks characterized by a narrow width and high abundance were mainly generated in the early-to-mid-stage of wetting-drying cycles. The main cracks, characterized by their wide width and less abundance, were mainly formed in the mid-to-late stage of wetting-drying cycles. After seven wetting-drying cycles, the total length of cracks showed a "stair-step" increase and the surface crack ratio exhibited a trend of moving from rapid growth to stable growth and then to a slight decline. The cumulative damage degree showed a rapid increase to stable growth with an increase in the number of wetting-drying cycles. Grey relational analysis revealed that, compared to other surface crack indicators, radon exhalation rate was the most closely correlated with the surface crack ratio. With a constant amplitude of wetting-drying cycles, the radon exhalation rate underwent four stages as the number of wetting-drying cycles increased: stage I witnessed a rapid increase, stage II witnessed a rapid decrease, stage III witnessed a gradual increase, and stage IV witnessed a stable or even slight decrease. With a constant number of wetting-drying cycles, the radon exhalation rate correspondingly increased with the amplitude of wetting-drying cycles, particularly noticeable when the alternation amplitude was 30 ± 20%. From the early mid-stage to the late stage of wetting-drying cycles, the curves of the radon exhalation rate, surface crack ratio, and cumulative damage degree tended to be consistent, showing a gradual increase. The research provided in this study offers valuable insights into radon control measures and environmental assessments on the beach surface of uranium mill tailing impoundments.

Improving Adversarial Robustness of ECG Classification Based on Lipschitz Constraints and Channel Activation Suppression.

Deep neural networks (DNNs) are increasingly important in the medical diagnosis of electrocardiogram (ECG) signals. However, research has shown that DNNs are highly vulnerable to adversarial examples, which can be created by carefully crafted perturbations. This vulnerability can lead to potential medical accidents. This poses new challenges for the application of DNNs in the medical diagnosis of ECG signals. This paper proposes a novel network Channel Activation Suppression with Lipschitz Constraints Net (CASLCNet), which employs the Channel-wise Activation Suppressing (CAS) strategy to dynamically adjust the contribution of different channels to the class prediction and uses the 1-Lipschitz's ℓ∞ distance network as a robust classifier to reduce the impact of adversarial perturbations on the model itself in order to increase the adversarial robustness of the model. The experimental results demonstrate that CASLCNet achieves ACCrobust scores of 91.03% and 83.01% when subjected to PGD attacks on the MIT-BIH and CPSC2018 datasets, respectively, which proves that the proposed method in this paper enhances the model's adversarial robustness while maintaining a high accuracy rate.

Single-cell RNA sequencing profiles reveal cell type-specific transcriptional regulation networks conditioning fungal invasion in maize roots.

Stalk rot caused by Fusarium verticillioides (Fv) is one of the most destructive diseases in maize production. The defence response of root system to Fv invasion is important for plant growth and development. Dissection of root cell type-specific response to Fv infection and its underlying transcription regulatory networks will aid in understanding the defence mechanism of maize roots to Fv invasion. Here, we reported the transcriptomes of 29 217 single cells derived from root tips of two maize inbred lines inoculated with Fv and mock condition, and identified seven major cell types with 21 transcriptionally distinct cell clusters. Through the weighted gene co-expression network analysis, we identified 12 Fv-responsive regulatory modules from 4049 differentially expressed genes (DEGs) that were activated or repressed by Fv infection in these seven cell types. Using a machining-learning approach, we constructed six cell type-specific immune regulatory networks by integrating Fv-induced DEGs from the cell type-specific transcriptomes, 16 known maize disease-resistant genes, five experimentally validated genes (ZmWOX5b, ZmPIN1a, ZmPAL6, ZmCCoAOMT2, and ZmCOMT), and 42 QTL or QTN predicted genes that are associated with Fv resistance. Taken together, this study provides not only a global view of maize cell fate determination during root development but also insights into the immune regulatory networks in major cell types of maize root tips at single-cell resolution, thus laying the foundation for dissecting molecular mechanisms underlying disease resistance in maize.

Inverse design of coupled subwavelength dielectric resonators with targeted eigenfrequency and Q factor utilizing deep learning.

Subwavelength all-dielectric resonators supporting Mie resonances are promising building blocks in nanophotonics. The coupling of dielectric resonators facilitates advanced shaping of Mie resonances. However, coupled dielectric resonators with anisotropic geometry can only be designed by time-consuming simulation utilizing parameter scanning, hampering their applications in nanophotonics. Herein, we propose and demonstrate that a combination of two fully connected networks can effectively design coupled dielectric resonators with targeted eigenfrequency and Q factor. Typical examples are given for validating the proposed network, where the normalized deviation rates of eigenfrequency and Q factor are 0.39% and 1.29%, respectively. The proposed neutral network might become a useful tool in designing coupled dielectric resonators and beyond.

Bound valley edge states in the continuum.

Topological valley photonics provides a unique way to manipulate the flow of light. In general, valley edge states that exhibit unidirectional propagation and are immune to defects and disorders could be realized at the interface between two valley photonic crystals with opposite valley Chern numbers. Herein, by merging the physics of valley edge states and bound states in the continuum, we propose and numerically demonstrate a novel, to the best of our knowledge, concept of edge states termed bound valley edge states in the continuum, which enjoys the topological features of valley edge states, such as, unidirectional propagation and immunity to disorders, but are formed at the interface between air and a single valley photonic crystal. Our results not only provide an effective way to reduce the size of valley photonic structures but also facilitate new applications where the proposed concept of bound valley edge states in the continuum could be exploited for optical sensing and unidirectional waveguiding.

Transverse Kerker Effect for Dipole Sources.

Transverse Kerker effect is known by the directional scattering of an electromagnetic plane wave perpendicular to the propagation direction with nearly suppression of both forward and backward scattering. Compared with plane waves, localized electromagnetic emitters are more general sources in modern nanophotonics. As a typical example, manipulating the emission direction of a quantum dot is of vital importance for the investigation of on-chip quantum optics and quantum information processing. Herein, we introduce the concept of transverse Kerker effect for dipole sources utilizing a subwavelength dielectric antenna, where the radiative power of magnetic, electric, and more general chiral dipole emitters can be dominantly redirected along their dipole moments with nearly suppression of radiation perpendicular to the dipole moments. This type of transverse Kerker effect is also associated with Purcell enhancement mediated by electromagnetic multipolar resonances induced in the dielectric antenna. Analytical conditions of transverse Kerker effect are derived for the magnetic, electric, and chiral dipole emitters. We further provide microwave experiment validation for the magnetic dipole emitter. Our results provide new physical mechanisms to manipulate the emission properties of localized electromagnetic source which might facilitate the on-chip quantum optics and beyond.

Enhancing plant growth and biomass production by overexpression of GA20ox gene under control of a root preferential promoter.

Overexpression of GA20 oxidase gene has been a recent trend for improving plant growth and biomass. Constitutive expression of GA20ox has successfully improved plant growth and biomass in several plant species. However, the constitutive expression of this gene causes side-effects, such as reduced leaf size and stem diameter, etc. To avoid these effects, we identified and employed different tissue-specific promoters for GA20ox overexpression. In this study, we examined the utility of At1g promoter to drive the expression of GUS (ß-glucuronidase) reporter and AtGA20ox genes in tobacco and Melia azedarach. Histochemical GUS assays and quantitative real-time-PCR results in tobacco showed that At1g was a root-preferential promoter whose expression was particularly strong in root tips. The ectopic expression of AtGA20ox gene under the control of At1g promoter showed improved plant growth and biomass of both tobacco and M. azedarach transgenic plants. Stem length as well as stem and root fresh weight increased by up to 1.5-3 folds in transgenic tobacco and 2 folds in transgenic M. azedarach. Both tobacco and M. azedarach transgenic plants showed increases in root xylem width with xylem to phloem ratio over 150-200% as compared to WT plants. Importantly, no significant difference in leaf shape and size was observed between At1g::AtGA20ox transgenic and WT plants. These results demonstrate the great utility of At1g promoter, when driving AtGA20ox gene, for growth and biomass improvements in woody plants and potentially some other plant species.

Invisible Mie scatterer.

Dielectric Mie scatterers possessing simultaneously magnetic and electric resonances can be used to tailor scattering utilizing the interference among electromagnetic multipole moments. Cloaking for this type of Mie scatterer is important for various applications. However, the existing cloaking mechanisms mainly focus on the elimination of net electric dipole moments, which have not been generalized to a Mie scatterer with both magnetic and electric responses yet. Herein, we propose and experimentally demonstrate an invisible Mie scatterer utilizing a hybrid skin cloak. The hybrid mechanism relies on the realization of a magnetic analog of a plasmonic cloak and the electric anapole condition to eliminate the net magnetic and electric dipole moments simultaneously. Microwave experiments are provided to validate the proposal. Our results not only introduce a new concept of skin cloaking for electromagnetic scatterers, but also provide new insight for the invisibility and illusion of Mie scatterers.

GmKIX8-1 regulates organ size in soybean and is the causative gene for the major seed weight QTL qSw17-1.

Seed weight is one of the most important agronomic traits in soybean for yield improvement and food production. Several quantitative trait loci (QTLs) associated with the trait have been identified in soybean. However, the genes underlying the QTLs and their functions remain largely unknown. Using forward genetic methods and CRISPR/Cas9 gene editing, we identified and characterized the role of GmKIX8-1 in the control of organ size in soybean. GmKIX8-1 belongs to a family of KIX domain-containing proteins that negatively regulate cell proliferation in plants. Consistent with this predicted function, we found that loss-of-function GmKIX8-1 mutants showed a significant increase in the size of aerial plant organs, such as seeds and leaves. Likewise, the increase in organ size is due to increased cell proliferation, rather than cell expansion, and increased expression of CYCLIN D3;1-10. Lastly, molecular analysis of soybean germplasms harboring the qSw17-1 QTL for the big-seeded phenotype indicated that reduced expression of GmKIX8-1 is the genetic basis of the qSw17-1 phenotype.

Establishment and optimization of a hemp (Cannabis sativa L.) agroinfiltration system for gene expression and silencing studies.

Industrial hemp (Cannabis sativa L.) is a high-yielding annual crop primarily grown for fiber, seeds, and oil. Due to the phytochemical composition of hemp, there has been an increased interest in the market for nutraceuticals and dietary supplements for human health. Recent omics analysis has led to the elucidation of hemp candidate genes involved in the syntheses of specialized metabolites. However, a detailed study of these genes has not been undertaken due to the lack of a stable transformation system. We report for the first time an agroinfiltration system in hemp utilizing vacuum infiltration, which is an alternative method to stable transformation. A combination of 0.015% Silwett L-77, 5 mM ascorbic acid, and thirty second sonication followed by a 10-minute vacuum treatment resulted in the highest ß-glucuronidase expression in the leaf, male and female flowers, stem, and root tissues. The phytoene desaturase gene was silenced with a transient hairpin RNA expression, resulting in an albino phenotype in the leaves and the male and female flowers. This agroinfiltration system would be useful for overexpression and silencing studies of target genes to regulate the yield of specialized metabolites in hemp.

Demonstration of highly efficient dual gRNA CRISPR/Cas9 editing of the homeologous GmFAD2-1A and GmFAD2-1B genes to yield a high oleic, low linoleic and α-linolenic acid phenotype in soybean.

BACKGROUND: CRISPR/Cas9 gene editing is now revolutionizing the ability to effectively modify plant genomes in the absence of efficient homologous recombination mechanisms that exist in other organisms. However, soybean is allotetraploid and is commonly viewed as difficult and inefficient to transform. In this study, we demonstrate the utility of CRISPR/Cas9 gene editing in soybean at relatively high efficiency. This was shown by specifically targeting the Fatty Acid Desaturase 2 (GmFAD2) that converts the monounsaturated oleic acid (C18:1) to the polyunsaturated linoleic acid (C18:2), therefore, regulating the content of monounsaturated fats in soybean seeds. RESULTS: We designed two gRNAs to guide Cas9 to simultaneously cleave two sites, spaced 1Kb apart, within the second exons of GmFAD2-1A and GmFAD2-1B. In order to test whether the Cas9 and gRNAs would perform properly in transgenic soybean plants, we first tested the CRISPR construct we developed by transient hairy root transformation using Agrobacterium rhizogenesis strain K599. Once confirmed, we performed stable soybean transformation and characterized ten, randomly selected T0 events. Genotyping of CRISPR/Cas9 T0 transgenic lines detected a variety of mutations including large and small DNA deletions, insertions and inversions in the GmFAD2 genes. We detected CRISPR- edited DNA in all the tested T0 plants and 77.8% of the events transmitted the GmFAD2 mutant alleles to T1 progenies. More importantly, null mutants for both GmFAD2 genes were obtained in 40% of the T0 plants we genotyped. The fatty acid profile analysis of T1 seeds derived from CRISPR-edited plants homozygous for both GmFAD2 genes showed dramatic increases in oleic acid content to over 80%, whereas linoleic acid decreased to 1.3-1.7%. In addition, transgene-free high oleic soybean homozygous genotypes were created as early as the T1 generation. CONCLUSIONS: Overall, our data showed that dual gRNA CRISPR/Cas9 system offers a rapid and highly efficient method to simultaneously edit homeologous soybean genes, which can greatly facilitate breeding and gene discovery in this important crop plant.

Edit at will: Genotype independent plant transformation in the era of advanced genomics and genome editing.

The combination of advanced genomics, genome editing and plant transformation biology presents a powerful platform for basic plant research and crop improvement. Together these advances provide the tools to identify genes as targets for direct editing as single base pair changes, deletions, insertions and site specific homologous recombination. Recent breakthrough technologies using morphogenic regulators in plant transformation creates the ability to introduce reagents specific toward their identified targets and recover stably transformed and/or edited plants which are genotype independent. These technologies enable the possibility to alter a trait in any variety, without genetic disruption which would require subsequent extensive breeding, but rather to deliver the same variety with one trait changed. Regulatory issues regarding this technology will predicate how broadly these technologies will be implemented. In addition, education will play a crucial role for positive public acceptance. Taken together these technologies comprise a platform for advanced breeding which is an imperative for future world food security.

Quantitative Proteomic Analysis of Low Linolenic Acid Transgenic Soybean Reveals Perturbations of Fatty Acid Metabolic Pathways.

To understand the effect of fatty acid desaturase gene (GmFAD3) silencing on perturbation of fatty acid (FA) metabolic pathways, the changes are compared in protein profiling in control and low linolenic acid transgenic soybeans using tandem mass tag based mass spectrometry. Protein profiling of the transgenic line unveiled changes in several key enzymes of FA metabolism. This includes enzymes of lower abundance; fabH, fabF, and thioestrase associated with FA initiation, elongation, and desaturation processes and LOX1_5, ACOX, ACAA1, MFP2 associated with ß-oxidation of α-linolenic acids pathways. In addition, the GmFAD3 silencing results in a significant reduction in one of the major allergens, Gly m 4 (C6T3L5). These results are important for exploring how plants adjust in their biological processes when certain changes are induced in the genetic makeup. A complete understanding of these processes will aid researchers to alter genes for developing value-added soybeans.

Rapid and Efficient Genetic Transformation of Sorghum via Agrobacterium-Mediated Method.

Genetic transformation via Agrobacterium-mediated methodology has been used in many sorghum studies. However, the transformation efficiency still varies significantly due to high dependence on sorghum genotypes and technical expertise. In this article, we describe a sorghum transformation procedure in sufficient detail using a public genotype, P898012. This system utilizes a standard binary transgenic vector carrying the bar gene as a selectable marker and immature embryos as starting explants. Glufosinate is employed as the selective agent during callus and shoot induction. This procedure is relatively rapid, efficient, highly reproducible, and should be applicable for many other sorghum genotypes. © 2018 by John Wiley & Sons, Inc.

Morphogenic Regulator-Mediated Transformation of Maize Inbred B73.

Maize B73 is a reference genome and has long been a major resource for genetics and molecular biology research. We have developed an efficient B73 transformation protocol by enabling somatic embryogenesis through differential co-expression of maize morphogenic regulators BBM and WUS2. We describe a successful protocol that utilizes Agrobacterium tumefaciens strain AGL1 harboring binary vector PHP78891 that comprises a BBM and WUS2 expression cassette as well as a green fluorescent protein (GFP) reporter cassette. The PHP78891 vector also contains, within the T-DNA region, a CRE/lox recombination system flanking the CRE/BBM/WUS2 co-expression cassette driven by the desiccation inducible RAB17 promoter that allows removal of the BBM/WUS2 cassette. Introduction and co-expression of BBM and WUS2 induced direct somatic embryogenesis (SE) in non-regenerable maize B73 from immature embryo explants. Removal of the CRE/BBM/WUS2 cassette is essential to allow regeneration to fertile plants. The GFP expression cassette outside the lox excision sites is retained in the transgenic plant genome, allowing subsequent phenotypic analysis of calli and regenerated transgenic events. This transformation system enables a selectable marker-free transformation process by taking advantage of BBM/WUS2-induced SE as a developmental selection system. © 2018 by John Wiley & Sons, Inc.

Transformation of Recalcitrant Sorghum Varieties Facilitated by Baby Boom and Wuschel2.

Most reliable transformation protocols for cereal crops, including sorghum (Sorghum bicolor L. Moench), rely on the use of immature embryo explants to generate embryogenic callus cells that are then transformed using Agrobacterium- or particle-bombardment-mediated DNA delivery. Subsequent to DNA transfer, most protocols rely on selectable markers for the recovery of stably transformed callus that is then regenerated to produce T0 plants. However, these protocols require specific genotypes that are innately capable of efficient embryogenic callus initiation. Here, we describe a system that makes use of the differential expression of the morphogenic regulators Baby Boom (Bbm) and Wuschel2 (Wus2) to achieve transformation in varieties of sorghum typically recalcitrant to standard transformation methods. © 2018 by John Wiley & Sons, Inc.

Transcriptome Profiling of Plant Genes in Response to Agrobacterium tumefaciens-Mediated Transformation.

Agrobacterium tumefaciens is a plant pathogen that causes crown gall disease. During infection of the host plant, Agrobacterium transfers T-DNA from its Ti plasmid into the host cell, which can then be integrated into the host genome. This unique genetic transformation capability has been employed as the dominant technology for producing genetically modified plants for both basic research and biotechnological applications. Agrobacterium has been well studied as a disease-causing agent. The Agrobacterium-mediated transformation process involves early attachment of the bacterium to the host's surface, followed by transfer of T-DNA and virulence proteins into the plant cell. Throughout this process, the host plants exhibit dynamic gene expression patterns at each infection stage or in response to Agrobacterium strains with varying pathogenic capabilities. Shifting host gene expression patterns throughout the transformation process have effects on transformation frequency, host morphology, and metabolism. Thus, gene expression profiling during the Agrobacterium infection process can be an important approach to help elucidate the interaction between Agrobacterium and plants. This review highlights recent findings on host plant differential gene expression patterns in response to A. tumefaciens or related elicitor molecules.

Transcriptomic Analysis of Arabidopsis Seedlings in Response to an Agrobacterium-Mediated Transformation Process.

Agrobacterium tumefaciens is a plant pathogen that causes crown gall disease. This pathogen is capable of transferring the T-DNA from its Ti plasmid to the host cell and, then, integrating it into the host genome. To date, this genetic transformation ability has been harnessed as the dominant technology to produce genetically modified plants for both basic research and crop biotechnological applications. However, little is known about the interaction between Agrobacterium tumefaciens and host plants, especially the host responses to Agrobacterium infection and its associated factors. We employed RNA-seq to follow the time course of gene expression in Arabidopsis seedlings infected with either an avirulent or a virulent Agrobacterium strain. Gene Ontology analysis indicated many biological processes were involved in the Agrobacterium-mediated transformation process, including hormone signaling, defense response, cellular biosynthesis, and nucleic acid metabolism. RNAseq and quantitative reverse transcription-polymerase chain reaction results indicated that expression of genes involved in host plant growth and development were repressed but those involved in defense response were induced by Agrobacterium tumefaciens. Further analysis of the responses of transgenic Arabidopsis lines constitutively expressing either the VirE2 or VirE3 protein suggested Vir proteins act to enhance plant defense responses in addition to their known roles facilitating T-DNA transformation.

Selectable marker independent transformation of recalcitrant maize inbred B73 and sorghum P898012 mediated by morphogenic regulators BABY BOOM and WUSCHEL2.

KEY MESSAGE: Discriminatory co-expression of maize BBM and WUS transcriptional factor genes promoted somatic embryogenesis and efficient Agrobacterium -mediated transformation of recalcitrant maize inbred B73 and sorghum P898012 genotypes without use of a selectable marker gene. The use of morphogenic regulators to overcome barriers in plant transformation is a revolutionary breakthrough for basic plant science and crop applications. Current standard plant transformation systems are bottlenecks for genetic, genomic, and crop improvement studies. We investigated the differential use of co-expression of maize transcription factors BABY BOOM and WUSCHEL2 coupled with a desiccation inducible CRE/lox excision system to enable regeneration of stable transgenic recalcitrant maize inbred B73 and sorghum P898012 without a chemical selectable marker. The PHP78891 expression cassette contains CRE driven by the drought inducible maize RAB17M promoter with lox P sites which bracket the CRE, WUS, and BBM genes. A constitutive maize UBI M promoter directs a ZsGreen GFP expression cassette as a reporter outside of the excision sites and provides transient, transgenic, and developmental analysis. This was coupled with evidence for molecular integration and analysis of stable integration and desiccation inducible CRE-mediated excision. Agrobacterium-mediated transgenic introduction of this vector showed transient expression of GFP and induced somatic embryogenesis in maize B73 and sorghum P898012 explants. Subjection to desiccation stress in tissue culture enabled the excision of CRE, WUS, and BBM, leaving the UBI M::GFP cassette and allowing subsequent plant regeneration and GFP expression analysis. Stable GFP expression was observed in the early and late somatic embryos, young shoots, vegetative plant organs, and pollen. Transgene integration and expression of GFP positive T0 plants were also analyzed using PCR and Southern blots. Progeny segregation analysis of primary events confirmed correlation between functional GFP expression and presence of the GFP transgene in T1 plants generated from self pollinations, indicating good transgene inheritance. This study confirms and extends the use of morphogenic regulators to overcome transformation barriers.