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Orchid seeds are 'dust-like.' The seed coat is usually thin, with only one to a few cell layers. It originates from the integuments formed during ovule development. In orchids, the outer integument is primarily responsible for forming a mature seed coat. The inner integument usually fails to develop after fertilization, becomes compressed, and collapses over the expanding embryo. Hence, the seed coat is formed from the funiculus, chalaza, and outer integumentary cells. The outermost layer of the seed coat, the testa, is lignified, usually at the radial and inner tangential walls. The subepidermal thin-walled layer(s), the tegmen, subsequently cold, resulting in seeds having only a single layer of seed coat cells. In some species, cells of the inner integument remain alive with the ability to synthesize and accumulate lipidic and or phenolic compounds in their walls covering the embryo. This cover is called the 'carapace,' a protective shield contributing to the embryo's added protection. A developmental and functional perspective of the integuments and seed coat during seed development and germination is presented in this review.

Summary Increasing grain yield has always been the primary goal of crop breeding. KLUH/CYP78A5 has been shown to affect seed size in several plant species, but the relevant molecular mechanism is still unclear and there are no reports of this gene contributing to yield. Here, we demonstrate that modified expression of TaCYP78A5 can enhance wheat grain weight and grain yield per plant by accumulating auxin. TaCYP78A5 is highly expressed in maternal tissues, including ovary and seed coat during wheat development. The constitutive overexpression of TaCYP78A5 leads to significantly increased seed size and weight but not grain yield per plant due to the strengthening of apical dominance. However, localized overexpression of TaCYP78A5 in maternal integument enhances grain weight and grain yield per plant by 4.3%–18.8% and 9.6%–14.7%, respectively, in field trials. Transcriptome and hormone metabolome analyses reveal that TaCYP78A5 participates in auxin synthesis pathway and promotes auxin accumulation and cell wall remodelling in ovary. Phenotype investigation and cytological observation show that localized overexpression of TaCYP78A5 in ovary results in delayed flowering and prolonged proliferation of maternal integument cells, which promote grain enlargement. Moreover, naturally occurring variations in the promoter of TaCYP78A5‐2A contribute to thousand‐grain weight (TGW) and grain yield per plant of wheat;TaCYP78A5‐2A haplotype Ap‐HapII with higher activity is favourable for improving grain weight and grain yield per plant and has been positively selected in wheat breeding. Then, a functional marker of TaCYP78A5 haplotype Ap‐HapII is developed for marker‐assisted selection in wheat grain and yield improvement.

Background The continuously developing pesticide resistance is a great threat to agriculture and human health. Understanding the mechanisms of insecticide resistance is a key step in dealing with the phenomenon. Insect cuticle is recently documented to delay xenobiotic penetration which breaks the previous stereotype that cuticle is useless in insecticide resistance, while the underlying mechanism remains scarce. Results Here, we find the integument contributes over 40.0% to insecticide resistance via different insecticide delivery strategies in oriental fruit fly. A negative relationship exists between cuticle thickening and insecticide penetration in resistant/susceptible, also in field strains of oriental fruit fly which is a reason for integument-mediated resistance. Our investigations uncover a regulator of insecticide penetration that miR-994 mimic treatment causes cuticle thinning and increases susceptibility to malathion, whereas miR-994 inhibitor results in opposite phenotypes. The target of miR-994 is a most abundant cuticle protein (CPCFC) in resistant/susceptible integument expression profile, which possesses capability of chitin-binding and influences the cuticle thickness-mediated insecticide penetration. Our analyses find an upstream transcriptional regulatory signal of miR-994 cascade, long noncoding RNA ( lnc19419 ), that indirectly upregulates CPCFC in cuticle of the resistant strain by sponging miR-994. Thus, we elucidate the mechanism of cuticular competing endogenous RNAs for regulating insecticide penetration and demonstrate it also exists in field strain of oriental fruit fly. Conclusions We unveil a regulatory axis of lnc19419  ~ miR-994 ~  CPCFC on the cuticle thickness that leads to insecticide penetration resistance. These findings indicate that competing endogenous RNAs regulate insecticide resistance by modulating the cuticle thickness and provide insight into the resistance mechanism in insects.

Mythimna separata larvae exhibit both solitary and gregarious phases under low and high population density, respectively; furthermore, differences in morphology, behavior and physiology have been observed in the two phases. The integument plays an essential role in the fitness, general metabolism, communication, and survival of insects; however, differences in the integument ultrastructure and gene expression in the solitary and gregarious phases are largely unknown. In this study, the integument ultrastructure of larvae in the solitary and gregarious phases was compared, and transcriptome analysis was conducted to identify which genes were differentially expressed in the two phases. The results showed that the gregarious larvae had thicker integuments and more polygonal particles on the cuticle surface than solitary larvae. Using the Illumina HiSeq™ sequencing platform, 2774 differentially expressed genes (DEGs) were generated. Among these, many transcripts were identified with roles in the synthesis of fatty acids; structural components of the integument and the insecticide detoxification were differentially expressed in the integument of the two larval phases. qRT-PCR was used to validate expression patterns of the selected transcripts. This study provides a valuable resource for understanding the molecular basis of behavioral and physiological differences in the two phases of M. separata.

We use Arabidopsis thaliana as a model to investigate coordination of cell proliferation and cell elongation in the three components that develop side by side in the seed. Two of these, the embryo and its nurturing annex, the endosperm, are placed under zygotic control and develop within the seed integument placed under maternal control. We show that integument cell proliferation and endosperm growth are largely independent from each other. By contrast, prevention of cell elongation in the integument by the mutation transparent testa glabra2 (ttg2) restricts endosperm and seed growth. Conversely, endosperm growth controlled by the HAIKU (IKU) genetic pathway modulates integument cell elongation. Combinations of TTG2 defective seed integument with reduction of endosperm size by iku mutations identify integument cell elongation and endosperm growth as the primary regulators of seed size. Our results strongly suggest that a cross talk between maternal and zygotic controls represents the primary regulator of the coordinated control of seed size in Arabidopsis.

Background In eudicots, germination begins with water uptake by the quiescent dry seed and is greatly related to the permeability of micropyle enriched callose layers. Once imbibition starts, seeds undergo a cascade of physiological, biochemical, and molecular events to initiate cellular activities. However, the effects of callose on water uptake and following seed metabolic events during germination are largely unknown. Cotton ( Gossypium hirsutum ) is a eudicot plant with natural fiber and edible oil production for humans. Here, we addressed this question by examining the role of GhGLU19 , a gene encoding β-1,3-glucanase, in cotton seed germination. Results GhGLU19 belongs to subfamily B and was expressed predominately in imbibed seeds and early seedlings. Compared to wild type, GhGLU19 -suppressing and GhGLU19 -overexpressing transgenic cotton lines showed the higher and lower seed germination percentage, respectively. Callose was enriched more at inner integument (ii) than that in embryo and seed coat in cotton seeds. In GhGLU19 -suppressing lines, callose at ii of cotton seeds was greatly increased and brought about a prolonged water uptake process during imbibition. Both proteomic and transcriptomic analysis revealed that contrary to GhGLU19 -overexpressing lines, the glycolysis and pyruvate metabolism was decreased, and abscisic acid (ABA) biosynthesis related genes were downregulated in imbibed seeds of GhGLU19 -suppressing lines. Also, endogenous ABA was significantly decreased in GhGLU19 -suppressing line while increased in GhGLU19 -overexpressing line. Conclusions Our results demonstrate that suppression of GhGLU19 improves cotton seed germination via accumulating callose of inner integument, modulating glycolysis and pyruvate metabolism, and decreasing ABA biosynthesis. This study provides a potential way for improving germination percentage in cotton seed production, and other eudicot crops.

Summary Cotton fibre is a unicellular seed trichome, and lint fibre initials per seed as a factor determines fibre yield. However, the mechanisms controlling fibre initiation from ovule epidermis are not understood well enough. Here, with single‐cell RNA sequencing (scRNA‐seq), a total of 14 535 cells were identified from cotton ovule outer integument of Xu142_LF line at four developmental stages (1.5, 1, 0.5 days before anthesis and the day of anthesis). Three major cell types, fibre, non‐fibre epidermis and outer pigment layer were identified and then verified by RNA in situ hybridization. A comparative analysis on scRNA‐seq data between Xu142 and its fibreless mutant Xu142 fl further confirmed fibre cluster definition. The developmental trajectory of fibre cell was reconstructed, and fibre cell was identified differentiated at 1 day before anthesis. Gene regulatory networks at four stages revealed the spatiotemporal pattern of core transcription factors, and MYB25‐like and HOX3 were demonstrated played key roles as commanders in fibre differentiation and tip‐biased diffuse growth respectively. A model for early development of a single fibre cell was proposed here, which sheds light on further deciphering mechanism of plant trichome and the improvement of cotton fibre yield. With scRNA‐seq, cotton fibre cell was identified differentiated at −1 DPA. It further refines the spatiotemporal patterns of two command genes, MYB25‐like and HOX3, who determine fibre differentiation and tip‐biased diffuse growth respectively.

Animal integumentary coloration plays a crucial role in visual communication and camouflage, and varies extensively among and within species and populations. To understand the pressures underlying such diversity, it is essential to elucidate the mechanisms by which animals have created novel integumentary coloration. Colours can be produced by selective absorption of light by skin pigments, through light scattering by structured or unstructured tissues, or by a combination of pigments and nanostructures. In this review, we highlight our current understanding of the interactions between pigments and structural integumentary tissues and molecules. We analyse the available evidence suggesting that these combined mechanisms are capable of creating colours and optical properties unachievable by either mechanism alone, thereby effectively expanding the animal colour palette. Moreover, structural and pigmentary colour mechanisms frequently interact in unexpected and overlooked ways, suggesting that classification of colours as being of any particular type may be difficult. Finally, we discuss how these mixtures are useful for investigating the largely unknown genetic, developmental and physical processes generating phenotypic diversity. This article is part of the themed issue ‘Animal coloration: production, perception, function and application’.

IntroductionOutside of procedural adverse events, there is little understanding of wider patient safety incidents (PSIs) in endoscopy. The aim of this study was to quantify endoscopy PSIs and identify their contributory human factors utilising a national data set.MethodsData were extracted from the National Reporting and Learning System (NRLS) which records staff-reported safety incidents in England and Wales. Two independent coders with backgrounds in safety and qualitative analysis coded data using a hybrid thematic analysis approach. The Human Factors Analysis and Classification System (HFACS) was applied to code contributory factors.ResultsFrom 2017-2019, 1811 endoscopy-related PSIs were identified, of which 629 were procedural adverse events (pAEs; directly related to procedure), 539 were non-procedural adverse events (nAEs; any incident not directly related to a procedure) and 16 were ‘never’ events. Inter-coder reliability was substantial (kappa 0.77). 842 human factors codes were identified across four levels: acts, preconditions, supervision and organisational influences. Decision-based errors were the most common acts (>40%) across categories (table 1). Patient factors were significant contributors in pAEs (74.5%) and co-ordination, communication (33.5 – 66.7%) and situational (27.1%) factors were key contributors in nAEs and never events.ConclusionsThis is the first overview of national-level endoscopy safety incident data and demonstrates the role human factors play in PSI development. These findings should inform patient safety improvement strategies in endoscopy.Abstract P194 Table 1 Procedural AEs Non-procedural AEs Never events Distribution 629/1181 (53.3%) 539/1181 (45.6%) 16/1181 (1.4%) Categories 1) Instrumental (49.8%)• Bleeding (15%)• Other (7.7%)• Cardiovascular (6.2%)• Pulmonary (4.3%)• Pancreatitis (4.0%)• Pain (3.7%)• Drug reaction (2.7%)• Infection (2.4%)• Integument (2.2%)• Thromboembolic (2.1%) • Follow up & surveillance (23.4%)• Access & booking (19.7%)• Quality (17.3%)• Specimens/histopathology (11.3%)• Peri-endoscopy care (7.4%)• Staffing, environment, infrastructure (5.2%)• Patient harm or injury (4.6%)• Communication (3.7%)• Equipment (3.3%)• Documentation (1.5%)• Consent (1.5%)• Decontamination (1.1%) • Wrong patient (62.5%)• Wrong site (37.5%) HFACS levels 1 Decision-based errors (43.6%) Decision-based errors (51.8%)Routine non-concordance (23.8%) Decision-based errors (58.3%)Skill-based errors (41.7%) 2 Patient (74.5%)Co-ordination and communication (16.5%) Communication (33.5%)Situational (27.1%) Co-ordination and communication (66.7%) 3 Planning (58.8%) 4 Organisational processes (42.3%)

RNA interference (RNAi) has been recognized as a novel and safe strategy in pest management due to its high sequence-dependent specificity. However, the existing dsRNA delivery methods largely restrict the application of the RNAi-based pest management strategy; thus, we previously constructed a nanocarrier-based transdermal dsRNA delivery system on the soybean aphid Aphis glycines with the help of nanocarrier and detergent. In the current study, we improved our transdermal dsRNA delivery system with a smaller and cheaper nanocarrier to investigate the efficacy of spraying aphid-infested soybean seedlings to apply our RNA pesticide. A dsRNA/nanocarrier/detergent formulation was performed, and the dsRNA could penetrate the aphid body wall within 4 h with the help of nanocarrier through the topical application. Four potential RNAi target genes ( TREH , ATPD , ATPE and CHS1 ) were selected and cloned, and their dsRNA fragments were synthesized and tested through the transdermal dsRNA delivery system. The delivered dsRNA efficiently silenced the target gene expression with the knockdown effects ranging from 86.86 to 58.87% and resulted in a high mortality up to 81.67% (dsATPD + dsATPE) through the topical application, when through the spray method, with the highest percent mortality of 78.50% (dsATPD + dsCHS1). Our novel transdermal dsRNA delivery system not only provides a powerful tool for gene functional analysis in laboratory, but also shows a great potential for the pest management in the field, which will promote the practice and development of RNAi-based pest management.