Determination of chlordimeform and its metabolite residue in milk by gas chromatography-tandem mass spectrometry.

Herein, the determination of chlordimeform and its metabolite 4-chloro-2-methylaniline residue in milk was performed for the first time using gas chromatography-tandem mass spectrometry (GC-MS/MS). Samples were extracted using acetonitrile, and cleaned using a quick, easy, cheap, effective, rugged, and safe (QuEChERS) method. Separation was performed using the DB-17 MS column. It was detected in a selected reaction monitoring (SRM) mode and quantified using a matrix-matched isotope internal standard method. Under optimal conditions, a good linear relationship was observed in the concentration range of 10-200 µg/kg. The limit of quantitation was 10.0 µg/kg. The spiked recoveries for the target substance ranged from 84.5 % to 107.3 %, with relative standard deviations (RSD) of <7.2 %. The spiked samples were further confirmed by gas chromatography-quadrupole-Orbitrap high-resolution mass spectrometry (GC-Orbitrap HRMS). The combined method resulted in high accuracy and sensitivity and was suitable for the determination of chlordimeform and its metabolite 4-chloro-2-methylaniline residue in milk.

Long term impact of electrical resistance heating on soil bacterial community based on a field test.

Thermal remediation is an effective technology for organic contaminant remediation. However, the application of thermal remediation may have negative effects on soil properties and ecological functions, which requires further investigation. Based on a pilot test of electrical resistance heating remediation (ERH), soil samples were collected at different locations after heating for 116 days. Most soil physicochemical properties were less affected by the heating temperature difference. Application of high temperature increased microbial abundance but inhibited alpha diversity of the bacterial community. More significant changes in microbial communities were observed at temperatures above 60 °C. The genera mainly affected by heating temperature included Flavobacteria, Brockia, and S085, while the increase in temperature also inhibited the abundance of nitrochlorobenzene functional genes. At 140 days after the end of the pilot test, the bacterial community affected by thermal remediation could recover effectively, and the recovery of the bacterial community was not affected by temperature difference during the heating period. This study provides valuable field evidence of the long term impact of soil ERH treatment on soil properties and microbial communities, and provides further references for optimization of remediation performance with coupled technologies.

Ethylene induced AcNAC3 and AcNAC4 take part in ethylene synthesis through mediating AcACO1 during kiwifruit (Actinidia chinensis) ripening.

BACKGROUND: Ethylene plays a vital role in the ripening process of kiwifruit. A terrific amount of transcription factors (TFs) have been shown to regulate ethylene synthesis in various fruits. RESULTS: In this research, two new NAC TFs, named AcNAC3 and AcNAC4, were isolated from kiwifruit, which belonged to NAM subfamily. Bioinformatics analysis showed that both AcNAC3 and AcNAC4 were hydrophilic proteins with similar three-dimensional structures. The expression levels of AcNAC3, AcNAC4 and AcACO1 increased during kiwifruit ripening, as well as were induced by ethylene and repressed by 1-methylcyclopropene (1-MCP). Correlation analysis exhibited that ethylene production was positively correlated with the expression levels of AcNAC3, AcNAC4 and AcACO1. Moreover, both AcNAC3 and AcNAC4 acted as transcriptional activators and could bind to and activate AcACO1 promoter. CONCLUSION: All results unveiled that the ethylene-induced AcNAC3 and AcNAC4 were transcriptional activators, and might participate in kiwifruit ripening and ethylene biosynthesis through activating AcACO1, providing a new insight of ethylene synthetic regulation during kiwifruit ripening. © 2024 Society of Chemical Industry.

Two NAC transcription factors regulated fruit softening through activating xyloglucan endotransglucosylase/hydrolase genes during kiwifruit ripening.

Kiwifruit is a climacteric fruit that is prone to ripening and softening. Understanding molecular regulatory mechanism of kiwifruit softening, is helpful to ensure long-term storage of fruit. In the study, two NAC TFs and two XTH genes were isolated from kiwifruit. Phylogenetic tree showed that both AcNAC1 and AcNAC2 belonged to NAP subfamily, AcXTH1 belong to I subfamily, and AcXTH2 belong to III subfamily. Bioinformatics analysis predicted that AcNAC1 and AcNAC2 possessed similar three-dimensional structural, and belonged to hydrophilic proteins. AcXTH1 and AcXTH2 were hydrophilic proteins and contained signal peptides. AcXTH1 had a transmembrane structure, but AcXTH2 did not. qRT-PCR results showed that AcNAC1, AcNAC2, AcXTH1 and AcXTH2 were increased during kiwifruit ripening. Correlation analysis showed that kiwifruit softening was closely related to endotransglucosylase/hydrolase genes and NAC TFs, as well as there was also a close relationship between AcXTHs and AcNACs. Moreover, both AcNAC1 and AcNAC2 were transcriptional activators located in nucleus, which bound to and activated the promoters of AcXTH1 and AcXTH2. In shortly, we proved that the roles of NAC TFs in mediating fruit softening during kiwifruit ripening. Altogether, our results clarified that AcNAC1 and AcNAC2 were transcriptional activators, and took part in kiwifruit ripening and softening through activating endotransglucosylase/hydrolase genes, providing a new insight of fruit softening network in kiwifruit ripening.

Application of Single-Cell Assay for Transposase-Accessible Chromatin with High Throughput Sequencing in Plant Science: Advances, Technical Challenges, and Prospects.

The Single-cell Assay for Transposase-Accessible Chromatin with high throughput sequencing (scATAC-seq) has gained increasing popularity in recent years, allowing for chromatin accessibility to be deciphered and gene regulatory networks (GRNs) to be inferred at single-cell resolution. This cutting-edge technology now enables the genome-wide profiling of chromatin accessibility at the cellular level and the capturing of cell-type-specific cis-regulatory elements (CREs) that are masked by cellular heterogeneity in bulk assays. Additionally, it can also facilitate the identification of rare and new cell types based on differences in chromatin accessibility and the charting of cellular developmental trajectories within lineage-related cell clusters. Due to technical challenges and limitations, the data generated from scATAC-seq exhibit unique features, often characterized by high sparsity and noise, even within the same cell type. To address these challenges, various bioinformatic tools have been developed. Furthermore, the application of scATAC-seq in plant science is still in its infancy, with most research focusing on root tissues and model plant species. In this review, we provide an overview of recent progress in scATAC-seq and its application across various fields. We first conduct scATAC-seq in plant science. Next, we highlight the current challenges of scATAC-seq in plant science and major strategies for cell type annotation. Finally, we outline several future directions to exploit scATAC-seq technologies to address critical challenges in plant science, ranging from plant ENCODE(The Encyclopedia of DNA Elements) project construction to GRN inference, to deepen our understanding of the roles of CREs in plant biology.

Field test of thermally activated persulfate for remediation of PFASs co-contaminated with chlorinated aliphatic hydrocarbons in groundwater.

The co-occurrence of per- and polyfluoroalkyl substances (PFASs) and chlorinated aliphatic hydrocarbons (CAHs) in groundwater has drawn increased attention in recent years. No studies have been conducted concerning the oxidative degradation of PFASs and/or CAHs by in situ thermally activated persulfate (TAP) in groundwater, primarily due to the difficulty in cost-effectively achieving the desired temperature in the field. In this study, the effects and mechanisms of PFASs degradation by in situ TAP at a site with PFASs and CAHs co-contaminants were investigated. The target temperature of 40.0-70.0 °C was achieved in groundwater, and persulfate was effectively distributed in the demonstration area - the combination of which ensured the degradation of PFASs and CAHs co-contaminants by in situ TAP. It was demonstrated that the reductions of perfluoroalkyl carboxylic acids (PFCAs) concentration in all monitoring wells were in the range of 43.7 %-66.0 % by in situ TAP compared to those maximum rebound values in groundwater, whereas no effective perfluoroalkane sulfonic acids (PFSAs) degradation was observed. The conversion of perfluoroalkyl acids (PFAAs) precursors was one of the main factors leading to the increase in PFCAs concentrations in groundwater during in situ TAP. CAHs were effectively degraded in most monitoring wells, and furthermore, no inhibitory effects of CAHs and Cl- on the degradation of PFASs were observed due to the presence of sufficient persulfate. Additionally, there were significant increases in SO42- concentrations and reductions of pH values in groundwater due to in situ TAP, warranting their long-term monitoring in groundwater. The integrated field and laboratory investigations demonstrated that the reductions in PFCAs and CAHs concentrations can be achieved by the oxidative degradation of in situ TAP.

GhTPPA_2 enhancement of tobacco sugar accumulation and drought tolerance.

Trehalose metabolism plays an important role in plant growth and response to abiotic stress. Trehalose-6-phosphate (Tre6P) can help regulate sugar homeostasis and act as an indication signal for intracellular sugar levels. Crop productivity can be greatly increased by altering the metabolic level of endogenous trehalose in plants, which can optimize the source-sink connection. In this study, the upland cotton GhTPP protein family was first homologously compared and 24 GhTPP genes were found. Transcriptome analysis revealed that GhTPP members had obvious tissue expression specificity. Among them, GhTPPA_2 (Gh_A12G223300.1) was predominantly expressed in leaves and bolls. The results of subcellular localization showed that GhTPPA_2 is localized in the chloroplast. Via PlantCare, we analyzed the promoters and found that the expression of GhTPPA_2 may be induced by light, abiotic stress, and hormones such as abscisic acid, ethylene, salicylic acid and jasmonic acid. In addition, GhTPPA_2 was overexpressed (TPPAoe) in tobacco, and we found that the TPPase activity of TPPAoe tobacco increased by 66 %. Soluble sugar content increased by 39 % and starch content increased by 27 %. Whereas, the transgenic tobacco had obvious growth advantages under 100 mM mannitol stress. Transcriptome sequencing results showed that the differential genes between TPPAoe and control were considerably enriched in functions related to photosynthesis, phosphate group metabolism, and carbohydrate metabolism. This study shows that GhTPPA_2 is involved in regulating sugar metabolism, improving soluble sugar accumulation and drought stress tolerance of tobacco, which provides theoretical basis for research on high yield and drought tolerance of crops.

Natural variation in Beauty Mark is associated with UV-based geographical adaptation in Gossypium species.

BACKGROUND: Anthocyanins, a class of specialized metabolites that are ubiquitous among plant species, have attracted a great deal of attention from plant biologists due to their chemical diversity. They confer purple, pink, and blue colors that attract pollinators, protect plants from ultraviolet (UV) radiation, and scavenge reactive oxygen species (ROS) to facilitate plant survival during abiotic stress. In a previous study, we identified Beauty Mark (BM) in Gossypium barbadense as an activator of the anthocyanin biosynthesis pathway; this gene also directly led to the formation of a pollinator-attracting purple spot. RESULTS: Here, we found that a single nucleotide polymorphism (SNP) (C/T) within the BM coding sequence was responsible for variations in this trait. Transient expression assays of BM from G. barbadense and G. hirsutum in Nicotiana benthamiana using luciferase reporter gene also suggested that SNPs in the coding sequence could be responsible for the absent beauty mark phenotype observed in G. hirsutum. We next demonstrated that the beauty mark and UV floral patterns are associated phenotypes and that UV exposure resulted in increased ROS generation in floral tissues; BM thus contributed to ROS scavenging in G. barbadense and wild cotton plants with flowers containing the beauty mark. Furthermore, a nucleotide diversity analysis and Tajima's D Test suggested that there have been strong selective sweeps in the GhBM locus during G. hirsutum domestication. CONCLUSIONS: Taken together, these results suggest that cotton species differ in their approaches to absorbing or reflecting UV light and thus exhibit variations in floral anthocyanin biosynthesis to scavenge reactive ROS; furthermore, these traits are related to the geographic distribution of cotton species.

UAV-based time-series phenotyping reveals the genetic basis of plant height in upland cotton.

Plant height (PH) is an important agronomic trait affecting crop architecture, biomass, resistance to lodging and mechanical harvesting. Elucidating the genetic governance of plant height is crucial because of the global demand for high crop yields. However, during the rapid growth period of plants the PH changes a lot on a daily basis, which makes it difficult to accurately phenotype the trait by hand on a large scale. In this study, an unmanned aerial vehicle (UAV)-based remote-sensing phenotyping platform was applied to obtain time-series PHs of 320 upland cotton accessions in three different field trials. The results showed that the PHs obtained from UAV images were significantly correlated with ground-based manual measurements, for three trials (R2 = 0.96, 0.95 and 0.96). Two genetic loci on chromosomes A01 and A11 associated with PH were identified by genome-wide association studies (GWAS). GhUBP15 and GhCUL1 were identified to influence PH in further analysis. We obtained a time series of PH values for three field conditions based on remote sensing with UAV. The key genes identified in this study are of great value for the breeding of ideal plant architecture in cotton.

Preparation and Properties of Organically Modified Na-Montmorillonite.

This study investigates the montmorillonite (MMT) content, rotational viscosity, and colloidal index of sodium montmorillonite (Na-MMT) as a function of the sodium agent dosage, reaction time, reaction temperature, and stirring time. Na-MMT was modified using different octadecyl trimethyl ammonium chloride (OTAC) dosages under optimal sodification conditions. The organically modified MMT products were characterized via infrared spectroscopy, X-ray diffraction, thermogravimetric analysis, and scanning electron microscopy. The results show that the Na-MMT with good properties (i.e., the maximum rotational viscosity and highest Na-MMT content with no decrease in the colloid index) was obtained at a 2.8% sodium carbonate dosage (measured based on the MMT mass), a temperature of 25 °C, and a reaction time of two hours. Upon organic modification of the optimized Na-MMT, OTAC entered the NA-MMT interlayer, and the contact angle was increased from 20.0° to 61.4°, the layer spacing was increased from 1.58 to 2.47 nm, and the thermal stability was conspicuously increased. Thus, MMT and Na-MMT were modified by the OTAC modifier.

Antimicrobial activity of gamma-poly (glutamic acid), a preservative coating for cherries.

We investigated the minimum inhibitory concentration (MIC), antibacterial activity, and preservation ability of four molar masses of γ-polyglutamic acid (PGA) against Escherichia coli, Bacillus subtilis, and yeast. The antibacterial mechanism was determined based on the cell structure, membrane permeability, and microscopic morphology of the microorganisms. We then measured the weight loss, decay rate, total acid, catalase activity, peroxidase activity, and malondialdehyde content toward the possible use of PGA as a preservative coating for cherries. When the molar mass was greater than 700 kDa, the MIC for Escherichia coli and Bacillus subtilis was less than 2.5 mg/mL. The mechanism of action of the four molar masses of PGA was different with respect to the three microbial species, but a higher molar mass of PGA corresponded to stronger inhibition against the microbes. PGA of 2000 kDa molar mass damaged the microbial cellular structure, resulting in excretion of alkaline phosphatase, but PGA of 1.5 kDa molar mass affected the membrane permeability and the amount of soluble sugar. Scanning electron microscopy indicated the inhibitory effect of PGA. The antibacterial mechanism of PGA was related to the molar mass of PGA and the microbial membrane structure. Compared with the control, a PGA coating effectively inhibit the spoilage rate, delay the ripening, and prolong the shelf life of cherries.

Genome-wide identification and expression analysis of the cotton patatin-related phospholipase A genes and response to stress tolerance.

MAIN CONCLUSION: Patatin-related phospholipase A genes were involved in the response of Gossypium hirsutum to drought and salt tolerance. pPLA (patatin-related phospholipase A) is a key enzyme that catalyzes the initial step of lipid hydrolysis, which is involved in biological processes, such as drought, salt stress, and freezing injury. However, a comprehensive analysis of the pPLA gene family in cotton, especially the role of pPLA in the response to drought and salt tolerance, has not been reported so far. A total of 33 pPLA genes were identified in this study using a genome-wide search approach, and phylogenetic analysis classified these genes into three groups. These genes are unevenly distributed on the 26 chromosomes of cotton, and most of them contain a few introns. The results of the collinear analysis showed that G. hirsutum contained 1-5 copies of each pPLA gene found in G. arboreum and G. raimondii. The subcellular localization analysis of Gh_D08G061200 showed that the protein was localized in the nucleus. In addition, analysis of published upland cotton transcriptome data revealed that six GhPLA genes were expressed in various tissues and organs. Two genes (Gh_A04G142100.1 and Gh_D04G181000.1) were highly expressed in all tissues under normal conditions, showing the expression characteristics of housekeeping genes. Under different drought and salt tolerance stresses, we detected four genes with different expression levels. This study helps to clarify the role of pPLA in the response to drought and salt tolerance.

Genetic and transcriptome analysis of a cotton leaf variegation mutant.

In eukaryotic photosynthetic organisms, chloroplast is not only a site for photosynthesis, but it also have a vital role in signal transduction mechanisms. Plants exhibit various colors in nature with various mutants induced by EMS, whose traits are regulated by developmental and environmental factors, making them ideal for studying the regulation of chloroplast development. In this study, the cotton leaf variegated mutant (VAR) induced by EMS was used for this experiment. Genetic analysis revealed that VAR phenotype was a dominant mutation and by performing freehand section inspection, it was noticed that the vascular bundles of VAR were smaller. Chloroplast ultrastructure showed that the stacking of grana thylakoid was thinner and the starch granules were increased significantly in VAR comparedto wild type (WT). Transcriptome analysis found that the KEGG was enriched in photosynthesis pathway, and GO was abundant in zinc ion transmembrane transport, electron transporter and cation binding terms. In addition, GhFTSH5 expression in VAR was significantly higher than WT and the promoter sequence of GhFTSH5 had differences. The results showed that the VAR plant had altered GhFTSH5 expression and disrupted chloroplast structure, which in turn affects plant photosynthesis. More importantly, this study lays a foundation for further analyzing molecular mechanism of cotton variegated phenotypes.

Constructing a Surface Multi-cationic Heterojunction for CsPbI1.5Br1.5 Perovskite Solar Cells with Efficiency beyond 14.

All-inorganic CsPbI1.5Br1.5 perovskite solar cells are considered as top cell candidates for tandem cells as a result of their excellent thermal stability and photoelectric performance. However, their power conversion efficiencies (PCEs) are still low and far below the theoretical limit mainly as a result of the severe non-radiative recombination and optical loss. Herein, we introduce an versatile method to construct a surface multi-cationic heterojunction to achieve an efficient and stable CsPbI1.5Br1.5 perovskite solar cell. By precisely controlling the content of FA+ and MA+ on PbBr2-rich perovskite films, a high-quality heterojunction layer is formed to help effectively passivate the surface defects and reduce the optical loss of the CsPbI1.5Br1.5 perovskite. In addition, the incorporation of a heterojunction layer can also improve energy-level alignment and reduce interfacial charge recombination loss. As a result, the champion device with the incorporation of SMH exhibits a PCE of 14.11%, which presents the highest reported efficiency for inorganic CsPbI1.5Br1.5 solar cells thus far while retaining 85% of the initial efficiency after 1000 h of storage without encapsulation.

Effect of androgen on bone metabolism in hyperuricemic rats.

Introduction: This study aimed to investigate the effect of androgen on bone metabolism in hyperuricemic rats. Material and methods: Forty male Wistar rats were randomly divided into four groups: sham operation group, simple hyperuricemic group, hyperuricemic castration group, and simple castration group. A rat model of chronic hyperuricemia was established using potassium oxonate and ethambutol. Blood was sampled from the vena angularis at week 0, 4, 6, 8 and 12 after surgery to detect for uric acid, calcium, phosphorus and alkaline phosphatase, and investigate the effect of androgen on bone metabolism in hyperuricemic rats. Results: From the 4th week, compared with the sham operation group, the differences in uric acid levels between the simple hyperuricemic group and hyperuricemia castration group were statistically significant (p < 0.05), suggesting the successful establishment of the model of hyperuricemia. At the 6th week, uric acid levels decreased in the two hyperuricemic groups, and the difference from the sham operation group decreased (p = 0.05), showing that the modeling method had deteriorated. At the 8th week, the differences in uric acid levels between the two castration groups and sham operation group were statistically significant (p < 0.05). At the 12th week, the differences in serum levels of phosphorus between the simple hyperuricemic group and hyperuricemic castration group were statistically significant (p < 0.05). Conclusions: Androgen can induce bone metabolism changes in rats with hyperuricemia.

Integrating advancements in root phenotyping and genome-wide association studies to open the root genetics gateway.

Plant adaptation to challenging environmental conditions around the world has made root growth and development an important research area for plant breeders and scientists. Targeted manipulation of root system architecture (RSA) to increase water and nutrient use efficiency can minimize the adverse effects of climate change on crop production. However, phenotyping of RSA is a major bottleneck since the roots are hidden in the soil. Recently the development of 2- and 3D root imaging techniques combined with the genome-wide association studies (GWASs) have opened up new research tools to identify the genetic basis of RSA. These approaches provide a comprehensive understanding of the RSA, by accelerating the identification and characterization of genes involved in root growth and development. This review summarizes the latest developments in phenotyping techniques and GWAS for RSA, which are used to map important genes regulating various aspects of RSA under varying environmental conditions. Furthermore, we discussed about the state-of-the-art image analysis tools integrated with various phenotyping platforms for investigating and quantifying root traits with the highest phenotypic plasticity in both artificial and natural environments which were used for large scale association mapping studies, leading to the identification of RSA phenotypes and their underlying genetics with the greatest potential for RSA improvement. In addition, challenges in root phenotyping and GWAS are also highlighted, along with future research directions employing machine learning and pan-genomics approaches.

A Rapid and Efficient Method for Isolation and Transformation of Cotton Callus Protoplast.

Protoplasts, which lack cell walls, are ideal research materials for genetic engineering. They are commonly employed in fusion (they can be used for more distant somatic cell fusion to obtain somatic hybrids), genetic transformation, plant regeneration, and other applications. Cotton is grown throughout the world and is the most economically important crop globally. It is therefore critical to study successful extraction and transformation efficiency of cotton protoplasts. In the present study, a cotton callus protoplast extraction method was tested to optimize the ratio of enzymes (cellulase, pectinase, macerozyme R-10, and hemicellulase) used in the procedure. The optimized ratio significantly increased the quantity and activity of protoplasts extracted. We showed that when enzyme concentrations of 1.5% cellulase and 1.5% pectinase, and either 1.5% or 0.5% macerozyme and 0.5% hemicellulase were used, one can obtain increasingly stable protoplasts. We successfully obtained fluorescent protoplasts by transiently expressing fluorescent proteins in the isolated protoplasts. The protoplasts were determined to be suitable for use in further experimental studies. We also studied the influence of plasmid concentration and transformation time on protoplast transformation efficiency. When the plasmid concentration reaches 16 µg and the transformation time is controlled within 12-16 h, the best transformation efficiency can be obtained. In summary, this study presents efficient extraction and transformation techniques for cotton protoplasts.

Overexpression of LT, an Oncoprotein Derived from the Polyomavirus SV40, Promotes Somatic Embryogenesis in Cotton.

Although genetic transformation has opened up a new era for cotton molecular breeding, it still suffers from the limitation problem of long transformation periods, which slows down the generation of new cotton germplasms. In this study, LT gene (SV40 large T antigen), which promotes the transformation efficiency of animal cells, was codon-optimized. Its overexpression vector was transformed into cotton. It was observed that EC (embryogenic callus) formation period was 33% shorter and transformation efficiency was slightly higher in the LT T0 generation than that of control. RNA-seq data of NEC (non-embryonic callus) and EC from LT and control revealed that more DEGs (differential expression genes) in NEC were identified than that of EC, indicating LT mainly functioned in NEC. Further KEGG, GO, and transcription factor analyses showed that DEGs were significantly enriched in brassinosteroid biosynthesis pathways and that bHLH, MYB, and AP2/ERF were the top three gene families, which are involved in EC formation. In addition, the key genes related to the auxin pathway were differentially expressed only in LT overexpression NEC, which caused early response, biosynthesis, and transportation of the hormone, resulting in EC earlier formation. In summary, the results demonstrated that LT can promote somatic embryogenesis in cotton, which provides a new strategy for improving cotton transformation and shortening EC formation time.

Multi-Dimensional Molecular Regulation of Trichome Development in Arabidopsis and Cotton.

Plant trichomes are specialized epidermal cells that are widely distributed on plant aerial tissues. The initiation and progression of trichomes are controlled in a coordinated sequence of multiple molecular events. During the past decade, major breakthroughs in the molecular understanding of trichome development were achieved through the characterization of various trichomes defective mutants and trichome-associated genes, which revealed a highly complex molecular regulatory network underlying plant trichome development. This review focuses on the recent millstone in plant trichomes research obtained using genetic and molecular studies, as well as 'omics' analyses in model plant Arabidopsis and fiber crop cotton. In particular, we discuss the latest understanding and insights into the underlying molecular mechanisms of trichomes formation at multiple dimensions, including at the chromatin, transcriptional, post-transcriptional, and post-translational levels. We summarize that the integration of multi-dimensional trichome-associated genes will enable us to systematically understand the molecular regulation network that landscapes the development of the plant trichomes. These advances will enable us to address the unresolved questions regarding the molecular crosstalk that coordinate concurrent and ordered the changes in cotton fiber initiation and progression, together with their possible implications for genetic improvement of cotton fiber.

Insights Into MicroRNA-Mediated Regulation of Flowering Time in Cotton Through Small RNA Sequencing.

The timing of flowering is a key determinant for plant reproductive. It has been demonstrated that microRNAs (miRNAs) play an important role in transition from the vegetative to reproductive stage in cotton; however, knowledge remains limited about the regulatory role of miRNAs involved in flowering time regulation in cotton. To elucidate the molecular basis of miRNAs in response to flowering time in cotton, we performed high-throughput small RNA sequencing at the fifth true leaf stage. We identified 56 and 43 miRNAs that were significantly up- and downregulated in two elite early flowering cultivars (EFC) compared with two late flowering cultivars (LFC), respectively. The miRNA targets by RNA sequencing analysis showed that GhSPL4 in SBP transcription factor family targeted by GhmiR156 was significantly upregulated in EFCs. Co-expression regulatory network analysis (WGCNA) revealed that GhSOC1, GhAP1, GhFD, GhCOL3, and GhAGL16 act as node genes in the auxin- and gibberellin-mediated flowering time regulatory networks in cotton. Therefore, elucidation of miRNA-mediated flowering time regulatory network will contribute to our understanding of molecular mechanisms underlying flowering time in cotton.