Transcriptional Landscape and Dynamics Involved in Sugar and Acid Accumulation during Apple Fruit Development.

In fleshy fruit, sugars and acids are central components of fruit flavor and quality. To date, the mechanisms underlying transcriptional regulation of sugar and acid during fruit development remain largely unknown. Here, we combined ATAC-seq with RNA-seq to investigate the genome-wide chromatin accessibility and to identify putative transcription factors related to sugar and acid accumulation during apple (Malus domestica) fruit development. By integrating the differentially accessible regions (DARs) and differentially expressed genes (DEGs), we generated a global dataset of promoter-accessibility- and expression-increased genes (PEIGs). Using this strategy, we constructed a transcriptional regulatory network enabling screening for key transcription factors and target genes involved in sugar and acid accumulation. Among these transcription factors, five fruit-specific Dof (DNA binding with one finger) genes were selected to confirm their regulatory effects, and our results showed that they could affect sugar or acid concentration by regulating the expression of sugar or acid metabolism-related genes in apple fruits. Our transcriptional regulatory network provides a suitable platform to identify candidate genes that control sugar and acid accumulation. Meanwhile, our dataset will aid in analyzing other characteristics of apple fruit that have not been illuminated previously. Overall, these findings support a better understanding of the regulatory dynamics during apple fruit development and lay a foundation for quality improvement of apple.

The MdCBF1/2-MdTST1/2 module regulates sugar accumulation in response to low temperature in apple.

Soluble sugar content is a key component in controlling fruit flavor, and its accumulation in fruit is largely determined by sugar metabolism and transportation. When the diurnal temperature range is greater, the fleshy fruits accumulated more soluble sugars and become more sweeter. However, the molecular mechanism underlying this response remains largely unknown. In this study, we verified that low-temperature treatment promoted soluble sugar accumulation in apple fruit and found that this was due to the upregulation of the Tonoplast Sugar Transporter genes MdTST1/2. A combined strategy using assay for transposase-accessible chromatin (ATAC) sequencing and gene expression and cis-acting elements analyses, we identified two C-repeat Binding Factors, MdCBF1 and MdCBF2, that were induced by low temperature and that might be upstream transcription factors of MdTST1/2. Further studies established that MdCBF1/2 could bind to the promoters of MdTST1/2 and activate their expression. Overexpression of MdCBF1 or MdCBF2 in apple calli and fruit significantly upregulated MdTST1/2 expression and increased the concentrations of glucose, fructose, and sucrose. Suppression of MdTST1 and/or MdTST2 in an MdCBF1/2-overexpression background abolished the positive effect of MdCBF1/2 on sugar accumulation. In addition, simultaneous silencing of MdCBF1/2 downregulated MdTST1/2 expression and apple fruits failed to accumulate more sugars under low-temperature conditions, indicating that MdCBF1/2-mediated sugar accumulation was dependent on MdTST1/2 expression. Hence, we concluded that the MdCBF1/2-MdTST1/2 module is crucial for sugar accumulation in apples in response to low temperatures. Our findings provide mechanistic components coordinating the relationship between low temperature and sugar accumulation as well as new avenues to improve fruit quality.

Biodegradation of Imazethapyr by Bacterial Strain IM9601 Isolated from Agricultural Soil.

The widespread utilization of the herbicide imazethapyr presents significant challenges to crop rotation and results in detrimental soil degradation issues. Bacterial biodegradation has emerged as a promising and eco-friendly approach for mitigating pesticide residues contamination in the environment. In this study, a novel bacterium, identified as Brevibacterium sp. IM9601, was isolated and characterized based on morphological, physiological, and biochemical characteristics, as well as 16S rRNA gene sequence. This strain exhibited the ability to utilize imazethapyr as its sole carbon source for growth. Response surface methodology (RSM) was applied to optimize the degradation conditions. The most favorable conditions were determined to be a temperature of 27 °C, pH of 6.0, and an initial inoculum with a final OD600 of 0.15. Under these optimized condition, bacterial strain IM9601 exhibited substantial imazethapyr degradation, with removal rates of 90.08 and 87.05% for initial imazethapyr concentrations of 50 and 100 mg L-1, respectively, achieved within a 5-day incubation period. This investigation highlights imazethapyr-degrading capabilities of the Brevibacterium genus bacterial strain IM9601, marking it as a potentially novel and effective solution for addressing the environmental pollution resulting from the usage of imazethapyr. The study contributes to the growing body of research on bioremediation approaches, offering a sustainable and environmentally friendly method for mitigating the adverse impacts of herbicide contamination in agricultural settings.

The SnRK2.3-AREB1-TST1/2 cascade activated by cytosolic glucose regulates sugar accumulation across tonoplasts in apple and tomato.

Soluble sugars are the core components of fruit quality, and the degree of sugar accumulation is largely determined by tonoplast-localized sugar transporters. We previously showed that two classes of tonoplast sugar transporters, MdERDL6 and MdTST1/2, coordinately regulate sugar accumulation in vacuoles. However, the mechanism underlying this coordination remains unknown. Here we discovered that two transcription factors, MdAREB1.1/1.2, regulate MdTST1/2 expression by binding their promoters in apple. The enhanced MdAREB1.1/1.2 expression in MdERDL6-1-overexpression plants resulted in an increase in MdTST1/2 expression and sugar concentration. Further studies established that MdSnRK2.3, whose expression could be regulated by expressing MdERDL6-1, could interact with and phosphorylate MdAREB1.1/1.2, thereby promoting the MdAREB1.1/1.2-mediated transcriptional activation of MdTST1/2. Finally, the orthologous SlAREB1.2 and SlSnRK2.3 exhibited similar functions in tomato fruit as in their apple counterparts. Together, our findings provide insights into the regulatory mechanism of tonoplast sugar transport exerted by SnRK2.3-AREB1-TST1/2 for fruit sugar accumulation.

Transcriptome Profiling and Identification of the Candidate Genes Involved in Early Ripening in Ziziphus Jujuba.

The early ripening jujube is an immensely popular fresh fruit due to its high commercial value as well as rich nutrition. However, little is known about the mechanism of jujube fruit's ripening. In this study, the transcriptome profiles were comprehensively analyzed between the 'Lingwu Changzao' jujube and its early-ripening mutant during the fruit development and maturity. A total of 5,376 and 762 differentially expressed genes (DEGs) were presented at 80 and 90 days after the flowering of the jujube fruit, respectively. Furthermore, 521 common DEGs were identified as candidate genes that might be associated with the fruit's early ripening. Our findings demonstrated that in a non-climacteric jujube fruit, abscisic acid (ABA) was more greatly involved in fruit ripening than ethylene. Meanwhile, the fruit ripening of the early-ripening mutant was regulated by eight promotors of DEGs related to glucose and fructose, seven repressors of DEGs related to brassinosteroid signal transduction, and a series of transcription factor genes (MYB, Bhlh, and ERF). Additionally, the expression of 20 candidate DEGs was further validated by real-time PCR during the late fruit maturation stage. Collectively, the present study sheds light on the metabolic mechanism of the fruit's early ripening and provides valuable candidate genes for the early-ripening mutant's breeding.

F-box protein MdAMR1L1 regulates ascorbate biosynthesis in apple by modulating GDP-mannose pyrophosphorylase.

Ascorbate (Asc) is an important antioxidant in plants and humans that plays key roles in various physiological processes. Understanding the regulation of Asc content in fruit plants is important for improving plant resiliency and optimizing Asc in food. Here, we found that both the transcript level and protein abundance of Asc Mannose pathway Regulator 1 Like 1 (MdAMR1L1) was negatively associated with Asc levels during the development of apple (Malus × domestica) fruit. The overexpression or silencing of MdAMR1L1 in apple indicated that MdAMR1L1 negatively regulated Asc levels. However, in the leaves of MdAMR1L1-overexpressing apple lines, the transcript levels of the Asc synthesis gene Guanosine diphosphate-mannose pyrophosphorylase MdGMP1 were increased, while its protein levels and enzyme activity were reduced. This occurred because the MdAMR1L1 protein interacted with MdGMP1 and promoted its degradation via the ubiquitination pathway to inhibit Asc synthesis at the post-translational level. MdERF98, an apple ethylene response factor, whose transcription was modulated by Asc level, is directly bound to the promoter of MdGMP1 to promote the transcription of MdGMP1. These findings provide insights into the regulatory mechanism of Asc biosynthesis in apples and revealed potential opportunities to improve fruit Asc levels.

CircRNA casein kinase 1 gamma 1 (circ-CSNK1G1) plays carcinogenic effects in thyroid cancer by acting as miR-149-5p sponge and relieving the suppression of miR-149-5p on mitogen-activated protein kinase 1 (MAPK1).

BACKGROUND: The initiation and development of thyroid cancer may be associated with the deregulation of circular RNAs (circRNAs). The purpose of this work was to explore the role of circRNA casein kinase 1 gamma 1 (circ-CSNK1G1) in thyroid cancer. METHODS: The expression of circ-CSNK1G1, miR-149-5p, and mitogen-activated protein kinase 1 (MAPK1) was concluded using quantitative real-time PCR (qPCR), and the expression of MAPK1 protein was detected by Western blot assay. Cell viability was monitored by CCK-8 assay. Cell proliferation was determined by colony formation assay and EdU assay. Cell apoptosis and cycle were checked by flow cytometry assay. Cell invasion was determined by transwell assay. The predicted binding relationship between miR-149-5p and circ-CSNK1G1 or MAPK1 was verified by dual-luciferase reporter assay. The role of circ-CSNK1G1 in vivo was determined by establishing animal models. RESULTS: The present work discovered the upregulation of circ-CSNK1G1 in tumor tissues of thyroid cancer. In function, circ-CSNK1G1 knockdown inhibited proliferation, survival, and invasion in cancer cells, and tumor growth in mouse models. MiR-149-5p was a target of circ-CSNK1G1, and the anti-tumor effects of circ-CSNK1G1 knockdown were abolished by miR-149-5p downregulation. In addition, miR-149-5p directly targeted MAPK1, and miR-149-5p restoration-inhibited cell proliferation and invasion were recovered by MAPK1 overexpression. CONCLUSION: Circ-CSNK1G1 acted as miR-149-5p to relieve the inhibition of miR-149-5p on MAPK1, thus promoting the malignant development of thyroid cancer.

Factors Influencing the Hydration, Dimensional Stability, and Strength Development of the OPC-CSA-Anhydrite Ternary System.

Due to the advantages of high early strength and rapid setting, ternary systems consisting of ordinary Portland clinker (OPC), calcium sulphoaluminate (CSA) clinker, and anhydrite have broad application prospects. However, further studies need to be undertaken to find a more optimal mixing proportion of this ternary binder in order to meet basic performance requirements. In this paper, isothermal calorimetric tests, chemical shrinkage tests, drying shrinkage tests, and compressive strength tests were carried out to systematically identify the effect of the OPC/CSA ratio and anhydrite dosage on the hydration, mechanical property development, and dimensional stability of ternary binders. It was found that a higher CSA content leads to a higher cumulative hydration heat, a shorter acceleration period, and a delayed induction period, which can be ascribed to the retardation of C3S at a high aluminate concentration. However, a higher addition of anhydrite can retard the main peak of hydration despite promoting the intermediate peak and improving the hydration reaction rate. The drying shrinkage of blends decreases first along with the CSA proportion and then increases. Moreover, a higher anhydrite content mitigates the drying shrinkage and hinders the strength development. Finally, considering the properties of both the fresh and hardened binder, the ternary blends with 5% anhydrite and OPC/CSA ratios ranging from 3/7 to 2/8 were identified as most suitable for applications that require a high early strength, stable late strength, and small level of shrinkage.

Combined Profiling of Transcriptome and DNA Methylome Reveal Genes Involved in Accumulation of Soluble Sugars and Organic Acid in Apple Fruits.

Organic acids and soluble sugars are the major determinants of fruit organoleptic quality. Additionally, DNA methylation has crucial regulatory effects on various processes. However, the epigenetic modifications in the regulation of organic acid and soluble sugar accumulation in apple fruits remain uncharacterized. In this study, DNA methylation and the transcriptome were compared between 'Honeycrisp' and 'Qinguan' mature fruits, which differ significantly regarding soluble sugar and organic acid contents. In both 'Honeycrisp' and 'Qinguan' mature fruits, the CG context had the highest level of DNA methylation, and then CHG and CHH contexts. The number and distribution of differentially methylated regions (DMRs) varied among genic regions and transposable elements. The DNA methylation levels in all three contexts in the DMRs were significantly higher in 'Honeycrisp' mature fruits than in 'Qinguan' mature fruits. A combined methylation and transcriptome analysis revealed a negative correlation between methylation levels and gene expression in DMRs in promoters and gene bodies in the CG and CHG contexts and in gene bodies in the CHH context. Two candidate genes (MdTSTa and MdMa11), which encode tonoplast-localized proteins, potentially associated with fruit soluble sugar contents and acidity were identified based on expression and DNA methylation levels. Overexpression of MdTSTa in tomato increased the fruit soluble sugar content. Moreover, transient expression of MdMa11 in tobacco leaves significantly decreased the pH value. Our results reflect the diversity in epigenetic modifications influencing gene expression and will facilitate further elucidating the complex mechanism underlying fruit soluble sugar and organic acid accumulation.

Nitrogen Levels Regulate Sugar Metabolism and Transport in the Shoot Tips of Crabapple Plants.

To comprehensively understand the responses of carbohydrate metabolism and transport to different levels of nitrogen supply in growing shoot tips of crabapple (Malus hupehensis Rehd), enzyme activities and related genes involved in the sugar metabolism pathway were investigated. The nitrogen and chlorophyll content of plants increased with increasing nitrogen supply. High nitrogen application increased the net photosynthesis rate and the growth rate of shoot tips but decreased the synthesis capability of sucrose and sorbitol in mature leaves. However, the shoot tips of plants under high-nitrogen treatment had higher contents of sucrose and sorbitol than did those under low-nitrogen treatment, while the activity of sucrose phosphate synthase and aldose-6-phosphate was increased and the transporters MdSOT and MdSUT were up-regulated. Moreover, the activities of enzymes involved in sucrose and hexose metabolism (including sucrose synthase, fructokinase, and hexokinase) were enhanced in the shoot tips of plants under high-nitrogen conditions, and the expression levels of MdSUSY3 and MdHK1 were significantly up-regulated. These findings indicate that a high nitrogen supply increases the metabolic capacity of assimilatory substances in shoot tips, accelerates the efficiency of sugar utilization and eventually leads to a rapid increase in the growth of shoot tips. Our results highlight that high nitrogen increases the capacity of sugar unloading and metabolic utilization in growing shoot tissues.

MdERDL6-mediated glucose efflux to the cytosol promotes sugar accumulation in the vacuole through up-regulating TSTs in apple and tomato.

Sugar transport across tonoplasts is essential for maintaining cellular sugar homeostasis and metabolic balance in plant cells. It remains unclear, however, how this process is regulated among different classes of sugar transporters. Here, we identified a tonoplast H+/glucose symporter, MdERDL6-1, from apples, which was highly expressed in fruits and exhibited expression patterns similar to those of the tonoplast H+/sugar antiporters MdTST1 and MdTST2. Overexpression of MdERDL6-1 unexpectedly increased not only glucose (Glc) concentration but also that of fructose (Fru) and sucrose (Suc) in transgenic apple and tomato leaves and fruits. RNA sequencing (RNA-seq) and expression analyses showed an up-regulation of TST1 and TST2 in the transgenic apple and tomato lines overexpressing MdERDL6-1 Further studies established that the increased sugar concentration in the transgenic lines correlated with up-regulation of TST1 and TST2 expression. Suppression or knockout of SlTST1 and SlTST2 in the MdERDL6-1-overexpressed tomato background reduced or abolished the positive effect of MdERDL6-1 on sugar accumulation, respectively. The findings demonstrate a regulation of TST1 and TST2 by MdERDL6-1, in which Glc exported by MdERDL6-1 from vacuole up-regulates TST1 and TST2 to import sugars from cytosol to vacuole for accumulation to high concentrations. The results provide insight into the regulatory mechanism of sugar accumulation in vacuoles mediated by the coordinated action of two classes of tonoplast sugar transporters.

Stripped Electrode Based Electrowetting-on-Dielectric Digital Microfluidics for Precise and Controllable Parallel Microdrop Generation.

Microdrop generation with excellent controllability and volume precision is of paramount significance for a large variety of microfluidic applications. In this work, we propose a new configuration comprising only stripped electrodes of rectangular shape for the closed electrowetting-on-dielectric digital microfluidic (EWOD DMF) system and investigate its parallel microdrop generation outcomes via a numerical approach. The microfluidic droplet motion is solved by a finite-volume scheme on a fixed computational domain. The numerical model is verified by an experimental study of microdrop production from an EWOD DMF device with three different electrode designs. After model verification, we examine the influences of the equilibrium contact angle and the spacing of the microchannel on stripped electrode based microdrop generation outcomes and discover five different regimes including the phenomena of satellite droplet formation and separation cessation. Despite the various generation outcomes, the daughter droplet size is found to vary linearly with a dimensionless EWOD parameter κ*. More importantly, for all successful generations, the deviation of the daughter droplet size from that of the stripped electrode is smaller than 3.5%, which even reaches zero in proper conditions. This new configuration can be utilized as a convenient alternative for electrowetting-induced parallel microdrop production with excellent precision and controllability.

Complete Genome Sequence of Pseudomonas syringae pv. lapsa Strain ATCC 10859, Isolated from Infected Wheat.

Pseudomonas syringae pv. lapsa is a pathovar of Pseudomonas syringae that can infect wheat. The complete genome of P. syringae pv. lapsa strain ATCC 10859 contains a 5,918,899-bp circular chromosome with 4,973 coding sequences, 16 rRNAs, 69 tRNAs, and an average GC content of 59.13%. The analysis of this genome revealed several gene clusters that are related to pathogenesis and virulence.

[Efficacy observation of treating diabetic nephropathy by shenshuaining granule combined telmisartan tablet].

OBJECTIVE: To observe the effect of Shenshuaining Granule (SG) combined telmisartan on serum creatinine (SCr) levels and urinary albumin contents in diabetic nephropathy (DN) patients, and to explore its efficacy. METHODS: Totally 204 DN patients were recruited, and further assigned to 3 groups, i.e., the early DN group, the clinical stage of DN with normal renal function group, the clinical stage of DN with insufficient renal function group. Patients in the same group were randomly allocated to the telmisartan treatment group, the SG treatment group, and the combination of SG and telmisartan treatment group, 68 in each group. Patients in the telmisartan treatment group took telmisartan tablet, 80 mg per day, once daily. Those in the SG treatment group took SG, 5 g each time, 3 times per day. Those in the combination of SG and telmisartan treatment group took telmisartan tablet (80 mg per day, once daily) and SG (5 g each time, 3 times per day). The therapeutic course for all was 3 successive months. SCr levels, serum urea nitrogen (BUN),24 h urine microalbumin (24 h U-MA) were detected before and after treatment. Results In three different treatment groups, 24 h U-MA decreased after treatment in the telmisartan treatment group; SCr and BUN decreased after treatment in the SG treatment group; and 24 h U-MA, SCr and BUN decreased after treatment in the combination of SG and telmisartan treatment group (P<0.05). In the clinical stage of DN with insufficient renal function group, SCr obviously increased after treatment in the telmisartan treatment group (P <0. 05). In the 3 DN stages, SCr and 24 h U-MA obviously decreased in the combination of SG and telmisartan treatment group, when compared with the telmisartan treatment group and the SG treatment group (P<0.05). Compared with the telmisartan treatment group, SCr and BUN obviously decreased in the SG treatment group, but 24 h U-MA quantitation obviously increased (P<0.05). BUN obviously decreased in the combination of SG and telmisartan treatment group (P<0. 05). CONCLUSION: The combination of SG and telmisartan could decrease urinary albumin, and stabilize SCr levels.

Chromogenic and fluorescent 'turn-on' chemodosimeter for fluoride based on a F(-)-triggered cascade reaction.

We developed a new chromogenic and fluorescent 'turn-on' chemodosimeter 1 based on a F(-)-triggered cascade reaction. This system displayed significant changes in UV/vis absorption and fluorescence emission intensities selectively for F(-) over other anions in a mixture of CH3CN/H2O(95 : 5, v/v) and in acetonitrile.