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The blueberry root-associated microbiome plays an essential role in its adaptation to acidic soils and in limiting the uptake of soil nutrients by its poor root system. Studies on the interactions of the root-associated microbiome in the various root niches may deepen our understanding of the beneficial effects in this particular habitat.

Blackberry shrubs are economically important for their production of small, pulped fruits. This species has attracted much attention because of the unique flavor of its fruits and their rich nutritional and medicinal value. In this study, microRNAs (miRNAs) and their target genes related to flavonoids and anthocyanins in blackberry fruits during ripening were analyzed and identified by small RNA and degradome sequencing technology, and the expression levels of key miRNAs in unripe and ripe blackberry fruits were verified via the RT-qPCR. A total of 258 known miRNAs were identified. Eighty differentially expressed miRNAs (DEMs) were detected in the fruits of the ripe group compared with those of the unripe group. Differentially expressed miR828-x/miR828-z and unigene0086056 (unknown function) were coexpressed. Moreover, miR858 had the most target genes for the synthesis of flavonoids and anthocyanins. Taken together, these results provide important value for improving the quality of blackberry fruits and breeding blackberry plants that produce high-flavonoid fruits for the pharmaceutical industry.

Flavonoids are important secondary metabolites with extensive pharmacological functions. Ginkgo biloba L. (ginkgo) has attracted extensive attention because of its high flavonoid medicinal value. However, little is understood about ginkgo flavonol biosynthesis. Herein, we cloned the full-length gingko GbFLSa gene (1314 bp), which encodes a 363 amino acid protein that has a typical 2-oxoglutarate (2OG)-Fe(II) oxygenase region. Recombinant GbFLSa protein with a molecular mass of 41 kDa was expressed in Escherichia coli BL21(DE3). The protein was localized to the cytoplasm. Moreover, proanthocyanins, including catechin, epicatechin, epigallocatechin and gallocatechin, were significantly less abundant in transgenic poplar than in nontransgenic (CK) plants. In addition, dihydroflavonol 4-reductase, anthocyanidin synthase and leucoanthocyanidin reductase expression levels were significantly lower than those of their CK counterparts. GbFLSa thus encodes a functional protein that might negatively regulate proanthocyanin biosynthesis. This study helps elucidate the role of GbFLSa in plant metabolism and the potential molecular mechanism of flavonoid biosynthesis.

BACKGROUND: The softness of blackberry fruits limits their postharvest shelf-life and commercial use, and abscisic acid (ABA) is considered one of the key hormones involved in fruit ripening. OBJECTIVE: This study aimed to explore the underlying physiological and molecular actions of ABA on blackberry fruit ripening and softening. METHODS: Various physiological indices of and plant hormone levels in treated and untreated blackberry fruits were determined simultaneously. The differentially expressed genes (DEGs) were analyzed by RNA-sequencing, and their expression profiles were detected. The ripening mechanism was elucidated by UHPLC-MS using two groups of fruits at 28 d. RESULTS: After 25 d, the ABA concentration and polygalacturonase (PG) and beta-1,4-endoglucanase (EG) activities in ABA-treated fruits were significantly higher than those in untreated fruits. Large differences in the expression profiles were detected at 28 d. The expression of DEGs related to cell wall softening and ABA synthesis was largely triggered after 25 or 28 d. Sixty-nine differentially accumulated metabolites were ultimately annotated as related to fruit ripening. CONCLUSIONS: ABA stimulates blackberry fruit ripening by promoting cell wall enzyme activities, the expression of various ripening-related genes and metabolite accumulation.

Compared with the traditional organic-inorganic hybrid perovskite, CsPbI3 is considered to be a better solar photovoltaic absorption material. However, under environmental conditions, it will undergo the phase transition from α...

Blackberry (Rubus spp.) fruit, contains high levels of anthocyanins in the pulp, is a good source of antioxidants. However, the molecular mechanisms underlying anthocyanin accumulation remains elusive. To improve our understanding of this process, we used RNA-seq to evaluate the transcriptional regulation of plant secondary metabolites, especially anthocyanin biosynthesis during fruit ripening. We constructed nine sequencing libraries, obtained 7.7 Gb clean bases and identified 50,563 annotated unigenes. Comparative transcriptome analysis revealed that 11,136, 33,268, and 31,913 genes were differentially expressed at Green vs. Red, Green vs. Black, and Red vs. Black, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment revealed that 35, 47, and 47 pathways were significantly enriched from these genes, respectively. We identified 58, 18, 24 and 48 differentially expressed genes (DEGs) involved in the anthocyanin biosynthesis, sucrose metabolism, abscisic acid (ABA), and ethylene (ETH) biosynthesis, respectively. Moreover, 66 DEGs of MYB were also identified in blackberry fruit. 24 DEGs from anthocyanin biosynthesis pathway and two DEGs of MYB were positively correlated with anthocyanin accumulation. Six DEGs of sucrose, six DEGs of ABA and seven DEGs of ETH were changed concomitantly with sucrose, ABA and ETH production. This study will provide an important information for understanding the molecular mechanism of anthocyanin accumulation during fruit ripening and a reference for related studies.

B-box (BBX) genes play important roles in plant growth, light morphogenesis, and environmental stress responses. Ginkgo (Ginkgo biloba L.) is known as a living fossil species that has a strong ability to adapt to environmental changes and tolerate harsh conditions. In this study, we chose this species to investigate the function of the GbBBX25 gene. We isolated the BBX gene from ginkgo and named it GbBBX25; this gene consists of an 819 bp open reading frame (ORF) that encodes 273 amino acids with two B-box domains but no CCT domain. GbBBX25 was localized in only the nucleus. The expression of GbBBX25 transcripts was observed in the leaves and was significantly enhanced under salt stress conditions. To further verify its function, we overexpressed the GbBBX25 gene in Populus davidiana × Populus bolleana and found that the transgenic Populus had greater soluble sugar levels and higher peroxidase (POD) activity in response to salt stress than nontransgenic (NT) Populus. Five genes related to salt stress were induced in transgenic plants with significantly higher expression levels than those in NT plants. This finding suggests that GbBBX25 improves the salt adaptation abilities of transgenic Populus and provides a scientific basis for related research.

As an emerging third-generation fruit, blackberry has high nutritional value and is rich in polyphenols, flavonoids and anthocyanins. Flavonoid biosynthesis and metabolism is a popular research topic, but no related details have been reported for blackberry. Based on previous transcriptome data from this research group, two blackberry flavonol synthase genes were identified in this study, and the encoded proteins were subjected to bioinformatics analysis. RuFLS1 and RuFLS2 are both hydrophobic acidic proteins belonging to the 2OG-Fe(II) dioxygenase superfamily. RuFLS2 was expressed at 27.93-fold higher levels than RuFLS1 in red-purple fruit by RNA-seq analysis. Therefore, RuFLS2-overexpressing tobacco was selected for functional exploration. The identification of metabolites from transgenic tobacco showed significantly increased contents of flavonoids, such as apigenin 7-glucoside, kaempferol 3

Blackberry is an economically important shrub species of Rubus in the Rosaceae family. It is rich in phenolic compounds, which have many health effects and pharmaceutical value. The utilization of metabolites from various blackberry tissues is still in the primary stage of development, so investigating the metabolites in various tissues is of practical significance. In this study, nontargeted LC - MS metabolomics was used to identify and measure metabolites in the roots, stems, leaves and fruits (green, red, and black fruits) of blackberry "Chester". We found that 1,427 and 874 metabolites were annotated in the positive and negative ion modes (POS; NEG), respectively. Differentially abundant metabolites (DAMs) between the leaf and root groups were the most abundant (POS: 249; NEG: 141), and the DAMs between the green and red fruit groups were the least abundant (POS: 21; NEG: 14). Moreover, the DAMs in different fruit development stages were far less than those in different tissues. There were significant differences in flavonoid biosynthesis-related pathways among the comparison groups. Trend analysis showed that the profile 10 had the largest number of metabolites. This study provides a scientific basis for the classification and efficient utilization of resources in various tissues of blackberry plants and the directional development of blackberry products.

BACKGROUND: Black raspberry is a vital fruit crop with a high antioxidant function. MADS-box genes play an important role in the regulation of fruit development in angiosperms. OBJECTIVE: To understand the regulatory role of the MADS-box family, a total of 80 MADS-box genes were identified and analyzed. METHODS: The MADS-box genes in the black raspberry genome were analyzed using bioinformatics methods. Through an analysis of the promoter elements, the possible functions of different members of the family were predicted. The spatiotemporal expression patterns of members of the MADS-box family during black raspberry fruit development and ripening were systematically analyzed. RESULTS: The genes were classified into type I (Mα: 33; Mβ: 6; Mγ: 10) and type II (MIKC*: 2; MIKCC: 29) genes. We also obtained a complete overview of the RoMADS-box gene family through phylogenetic, gene structure, conserved motif, and cis element analyses. The relative expression analysis showed different expression patterns, and most RoMADS-box genes were more highly expressed in fruit than in other tissues of black raspberry. CONCLUSIONS: This finding indicates that the MADS-box gene family is involved in the regulation of fruit ripening processes in black raspberry.

This study presents a novel approach to computing all intersections between two Bézier curves using cubic hybrid clipping. Each intersection is represented by two strip intervals that contain an intersection. In each step, one curve is bounded by two fat lines, and the other is bounded by two cubic Bézier curves, clipping away the domain that does not contain the intersections. By selecting the moving control points of the cubic hybrid curves, better cubic polynomial bounds are obtained to make the proposed method more efficient. It was proved that the two strip intervals have second- and fourth-order convergence rates for transversal intersections. Experimental results show that the new algorithm is the most efficient among all existing curve/curve intersection approaches.

Fruit cracking tremendously damages the appearance of fruit, easily leads to pathogen invasion, greatly reduces the marketability and causes immense economic losses. The pivotal role of long non-coding RNAs (lncRNAs) in diverse biological processes has been confirmed, while the roles of lncRNAs underlying fruit cracking remain poorly understood. In this study, the incidence of fruit cracking was 7.26% under the bagging treatment, the control group was 38.11%, indicating that bagging considerably diminished the fruit cracking rate. LncRNA libraries for fruit cracking (FC), fruit non-cracking (FNC) and fruit non-cracking under bagging (FB) in pomegranate (

MicroRNA (miRNA) is an endogenous single-stranded noncoding small RNA with a base length of 18-30, which widely exists in eukaryotic cells. It plays an important role in plant growth and development, metabolic biosynthesis and stress response. Mature miRNAs are evolutionarily conserved and have both sequential and tissue-specific expression. This paper summarizes the production, action mechanisms, main technologies, research progress and biological functions of miRNAs. miRNA research includes miRNA discovery (genetic screening, direct cloning, bioinformatics prediction and high-throughput sequencing) and biological function analysis (overexpression, the expression or inhibition of target gene mutants). It was found that most miRNAs are involved in plant secondary metabolism biosynthesis and have great potential as molecular breeding tools. The same biological process can be regulated by different miRNAs. Interestingly, the role of the same miRNA in different plants may behave in completely opposite manner, and the mode of action of each miRNA is also unique. Moreover, miRNA interaction and miRNA self-regulation will also be the developmental direction in the future. However, miRNA target recognition and functional verification are still major challenges for the future. This study will provide a reference for further exploration and verification the specific biological functions and related applications of miRNAs in plants.

Detection of arterial stiffness is an important method to predict the occurrence of hypertension complications and to screen patients with high cardiovascular risk. In order to predict the damage of AASI to the renal function of patients with essential hypertension, the prediction of AASI based on stepwise Regression equation scanning for renal function damage in patients with essential hypertension is proposed. Measure the 24 h ambulatory blood pressure of the selected subjects, establish a linear Regression equation scanning, and calculate the slope of the straight line, and finally, the slope is AASI. According to the quartiles, AASI is divided into four parts: group I < 0.53 ( n = 49 ); 0.53 ≤ group II < 0.60 ( n = 51 ); 0.60 ≤ group III < 0.69 ( n = 48 ); group IV ≥ 0.69 ( n = 44 ). Experiment result shows the following: with the increase of AASI, cystatin (CysC) also increased significantly, while CysC-eGFR decreased significantly ( P < 0.05 ). Compared with groups I, II, and III, Scr and CysC in group IV increased ( P < 0.05 ), and Ccr, CysC-eGFR, and (CKD-EPI)-eGFR all decreased ( P < 0.05 ). AASI is positively correlated with CysC performance, and the correlation coefficient r is 0.637. It is negatively correlated with Ccr performance, and r is -0.361. It is negatively correlated with CysC-eGFR, and r is -0.698. And it is negatively correlated with (CKD-EPI)-eGFR, and r is -0.331. Age and 24 h PP also showed an increasing trend with the increase of AASI, and it suggests that age may be an influencing factor that promotes kidney damage caused by hypertension; it also suggests that AASI can be used as a new indicator of arterial compliance; AASI is linearly related to various indicators of renal damage and can be used as a predictive indicator of renal damage caused by essential hypertension; cystatin C and the estimated glomerular filtration rate CysC-eGFR based on cystatin C are better than other indicators reflecting glomerular filtration rate, more sensitively assess the degree of early renal damage. Obesity may also be a factor that promotes kidney damage caused by hypertension.

Blackberry and raspberry have high nutritional, health value, and are popular with consumers for their unique flavors. To explore the relationships between nutrient accumulation, antioxidant substance contents in blackberry and raspberry fruits, and fruit growth and development, seven

With the advancement of blueberry industrialization, cultivation measures for obtaining high-quality fruits and technologies for obtaining high levels of the main secondary metabolites have become inevitable requirements for further development of the blueberry industry. This study applied different shading treatments and found that the FT1 shading treatment yielded the largest values for the single fruit weight, solid longitudinal diameter and transverse diameter of blueberry fruit as well as the highest solidity-acid ratio and total phenol and vitamin C contents. Moreover, 470 known metabolites were obtained from blueberry fruits. Interestingly, the differentially abundant metabolites related to ABC transporters, pyrimidine metabolism, and purine metabolism pathways were commonly identified from the three comparisons, which indicated that these three metabolic pathways in blueberry fruits are vulnerable to shading treatment. This study provides a theoretical basis for the application of summer shading to improve the quality and antioxidant substances of small berries.

Blueberry belongs to the genus Vaccinium L. in the Ericaceae and is an economically important shrub that produces small berries that are rich in nutrients. There were differences in the appearance of blueberry leaves under different shade treatments. To explore the differences in metabolites in blueberry leaves under different shading treatments, nontargeted liquid chromatography–mass spectrometry (LC–MS) metabonomic analysis was performed. Different shade intensities resulted in significant differences in the contents of metabolites. A total of 6879 known metabolites were detected, including 750 significantly differentially expressed metabolites, including mainly lipids and lipid-like molecules and phenylpropanoid and polyketide superclass members. Based on a Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, the flavone and flavonol biosynthesis pathways were the most significantly enriched. The results of this study provide a reference and scientific basis for the establishment of a high-quality and high-yield shaded blueberry cultivation system.

Nitrogen (N) is an indispensable element for plant growth and development. To understand the regulation of underlying carbon (C) and N metabolism in blackberry plants, we performed integrated analyses of the physiology, metabolome and transcriptome. Blackberry plants were subjected to no N, nitrate (NO

With improving living standards, traditional blueberry planting modes cannot meet commercial demands, and blueberry cultivation with soilless substrate has become a popular solution in the blueberry industry. In this study, different soilless substrate treatments were found to markedly influence fruit appearance and intrinsic quality. The fruit in the 50:50 peat/pine bark (v/v) (FPB) treatment group had the maximum single fruit weight, largest vertical diameter, and brightest color, as well as the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) value, solid-acid ratio and anthocyanin content. The fruit in the 50:50 pine bark/rice husk (v/v) (FBR) treatment group had the highest total phenol and flavonoid levels, largest drip loss value, and lowest total pectin content and firmness value. Metabolomic analysis showed that flavonoid, carbohydrate, and carbohydrate conjugate, and amino acid, peptide, and analog levels were significantly different between groups. Fruit in the FPB group had the highest sucrose, D-fructose 1,6-bisphosphate, salidroside, tectorigenin, naringenin chalcone, trifolirhizin, and galangin contents. The increase in the relative expression of phenylalanine (Phe) promoted the synthesis of fruit polyphenols in the FBR group. Our results provide new insights into the effects of different substrates on the quality of blueberries and a reference for the soilless substrate cultivation of blueberries.

Ginkgo biloba L. leaves are a flavonoid resource for the pharmaceutical industry. The flavonoid 3'-hydroxylase (F3'H) is a key enzyme in the flavonoid biosynthesis pathway. However, the role of F3'H in flavonoid biosynthesis and metabolism is unclear. In this study, we characterized and functionally analyzed the ginkgo F3'H gene GbF3'H1 that encodes a protein of 520 amino acids. Expression profiling showed that GbF3'H1 was highly expressed in the leaves of ginkgo in September. Subcellular localization showed that GbF3'H1 occurred predominately in the cytoplasm. Transgenic poplars overexpressing GbF3'H1 had more red pigmentation in leaves than did wild-type (WT) plants. Furthermore, the concentrations of epigallocatechin, gallocatechin, and catechin in the downstream products synthesized by flavonoids were significantly higher in the transgenic plants than in the WT plants. These results indicate that the overexpression of GbF3'H1 enhances flavonoid production in transgenic plants and provides new insights into flavonoid biosynthesis and metabolism.

Afforestation and agroforestry both sequester atmospheric carbon (C) and store C in the soil. However, the response of soil nutrients and soil organic carbon (SOC) to these practices has not been fully examined, especially in deep soil layers. We therefore investigated the effects of afforestation and agroforestry on the nutrient contents and organic carbon (OC) stocks within a 100‐cm depth in eastern China. The soil nutrient contents and stoichiometry as well as the SOC contents and stocks exhibited different vertical patterns and varied among the five planting systems. The total nitrogen (TN), total phosphorus (TP), and SOC contents decreased dramatically at a 20‐cm depth, and the total potassium (TK) content exhibited a different pattern. Compared with adjacent croplands and nurseries, the forest plantation and agroforestry systems had higher SOC stocks, ranging from 42.9 to 111.6 t·hm⁻² in the whole soil profile. The SOC distribution among the depth gradient varied among planting systems, and the topsoil SOC stocks played crucial roles in total SOC storage within a depth of 100 cm in the cropland and nursery systems. The soil pH and bulk density (BD) were negatively correlated with the SOC content, and a significant positive linear relationship occurred between the TN and SOC contents and between the TP and SOC contents. We recommend that combinations of nitrogen (N) and phosphorus (P) fertilizers would benefit SOC storage and improve stabilization in agroforestry and plantation systems.

Copper indium gallium diselenide (CIGS) thin-film solar cells are fabricated through several deposition and annealing processes at high temperatures, which can generate significant thermal residual stress to solar cells. Moreover, since CIGS solar cells with flexible substrates are rollable and bendable, they are susceptible to mechanical stresses during these processes. In addition, partial shading (hotspot) can exert high heat on the CIGS solar cell. In this paper, we investigate the thermo-mechanical stress of each active layer of CIGS solar cells due to annealing, external bending, and hotspot using finite element method (FEM). We found that average stress of each active layer decreases and maximum stress in the cell increases when interface crack is introduced between cadmium sulfide (CdS)/CIGS. Our overarching goal is to quantify the relationship between the fabrication/operating process and the reliability of CIGS solar cells (energy release rate and internal stresses), which could improve the reliability of flexible solar cells. It is also found that lowering the annealing temperature can reduce the stresses in the cells and lowering CIGS thickness can reduce the delamination probability of CdS/CIGS interface. Finally, we investigate the effect of the crack length of the CdS/CIGS interface on the electrical performance of CIGS solar cells through FEM simulations. We found that as the crack size between CdS and CIGS layers increases, short-circuit current density decreases, while open-circuit voltage remains almost constant.

Ginkgo (Ginkgo biloba L.) is a tree valued for the high medicinal and nutritional value of its leaves and seeds. However, the metabolite profiles of ginkgo leaves and seeds and their changes during development have not been comprehensively analyzed, which hinders improvements in the utilization of ginkgo. A comprehensive and systematic untargeted LC-MS metabolomics analysis of the metabolites in ginkgo leaves (male and female) and seeds at two developmental stages identified 8146 known metabolites, which mainly included lipids and lipid-like molecules, phenylpropanoids and polyketides, organoheterocyclic compounds, organic acids and derivatives, organic oxygen compounds, and benzenoids. Some of the identified metabolites have known healthcare and food value, and some of the others are newly discovered metabolites with potential for new drug development. The small number of differential Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways between leaves of male and female gingko trees indicated that the developmental stage affected the metabolic pathways more significantly than sex. Among the flavonoid constituents of ginkgo, 653 flavonoid metabolites were identified, and these included some new flavonoid components, which confirmed that the developmental stage is a critical factor in secondary metabolite variations. This study illuminated the metabolites and medicinal and edible values of ginkgo leaves and seeds at different developmental stages and thus supports further effective utilization of ginkgo leaves and seeds.

Background: The 3rd-generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI), like osimertinib, provide marked clinical benefit for EGFR-mutant non-small cell lung cancer (NSCLC) patients with extended overall survival vs former EGFR TKIs (e.g., gefitinib). Approximate 10%-24% of NSCLC patients acquired C797S mutation when treated with osimertinib. Unfortunately, limited treatments are available for patients after osimertinib resistance. QLH11811 is a new generation EGFR TKI designed to target the EGFR with ex19del/L858R/T790M/C797S mutations. Here, we disclosed its preclinical data to support its clinical development in EGFR-mutant NSCLC. Methods: The inhibitory activity of QLH11811 on mutated and wild-type EGFR was tested in engineered cell lines and patient-derived organoid (PDO). The in vivo antitumor activity of QLH11811 was evaluated in the patient-derived xenograft (PDX) model with cis EGFR ex19del/T790M/C797S triple mutations, and the H1975 (cis EGFR L858R/T790M/C797S and cis EGFR ex19del/T790M/C797S), PC-9 (EGFR ex19del), and Ba/F3 (EGFR ex19del/C797S) cell line-derived xenograft (CDX) models. The pharmacokinetic (PK) profile was investigated in animals, and the human PK profile was projected using allometric scaling method. Results: QLH11811 displayed potent anti-proliferation activity against Ba/F3 (EGFR ex19del/T790M/C797S, L858R/T790M/C797S, ex19del/C797S, or L858R/C797S), PC-9 (EGFR ex19del/T790M/C797S), H1975 (EGFR L858R/T790M/C797S, L858R/T790M), H3255 (L858R) and HCC827 (ex19del) with IC50 of 2.6, 3.1, 2.4, 4.1, 51, 50, 27, 21, and 11 nM, respectively. QLH11811 also showed excellent selectivity when compared the above values with its IC50 against Ba/F3 (EGFR wild-type, 61 nM) and A431 (EGFR wild-type, 440nM). QLH11811 demonstrated excellent inhibitory activities against seven osimertinib-resistant PDO models. Daily oral QLH11811 significantly inhibited tumor growth at all doses tested (P Conclusion: The in vitro and in vivo preclinical data demonstrated QLH11811 is a highly potent and selective 4th-generation EGFR TKI with activity against the osimertinib-resistant NSCLC with EGFR C797S mutation. The preclinical PK data supported the efficacious dose of QLH11811 in human would be 103 mg. Citation Format: Shansong Zheng, Wei Deng, Qingmei Zheng, Yingying Yang, Na Li, Tan Pang, Xueying Feng, Simon Taylor, Lina Ma, Yaqiong Wu, Ziwei Zhao. QLH11811, a selective 4th-generation EGFR inhibitor for osimertinib-resistant EGFR-mutant NSCLC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5457.

The flavonoids in Ginkgo biloba L. (ginkgo) have important medicinal uses due to their antioxidant, antitumor, and blood circulation-promoting effects. However, the genetic mechanisms underlying flavonoid biosynthesis in ginkgo remain elusive. Flavonoid 3′, 5′-hydroxylase (F3′5′H) is an important enzyme in flavonoid synthesis. We detected a novel differentially expressed GbF3′5′H1 gene homologous to the F3′5′H enzyme involved in the flavonoid synthesis pathway through transcriptome sequencing. In this study, we characterized this gene, performed an expression analysis, and heterologously overexpressed GbF3′5′H1 in Populus. Our results showed that GbF3′5′H1 is abundant in the leaf and highly expressed during April. We also found four metabolites closely related to flavonoid biosynthesis. Importantly, the contents of 4′,5-dihydroxy-7-glucosyloxyflavanone, epicatechin, and gallocatechin were significantly higher in transgenic plants than in nontransgenic plants. Our findings revealed that the GbF3′5′H1 gene functions in the biosynthesis of flavonoid-related metabolites, suggesting that GbF3′5′H1 represents a prime candidate for future studies (e.g., gene-editing) aiming to optimize ginkgo flavonoid production, especially that of flavan-3-ols.

Chilo suppressalis Walker (Lepidoptera: Crambidae) is a widely destructive pest occurring in rice, particularly in the rice-growing regions of Asia. In recent years, C. suppressalis has developed resistance to several insecticides because of the extensive use of insecticides. The resistance levels to four insecticides were determined among populations from different regions of Sichuan Province, China, using a drop-method bioassay. Based on LC50 values of a laboratory susceptible strain, all field populations showed moderate level of resistance to triazophos (23.9- to 83.5-fold) and were either susceptible or had a low level of resistance to abamectin (2.1- to 5.8-fold). All field-collected populations had a low or moderate level of resistance to chlorpyrifos (1.7- to 47.1-fold) and monosultap (2.7- to 13.5-fold). The synergism experiment indicated that the resistance of the XW19 to triazophos may be associated with cytochrome P450 monooxygenases (P450s), with the highest synergistic ratio (SR) of 3.05-fold and increased ratio (IR) of 2.28-fold for piperonylbutoxide (PBO). The P450 activity of the TJ19 population was the greatest among the six field populations. Moreover, the relative expression levels of four resistance-related P450 genes were detected with qRT-PCR, and the results indicated that CYP324A12, CYP321F3 and CYP9A68 were overexpressed in the resistant population, especially in the XW19 population (by 1.2-, 3.4 -, and 18.0-fold, respectively). In addition, the relative expression levels of CYP9A68 among the CZ19 and TJ19 populations were also enhanced 10.5- and 24.9-fold, respectively. These results suggested that CYP324A12, CYP321F3 and CYP9A68 may be related to the resistance development of C. suppressalis to triazophos.

In this study, Zn/Cu-bearing smelting slag was recycled via an integrated acid dissolution and hematite precipitation method. The slag was dissolved in nitric acid to generate an acid solution containing 23.5 g/L Fe, 4.45 g/L Zn and 2.81 g/L Cu, which was subjected to hydrothermal treatment with the addition of levulinic acid (LA). More than 99.95% of the initial Fe content was removed as hematite particles with diameters of approximately 200 nm, and the residual Fe concentration in the acid was 0.43 mg/L. The generated hematite contained 97.3% Fe2O3, 0.64% ZnO and 0.58% CuO. Greater than 99% of the initial Zn and Cu was retained in the acid and further precipitated as Zn/Cu-bearing solids by adjusting the solution pH to 9. The precipitated Zn/Cu-bearing solids contained 33.6% Zn and 21.7% Cu, whereas the Fe content was less than 0.2%. This paper is the first report of an environmentally friendly approach for recycling smelting slag without generating any hazardous waste.

Based on Ginkgo biloba transcriptome data, 22 SNP loci using high-resolution melting curve technology were validated and genetic diversity was analyzed in three populations. Ginkgo biloba (Ginkgo) is a long-lived dioecious gymnosperm with unique morphological characteristics and play a significant role in evolutionary relationship research. In this study, we used Illumina paired-end RNA sequencing technology for facilitating the gene discovery and single nucleotide polymorphism (SNP) development in Ginkgo. We collected 44.08 G clean bases from six transcriptome datasets. The transcriptome data generated 98,919 unigenes among which 42,667 (43.13%) were successfully annotated. A total of 139,854 putative SNPs were identified; most of the SNPs were transition-type with the nucleotide transitions C–T or A–G. Further, 54532 (38.99%) SNPs were found in protein-coding regions: 23483 (43.06%) were synonymous and 31049 (56.94%) were nonsynonymous. 22 SNPs were subjected to PCR amplification and Sanger sequencing, and all of them were validated. To test the practicability of identified SNPs, these validated SNPs were also assessed by genotyping three natural populations with 84 individuals by high-resolution melting curve (HRM) analysis. Observed and expected heterozygosity varied from 0.0119 to 0.9643 and from 0.0581 to 0.5024, respectively. HRM technology was first applied for the SNP genotyping in Ginkgo. SNPs identified by RNA-Seq provided a useful resource for genetic and genomic studies in Ginkgo. Moreover, the collection of nonsynonymous SNPs annotated with their predicted functional effects also provided a valuable asset for further discovery of genes, identification of gene variants, and development of functional markers.

Blackberry and raspberry have high nutritional, health value, and are popular with consumers for their unique flavors. To explore the relationships between nutrient accumulation, antioxidant substance contents in blackberry and raspberry fruits, and fruit growth and development, seven Rubus cultivars were selected, and contents of the main active substance were determined. “Clode Summit” had the highest soluble sugar and fructose contents, “Chester”—the highest total phenol content, and “Bristol’—the highest anthocyanin content. Generally, the contents of flavonoids and total phenols showed a downward trend with the development of fruit in seven Rubus cultivars, and the content of anthocyanins increased rapidly in the later stage of development. Pearson correlation analysis showed extremely significant correlation between antioxidant activity and the contents of vitamin E, total phenols, and flavonoids. Flavonoids were extremely significantly positively correlated with the content of total phenols, and the contents of flavonoids and anthocyanins in various cultivars were highly negatively correlated. Considering the different nutritional ingredients and active antioxidant substance contents, “Clode Summit”, “Bristol”, and “Chester” are recommended for raw consumption, processing, and medicinal purposes, respectively. These results provide a reference for comparing the main active substance contents in different Rubus cultivars and their changes across fruit development stages.

Blackberry fruit is rich in anthocyanins, showing incomparable nutritional and health value. Anthocyanins are flavonoids that are downstream products of the flavonoid biosynthesis pathway. They are important secondary metabolites with antioxidant and anticancer functions. Therefore, we performed combined transcriptome and metabolome analyses of unripe and ripe blackberry fruits to identify the molecular mechanisms of flavonoid biosynthesis. Herein, we identified 997 differentially expressed metabolites (DEMs) in positive-ion mode, 411 DEMs in negative-ion mode, and 17,566 differentially expressed genes (DEGs). Eight DEMs and 16 DEGs were annotated to the flavonoid biosynthesis pathway. Four DEGs related to flavonoids (unigene 0028088, unigene 0039337, unigene 0063209, and unigene 0014842) were positively correlated with related flavonoid DEMs. Interestingly, 8 DEGs and 11 flavonoid-related DEMs showed significant negative correlations. This study lays a foundation for further research on the biosynthesis and metabolism of flavonoid, providing a reference for the future breeding and cultivation of excellent high-flavonoid germplasm resources and the blackberry-related pharmaceutical industry.

Agroforestry provides opportunities to decrease the levels of carbon dioxide (CO2) released into the atmosphere by increasing the carbon (C) stored in agricultural systems. In agroforestry systems, soil C pools serve as the most important and stable C sink, but there is limited information on the vertical and seasonal variations of soil C pools. In this study, the vertical and seasonal variations of soil organic C (SOC) and its labile pools were measured in five planting systems: a pure ginkgo (Gingko biloba. L) planting system, a pure wheat (Triticum aestivum L.) field, a pure metasequoia (Metasequoia glyptostroboides Hu et Cheng) seedling system, a ginkgo and wheat agroforestry system, and a ginkgo and metasequoia seedling agroforestry system. Among these systems, the ginkgo and wheat system had a significantly higher SOC content than the other systems throughout the year, particularly at depths of 0–10 cm and 10–20 cm. Additionally, the pure ginkgo and pure metasequoia systems had lower SOC contents than the other planting systems, and this decrease was attributed to the relatively limited tree litter input and lower fine root biomass. Microbial biomass C (MBC) and soil readily oxidizable C (ROC) exhibited similar vertical and seasonal variations and reached minimum values in winter. The highest MBC and ROC contents were observed in the ginkgo and wheat system at a depth of 0–10 cm, i.e., 127.3 mg kg−1 and 4.49 g kg−1, respectively. The highest water-soluble organic carbon (WSOC) content was observed in summer at a depth of 0–10 cm, i.e., 472.2 mg kg−1. A Pearson correlation analysis indicated that soil properties were significantly correlated with SOC and labile C fractions. The results suggested that an agroforestry system resulted in a greater increase in the soil C sink; in particular, the ginkgo and wheat system achieved the best results. Basic soil properties played key roles in soil carbon formation. These results provide important information about SOC and labile C fraction dynamics resulting from planting systems and depth variations and strengthen our understanding of soil C sequestration in agroforestry systems.

Ginkgo biloba breeding commonly concentrates on the selection of superior trees that have high flavonoid contents. The flavonoids present in Ginkgo leaves have strong medicinal implications, including anti-dengue, anti-HIV, anticancer, antioxidant and anti-inflammatory properties. Flavonoids play important roles in plant immune responses; however, the molecular mechanisms underlying these interesting responses remain unclear. To obtain a comprehensive understanding, we performed the transcriptome sequencing of Ginkgo with different flavonoid contents. Using an Illumina sequencing platform, we obtained approximately 533,952,528 clean reads. After the sequences were filtered and assembled, the transcriptome data generated 37,625 unigenes, of which 21,472 (57.07%) were successfully annotated in five public databases. Among those genes, many candidates were involved in flavonoid biosynthesis, transport and regulation. Expression profiles were generated, and 457 genes were found to be significantly differentially expressed between the Sample_GB_FH1/2/3 (FH) and Sample_GB_FL1/2/3 (FL) libraries; 246 (53.83%) genes were up-regulated, and 211 (46.17%) were down-regulated. These genes included 14 genes that were enriched in flavonoid transport, 1 MYB gene that encoded a putative transcription factor (TF), and 1 dihydroflavonol reductase (DFR) gene that was involved in the flavonoid pathway. Our results provide comprehensive gene expression information about the Ginkgo transcriptome and can facilitate our understanding of the molecular mechanisms of flavonoid development in Ginkgo. Furthermore, our results markedly expand both the available Ginkgo genetic library and analyses of the species and provide valuable information to the Ginkgo-related pharmaceutical industry.

The response of soil fauna to the litter decomposition process has received considerable attention, but this effect has not been fully examined in agroforestry systems. A 1-year in situ decomposition experiment was carried out in a pure ginkgo plantation and two ginkgo agroforestry systems using a litterbag method (11 different treatments were tested in three systems). We found that the application of different organic materials (crop residues) produced positive effects on the number of soil fauna in the ginkgo planting systems; the mixture of ginkgo leaves and corn leaves was the best performing treatment. Collembola and Acarina were the predominant groups in the litter bags and were mainly responsible for the differences among the treatments. Litter mixing promoted the abundance, richness, and diversity of soil fauna, and significant differences regarding the Shannon–Wiener index of the soil fauna were observed among the 11 treatments in July. Significantly higher soil MBC (microbial biomass carbon) and MBN (microbial biomass nitrogen) were observed in agroforestry systems than in pure ginkgo plantations. These results suggest that the practice of intercrop residue application plays an important role in enhancing soil ecosystem function in ginkgo agroforestry systems and may ultimately contribute to sustainable intercrop production, soil fertility, and local economic diversity.

Spodoptera litura (Fabricius) is a widely distributed worldwide insect pest, particularly of vegetable crops. To determine the sensitivity of S. litura to insecticides, the toxicities of beta-cypermethrin, chlorpyrifos, emamectin benzoate, methoxyfenozide, and indoxacarb were tested from 2014 to 2016 using a diet incorporation assay on populations of S. litura collected from different districts in Sichuan Province and a susceptible strain (Lab-HN). Compared to the Lab-HN strain, field populations showed high levels of resistance to beta-cypermethrin (267.3- to 1789.2-fold). Field populations exhibited low or moderate resistance to chlorpyrifos in 2014 (8.9- to 24.4-fold) and high levels in 2016 (440.5- to 950.5-fold). All populations collected in 2014 and 2016 were susceptible or had a low or moderate level of resistance to indoxacarb (2.2- to 31.0-fold). No field populations exhibited resistance to emamectin benzoate or methoxyfenozide (0.7- to 1.5-fold and 0.9- to 1.4- fold) in 2014, whereas the resistance in 2016 increased to 29.8- to 55.2-fold and 38.1- to 59.0- fold, respectively. A synergist experiment indicated that the Lab-HN strain and field-collected population from Shuangliu in 2016 (SL16 population) to beta-cypermethrin may be related to cytochrome P450 monooxygenases (P450s), with the highest synergism ratios (SR) of piperonyl butoxide (PBO) of 34.4- and 73.3-fold, but not to carboxylesterase (CarE) or glutathione-S- transferase (GST). We detected the activities of detoxification enzymes and found that the ethoxycoumarin O-deethylase activities were also the strongest in the Lab-HN strain and the SL16 population with the highest SR values of 2.6- and 4.1-fold, respectively. Non-denaturing polyacrylamide gel electrophoresis (native PAGE) indicated that the inhibition of in vivo P450 activity caused by PBO was due to the binding of the synergist to the E5 or E6 isozyme, which were absent in the Lab-HN strain.

Background Transitions from traditional cropping systems to afforestation and agroforestry practices are the most common patterns of land use change in China, but little is known about how soil properties and bacterial communities are altered. This study compared four afforestation and agroforestry practices that had been converted from corn and wheat rotation systems for several years with adjacent legacy corn and wheat rotation systems to test the impacts of these land conversions on soil properties and bacterial communities. Results The results revealed that afforestation and agroforestry practices significantly increased soil organic carbon (SOC) and total nitrogen (TN) levels compared with those in the legacy cropping systems, and the ginkgo (Ginkgo biloba L.)-wheat (Triticum aestivum L.) system had the highest SOC and TN contents. Afforestation and agroforestry practices increased the number of unique bacterial operational taxonomic units (OTUs) in the topsoil and subsoil, and the metasequoia (Metasequoia glyptostroboides Hu et Cheng) system had the highest diversity and richness indices. Land conversion had a greater effect on the bacterial community beta diversity in the topsoil than in the subsoil. Redundancy analysis (RDA) showed that SOC, TN, and total phosphorus (TP) were the most important factors explaining variations in the bacterial community structure. The bacterial phyla Proteobacteria, Acidobacteria, Chloroflexi, Actinobacteria, and Nitrospirae dominated both the topsoil and the subsoil, and Proteobacteria was the most abundant. Alphaproteobacteria and Betaproteobacteria were the dominant classes. Only the relative abundance of Actinobacteria was significantly different among land use patterns in both the topsoil and the subsoil, and Nitrospirae was significantly different among land use patterns only in the topsoil, with higher relative abundance in the gingko and metasequoia systems, while other bacterial phyla were not markedly affected by land conversion. Conclusion The results demonstrate that afforestation and agroforestry practices have significant impacts on the soil bacterial community and may be key factors influencing taxonomic shifts and functional variations in bacterial communities, especially in the topsoil.

Blackberry and raspberry flavonoid compounds, including anthocyanins, are known for their nutritional and health benefits. Five representative Rubus fruits (blackberry, red raspberry, yellow raspberry, black raspberry, Rubus hybrid ‘Young’ fruit) were examined. The yellow raspberry ‘Colde Summit’ had the brightest color. The Rubus hybrid ‘Young’ had the highest brix and ‘Colde Summit’ the lowest solidity-acid ratio. In total, 2118 and 1089 known metabolites were identified from five representative Rubus fruits by LC-MS in positive and negative ion modes. Isoquinoline alkaloid biosynthesis was the most enriched pathway in the biosynthesis of other secondary metabolites, followed by flavonoid and anthocyanin biosynthesis. Fewer pathways differed between red raspberry and yellow raspberry than among other Rubus fruits. Significant differences among groups occurred in cyanidin-3-sambubioside content in the anthocyanin pathway, which may be predominantly responsible for color differences. This study provides a theoretical basis for the comprehensive utilization and quality improvement of Rubus.

Litter decomposition is a crucial biogeochemical process linking nutrient cycling and carbon (C) storage in ecosystems, but few studies have investigated this process in agroforestry systems, where tree leaf litter is mixed with intercrop residues. A 360-day in situ litter bag decomposition experiment was conducted in three ginkgo (Ginkgo biloba L.) plantation systems (a ginkgo-corn (Zea mays L.)-wheat (Triticum aestivum L.) system, ginkgo-rape (Brassica napus L.)-soybean (Glycine max (L.) Merr.) system, and pure ginkgo system). Ginkgo leaves decomposed fastest in the ginkgo-corn-wheat system, followed by the ginkgo-soybean-rape system, and the pure ginkgo system. Among all litter species, corn leaves and a ginkgo-corn mixture in the ginkgo-corn-wheat system decomposed fastest and wheat straw most slowly. The Olson’s litter exponential decay model showed the same results; approximately 9 months and slightly less than 27 months was required to decompose 50 and 95% of the litter, respectively. Compared to single-species litter, mixed litters accelerated litter decomposition, except for the ginkgo-wheat mixture. Litter nitrogen (N) loss varied dramatically among litter species during the 360-day in situ incubation. The agroforestry system, litter quality, and mixed effects play important roles in litter decomposition. The Ca content, organic carbon, and living vegetation should be taken into account when studying litter decomposition in agroforestry. Analysis during the litter decomposition process clearly indicated that litter N loss changes dramatically.

Salt-sensitive hypertension is associated with severe organ damage. Generating oxygen radicals is an integral component of salt-induced kidney damage, and activated leukocytes are important in oxygen radical biosynthesis. We hypothesized that a high-salt diet causes the upregulation of immune-related mechanisms, thereby contributing to the susceptibility of Dahl salt-sensitive rats to hypertensive kidney damage. For verifying the hypothesis, we investigated leukocytes adhering to retinal vessels when Dahl salt-sensitive rats were challenged with a high-salt (8% NaCl) diet using acridine orange fluoroscopy and a scanning laser ophthalmoscope. The high-salt diet increased leukocyte adhesion after 3 days and was associated with a significant increase in mRNA biosynthesis of monocyte chemotactic protein-1 and intercellular adhesion molecule-1 (ICAM-1) -related molecules in the kidney. Losartan treatment did not affect increased leukocyte adhesion during the early, pre-hypertensive phase of high salt loading; however, losartan attenuated the adhesion of leukocytes during the hypertensive stage. Moreover, the inhibition of leukocyte adhesion in the pre-hypertensive stage by anti-CD18 antibodies decreased tethering of leukocytes and was associated with the attenuation of functional and morphological kidney damage without affecting blood pressure elevation. In conclusion, a high-salt challenge rapidly increased leukocyte adhesion through the over-expression of ICAM-1. Increased leukocyte adhesion in the pre-hypertensive stage is responsible for subsequent kidney damage in Dahl salt-sensitive rats. Immune system involvement may be a key component that initiates kidney damage in a genetic model of salt-induced hypertension.

Additional file 1: Table S1. Primer pairs for quantitative real-time PCR. Table S2. Results of the comparison of sequences to the reference genome.

Surface sediment samples were collected from a source water reservoir in Zhejiang Province, East of China to investigate pollution characteristics and potential ecological risk of heavy metals. The BCR sequential extraction method was used to determine the four chemical fractions of heavy metals such as acid soluble, easily reducible, easily oxidizable and residual fractions. The heavy metals pollution and potential ecological risk were evaluated systematically using geoaccumulation index (Igeo) and Hakanson potential ecological risk index (H′). The results showed that the sampling sites from the estuaries of tributary flowing through downtowns and heavy industrial parks showed significantly (p

The white-backed planthopper, Sogatella furcifera (Horvath) (Hemiptera, Delphacidae), is an energetic rice insect pest in rice production or rice-growing areas. Due to excessive use of the chemical insecticide, S. furcifera has produced the high resistance to some frequently used insecticides. In this paper, the resistance levels of S. furcifera from the eight different areas of Sichuan Province against the five chemicals were monitored by using the rice seedling dipping during 2017–2018 to understand the resistance levels. The results showed that most of all populations have developed low or moderate level of resistance for chlorpyrifos (3.4 to 44.3-fold) and thiamethoxam (3.9- to 15.5-fold), the populations in the LS (1.7 to 5.4- fold)and WS (1.6 to 5.0- fold) regions were still sensitive or low resistance levels compared with other local populations. Almost all populations displayed the susceptible to imidacloprid (0.9- to 5.0-fold), buprofezin (0.9- to 4.3-fold) or low levels of resistance to pymetrozine (1.5- to 6.8-fold). The synergism experiment indicated that P450 enzymes may be important contributed to the metabolic detoxification of chlorpyrifos. The cross-resistance bioassay showed that there was no cross-resistance between chlorpyrifos and triflumezopyrim, but for sulfoxaflor, in the XY17 population. The relative expression level of twelve insecticide resistant-related P450 genes were analyzed by using qRT-PCR and found that CYP4C77, CYP418A1, CYP418A2, CYP408A3 and CYP6ER4 were significantly more expressed in the 3rd-instar nymph of the XY17 and XY18 field populations. To determine the main resistant-related P450 gene for chlorpyrifos, the relative expression level of five P450 genes were detected by using qRT-PCR from the G2 and G4 generation of XY17 under the pressure with LC50 of chlorpyrifos. The results showed that CYP6ER4 was significantly up-regulated expression in XY17 G2 and G4 generations population over 700-fold (P

Cold-rolling sludge (CRS), a typical solid waste, is mass produced during strip cleaning and rolling. Given its low Fe concentration, CRS cannot be directly recycled as iron pellet for steel making, and it is commonly solidified before safety landfill. Herein, CRS was purified as commercial iron concentrate powder via a coupled alkaline hydrothermal and calcination route. The sludge contained 45.6 wt% Fe, and its major impurities included 6.2 wt% Al, 13.5 wt% carbon and 9.7 wt% halite. After the hydrothermal treatment, the contents of Al and carbon in the treated sludge decreased to 0.16 and 12.1 wt%, respectively, and halite was almost completely removed. Subsequently, the Fe content in the treated sludge increased to 60.6 wt%, which was higher than that obtained with conventional methods, e.g. direct washing (51.4 wt% Fe), alkaline washing (52.7 wt% Fe), Fenton oxidation (52.4 wt%) and direct calcination (53.7 wt%). The treated sludge was further calcined at 800 °C. The calcined product showed unchanged amounts of Al, carbon content that dropped to 3.5 wt% and Fe that steadily rose to up to 66.1 wt%. These findings enabled the calcined product to be used as a high-grade iron concentrate powder. NaOH concentration, hydrothermal temperature and time were optimised to 8 M, 160 °C and 6 h at the laboratory scale, respectively. The solid–liquid ratio and recycling of supernatant were limited to 1 and 4 at the pilot scale, respectively. Under optimal conditions, the supernatant was reused four times, and the obtained product contained 62.7 wt% Fe, which meets the commercial standard of iron concentrate powder (>60 wt%). This coupled route was considerably effective for recycling CRS as iron concentrate powder and was successfully employed at the pilot scale.