Fulvic acid alleviates cadmium-induced root growth inhibition by regulating antioxidant enzyme activity and carbon-nitrogen metabolism in apple seedlings.

Introduction: Substantial previous studies have reported that fulvic acid (FA) application plays an important role in Chinese agricultural production. However, little is known about the mechanisms for using FA to increase apple trees resistance to Cd toxicity. In order to clarify the mechanism underlying FA alleviation in Cd-induced growth inhibition in apple seedlings. Methods: Herein, we treated M9T337 seedlings to either 0 or 30 µM/L Cd together with 0 or 0.2 g/L FA and analyzed the root growth, antioxidant enzyme activities, carbon (C) assimilation, nitrogen (N) metabolism, and C and N transport. Results: The results presented that, compared with CK (without Cd addition or FA spraying application), Cd poisoning significantly inhibited the root growth of apple seedlings. However, this Cd-induced root growth inhibition was significantly alleviated by FA spraying relative to the Cd treatment (Cd addition alone). On the one hand, the mitigation of inhibition effects was due to the reduced oxidative damage by enhancing antioxdiant enzyme (SOD, POD, and CAT) activities in leaves and roots. On the other hand, this growth advantage demonstrated compared to the Cd treatment was found to be associated with the strengthen of photosynthetic performance and the elevation of C and N metabolism enzymes activities. Meanwhile, we also found that under Cd stress condition, the distribution of C and N nutrients in apple seedlings was optimised by FA spraying application relative to the Cd treatment, according to the results of 13C and 15N tracing. Conclusion: Conclusively, our results suggested that the inhibitory effect of Cd on apple seedlings root growth was alleviated by FA through regulating antioxdiant capacities and C and N metabolism.

Enhanced ferromagnetism, perpendicular magnetic anisotropy and high Curie temperature in the van der Waals semiconductor CrSeBr through strain and doping.

Two-dimensional (2D) intrinsic van der Waals ferromagnetic semiconductor (FMS) crystals with strong perpendicular magnetic anisotropy and high Curie temperature (TC) are highly desirable and hold great promise for applications in ultrahigh-speed spintronic devices. Here, we systematically investigated the effects of a biaxial strain ranging between -8% and +8% and doping with different charge carrier concentrations (≤0.7 electrons/holes per unit cell) on the electronic structure, magnetic properties, and TC of monolayer CrSeBr by combining first-principles calculations and Monte Carlo (MC) simulations. Our results demonstrate that the pristine CrSeBr monolayer possesses an intrinsic FMS character with a band gap as large as 1.03 eV, an in-plane magnetic anisotropy of 0.131 meV per unit cell, and a TC as high as 164 K. At a biaxial strain of only 0.8% and a hole density of 5.31 × 1013 cm-2, the easy magnetization axis direction transitions from in-plane to out-of-plane. More interestingly, the magnetic anisotropy energy and TC of monolayer CrSeBr are further enhanced to 1.882 meV per unit cell and 279 K, respectively, under application of a tensile biaxial strain of 8%, and the monolayer retains its semiconducting properties throughout the entire range of investigated strains. It was also found that upon doping monolayer CrSeBr with holes with a concentration of 0.7 holes per unit cell, the perpendicular magnetic anisotropy and TC are increased to 0.756 meV per cell and 235 K, respectively, and the system tends to become metallic. These findings will help to advance the application of 2D intrinsic ferromagnetic materials in spintronic devices.

PI3Kα inhibitor GNE-493 triggers antitumor immunity in murine lung cancer by inducing immunogenic cell death and activating T cells.

Phosphatidylinositol 3-kinase (PI3K) is frequently hyperactivated in cancer, playing pivotal roles in the pathophysiology of both malignant and immune cells. The impact of PI3K inhibitors on the tumor microenvironment (TME) within lung cancer remains largely unknown. In this study, we explored the regulatory effects of GNE-493, an innovative dual inhibitor of PI3K and mammalian target of rapamycin (mTOR), on the TME of lung cancer. First, through the analysis of The Cancer Genome Atlas-lung squamous cell carcinoma (LUSC) cohort, we found PIK3CA to be related to CD8 T cells, which may affect the overall survival rate of patients by affecting CD8 function. We herein demonstrated that GNE-493 can significantly inhibit tumor cell proliferation and promote cell apoptosis while increasing the expression of the immunogenic death-related molecules CRT and HSP70 using in vitro cell proliferation and apoptosis experiments on the murine KP lung cancer cell line and human A549 lung cancer cell line. Next, through the establishment of an orthotopic tumor model in vivo, it was found that after GNE-493 intervention, the infiltration of CD4+ and CD8+ T cells in mouse lung tumor was significantly increased, and the expression of CRT in tumors could be induced to increase. To explore the mechanisms underlying PI3K inhibition-induced changes in the TME, the gene expression differences of T cells in the control group versus GNE-493-treated KP tumors were analyzed by RNA-seq, and the main effector pathway of anti-tumor immunity was identified. The IFN/TNF family molecules were significantly upregulated after GNE-493 treatment. In summary, our findings indicate that GNE-493 promotes immunogenic cell death in lung cancer cells, and elucidates its regulatory impact on molecules associated with the adaptive immune response. Our study provides novel insights into how PI3K/mTOR inhibitors exert their activity by modulating the tumor-immune interaction.

The PpMYB75-PpDFR module reveals the difference between 'SR' and its bud variant 'RMHC' in peach red flesh.

'Red Meat Honey Crisp (RMHC)' has been widely cultivated by growers in recent years due to its early maturity, and red meat type characteristics. As a bud variant of 'Super Red (SR)' peach, red flesh is the most distinctive characteristic of 'Red Meat Honey Crisp (RMHC)'. However, the mechanism of red flesh formation in 'RMHC' remains unclear. In this study, 79 differentially produced metabolites were identified by metabolomics analysis. The anthocyanin content in 'RMHC' was significantly higher than that in 'SR' during the same period, such as cyanidin O-syringic acid and cyanidin 3-O-glucoside. Other flavonoids also increased during the formation of red flesh, including flavonols (6-hydroxykaempferol-7-O-glucoside, hyperin), flavanols (protocatechuic acid, (+)-gallocatechin), and flavonoids (chrysoeriol 5-O-hexoside, tricetin). In addition, transcriptomic analysis and RT-qPCR showed that the expression levels of the flavonoid synthesis pathway transcription factor MYB75 and some structural genes, such as PpDFR, PpCHS, PpC4H, and PpLDOX increased significantly in 'RMHC'. Subcellular localization analysis revealed that MYB75 was localized to the nucleus. Yeast single hybridization assays showed that MYB75 bound to the cis-acting element CCGTTG of the PpDFR promoter region. The MYB75-PpDFR regulatory network was identified to be a key pathway in the reddening of 'RMHC' flesh. Moreover, this is the first study to describe the cause for red meat reddening in 'RMHC' compared to 'SR' peaches using transcriptomics, metabolomics and molecular methods. Our study identified a key transcription factor involved in the regulation of the flavonoid synthetic pathway and contributes to peach breeding-related efforts as well as the identification of genes involved in color formation in other species.

Effects of Fulvic Acid on Growth and Nitrogen Utilization Efficiency in M9T337 Seedlings.

Both fulvic acid (FA) and nitrogen (N) play important roles in agricultural production in China. Plants typically show a higher nitrogen utilization efficiency (NUE) under FA application. However, the role of FA application in apple growth and NUE remains unclear. A hydroponic culture experiment was performed, and M9T337 seedlings (a dwarf apple rootstock) were used as the experimental subjects. The biomass, photosynthesis, accumulation, and distribution of photosynthates, N absorption and assimilation, and relative gene expression in the seedlings were examined after treatment with five different concentrations of FA (0, 60, 120, 180, and 240 mg·L-1, represented by CK, FA1, FA2, FA3, FA4, respectively). The results showed that the seedling dry weight and 15NUE were enhanced by FA, and both were highest under the FA2 (the concentration of fulvic acid is 120 mg·L-1) treatment. Further analysis revealed that under the FA2 treatment, the root morphology was optimized, and the root activity was relatively high. Compared with CK (control, the concentration of fulvic acid is 0 mg·L-1), the FA2 treatment strengthened photosynthesis, elevated the key enzyme activities related to C metabolism, upregulated the gene expression of sugar transport proteins, and increased the root sorbitol and sucrose contents, which suggested that the FA2 treatment optimally affected the root growth and N absorption because it enhanced photosynthate synthesis and the leaf-to-root translocation of photoassimilates. The seedlings in the FA2 treatment group also showed a significantly higher NO3- influx rate and NRT (nitrate transporter) gene expression in the roots. Moreover, relatively high N metabolism-related enzyme activities in the leaves and roots were also observed under the FA2 treatment. The isotope labeling results showed that the optimal FA2 supply not only promoted seedling 15N absorption but also optimized the distribution of C and N in the seedlings. These results suggested that an optimal FA supply (120 mg·L-1) enhanced seedling NUE by strengthening photoassimilate synthesis and transport from leaves to roots, regulating N absorption, assimilation, and distribution.

Hole-Doping-Induced Perpendicular Magnetic Anisotropy and High Curie Temperature in a CrSX (X = Cl, Br, I) Semiconductor Monolayer.

A large perpendicular magnetic anisotropy and a high Curie temperature (TC) are crucial for the application of two-dimensional (2D) intrinsic ferromagnets to spintronic devices. Here, we investigated the electronic and magnetic properties of carrier-doped Van der Waals layered CrSX (X = Cl, Br, I) ferromagnets using first-principles calculations. It was found that hole doping can increase the magnitude of the magnetic anisotropy energy (MAE) and change the orientation of the easy magnetization axis at small doping amounts of 2.37 × 1013, 3.98 × 1012, and 3.33 × 1012/cm2 for CrSCl, CrSBr, and CrSI monolayers, respectively. The maximum values of the MAE reach 57, 133, and 1597 µeV/u.c. for the critical hole-doped CrSCl, CrSBr, and CrSI with spin orientation along the (001) direction, respectively. Furthermore, the Fermi energy level of lightly hole-doped CrSX (X = Cl, Br, I) moves into the spin-up valence band, leading to the CrSX (X = Cl, Br, I) magnetic semiconductor monolayer becoming first a half-metal and then a metal. In addition, the TC can also be increased up to 305, 317, and 345 K for CrSCl, CrSBr, and CrSI monolayers at doping amounts of 5.94 × 1014, 5.78 × 1014, and 5.55 × 1014/cm2, respectively. These properties suggest that the hole-doping process can render 2D CrSX (X = Cl, Br, I) monolayers remarkable materials for application to electrically controlled spintronic devices.

Modulation of the assembly fashion among metal-organic frameworks for enantioretentive epoxide activation.

Highly enantioretentive alcoholysis of epoxides is an important way to synthesize enantiopure ß-alkoxy alcohols, which are irreplaceable intermediates demanded by biomedicines, fine chemicals and other industries. In this report, we exploit a series of Zr-based metal-organic frameworks (Zr-MOFs) as the catalysts to achieve high activity and enantioretentivity in the alcoholysis of styrene oxide via modulating their assembly fashions. It is explored that hcp-UiO-66 not only exhibits a ∼10 fold improved catalytic activity than both hxl-CAU-26 and fcc-UiO-66 of varied assemblies but also maintains superior product enantioretentivity. Theoretic calculations together with experimental proof discloses the origin of distinct catalytic activity caused by different assembly fashions. This assembly modulation strategy offers a potential protocol for seeking high-performance catalysts among MOFs by virtue of their rich polymorphisms.

MdSWEET23, a sucrose transporter from apple (Malus × domestica Borkh.), influences sugar metabolism and enhances cold tolerance in tomato.

Photosynthetic products in most fleshy fruits are unloaded via the apoplasmic pathway. Sugar transporters play an important role in the apoplasmic unloading pathway and are involved in sugar transport for fruit development. The MdSWEET23, cloned from ''Hanfu'' apple (Malus × domestica Borkh.) fruits, belongs to Clade III of the SWEET family. Subcellular localization revealed that MdSWEET23 is localized on the plasma membrane. ß-glucuronidase activity assays showed that MdSWEET23 was primarily expressed in the sepal and carpel vascular bundle of apple fruits. Heterologous expression assays in yeast showed that MdSWEET23 functions in sucrose transport. The overexpression of MdSWEET23 in the ''Orin" calli increased the soluble sugar content. The silencing of MdSWEET23 significantly reduced the contents of sucrose and sorbitol in apple fruits. Ectopic overexpression of MdSWEET23 in tomato altered sugar metabolism and distribution in leaves and fruits, causing a reduction in photosynthetic rates and plant height, enhanced cold stress tolerance, and increased the content of sucrose, fructose, and glucose in breaking color fruits, but did not increase sugar sink potency of tomato fruits.

Effects and Mechanism Analysis of Non-Bagging and Bagging Cultivation on the Growth and Content Change of Specific Substances of Fuji Apple Fruit.

The experiment was conducted at the Taidong Base of Shandong Institute of Pomology, Tai'an City, Shandong Province, China, from May to October 2022. Using Fuji Apple Tianhong.2/SH/Malus robusta (M. robusta) as experimental materials, the differences and mechanisms of the effects of non-bagging and bagging cultivation on the growth and changes in some substance content of Fuji apple fruits were studied. The results showed that compared with bagging, non-bagging cultivation increased single fruit weight and decreased fruit shape index, increased fruit sugar content, reduced acid content, and increased taste. It increased the content of vitamin C (VC) and protein in the fruit, increased the types and content of aromatic components in the fruit, significantly increased the activity of sugar- and acid-related enzymes, and improved the antioxidant capacity of the fruit. Compared to bagging cultivation, non-bagging cultivation improved the weight, taste (sugar acid ratio), and aroma of Fuji fruit, which is related to increasing the content of auxin (IAA), cytokinin (ZR), and salicylic acid (SA) and reducing the content of abscisic acid (ABA) in the fruit, as well as increasing the content of medium and trace elements calcium (Ca), iron (Fe), manganese (Mg), and boron (B). One of the mechanisms involved is the significant increase in gene expression related to phenylpropanoid biosynthesis, pentose and glucuronate interconversion, starch and sucrose metabolism, zeatin biosynthesis, microtubules, motor proteins microtubule movement, xyloglucan metabolic process, cell division, and peroxidase activity. In short, non-bagging cultivation is more conducive to improving the intrinsic quality and flavor development of Fuji apples, and one of the mechanisms is that non-bagging cultivation is beneficial for increasing the expression of related genes.

Granule Dendrobii suppresses chronic atrophic gastritis induced by N-methyl-N'-nitro-N-nitrosoguanidine by modulating the gastrointestinal bacteria in rats.

Chronic atrophic gastritis (CAG) is an important stage in the transformation of the normal gastric mucosa into gastric cancer. Granule Dendrobii (GD), a proprietary Chinese medicine, has proven clinical efficacy in treating CAG. GD might promote the reversal of precancerous lesions by improving them in CAG patients. However, the mechanism of GD in CAG treatment is relatively less understood. Here, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced CAG rats were treated with GD and its efficacy was evaluated by observing the changes in the rats' weight and the pathology of gastric tissues. The potential effect of GD on the bacteria was predicted and verified in the large and small intestines and stomachs of CAG rats using amplicon sequencing and RT-qPCR. The results showed that GD could ameliorate the symptoms of body weight loss in CAG rats. Hematoxylin-Eosin (HE) and Alcian Blue (AB) staining showed that GD significantly improved the pathological state of the gastric mucosa in CAG rats. The relative abundance (RA) of Lactobacillus and Turicibacter significantly decreased after GD intervention compared with that of the model group (P < 0.05), indicating that GD might improve CAG by regulating the RA of Lactobacillus and Turicibacter. These findings revealed that Lactobacillus and Turicibacter as bacteria agents associated with gastritis, have the potential to inhibit gastric cancer, especially Turicibacter maybe another pathogen of CAG besides Helicobacter pylori (HP), which is worthy of further study. Meanwhile, the findings provided new ideas and materials for the research and development of new CAG drugs.

Apple-marigold intercropping improves soil properties by changing soil metabolomics and bacterial community structures.

Marigold can protect crops against soil-borne diseases. However, the effects of intercropping with marigold on apple rhizosphere soils are not known. In this study, we investigated the metabolite profiles and bacterial community structures in rhizosphere soils of the apple-marigold intercropping system by high-throughput sequencing and soil metabolomics. The results show that intercropping marigold could significantly enhance soil moisture, nitrogen, and enzyme activities compared with clean tillage. The soil metabolite profiles and the soil bacterial community structures in the rhizosphere soils were different between the inter-and mono-cropping systems. Among nine metabolites, carbohydrates were more increased in the intercropping system than in the monocropping system. Pathway enrichment analysis revealed that the greatest differential, in terms of metabolic pathway, was starch and sucrose metabolism. Moreover, intercropping marigold significantly increased the relative abundance of plant growth promoting bacteria in rhizosphere soils, such as Rhizobiales, Pseudomonadales, and Bacillales. These results indicate that marigold intercropping positively affected the apple orchard's soil quality and may provide a new intercropping technique to improve soil fertility in orchards and promote plant growth.

Transcriptome and metabolome analysis of flavonol synthesis in apricot fruits.

Introduction: Apricot fruits are edible and serve as a source of medicinal compounds. Flavonols are important plant secondary metabolites that have antioxidant and antitumor effects and may promote cardiovascular health. Methods: The flavonoid content in three stages of the 'Kuijin' and the 'Katy' was observed, followed by the combination of metabolome and transcriptome analysis to explore the metabolic basis of flavonol synthesis. Results: The differences in the metabolite contents between stages (of the same cultivar) and between cultivars (at the same stage) revealed decreases in the flavonoid content as fruits developed (i.e., from 0.28 mg/g to 0.12 mg/g in 'Kuijin' and from 0.23 mg/g to 0.05 mg/g in 'Katy'). To decipher the regulation of flavonol synthesis in apricot (Prunus armeniaca L.), the metabolomes and transcriptomes of fruit pulp at three developmental stages of 'Kuijin' and the 'Katy' were analyzed. A total of 572 metabolites were detected in 'Kuijin' and the 'Katy' pulp, including 111 flavonoids. The higher flavonol content young 'Kuijin' fruits at 42 days after full bloom is mainly due to 10 types of flavonols. Three pairs of significant differences in flavonol content were identified. From these three comparison groups, three structural genes were strongly correlated with the levels of 10 types of flavonols (Pearson correlation coefficients > 0.8, p value < 0.05), including PARG09190, PARG15135, and PARG17939. The weighted gene co-expression network analysis showed that the turquoise module genes were highly correlated with flavonol contents (P < 0.01). There were 4897 genes in this module. Out of 4897 genes, 28 transcription factors are associated with 3 structural genes based on weight value. Two of the transcription factors are not only associated with PARG09190 but also with PARG15135, indicating their critical importance in the flavonols biosynthesis. The two TFs are PARG27864 and PARG10875. Discussion: These findings provide new insights into the biosynthesis of flavonols and may explain the significant differences in flavonoid content between the 'Kuijin' and the 'Katy' cultivars. Moreover, it will aid in genetic improvement to enhance the nutritional and health value of apricots.

Yiwei decoction promotes apoptosis of gastric cancer cells through spleen-derived exosomes.

Yiwei decoction (YWD) is a formula of traditional Chinese medicine (TCM) that is clinically effective for the prevention and treatment of gastric cancer recurrence and metastasis. According to the theory of TCM, YWD tonifies the body and strengthens the body's resistance to gastric cancer recurrence and metastasis potentially via the immune regulation of the spleen. The aims of the present study were to investigate whether YWD-treated spleen-derived exosomes in rats inhibit the proliferation of tumor cells, to elucidate the anticancer effects of YWD, and to provide evidence supporting the use of YWD as a new clinical treatment for gastric cancer. Spleen-derived exosomes were obtained by ultracentrifugation and identified by transmission electron microscopy, nanoparticle tracking analysis, and western blot analysis. The location of the exosomes in tumor cells was then determined by immunofluorescence staining. After tumor cells were treated with different concentrations of exosomes, the effect of exosomes on cell proliferation was determined by cell counting kit 8 (CCK8) and colony formation assays. Tumor cell apoptosis was detected by flow cytometry. Particle analysis and western blot analysis identified the material extracted from spleen tissue supernatant as exosomes. Immunofluorescence staining showed that spleen-derived exosomes were taken up by HGC-27 cells, and the CCK8 assay confirmed that the relative tumor inhibition rate of YWD-treated spleen-derived exosomes in the 30 µg/mL reached 70.78% compared to control exosomes in the 30 µg/mL (p < 0.05). Compared to control exosomes in the 30 µg/mL, the colony formation assay indicated that YWD-treated spleen-derived exosomes in the 30 µg/mL colonies have decreased by 99.03% (p < 0.01). Moreover, flow cytometry analysis showed that treatment with YWD-treated exosomes in the 30 µg/mL increased the apoptosis rate to 43.27%, which was significantly higher than that of the control group in the 30 µg/mL (25.91%) (p < 0.05). In conclusion, spleen-derived exosomes from YWD-treated animals inhibit the proliferation of HGC-27 cells via inducing apoptosis, suggesting that spleen-derived exosomes are involved in mediating the antitumor effect of YWD. These results demonstrated a novel exosome-mediated anticancer effect of YWD as a TCM formula, thereby supporting the use of YWD-treated exosomes as a new approach for the clinical treatment of gastric cancer.

Experimental evidence of shikonin as a novel intervention for anti-inflammatory effects.

Shikonin is a natural product with antioxidant and anti-inflammatory activities. The biological activity of shikonin is still not fully understood, as well as its association with innate immunity and immune and inflammatory bowel disease (IBD) in humans. In this study, the toxicity of shikonin on Raw264.7 cells was assayed by MTT, and polarization of inflammatory macrophages was determined by flow cytometry. The results showed that shikonin can inhibit the polarization of macrophages towards M1 type and significantly inhibited the production of NO in the concentration range of 0.5-1 µM. In addition, after treatment with shikonin, the production of IL-1ß and TNF-α was significantly decreased. After shikonin administration, the body weight loss and decrease of colon length were significantly suppressed in DSS-treated colitis C57BL/6 mice. The pro-inflammatory cytokines TNF-α and IL-1ß in colonic homogenate were significantly decreased. Shikonin treatment resulted in a notable improvement in the histopathological manifestations in DSS-treated animals at 25/50 mg/kg. Meanwhile, we found that shikonin can regulate differentiation of T helper 17 cell (Th17)/regulatory T cell (Treg), thereby regulating the balance of Th17/Treg cells and exerting an anti-inflammatory effect in IBD animal models. In conclusion, we found that shikonin protects against DSS-induced acute colitis by, among other things, reducing immune cell infiltration, polarizing macrophages, and regulating Th17/Treg differentiation, as well as by downregulating the release of inflammatory cytokines. These findings showed that shikonin can improve inflammation by affecting macrophage polarization. Our experimental data provide experimental evidence and theory basis for research on anti-inflammatory effects for the shikonin as health or functional food.

Exogenous 24-epibrassinolide promoted growth and nitrogen absorption and assimilation efficiency of apple seedlings under salt stress.

Introduction: High salinity significantly hampers global agricultural productivity. Plants typically undergo lower nitrogen utilization efficiency (NUE) under salt stress. As an active byproduct from brassinolide biosynthesis, 24-epibrassinolide (EBR) is involved in regulating the stress-treated plant N absorption and assimilation. However, the exogenous EBR application effects' on N absorption and assimilation in apple exposed to the salt-stressed condition remains unclear. Methods: We sprayed exogenous EBR (0.2 mg L-1) on apple dwarf rootstock (M9T337) seedlings (growing hydroponically) under salt (NaCl) stress in a growth chamber. We analyzed the seedling development, photosynthesis and its-mediated C fixation, N ( NO 3 - ) absorption and assimilation in reponse to exogenous EBR application under salt stress. Results: The findings demonstrated that NaCl stress greatly hampered seedlings' root growth and that exogenous EBR application obviously alleviated this growth suppression. Exogenous EBR-treated plants under NaCl stress displayed the more ideal root morphology and root activity, stronger salt stress tolerance and photosynthetic capacity as well as higher C- and N-assimilation enzyme activities, NO 3 - ion flow rate and nitrate transporter gene expression level than did untreated plants. Furthermore, the results of isotope labeling noted that exogenous EBR application also enhanced 13C-photoassimilate transport from leaves to roots and 15 NO 3 - transport from roots to leaves under NaCl stress. Conclusion: Our findings imply that exogenous EBR application, through strengthening photosynthesis, C- and N-assimilation enzyme activities, nitrate absorption and transport as well as synchronized optimizing the distribution of seedlings' C and N, has a fundamental role in improving NUE in apple rootstock seedlings under salt stress.

Inhibitory effect and underlying mechanism of cinnamon and clove essential oils on Botryosphaeria dothidea and Colletotrichum gloeosporioides causing rots in postharvest bagging-free apple fruits.

Bagging-free apple is more vulnerable to postharvest disease, which severely limits the cultivation pattern transformation of the apple industry in China. This study aimed to ascertain the dominant pathogens in postharvest bagging-free apples, to evaluate the efficacy of essential oil (EO) on inhibition of fungal growth, and to further clarify the molecular mechanism of this action. By morphological characteristics and rDNA sequence analyses, Botryosphaeria dothidea (B. dothidea) and Colletotrichum gloeosporioides (C. gloeosporioides) were identified as the main pathogens isolated from decayed bagging-free apples. Cinnamon and clove EO exhibited high inhibitory activities against mycelial growth both in vapor and contact phases under in vitro conditions. EO vapor at a concentration of 60 µL L-1 significantly reduced the incidence and lesion diameter of inoculated decay in vivo. Observations using a scanning electron microscope (SEM) and transmission electron microscope (TEM) revealed that EO changed the mycelial morphology and cellular ultrastructure and destroyed the integrity and structure of cell membranes and major organelles. Using RNA sequencing and bioinformatics, it was demonstrated that clove EO treatment impaired the cell membrane integrity and biological function via downregulating the genes involved in the membrane component and transmembrane transport. Simultaneously, a stronger binding affinity of trans-cinnamaldehyde and eugenol with CYP51 was assessed by in silico analysis, attenuating the activity of this ergosterol synthesis enzyme. Moreover, pronounced alternations in the oxidation/reduction reaction and critical materials metabolism of clove EO-treated C. gloeosporioides were also observed from transcriptomic data. Altogether, these findings contributed novel antimicrobial cellular and molecular mechanisms of EO, suggesting its potential use as a natural and useful preservative for controlling postharvest spoilage in bagging-free apples.

Genome-wide analysis of the 6B-INTERACTING PROTEIN1 gene family with functional characterization of MdSIP1-2 in Malus domestica.

Trihelix transcription factors consist of five subfamilies, including GT-1, GT-2, SH4, GTγ, and SIP1, which play important roles in the responses to biotic and abiotic stresses, however, seldom is known about the role of the SIP1 genes in apples. In this study, 12 MdSIP1 genes were first identified in apples by genome-wide analysis, and contained conserved MYB/SANT-like domains. Expression patterns analyses showed that the MdSIP1 genes had different tissue expression patterns, and different transcription levels in response to abiotic stresses, indicating that MdSIP1s may play multiple roles under various abiotic stresses. Among them, the MdSIP1-2 gene was cloned and ectopic transformed into Arabidopsis, and its biology function was identified. The subcellular localization showed that MdSIP1-2 protein was specifically localized in the nucleus, and that overexpression of MdSIP1-2 promoted the development of lateral roots, increased abscisic acid (ABA) sensitivity, and improved salt and drought tolerance. These findings suggested that MdSIP1-2 plays an important role in root development, ABA synthesis, and salt and drought stress tolerance. In conclusion, these results lay a solid foundation for determining the role of MdSIP1 in the growth and development and abiotic stress tolerance of apples.

Residual levels and dietary intake risk assessment of 11 pesticides in apricots from different ecological planting regions in China.

The frequent and massive use of pesticides has led to pesticide residues in apricot, threatening food safety and human health. A reliable and simple modified QuEChERS method with ultra-performance liquid chromatography-tandem mass spectrometry was developed for the simultaneous determination of 11 pesticides in apricot. Method validation indicated that satisfied linearity (R2 ≥ 0.9959), accuracy (recoveries of 72-119%), sensitivity (limits of detection, 0.03-0.30 µg/kg; limits of quantification, 0.13-1.00 µg/kg), and precision (relative standard deviations ≤ 11.9%), and matrix effects were 0.89-1.13. Apricot samples from different ecological regions in China were collected and tested using the proposed methods. Monitoring results were used to assess the dietary intake risk of Chinese populations of different ages and genders. Dietary risk assessment revealed that the risk quotients were 0.003-1.184% for different gender and age groups in China, indicating none unacceptable public health risk for general population. This work was thus significant in developing a simpler, more efficient and economical analysis method and food safety risks of the 11 pesticides on apricot and facilitated the establishment of maximum residue limits.