Selenium-Rich Black Soldier Fly Supplementation Enriches Serum Indexes and Egg Selenium Content in Laying Hens.

A certain amount of selenium (Se) is usually added to the diet of laying hens to improve the quality and nutritional value of eggs. The present study was carried out to investigate the effect of selenium-rich black soldier fly (BSF) supplementation in diets on laying production performance, egg quality, serum indexes, and egg selenium content of Hy-line variety brown laying hens. A total of 288 at 49-week-old healthy laying hens were divided into 3 treatment groups with 6 replicates per group and 16 hens per replicate using a single-factor completely randomized design. Treatments consisted of (1) control (basal diet without supplemental Se), (2) 0.30 mg/kg supplemental Se, (Se as sodium selenite, SS), and (3) 0.30 mg/kg supplemental Se (Se as selenium-rich black soldier fly, SE-BSF). Laying performance was not affected by dietary Se. There was no effect of selenium-rich BSF on egg quality (P > 0.05). The contents of malonaldehyde (MDA) were significantly reduced (P < 0.05). On the contrary, dietary Se supplementation increased the activity of superoxide dismutase (SOD, P < 0.05) and catalase (CAT, P < 0.05) and increased the concentration of reduced glutathione (P < 0.05). In addition, selenium-rich BSF supplementation significantly increased the Se content of eggs (P < 0.05). These results indicate that Se supplementation did not affect laying production performance and egg quality of laying hens, but the supplementation could improve antioxidant capacity and increased the Se content of eggs.

A comparison study on polysaccharides extracted from banana flower using different methods: Physicochemical characterization, and antioxidant and antihyperglycemic activities.

This work investigated and compared the physicochemical characteristics, and antioxidant and antihyperglycemic properties in vitro of polysaccharides from a single banana flower variety (BFPs) extracted by different methods. BFPs extracted using hot water (HWE), acidic (CAE), alkaline (AAE), enzymatic (EAE), ultrasonic (UAE) and hot water-alkaline (HAE) methods showed different chemical composition, monosaccharide composition, molecular weight, chain conformation and surface morphology, but similar infrared spectra characteristic, main glycosidic residues, crystalline internal and thermal stability, suggesting that six methods have diverse impacts on the degradation of BFPs without changing the main structure. Then, among six BFPs, the stronger antioxidant activity in vitro was found in BFP extracted by HAE, which was attributed to its maximum uronic acid content (21.67 %) and phenolic content (0.73 %), and moderate molecular weight (158.48 kDa). The highest arabinose and guluronic acid contents (18.59 % and 1.31 % in molar ratios, respectively) and the lowest uronic acid content (14.30 %) in BFP extracted by HWE contributed to its better α-glucosidase inhibitory activity in vitro (66.55 %). The data offered theoretical evidence for choosing suitable extraction methods to acquire BFPs with targeted biological activities for applications, in which HAE and HWE could serve as beneficial methods for preparing antioxidant BFP and antihyperglycemic BFP, respectively.

Tomato CYP94C1 inactivates bioactive JA-Ile to attenuate jasmonate-mediated defense during fruit ripening.

As the master regulators of the ET signaling pathway, EIL transcription factors directly activate the expression of CYP94C1 to inactivate bioactive JA-Ile, thereby attenuating JA-mediated defense during fruit ripening. Knockout of CYP94C1 improves tomato fruit resistance to necrotrophs without compromising fruit quality.

Nematocidal Potential of Phenolic Acids: A Phytochemical Seed-Coating Approach to Soybean Cyst Nematode Management.

Soybeans, one of the most valuable crops worldwide, are annually decimated by the soybean cyst nematode (SCN), Heterodera glycines, resulting in massive losses in soybean yields and economic revenue. Conventional agricultural pesticides are generally effective in the short term; however, they pose growing threats to human and environmental health; therefore, alternative SCN management strategies are urgently needed. Preliminary findings show that phenolic acids are significantly induced during SCN infection and exhibit effective nematocidal activities in vitro. However, it is unclear whether these effects occur in planta or elicit any negative effects on plant growth traits. Here, we employed a phytochemical-based seed coating application on soybean seeds using phenolic acid derivatives (4HBD; 2,3DHBA) at variable concentrations and examined SCN inhibition against two SCN types. Moreover, we also examined plant growth traits under non-infected or SCN infected conditions. Notably, 2,3DHBA significantly inhibited SCN abundance in Race 2-infected plants with increasingly higher chemical doses. Interestingly, neither compound negatively affected soybean growth traits in control or SCN-infected plants. Our findings suggest that a phytochemical-based approach could offer an effective, more environmentally friendly solution to facilitate current SCN management strategies and fast-track the development of biopesticides to sustainably manage devastating pests such as SCN.

Sucrose phosphate synthase 8 is required for the remobilization of carbon reserves in rice stems during grain filling.

Carbon reserve remobilization in stems is closely related to rice grain filling. Sucrose phosphate synthase (SPS) is highly associated with carbon reserve remobilization. In this study, we investigated the expression pattern of SPS genes in various rice tissues, and found that SPS8 is the major SPS isoform in rice stems during the grain-filling stage. We then constructed sps8 mutants using the CRISPR/Cas9 system. The SPS activity of the sps8 mutants was markedly reduced in the stems. In addition, the sps8 mutants exhibited significant starch accumulation in stems. 14C-labelling experiments revealed that the remobilization of non-structural carbohydrates from rice stems to grains was impaired in the sps8 mutants. In the sps8 mutants, grain filling was delayed and yield decreased by 15% due to a reduced percentage of ripened grains. RNA sequencing and quantitative PCR analyses indicated that the genes involved in starch synthesis and degradation were up-regulated in the sps8 mutant stems. In addition, the activity of the enzymes involved in starch synthesis and degradation was increased in the sps8 stems. These results demonstrate that SPS8 is required for carbon reserve remobilization from rice stems to grains, and that its absence may enhance 'futile cycles' of starch synthesis and degradation in rice stems.

Redesigning the tomato fruit shape for mechanized production.

Crop breeding for mechanized harvesting has driven modern agriculture. In tomato, machine harvesting for industrial processing varieties became the norm in the 1970s. However, fresh-market varieties whose fruits are suitable for mechanical harvesting are difficult to breed because of associated reduction in flavour and nutritional qualities. Here we report the cloning and functional characterization of fs8.1, which controls the elongated fruit shape and crush resistance of machine-harvestable processing tomatoes. FS8.1 encodes a non-canonical GT-2 factor that activates the expression of cell-cycle inhibitor genes through the formation of a transcriptional module with the canonical GT-2 factor SlGT-16. The fs8.1 mutation results in a lower inhibitory effect on the cell proliferation of the ovary wall, leading to elongated fruits with enhanced compression resistance. Our study provides a potential route for introducing the beneficial allele into fresh-market tomatoes without reducing quality, thereby facilitating mechanical harvesting.

Rapid generation of a tomato male sterility system and its feasible application in hybrid seed production.

KEY MESSAGE: A practical approach for the rapid generation and feasible application of green hypocotyl male-sterile (GHMS) tm6 dfr lines in tomato hybrid breeding was established. Male sterility enables reduced cost and high seed purity during hybrid seed production. However, progress toward its commercial application has been slow in tomato due to the disadvantages of most natural male-sterile mutants. Here, we developed a practical method for efficient tomato hybrid seed production using a male-sterile system with visible marker, which was rapidly generated by CRISPR/Cas9-mediated gene editing. Two closely linked genes, TM6 and DFR, which were reported to be candidates of ms15 (male sterile-15) and aw (anthocyanin without) locus, respectively, were knocked out simultaneously in two elite tomato inbred lines. Mutagenesis of both genes generated green hypocotyl male-sterile (GHMS) lines. The GHMS lines exhibited male sterility across different genetic backgrounds and environmental conditions. They also showed green hypocotyl due to defective anthocyanin accumulation, which serves as a reliable visible marker for selecting male-sterile plants at the seedling stage. We further proposed a strategy for multiplying the GHMS system and verified its high efficiency in stable male sterility propagation. Moreover, elite hybrid seeds were produced using GHMS system for potential side effects evaluation, and no adverse influences were found on seed yield, seed quality as well as important agronomic traits. This study provides a practical approach for the rapid generation and feasible application of male sterility in tomato hybrid breeding.

Genome-wide transcriptome analysis uncovers gene networks regulating fruit quality and volatile compounds in mango cultivar 'Tainong' during postharvest.

Mango is one of the most economically important fruit; however, the gene regulatory mechanism associated with ripening and quality changes during storage remains largely unclear. This study explored the relationship between transcriptome changes and postharvest mango quality. Fruit quality patterns and volatile components were obtained using headspace gas chromatography and ion-mobility spectrometry (HS-GC-IMS). The changes in mango peel and pulp transcriptome were analyzed during four stages (pre-harvesting, harvesting, maturity, and overripe stages). Based on the temporal analysis, multiple genes involved in the biosynthesis of secondary metabolites were upregulated in both the peel and pulp during the mango ripening process. Moreover, cysteine and methionine metabolism related to ethylene synthesis were upregulated in the pulp over time. Weighted gene co-expression network analysis (WGCNA) further showed that the pathways of pyruvate metabolism, citrate cycle, propionate metabolism, autophagy, and SNARE interactions in vesicular transport were positively correlated with the ripening process. Finally, a regulatory network of important pathways from pulp to peel was constructed during the postharvest storage of mango fruit. The above findings provide a global insight into the molecular regulation mechanisms of postharvest mango quality and flavor changes.

Degradation of Aflatoxin B1 in Peanut Oil by Ultraviolet-LED Cold-Light Irradiation and Structure Elucidation of the Degradation Products.

This study aimed to determine the efficiency of ultraviolet (UV)-LED cold light treatment on the degradation of aflatoxin (AF)B1 in peanut oils. The peanut oil samples obtained from different places in China and abroad were determined for AFB1 degradation efficiency of the UV-LED cold-light irradiation method. The degradation products were analyzed by ultra-high performance liquid chromatography coupled to quadrupole orbitrap high-resolution mass spectrometry (UPLC-Q-Exactive MS). The results indicated that the AFB1 content in all peanut oil samples decreased rapidly after 5 min of irradiation. Four main photodegradation products (C18H16O7, C17H14O7, C17H14O7, and C17H14O8) were identified using the established LC-MS method. Their chemical structures were postulated based on the LC-MS data. Also, the degradation pathways were proposed based on the data obtained. Oxidation and reduction reactions were mainly responsible for AFB1-decomposition. The reactions occurred at the furan and lactone rings. These findings demonstrated that UV-LED cold-light irradiation was an effective method for treating AFB1- contaminated peanut oil.

CRISPR/LbCas12a-Mediated Genome Editing in Soybean.

Currently methods for generating soybean edited lines are time-consuming, inefficient, and limited to certain genotypes. Here we describe a fast and highly efficient genome editing method based on CRISPR-Cas12a nuclease system in soybean. The method uses Agrobacterium-mediated transformation to deliver editing constructs and uses aadA or ALS genes as selectable marker. It only takes about 45 days to obtain greenhouse-ready edited plants at higher than 30% transformation efficiency and 50% editing rate. The method is applicable to other selectable markers including EPSPS and has low transgene chimera rate. The method is also genotype-flexible and has been applied to genome editing of several elite soybean varieties.

Recoloring tomato fruit by CRISPR/Cas9-mediated multiplex gene editing.

Fruit color is an important horticultural trait, which greatly affects consumer preferences. In tomato, fruit color is determined by the accumulation of different pigments, such as carotenoids in the pericarp and flavonoids in the peel, along with the degradation of chlorophyll during fruit ripening. Since fruit color is a multigenic trait, it takes years to introgress all color-related genes in a single genetic background via traditional crossbreeding, and the avoidance of linkage drag during this process is difficult. Here, we proposed a rapid breeding strategy to generate tomato lines with different colored fruits from red-fruited materials by CRISPR/Cas9-mediated multiplex gene editing of three fruit color-related genes (PSY1, MYB12, and SGR1). Using this strategy, the red-fruited cultivar 'Ailsa Craig' has been engineered to a series of tomato genotypes with different fruit colors, including yellow, brown, pink, light-yellow, pink-brown, yellow-green, and light green. Compared with traditional crossbreeding, this strategy requires less time and can obtain transgene-free plants with different colored fruits in less than 1 year. Most importantly, it does not alter other important agronomic traits, like yield and fruit quality. Our strategy has great practical potential for tomato breeding and serves as a reference for improving multigene-controlled traits of horticultural crops.

Effects of different extraction methods on the structural, antioxidant and hypoglycemic properties of red pitaya stem polysaccharide.

Processing conditions can change the compositions and microstructures of polysaccharides, resulting in favorable and unfavorable effects on their chemical characteristics and bioactivites. Here, this study comparatively evaluated the effects of the commonly used hot water, alkaline, acidic, enzymatic, ultrasonic and hot water-alkaline extractions on the structural features and antioxidant and hypoglycemic properties of pitaya stem polysaccharides. Nuclear magnetic resonance spectroscopy showed six polysaccharides had similar glycosyl types. Scanning electron microscopy exhibited the surface morphology of the extracted six polysaccharides differed significantly. Polysaccharide obtained by hot water showed better antioxidant and hypoglycemic properties than that of the other polysaccharides. These data suggested that alkaline, acidic, enzymatic, ultrasonic and hot water-alkaline extractions have various influences on the degradation of polysaccharides without varying the major structure in comparison with hot water extraction. Additionally, monosaccharide composition and molecular weight of polysaccharides are two chief factors affecting the bioactivity of pitaya stem polysaccharides.

Tomato MED25 regulates fruit ripening by interacting with EIN3-like transcription factors.

Fruit ripening relies on the precise spatiotemporal control of RNA polymerase II (Pol II)-dependent gene transcription, and the evolutionarily conserved Mediator (MED) coactivator complex plays an essential role in this process. In tomato (Solanum lycopersicum), a model climacteric fruit, ripening is tightly coordinated by ethylene and several key transcription factors. However, the mechanism underlying the transmission of context-specific regulatory signals from these ripening-related transcription factors to the Pol II transcription machinery remains unknown. Here, we report the mechanistic function of MED25, a subunit of the plant Mediator transcriptional coactivator complex, in controlling the ethylene-mediated transcriptional program during fruit ripening. Multiple lines of evidence indicate that MED25 physically interacts with the master transcription factors of the ETHYLENE-INSENSITIVE 3 (EIN3)/EIN3-LIKE (EIL) family, thereby playing an essential role in pre-initiation complex formation during ethylene-induced gene transcription. We also show that MED25 forms a transcriptional module with EIL1 to regulate the expression of ripening-related regulatory as well as structural genes through promoter binding. Furthermore, the EIL1-MED25 module orchestrates both positive and negative feedback transcriptional circuits, along with its downstream regulators, to fine-tune ethylene homeostasis during fruit ripening.

Betacyanins and Anthocyanins in Pulp and Peel of Red Pitaya (Hylocereus polyrhizus cv. Jindu), Inhibition of Oxidative Stress, Lipid Reducing, and Cytotoxic Effects.

This study aimed to promote red pitaya fruit parts as alternate sources of nutraceuticals. The red pitaya of Chinese origin was determined for its in vitro efficacy, where the fruit extracts were evaluated based on the selected antioxidative properties, lipid-reducing capacity, and cytotoxicity. The betanin, total betacyanins, total anthocyanins, and DPPH radical scavenging activity of the red pitaya pulp and peel extracts were determined by spectrophotometric analyses. Cell culture assays were used to examine in vitro efficacy and cytotoxicity of the pitaya extracts. The result showed that red pitaya peel extract had a higher total betacyanins and total anthocyanins content than the pulp extract, but the peel extract had a lower DPPH radical scavenging effect than the pulp extract. The red pitaya extracts also had a protective effect in reducing oxidative stress, especially the peel extract. All fruit samples had a low anticancer potential except for betanin and anthocyanin standards. The protective effect of pitaya peel could be attributed to betacyanins and anthocyanins. Both pulp and peel extracts had a weak anticancer effect because these extracts contained polysaccharides and other phytochemicals that were not cytotoxic. As the peel extract of red pitaya was not cytotoxic, it is a potent source of betacyanins for reducing oxidative stress.

Systematic analysis of the sugar accumulation mechanism in sucrose- and hexose- accumulating cherry tomato fruits.

BACKGROUND: Sugar content is an important indicator of fruit quality. Except for a few wild tomato species that accumulate sucrose in the fruits, most cultivated tomato species accumulate hexose. Although several studies have focused on wild sucrose-accumulating tomato, the sucrose accumulation mechanism is still unclear. RESULTS: Here, two homozygous inbred cherry tomato lines ('TB0023' and 'TB0278', which accumulated sucrose and hexose, respectively) were selected to analyze the sugar accumulation mechanism. Carbohydrate analysis, cytological observation, gene expression and enzyme activity analysis and proteomics methods were used in this study. The results indicated that glucose and fructose were absolutely dominant in the soluble sugar content of hexose-accumulating cherry tomato fruit, while sucrose and a certain proportion of hexose were the main forms of soluble sugar in sucrose-accumulating cherry tomato fruit. The phloem unloading pathway of the hexose-accumulating cherry tomato fruit switched from symplastic to apoplastic during fruit development, and the sucrose-accumulating cherry tomato probably had a mixed unloading pathway involving the symplastic and apoplastic. High activity of acid invertase (AI), sucrose phosphate synthase (SPS), sucrose synthase (SS) and sugar transporters LeSUT1, SlSWEET2a and SlSWEET12c were important factors for hexose accumulation in the hexose-accumulating cherry tomato fruit, while LeSUT2, SPS, SS, SlSWEET1b, SlSWEET5b, SlSWEET11b, SlSWEET7a, SlSWEET14 were responsible for solute sugar accumulation in the sucrose-accumulating cherry tomato. CONCLUSIONS: This study provides detailed evidence for elucidation of the tomato sugar accumulation mechanism from the perspective of cell structure, physiology and molecular biology, providing a theoretical basis for the improvement of tomato quality and aiding the utilization of tomato genetic resources.

Rare and Common Variants in COL4A1 in Chinese Patients With Intracerebral Hemorrhage.

Here, we screened the COL4A1 variants in Chinese intracerebral hemorrhage (ICH) patients to summarize the relationship between the variants and clinical characteristics. Targeted sequencing of a 65-gene panel including COL4A1 was performed to detect all the coding regions and ±10-bp splicing sites. In total, 568 patients were included. Regarding rare nonsynonymous variants with a minor allele frequency (MAF) <0.5%, 6 missense variants and five suspicious splice site variants, absent in 573 healthy controls, were found in 11 patients. The subgroup carrying rare variants did not show specific phenotype compared with non-variant carriers. For the single nucleotide polymorphism (SNP) loci with an MAF> 5%, we did not find a significant association between the allele or genotype distribution of the SNP loci and the risk of ICH. Rs3742207 was nominally associated with death at 1-year follow-up (p = 0.02027, OR 1.857, 95% CI 1.101-3.133) after adjusted by age, hypertension history, hematoma volume and recurrent ICH history. Nevertheless, after the Bonferroni correction, the association was no longer significant. In conclusion, rare nonsynonymous variants in COL4A1 were identified in 1.94% (11/568) of Chinese ICH patients, while rs3742207 maybe indicate a worse prognosis of ICH.

Alternative transcription and feedback regulation suggest that SlIDI1 is involved in tomato carotenoid synthesis in a complex way.

Carotenoid pigments confer photoprotection and visual attraction and serve as precursors for many important signaling molecules. Herein, the orange-fruited phenotype of a tomato elite inbred line resulting from sharply reduced carotenoid levels and an increased ß-carotene-to-lycopene ratio in fruit was shown to be controlled by a single recessive gene, oft3. BSA-Seq combined with fine mapping delimited the oft3 gene to a 71.23 kb interval on chromosome 4, including eight genes. Finally, the oft3 candidate gene SlIDI1, harboring a 116 bp deletion mutation, was identified by genome sequence analysis. Further functional complementation and CRISPR-Cas9 knockout experiments confirmed that SlIDI1 was the gene underlying the oft3 locus. qRT-PCR analysis revealed that the expression of SlIDI1 was highest in flowers and fruit and increased with fruit ripening or flower maturation. SlIDI1 simultaneously produced long and short transcripts by alternative transcription initiation and alternative splicing. Green fluorescent protein fusion expression revealed that the long isoform was mainly localized in plastids and that an N-terminal 59-amino acid extension sequence was responsible for plastid targeting. Short transcripts were identified in leaves and fruit by 5' RACE and in fruit by 3' RACE, which produced corresponding proteins lacking transit peptides and/or putative peroxisome targeting sequences, respectively. In SlIDI1 mutant fruit, SlBCH1 transcription involved in ß-carotenoid catabolism was obviously suppressed, which may be responsible for the higher ß-carotene-to-lycopene ratio and suggested potential feedback regulatory mechanisms involved in carotenoid pathway flux.

Development of Green Banana Fruit Wines: Chemical Compositions and In Vitro Antioxidative Activities.

This study aimed to develop functional fruit wines using whole fruit, pulp, and peels from green bananas. The boiled banana homogenates were mixed with cane sugar before wine fermentation. Quality parameters, phenolic compounds, flavor components, and antioxidative properties of the green banana peel wine (GBPW), green banana pulp wine (GBMW), and whole banana wine (GBW) were determined. High-performance liquid chromatography was used to determine the phytochemical compounds in three wines, and the flavor components were further analyzed using headspace solid-phase microextraction combined with gas chromatography-mass spectrometry. The flavor components and in vitro antioxidant activities were, respectively, determined using the relative odor activity value and the orthogonal projections on latent structure discrimination analysis (OPLS-DA). In vitro antioxidative capacities for these wines were evaluated using antioxidant chemical assays and cell culture methods. The total phenolic and total tannin content of the GBPW, GBMW, and GBW showed reducing trends with increasing fermentation days, whereas the total flavonoid content of the wine samples exhibited downward trends. The antioxidant capacities of the three wine samples were higher than those of the raw fruit samples, except for the metal chelation rate (%). Additionally, the main flavor component in the wine samples was 3-methyl-1-butanol. Its percentages in the GBPW, GBMW, and GBW were 72.02%, 54.04%, and 76.49%, respectively. The OPLS-DA results indicated that the three wines presented significantly different antioxidant activities. The cell-culture-based antioxidant analysis showed that these wine samples had protective effects against the oxidative stress of the 3T3-L1 preadipocytes induced by hydrogen peroxide. This study provided a theoretical basis for defining the antioxidant characteristics of banana wines and expanding novel channels for using banana peels to develop nutraceuticals.

Novel Antioxidant and Hypoglycemic Water-Soluble Polysaccharides from Jasmine Tea.

There have been few studies dealing with chemical elucidation and pharmacological potentials of water-soluble polysaccharides from jasmine tea, limiting their use in functional foods. In this study, water-soluble polysaccharides (named as JSP) were extracted from Jasminum sambac (L.) Aiton tea and fractionated to afford two sub-fractions (JSP-1 and JSP-2). The main structural characteristics of novel JSP sub-fractions were determined by high performance gel permeation chromatography, ultra-performance liquid chromatography-tandem mass spectrometry, Fourier transform infrared, and nuclear magnetic resonance analysis. Physiologically, the abilities of JSP-1 and JSP-2 to reduce ferric ions, scavenge DPPH and hydroxyl radicals, as well as protect islet cells were confirmed in vitro. JSP-1 exhibited better antioxidant and hypoglycemic activities than JSP-2. The molecular weights of JSP-1 and JSP-2 were 18.4 kDa and 14.1 kDa, respectively. JSP-1 was made up of glucose, galactose, rhamnose, xylose, arabinose, and galacturonic acid with molar ratios 1.14:4.69:1.00:9.92:13.79:4.09, whereas JSP-2 with a triple helical structure was composed of galactose, rhamnose, xylose, arabinose, and galacturonic acid as 3.80:1.00:8.27:11.85:5.05 of molar ratios. JSP-1 contains →1)-α-Galƒ-(3→, →1)-α-Galƒ-(2→, →1)-α-Araƒ-(5→, →1)-α-Araƒ-(3→, →1)-α-Araƒ-(3,5→, →1)-ß-Xylp-(2→ and →1)-ß-Xylp-(3→ residues in the backbone. These results open up new pharmacological prospects for the water-soluble polysaccharides extracted from jasmine tea.