Integrating genes and metabolites: unraveling mango's drought resilience mechanisms.

BACKGROUND: Mango (Mangifera indica L.) faces escalating challenges from increasing drought stress due to erratic climate patterns, threatening yields, and quality. Understanding mango's drought response mechanisms is pivotal for resilience and food security. RESULTS: Our RNA-seq analyses unveil 12,752 differentially expressed genes linked to stress signaling, hormone regulation, and osmotic adjustment. Weighted Gene Co-expression Network Analysis identified three essential genes-WRKY transcription factor 3, polyamine oxidase 4, and protein MEI2-like 1-as drought defense components. WRKY3 having a role in stress signaling and defense validates its importance. Polyamine oxidase 4, vital in stress adaptation, enhances drought defense. Protein MEI2-like 1's significance emerges, hinting at novel roles in stress responses. Metabolite profiling illuminated Mango's metabolic responses to drought stress by presenting 990 differentially abundant metabolites, mainly related to amino acids, phenolic acids, and flavonoids, contributing to a deeper understanding of adaptation strategies. The integration between genes and metabolites provided valuable insights by revealing the correlation of WRKY3, polyamine oxidase 4 and MEI2-like 1 with amino acids, D-sphingnosine and 2,5-Dimethyl pyrazine. CONCLUSIONS: This study provides insights into mango's adaptive tactics, guiding future research for fortified crop resilience and sustainable agriculture. Harnessing key genes and metabolites holds promise for innovative strategies enhancing drought tolerance in mango cultivation, contributing to global food security efforts.

Precise Correction of Lhcgr Mutation in Stem Leydig Cells by Prime Editing Rescues Hereditary Primary Hypogonadism in Mice.

Hereditary primary hypogonadism (HPH), caused by gene mutation related to testosterone synthesis in Leydig cells, usually impairs male sexual development and spermatogenesis. Genetically corrected stem Leydig cells (SLCs) transplantation may provide a new approach for treating HPH. Here, a novel nonsense-point-mutation mouse model (LhcgrW495X ) is first generated based on a gene mutation relative to HPH patients. To verify the efficacy and feasibility of SLCs transplantation in treating HPH, wild-type SLCs are transplanted into LhcgrW495X mice, in which SLCs obviously rescue HPH phenotypes. Through comparing several editing strategies, optimized PE2 protein (PEmax) system is identified as an efficient and precise approach to correct the pathogenic point mutation in Lhcgr. Furthermore, delivering intein-split PEmax system via lentivirus successfully corrects the mutation in SLCs from LhcgrW495X mice ex vivo. Gene-corrected SLCs from LhcgrW495X mice exert ability to differentiate into functional Leydig cells in vitro. Notably, the transplantation of gene-corrected SLCs effectively regenerates Leydig cells, recovers testosterone production, restarts sexual development, rescues spermatogenesis, and produces fertile offspring in LhcgrW495X mice. Altogether, these results suggest that PE-based gene editing in SLCs ex vivo is a promising strategy for HPH therapy and is potentially leveraged to address more hereditary diseases in reproductive system.

Green One-Pot Syntheses of 2-Sulfoximidoyl-3,6-dibromo Indoles Using N-Br Sulfoximines as Both Brominating and Sulfoximinating Reagents.

A green one-pot 2,3,6-trifunctionalization of N-alkyl/aryl indoles was achieved by adding three equivalents of N-Br sulfoximine to the indole solution. A variety of 2-sulfoximidoyl-3,6-dibromo indoles were prepared with 38-94% yields using N-Br sulfoximines as both brominating and sulfoximinating reagents. Based on the results of controlled experiments, we propose that a radical substitution involving 3,6-dibromination and 2-sulfoximination occurs in the reaction process. This is first time that 2,3,6-trifunctionalization of indole in one pot has been achieved.

Genome-Wide Characterization, Identification and Expression Profile of MYB Transcription Factor Gene Family during Abiotic and Biotic Stresses in Mango (Mangifera indica).

Mango (Mangifera indica) is an economically important fruit tree, and is cultivated in tropical, subtropical, and dry-hot valley areas around the world. Mango fruits have high nutritional value, and are mainly consumed fresh and used for commercial purposes. Mango is affected by various environmental factors during its growth and development. The MYB transcription factors participates in various physiological activities of plants, such as phytohormone signal transduction and disease resistance. In this study, 54 MiMYB transcription factors were identified in the mango genome (371.6 Mb). A phylogenetic tree was drawn based on the amino acid sequences of 222 MYB proteins of mango and Arabidopsis. The phylogenetic tree showed that the members of the mango MYB gene family were divided into 7 group, including Groups 1, -3, -4, -5, -6, -8, and -9. Ka/Ks ratios generally indicated that the MiMYBs of mango were affected by negative or positive selection. Quantitative real-time PCR showed that the transcription levels of MiMYBs were different under abiotic and biotic stresses, including salicylic acid, methyl jasmonate, and H2O2 treatments, and Colletotrichum gloeosporioides and Xanthomonas campestris pv. mangiferaeindicae infection, respectively. The transcript levels of MiMYB5, -35, -36, and -54 simultaneously responded positively to early treatments with salicylic acid, methyl jasmonate, and H2O2. The transcript level of MiMYB54 was activated by pathogenic fungal and bacterial infection. These results are beneficial for future interested researchers aiming to understand the biological functions and molecular mechanisms of MiMYB genes.

AAV-mediated gene therapy produces fertile offspring in the Lhcgr-deficient mouse model of Leydig cell failure.

Leydig cell failure (LCF) caused by gene mutation results in testosterone deficiency and infertility. Serum testosterone levels can be recovered via testosterone replacement; however, established therapies have shown limited success in restoring fertility. Here, we use a luteinizing hormone/choriogonadotrophin receptor (Lhcgr)-deficient mouse model of LCF to investigate the feasibility of gene therapy for restoring testosterone production and fertility. We screen several adeno-associated virus (AAV) serotypes and identify AAV8 as an efficient vector to drive exogenous Lhcgr expression in progenitor Leydig cells through interstitial injection. We observe considerable testosterone recovery and Leydig cell maturation after AAV8-Lhcgr treatment in pubertal Lhcgr-/- mice. Of note, this gene therapy partially recovers sexual development, substantially restores spermatogenesis, and effectively produces fertile offspring. Furthermore, these favorable effects can be reproduced in adult Lhcgr-/- mice. Our proof-of-concept experiments in the mouse model demonstrate that AAV-mediated gene therapy may represent a promising therapeutic approach for patients with LCF.

Ecosystem impact and dietary exposure of polychlorinated biphenyls (PCBs) and heavy metals in Chinese mitten crabs (Eriocheir sinensis) and their farming areas in Jiangsu, China.

This study investigated the presence of 18 dioxin-like and non-dioxin-like polychlorinated biphenyls (dl- and ndl-PCBs), heavy metals (Cd, Hg, Pb, and As) in Chinese mitten crabs (Eriocheir sinensis) and their distribution in Jiangsu, China. Risk assessment and source apportionment were employed for evaluating the eco-toxicological impact and human exposure. It was found that the compositions of PCBs varied spatially, suggesting different sources of pollutants, whilst PCB 28, 105, 114, and 126 were consistently found in all sample types, suggesting a common pollution source remained, and the bio-accumulation process was in effect. The total PCBs in sediment were found much higher than in water, and brown meat had the highest and most diverse PCB congeners among all tissues. The presence of heavy metals was found in all samples in descending order of As>Cd>Pb>Hg and in the order of shell>brown meat>white meat>gill for crabs. The results of risk assessment indicated that the potential carcinogenic and non-carcinogenic risks were within the acceptable range for long-term consumption of the crabs overall. However, the highest toxic equivalent (TEQ), carcinogenic, and non-carcinogenic risks were all recorded in Location C, where dl-PCB 126, 169, and As contributed to the majority of the risks. The ecological risk posed by all HMs was low, but cases of serious point source pollution have been found in the investigated regions, and risks caused by Cd individually should raise concerns. Source apportionment study revealed that the contaminants mostly originated from anthropogenic activities. Natural deposition and transportation played an important role as well.

Sensitive Immunochromatographic Assay Using Highly Luminescent Quantum Dot Nanobeads as Tracer for the Detection of Cyproheptadine Hydrochloride in Animal-Derived Food.

Cyproheptadine hydrochloride (CYP), used as human and veterinary drug, has been used illegally as feed additive for food-producing animals, which could remain in food and jeopardize human health. There is a need for on-site detection of CYP residue in animal-derived food. In this study, a hapten was designed, and a specific monoclonal antibody (mAb) was developed to detect CYP with an IC50 of 1.38 ng/mL and negligible cross-reactivity (CR) for other analogs. Forthermore, a high sensitive immunochromatographic assay (QBs-ICA) was developed using quantum dot nanobeads as reporters. The assay showed the linear detection range (IC20-IC80) of 0.03-0.52 ng/mL, the limit of detection (LOD) and visual detection limit (VDL) reached to 0.01 and 0.625 ng/mL, respectively. Spiked recovery study in pig urine and pork confirmed that the QBs-ICA was applicable for on-site testing. This assay showed better sensitivity and speedy than the reported instrumental analysis and immunoassays.

Development of a fast-responsive and turn on fluorescent probe with large Stokes shift for specific detection of cysteine in vivo.

Cysteine has a great effect on the physiological and pathological processes, which could bring out various diseases such as skin lesions, edema, hair depigmentation, Alzheimer's, Parkinson's, and liver damage due to the abnormal concentrations of cysteine. Therefore, it is of great impoatance to develop a method for imaging Cys. Herein, a novel fluorescent probe was developed for imaging Cys in vivo specially. This turn-on probe exhibited favorable advantages including large Stokes shift (90 nm), fast response (10 min), good selectivity, low cytotoxicity and so on. Furthermore, the probe could be applied to monitoring cysteine in living HeLa cells, which indicates that this turn-on probe could penetrate viable cell membranes and image Cys over other analystes especially HCy and GSH.

Iodine-catalyzed guanylation of amines with N,N'-di-Boc-thiourea.

Herein, we report that iodine-catalyzed guanylation of primary amines can be accomplished with N,N'-di-Boc-thiourea and TBHP to afford the corresponding guanidines in 40-99% yields. Oxidation of the HI byproduct by TBHP eliminates the need for an extra base to prevent the protonation of substrates and makes the reaction especially useful for both electronically and sterically deactivated primary anilines.

A highly diastereoselective decarboxylative mannich reaction of ß-keto acids with optically active N-sulfinyl α-imino esters.

A range of protected γ-oxo-α-amino esters have been prepared in a highly regio- and stereoselective manner through the decarboxylative Mannich reaction of ß-keto acids with optically active N-tert-butanesulfinyl α-imino esters in the presence of 3 mol % La(OTf)(3) or 5 mol % Y(OTf)(3) at 20 °C. Preliminary mechanistic studies indicate that the reaction proceeds through imine addition followed by decarboxylation.

Catalytic decarboxylative alkylation of ß-keto acids with sulfonamides via the cleavage of carbon-nitrogen and carbon-carbon bonds.

An efficient decarboxylative alkylation reaction of ß-keto acids with N-benzylic or N-allylic sulfonamides has been developed, for the first time, through sequential cleavage of carbon-nitrogen and carbon-carbon bonds in the presence of 10 mol% of FeCl(3).

Phylogenetic analysis of methanogens in the pig feces.

In order to assess methanogen diversity in feces of pigs, archaeal 16S rRNA gene clone libraries were constructed from feces of the pig. After the amplification by PCR using primers Met86F and Met1340R, equal quantities of PCR products from each of the five pigs were mixed together and used to construct the library. Sequence analysis showed that the 74 clones were divided into ten phylotypes as defined by RFLP analysis. Phylogenetic analysis showed that three phylotypes were most closely affiliated with the genus Methanobrevibacter (46% of clones). The library comprised 55.4% unidentified euryarchaeal clones. Three phylotypes (LMG4, LMG6, LMG8) were not closely related to any known Euryarchaeota sequences. The phylogenetic analysis indicated that the archaea found in the libraries were all clustered into the Euryarchaeota. The data from the phylogenetic tree showed that those sequences belonged to three monophyletic groups. Phylotypes LGM2 and LGM7 grouped within the genus Methanobrevibacter. Phylotypes LGM4, LGM6, LGM8 and LGM9 grouped within the genus Methanosphaera. Other phylotypes grouped together, and formed a distantly related sister group to Aciduliprofundum boonei and species of the Thermoplasmatales including Thermoplasma volcanium and Thermoplasm acidophilum. Our results showed that methanogens belonging to the genus Methanobrevibacter were predominant in pig feces, and that many unique unknown archaea sequences were also found in the library. Nevertheless, whether these unique sequences represent new taxonomic groups and their role in the pig gut need further investigation.

[Methanogen diversity in the piglet colon and its correlation with environmental factors].

OBJECTIVE: We investigated the correlation between the methanogen diversity in the piglet colon and the environmental factor (weaning stress, diet type and age). METHODS: The colonic contents of piglets at 7, 14, 21, 28 and 35 days old were collected for the determination of volatile fatty acids and total DNA extraction. DNA was subject to PCR-DGGE (Denaturing gradient gel electrophoresis) analysis and interesting bands were excised and sequenced. Redundancy analysis (RDA) was performed for the correlation between methanogen diversity and environmental factors. RESULTS: As the piglets grew, acetate, propionate and the total volatile fatty acid production increased significantly (P < 0.05), but butyrate concentration remained stable (P > 0.05). The similarity indices of DGGE profiles was higher during the period of the 7 day to 24 day after birth, with grouped in one cluster, and the samples from the 35 days were dropped into another cluster. DGGE analysis showed three dominant bands appeared in samples of 35 d old, with their 16S rRNA gene sequences closely related to Methanobacteria and a novel group of uncultivated Achaean. Redundancy analysis (RDA) showed that age has the largest relevance to the community structure of bacteria in colonic samples, followed by the diet type. CONCLUSION: The results indicated that the methanogen community in the piglet colon was stable during the first 24 days after birth, and became more diverse at 35 days of age. The methanogens community was mainly affected by age and the diet type.

Catalyst-free alkylation of sulfinic acids with sulfonamides via sp(3) C-N bond cleavage at room temperature.

An unprecedented catalyst-free alkylation of sulfinic acids with sulfonamides has been developed via sp(3) C-N bond cleavage at room temperature. In the absence of external catalysts and additives, a wide variety of N-benzylic and N-allylic sulfonamides couple with sulfinic acids to give structurally diversified sulfones in moderate to excellent yields. Furthermore, the reaction of N-(2-acyl)allylic sulfonamides with sulfinic acids provides a convenient access to trisubstituted allyl sulfones with exclusive Z selectivity.

Selective benzylic and allylic alkylation of protic nucleophiles with sulfonamides through double Lewis acid catalyzed cleavage of sp3 carbon-nitrogen bonds.

The acid-catalyzed benzylic and allylic alkylation of protic nucleophiles is fundamentally important for the formation of carbon-carbon and carbon-heteroatom bonds, and it is a formidable challenge for benzylic and allylic amine derivatives to be used as the alkylating agents. Herein we report a highly efficient benzylic and allylic alkylation of protic carbon and sulfur nucleophiles with sulfonamides through double Lewis acid catalyzed cleavage of sp(3) carbon-nitrogen bonds at room temperature. In the presence of a catalytic amount of inexpensive ZnCl(2)-TMSCl (TMSCl: chlorotrimethylsilane), 1,3-diketones, beta-keto esters, beta-keto amides, malononitrile, aromatic compounds, thiols, and thioacetic acid can couple with a broad range of tosyl-activated benzylic and allylic amines to give diversely functionalized products in good to excellent yields and with high regioselectivity. Furthermore, the cross-coupling reaction of 1,3-dicarbonyl compounds with benzylic propargylic amine derivatives has been successfully applied to the one-step synthesis of polysubstituted furans and benzofurans.

Catalytic selective bis-arylation of imines with anisole, phenol, thioanisole and analogues.

The first highly efficient double Friedel-Crafts reaction of N-tosyl imines with anisole, phenol, thioanisole and analogues has been developed to produce the corresponding symmetric diarylmethanes and triarylmethanes with high regioselectivity in the presence of a catalytic amount of Bi(2)(SO(4))(3)-TMSCl at room temperature.