Wide-ranging research efforts have been directed to prioritize scientific and technological inventions for healthcare monitoring. In recent years, the effective utilization of functional nanomaterials in various electroanalytical measurements realized a rapid, sensitive, and selective detection and monitoring of a wide range of biomarkers in body fluids. Owing to good biocompatibility, high organic capturing ability, strong electrocatalytic activity, and high robustness, transition metal oxide-derived nanocomposites have led to enhancements in sensing performances. The aim of the present review is to describe key advancements of transition metal oxide nanomaterials and nanocomposites-based electrochemical sensors, along with current challenges and prospects towards the development of a highly durable and reliable detection of biomarkers. Moreover, the preparation of nanomaterials, electrode fabrication, sensing mechanism, electrode-bio interface, and performance of metal oxides nanomaterials and nanocomposite-based sensor platforms will be described. Owing to their strong electrocatalytic activity, low price, high organic catching aptitude, reduced dimension, large surface-to-volume ratio, and high degree of crystalline nature, metal oxide nanocomposites have been extensively employed and are already established as an active electrocatalyst for sensing various emergent biomarkers ,2,3. TypHowever, because of their high surface energy, the ultra-small-sized nanoparticles are frequently less durable and effortlessly aggregate into larger particles, which may have an effect on the reduction of the specific surface area ,11,12. TOver the last decade, numerous metal oxide nanocomposites such as one-dimension (1D), two-dimension (2D), and three-dimension (3D) have exhibited outstanding catalytic and sensing properties . In the Previous research studies and reviews have reported that precious metal nanomaterials dispersed on numerous metal oxide supports as metal nanocomposites, which are catalytically more active for various electrochemical reactions ,20,21,22The recent investigations appear to indicate that small metal clusters in the heterostructures are the real active source for catalytic reactions. These tiny Au clusters possess low-coordinate Au atoms in comparison to bulk Au poly-crystal surfaces and take higher-energy d states, which are easily reactive and absorb/activate oxygen moieties . In receThe use of metal oxide nanomaterials and their nanocomposites has become increasingly predominant, owing to the inadequate approaches to upsurge the activity of an oxide containing a single metal ,31. The In recent years, significant technological developments have been developed in the practice of pure metal oxides and multi-metal oxides nanomaterials in electroanalytical sciences due to their quantum confinement effect, their high surface area to volume ratios, biocompatibility, chemical constancy, surface action, and adaptable electron transport possessions ,36,37. S−2 mM−1, and a broad sensing range of 7.5 µM–3 mM. This electrochemical acetaminophen sensor also showed good selectivity in the presence of interfering species and good durability (up to 3000 s) and practical analysis of serum and tablet samples. Yang et al. reported a NiO nanomaterials-based enzyme-free electrochemical detection of glucose [−1 cm−2, a linear detection range from 4 μM to 7.5 mM, and good stability. Ahmad and co-workers recently developed an enzyme-free electrochemical uric acid sensor based on nano-berry morphological structured cobalt oxide (Co3O4) nanostructures with a low detection limit of ~2.4 µM (linear range of 5–3000 μM) and high sensitivity of 206 μA mM−1 cm−2 [For instance, recently, Annandurai et al. developed a nickel oxide (NiO) nanoparticles-based electrochemical sensor platform for the detection of 4-acetaminophen in tablets and human blood serum samples . Nickel  glucose . The preM−1 cm−2 . 2O2) was developed by Morris et al. based on iron oxide (Fe3O4) nanodots modified with an indium tin oxide (ITO) electrode [−3–6.5 × 10−3). The sensor response reached a steady state signal within 5 s at the Fe3O4 nanodots/ITO electrode because of its large electrochemically active surface area and strong adsorption capability for immobilizing H2O2 on the electrode surface. The swift electron transfer among Fe2+ and Fe3+ ions created Fe3O4 nanodots as an exceptional electrical conductor. The as-developed Fe3O4 nanodots/ITO sensor showed good selectivity in the presence of various electrochemically active substances  and inorganic substances  and stability (more than 12 days). An enzyme-mimic hydrogen peroxide  comprises an active pair of electron-hole with a large volume of active surface area, whereas thionine could enhance the efficacy of an electron transfer process among the enzyme and the electrode. The fabricated biosensor platform showed a LOD of ~1.8 nM with a wide sensing range (10–1000 µM) and high sensitivity of ~70.2 µA µM−1 cm−2 towards the detection of cholesterol. Numerous pure metal oxides nanomaterials derived from electrochemical sensors and biosensor platforms are reported for sensing and determining various biomarkers such as glucose, lactic acid, dopamine, ascorbic acid, etc. Owing to their biocompatibility, wider operating potential window, lower charging/background current, and electrochemical redox characteristics, mixed transition metal oxide nanomaterials are becoming more interesting and receiving more and more attention in electrochemical and bio-electrochemical sensing applications.Recently, Yan et al. demonstrated another interesting electrochemical cholesterol biosensor using thionine and Cuaterials . In this2O4, where M = Mn, Ni, Fe, Cu, and Zn) exhibited interesting projections due to their outstanding catalytic and physicochemical properties [2O4) nanostructured electrode materials have been developed with different morphologies such as nanoflowers, nanosheets, nanorods, nanowires, etc., towards the sensitive detection of various biomarkers [2O4 nanoflowers as an enzyme-free electrocatalyst towards the direct oxidation of glucose and lactic acid under an alkaline electrolyte  nanoparticles based on non-enzymatic electrochemical detection of ascorbic acid in real pharmaceutical samples of Vitamin-C tablets and drinks [The improved electrocatalytic activity of the NiCod drinks . The as-2+/Ni3+), and ZnO and CoO provided exceptional chemical and mechanical stability. Other reliable electrochemical dopamine sensor-based mixed Al–Mn0.65–Cr1.76-oxide nanomaterials were developed by Alam et al. [0.65–Cr1.76-oxide nanomaterials sensor exhibited high sensitivity (~55.8 μA μM−1 cm−2), low detection limit (~96.8 pM), wide linear range (0.1 nM–0.01 mM), good selectivity, and long-term stability towards the detection of dopamine. Enzyme-free simultaneous sensing of acetylcholine and ascorbic acid using mixed ZnO/CuO nanoleaves oxides [−1cm−2 for acetylcholine and 94.94 pAμM−1cm−2 for ascorbic acid). Various mixed metal oxides nanomaterials-based electrochemical biosensor platforms were demonstrated towards the detection of emerging biomarkers [Durai et al. reported mixed ternary NiCoZnO nanorods modified with glassy carbon electrodes for sensing dopamine in biofluids with a detection limit of 0.01 nM and a linear range from 1.0 nM to 0.5 µM . The devm et al. . The fabs oxides . The simomarkers ,58,59. I2O4 spinel oxides and their composite nanomaterials have presented favourable sensing abilities, but there are still various routes to discover for practical sensor applications. For instance, two-dimensional MXenes, transition metal chalcogenides, phosphorous, etc., can be effectively employed to develop heterojunctions towards the detection of various biomarkers [Most of the transition pure and mixed metal oxides nanomaterials may be employed as the sensor platforms for non-enzymatic sensing of glucose, lactic acid, hydrogen peroxide, dopamine, etc. . Due to omarkers ,61,62,63Owing to synergistic interactions, multiple active sites, and intrinsic catalytic activity, the nanocomposites of transition metal oxides with metal  nanoparticles, polymeric materials, and carbon (carbon nanotubes (CNTs), graphene, and fullerene) derived nanomaterials offer amplified electrochemical properties in comparison to the original constituents ,65,66,672O4 exhibited a low detection limit (5.8 μM) and high sensitivity (44.86 µA µM−1 cm−2)) towards sensing glucose compared to Ag-dispersed NiCo2O4 nanostructures [2O4 than Ag nanoparticles, Au–NiCo2O4 led more conductively than Ag–NiCo2O4. Besides the sensing glucose, a variety of electrochemical sensors based on metal/metal oxide nanocomposites and binary metal oxide/metal oxide nanocomposites have been demonstrated for the detection of various biomarkers, including hydrogen peroxide, lactic acid, dopamine, NADH, ascorbic acid, paracetamol, cysteine, etc. [It has been reported that Au-dispersed NiCoructures . Accordine, etc. ,70,71. T2O3) nanostructures [2O3 nanocomposites electrode delivered a low detection limit of 102 pM and a wide linear range of 0.001–0.900 μM. In addition to PANI polymeric materials, polypyrrole (PPy) and poly (PEDOT) were successfully integrated with metal oxide nanomaterials for sensing biomarkers for improving their electrical conductivity [Recent studies showed that PANI-derived electrochemical sensor practices have certain characteristic demerits including poor selectivity and reproducibility. PANI is often integrated with metal nanoparticles and metal oxide nanomaterials to overcome those challenges. Over the past decade, numerous transition metal oxide nanocomposites with polymeric materials have fascinated the field of electro-analysis ,66,72. Pructures . The as-uctivity ,74,75. Owing to a wide operating potential window, strong surface chemistry, lower price, and chemical inertness, various carbon-derived nanocomposites of metal oxides with graphite, carbon paste (CP), carbon nanotubes (CNTs), graphene (GR), carbon fibers (CFs), activate porous carbon, and carbon nanodots are usually utilized as the sensing materials for the sensitive detection of a variety of biomarkers ,77,78,793N4-SWCNTs), reduced graphene oxide (rGO), and an electrodeposited cyclodextrin-metal organic framework (CD-MOF) [3N4-SWCNTs/rGO/CD-MOF nanocomposites attained the sensitive and selective detection of Bn with a broad linear range of 1.0 nM–0.5 µM and low LOD of 0.46 nM with a sensitivity of 220 A/M  sensor was established by Zhao et al. based on graphitized carbon-nitride single-walled carbon nanotube nanocomposites (C(CD-MOF) . The res 220 A/M . In addiTwo-dimensional (2D) transition metal (TM) carbides, nitrides, and carbonitride, also termed as MXenes, have been considered to be promising materials with the integration of metal oxides nanocomposites for constructing sensors systems . In genePark and co-workers demonstrated an electrochemical biosensor platform based on nickel oxide (NiO)–reduced graphene oxide (rGO)/MXene nanocomposites for sensing active influenza viruses (H1N1 and H5N2) and viral proteins . The det3O4 nanocomposites-based electrochemical sensor platforms for detecting dopamine in the presence of ascorbic acid (AA) and serotonin (SE) under physiological pH 7.0 [In addition, metal oxides dispersed on conducting polymers-based nanocomposites have been employed as the sensing electrode materials for biomarkers ,89. A val pH 7.0 . The as-l pH 7.0 ,92,93. −1, a broad sensing range (0.01–100 ng mL−1), good specificity, and practicability. The effectual design of PEC sensing materials with high sensitivity and selectivity, non-toxic, low-energy excitation, and low expense is a central challenge.The design of a PEC sensor is also considered to be a new detection approach, which has attracted more attention ,93. In t−1 cm−2 for glucose and a linear range (100 nM–88.0 μM) with a LOD of ~27.0 nM and a sensitivity of ~6.19 mA μM−1 cm−2 , gold (Au), platinum (Pt), boron-doped diamond (BDD), etc., have been established for detecting biomolecules ,95,96. FM−1 cm−2 . The as-The design and establishment of high-sensing performance of durable and less-expensive sensing elements are of enormous importance for the growth and practical applications of electrochemical sensors and biosensor systems. Over the recent years, a lot of advanced and functional nanocomposites of metal oxides materials have been reported with high sensitivity, good specificity and reproducibility, and high practicability . The eleIn recent years, metal oxides nanocomposites have been attractive in various electrochemical technologies, in particular, in biosensors and bioelectronics. Especially, due to the characteristic electronic structure, high catalytic sites, and maximum atom utilization efficiency, single-atom catalysts (SAC) based on iron, nickel, and copper oxides are considered to be promising sensing elements. The high utilization of metal atoms in the electrode materials makes them deliver an outstanding sensing performance with lower consumption and a reduction of the cost of the electrode materials. In general, SACs are often dispersion on carbon nanomaterials or polymeric nanomaterials. The high sensing performance of the detection elements based on metal oxides nanocomposites is mainly attributed to various features such as size, shape or morphology, conducting support, coordination site, composition, electron confinement, and distance between the inter-particles. Transition metal oxide-derived nanomaterials and their nanocomposites provide an excessive potential for the progress of high-performance electrochemical sensors and biosensor platforms. Owing to improved detection performance with rapid response, low detection limit, high sensitivity, and potential selectivity in the presence of a real complex environment and robust analysis, metal oxides and their nanocomposites established widespread consideration. The excellent characteristics of metal oxides-based nanocomposites make them attractive as sensing elements with high surface area to volume ratios, biocompatibility, chemical stability, surface reaction activity, and adjustable electron transport properties. The unique electronic and catalytic properties of metal oxide-based nanocomposites are therefore able to increase the sensitivity, selectivity, and stability of the sensor platforms towards the detection of biomarkers. The present review offers major advancements in transition metal oxide nanomaterials and their nanocomposites-based electrochemical sensor and biosensor platforms for healthcare monitoring. Subsequently, the facile sensing electrode fabrication, catalytic activity, reaction mechanism, hetero-structured active sites, and surface engineering for enhancing transducer performance are described. Transition metal oxides-based nanocomposites with numerous 1D- or 2D- or 3D-nanostructures, including nanowires, nanotubes, nanoribbons, nanosheets, and nanodots are enduring to be at the lead of nanoscience and nanotechnology for electrocatalytic and sensor applications due to their unique physicochemical and electrochemical characteristics, which are associated to their dimensional anisotropy. The versatile utilization of nanostructured metal oxides and their nanocomposites in efficient electrocatalytic sensor applications has established worldwide concern and amplified investigative attention in commercial analytical applications towards clinical and biomedical diagnostics and healthcare monitoring. The enhancement of their overall sensing recital, primarily including the activity, selectivity, and solidity, is suggestively contingent on the progress of new composite materials employed as sensing elements. It is commonly understood that surface/interface reactions are crucial in several electrochemical processes, from electrochemical processes on an electrode to heterogeneous catalytic reactions. Hence, researchers prefer to synthesize smaller size and hetero-structured nanocomposites materials to ensure that larger active surfaces and multiple active sites are exposed in the selective electrocatalytic reactions. In-depth explorations are required to address challenges, including stabilization of multiple active sites, optimizing the adsorption of analytes and adsorbed product intermediates, reusable sensors platforms, etc. In recent years, heterostructures or nanocomposites of catalytically active sites dispersed on a durable matrix without allowing for agglomeration have emerged and appeared as excellent candidates to attain sensitive and selective detection of biomarkers via stabilization and tuning of electrochemically active sites. Finally, it is believed that further investigations may be studied for correlating the relationship between morphological/crystalline structures of the metal oxides nanocomposites and the reaction mechanism of analytes. Consequently, an organized inspection of the effect of morphological and crystalline structures, coordination environment, synergistic effects of multiple active elements, double layer interface, etc., of the transition metal oxides and their nanocomposites on the electrocatalytic and sensing performance towards clinical, biomedical, healthcare monitoring, and biological applications desires examination.


Future reproduction methods for boosting crucial traits will likely be influenced by the proportion of genetic and phenotypic variances. Using PCR-DNA sequencing, the variations in the nucleotide sequences of the immunological genes between healthy buffaloes and buffaloes suffering from inflammatory reproductive disorders were discovered. When buffaloes experienced inflammatory reproductive disorders, they were more likely to express the examined genes. The molecular alterations may, therefore, provide crucial information about the relationships between the physiologies of the various reproductive pathways and help diagnose inflammatory postpartum disorders.IKBKG, LGALS, IL1B, CCL2, RANTES, MASP2, HMGB1, and S-LZ) genes’ nucleotide sequence differences between healthy buffaloes and buffaloes affected by inflammatory reproductive diseases were found by employing PCR-DNA sequencing. According to Fisher’s exact test (p ˂ 0.01), there were noticeably different probabilities of all major nucleotide changes spreading among buffalo groups with and without reproductive problems. Buffaloes were significantly more likely to express the examined genes when they had inflammatory reproductive diseases. The outcomes might support the significance of these markers’ nucleotide variations and gene expression patterns as indicators of the prevalence of inflammatory reproductive disorders and provide a workable buffalo management policy.The immunological genes that may interact with inflammatory postpartum diseases in Italian buffaloes were examined in this study. A total number of 120 female Italian buffaloes  were employed. Each buffalo’s jugular vein was pierced to get five milliliters of blood. To obtain whole blood and extract DNA and RNA, the blood was placed within tubes containing sodium fluoride or EDTA anticoagulants. The immunological ( The postpartum period, or the first 45 days following delivery, is the most stressful time due to decreased feed intake, endocrine and metabolic changes during parturition and lactation, and usually a multitude of physiological stressors with immunological roots accompanying it . If thesBuffaloes are predisposed to inflammatory reproductive disorders after delivery since the majority of adult buffalo problems frequently manifest within the first month ,9,10. InIn mammals, there are two types of immunity: innate immunity and adaptive immunity. Both vertebrates and invertebrates have innate immunity, whereas only vertebrates have adaptive immunity . ExampleGiven that all evolution results from genetic variants, describing the genetic makeup of a population necessitates an understanding of that group’s evolutionary history ,23. AddiAccording to our current understanding, there is little data on the immunological changes and genetic variations linked to inflammatory postpartum disorders in buffalo, and there are also few reliable diagnostic and prognostic tools to help us improve our preventative and therapeutic measures. Furthermore, it is important to emphasize that more research is necessary to identify any relevant immunological indicators associated with reproductive disorders that determine whether buffalo are resistant to or susceptible to these diseases. In this study, real-time PCR and PCR-DNA sequencing techniques were used to assess the efficiency of putative immune genes as candidates for inflammatory reproductive disease incidence prediction and tracking in Italian buffaloes.The 120 Italian buffaloes used in this study were grown on a private farm in the Delta region of northern Egypt, 60 of which had inflammatory reproductive problems, and the remaining 60 appeared to be healthy. The investigated female buffaloes were aged 4.5 years and in the second lactation season with an average milk yield of 15 ± 2.7 kg. Researched animals were grown in a commercial dairy herd with approximately 300 dairy buffaloes. The healthy group had a history of regular reproductive performance . The buffaloes in the second group exhibit mastitis  and endometritis . Each buffalo’s jugular vein was pierced to get five milliliters of blood. The samples were placed into tubes encompassing anticoagulants (EDTA or sodium fluoride) in order to collect entire blood and extract DNA and RNA. All animal management techniques, test experimental collection and sample eradication were carried out under the direction of the Veterinary Medical School of the University of Sadat City (Code VUSC-027-1-23) in accordance with IACUC guidelines.IKBKG, LGALS, IL1B, CCL2, RANTES, MASP2, HMGB1, and S-LZ) were amplified. The Bubalus bubalis genome, which was accessible in GenBank, was utilized to create the oligonucleotide sequences for amplification. The primers used during the PCR are listed in The genetic material JET complete blood genomic DNA isolation kit and the manufacturer’s instructions  were used to extract DNA from the genome using total blood. DNA with good purity and concentration was taken into consideration using Nanodrop. PCR was conducted on a conserved part, i.e., exon of investigated genes that enables accurate molecular characterization of genes and deciphers physiological differences in resistance/susceptibility to inflammatory postpartum disorders. Therefore, parts of coding sites (CDS) for immunity-related genes . At a starting temperature of 94 °C for unwinding, the PCR combinations were used for five minutes. The 35 cycles consisted of 30-s rounds 30-s rounds of denaturation at 94 °C for 1 min, one-minute annealing cycles depending on the temperature range shown in Preceding DNA sequencing, kits from Jena Bioscience # pp-201s/Munich of Hamburg, Germany, presented techniques to purify PCR in order to remove primer dimmers, non-specific bands, and other impurities and provide the desired cloned PCR product for the projected scope . When asChromas 1.45 and BLAST 2.0 were the tools used for examining the DNA analysis findings . Compari® ND-1000 spectrophotometer. The producer’s method was used to create the whole nucleic acid in each sample . SYBR Green PCR Master Mix and quantitative RT-PCR  were used to assess the immune genes’ expression profiles. SYBR Green PCR Master Mix  has been used to determine the relative amount of mRNA that each sample possesses.The RNA was completely isolated using the Trizol solution per the manufacturer’s instructions from the blood samples collected from the studied buffaloes . We quantified and confirmed the amount of RNA that was isolated using a NanoDropBubalus bubalis genome assembly (NDDB_SH_1) from GenBank , the significant distribution of SNPs for the identified genes was observed between the buffaloes under study. The t-test was used to determine the statistical significance of the differences in the expression profile of immune genes between normal and inflammatory reproductive illness-affected buffaloes. Mean and standard error (mean ± SE) were used to present the results. Differences were deemed significant at p < 0.05. Using Fisher’s exact test analysis (p < 0.01). All of the reproductive markers under research had exonic region mutations, which resulted in different coding DNA sequences in normal buffaloes versus inflammatory reproductive disorder-affected ones. The immunological indicators that were being examined all had the exonic region modifications shown in SNP variations in amplified DNA nucleotides associated with inflammatory reproductive disorders were found in the results of PCR-DNA sequencing on both normal and affected buffaloes for the IKBKG (412-bp), LGALS (395-bp), IL1B (358-bp), CCL2 (347-bp), RANTES (399-bp), MASP2 (513-bp), HMGB1 (540-bp), and S-LZ (455-bp) genes. The DNA sequence differences between the GenBank-sourced reference gene nucleotide sequence and the reproductive indicators examined in the studied buffaloes were utilized to verify all of the discovered SNPs . Table 3IKBKG gene (412-bp) contained two non-synonymous SNPs, C78G and G328A, which caused the amino acids T26M and G110S to be substituted, respectively. For the LGALS gene (395-bp), synonymous mutation 64N occurred as a result of T192C SNP. The nucleotide sequence of the IL1B gene (359-bp) revealed four recurrent non-synonymous SNPs: T18M, V42A, L69S, and S104L, resulting from C53T, T125C, T205C, and C311T SNPs, respectively. One recurrent SNP in the CCL2 gene (347-bp) was discovered using DNA sequencing; C201T resulted in synonymous mutation, 67P. For the RANTES gene (399-bp), three observed recurrent synonymous SNPs, G96C, A141G, and C370T, resulted in 32T, 47S, and 124L amino acids, respectively. One recurrent SNP was found when the MASP2 gene (343-bp) was sequenced, where synonymous mutation 155A occurred as a result of T465A SNP. Three recurrent SNPs in the HMGB1 gene (540-bp) were discovered using DNA sequencing: C227A, C275T, and T407C, which resulted in non-synonymous mutations, S76L, P92L, and F136S, respectively. Three recurrent SNPs in the S-LZ gene (455-bp) were discovered using DNA sequencing; two of these, G33A and A222G, resulted in synonymous mutations, 11R and 74L, respectively. In comparison, the non-synonymous mutation caused by the G254A SNP led to the substitution of the amino acid R85K.The IKBKG, LGALS, IL1B, CCL2, RANTES, MASP2, HMGB1, and S-LZ were noticeably upregulated (CCL2 (2.21 ± 0.18), and the lowest level was found for LGALS (1.47 ± 0.15). Among all the genes analyzed in the normal buffaloes, the gene with the greatest possible mRNA level was found for S-LZ (0.65 ± 0.06), while the lowermost level was found for IKBKG (0.46 ± 0.15).In inflammatory reproductive disorder-affected buffaloes, expression levels of the genes egulated . The higIKBKG, LGALS, IL1B, CCL2, RANTES, MASP2, HMGB1, and S-LZ) genes in inflammatory reproductive disorder-affected and healthy Italian buffaloes were identified in this study utilizing sequenced amplified PCR products. The findings show that there are differences between the SNPs involving the healthy and inflammatory reproductive disorder-affected buffaloes. The assessed buffaloes had a considerable nucleotide polymorphism dispersion (p < 0.01), according to the Fisher’s exact test. It is important to emphasize that the polymorphisms found and made available in this context give new data for the evaluated indicators when compared to the pertinent datasets collected from GenBank.Knowing the genes, fundamental mutations, and communications with other variables that confer resistant animals is crucial for the efficient exploitation of disease-resistant cattle or the complete elimination of diseased animals . The immIKBKG gene (412-bp). According to the results of the basal local alignment search tool (BLAST), the cattle database’s altered mutated bases are conserved (GenBank accession number XM_027535016). Additionally, databases for sheep and goats both have the G328A SNP . A synonymous mutation for the LGALS gene (395-bp) was brought on by the T192C SNP. Cattle (GenBank accession number XM_027540847.1) retain the altered base. Four recurring non-synonymous SNPs, C53T, T125C, T205C, and C311T SNPs, were identified in the 359-bp nucleotide sequence of the IL1B gene. The identified SNPs were found to be conserved in other closely related species by nucleotide sequence alignment. For example, the goat has a conserved C53T SNP (GenBank accession number DQ837160.1), the cattle have a conserved T205C SNP (GenBank accession number EU276067.1), and the Syncerus caffer  has a conserved C311T mutated base (GenBank accession number AB571651.1). DNA sequencing led to the discovery of one recurrent SNP (in the CCL2 gene 347-bp): C201T. When compared to the genome of cattle, the changed nucleotide was shown to be conserved (GenBank accession number EU276059.1).Our findings demonstrated the presence of two nonsynonymous SNPs, C78G and G328A, in the RANTES gene (399 bp). In the genome of cattle, the three altered nucleotides are conserved . The A141G SNP was also discovered to be conserved in databases for sheep and goats, with the GenBank accession numbers XM_027975305.2 and XM_005693201.3, respectively. The recurrent T465A SNP for the MASP2 gene (343-bp) sequence was reported in nucleotide sequences from sheep and cattle, respectively, with the GenBank accession codes CP011908.1 and CP027092.1. C227A, C275T, and T407C are three recurrent SNPs that were found using DNA sequencing in the 540-bp HMGB1 gene. By comparing the three detected SNPs to cattle and sheep DNA sequences stored in GenBank under accession numbers CP027087.1, XM_042254827.1, CP011895.1, and LR962873.1, it was discovered that they were identical. However, only the C227A and T407C SNPs (GenBank accession numbers XM_018056617.1 and XM_018056617.1) were linked to the goat sequence. DNA sequencing was used to identify three recurrent SNPs in the S-LZ gene (455 bp), two of which, G33A and A222G, led to synonymous alterations. While the G254A SNP-caused non-synonymous mutation. The sheep GenBank sequence with accession numbers AH008120.2 and CP011908.1 matched the three newly identified SNPs. While the goat sequence shared the identical changed base for the G254A SNP (GenBank under accession number KC954666.1), the G33A was discovered to be aligned with the cattle database with the GeBank number M95099.1. It should be noted that the A222G SNP has the identical altered base in both cattle and goats, with accession numbers that may be found in GenBank: MF996700.1 and KC526216.1, respectively. The conservation behavior in the altered bases may be attributed to close relatedness among ruminant species, where genetic resource conservation programs are contributing to an increase in the numbers and to the preservation of valuable gene reservoirs [Bubalus bubalis that were used in our work and were published in PubMed. The variance of the immune  markers and how they relate to inflammatory reproductive disorders in Italian buffaloes have not, to our knowledge, been the subject of any prior research. The candidate gene method has, however, been utilized to assess the veracity of postpartum problems in buffalo. For instance, sequence analysis variations of the TLR2 and TLR4 genes were linked to endometritis and mastitis in river buffalo [Similarly, when comparing our DNA sequencing results with those of closely related species, three recurrent synonymous SNPs, G96C, A141G, and C370T, were found in the servoirs . AnotherTES gene 9 bp. In TLR4, IL10, TLR7, NCF4, TNF-α, and LITAF) genes, there were changes in the nucleotide sequence between endometritis-affected cows and healthy ones. In addition, using PCR-DNA sequencing of immunological  markers from the investigated Holstein and Brown Swiss dairy cows, nucleotide sequence alterations in the form of SNPs related to mastitis tolerance/susceptibility were found [LTF gene was linked to mastitis resistance [TLR4 gene polymorphisms and mastitis susceptibility were found to significantly correlate in sheep [TLR2 gene. In Barki sheep, TLR4 gene SNPs were likewise linked to postpartum disorder risk [In order to better understand the resistance and vulnerability of inflammatory reproductive diseases in livestock, genetic polymorphisms of immunological markers for postpartum endometritis were examined in Holstein dairy cows . Using Pre found . In cowssistance ,46,47. Tin sheep  and cattin sheep ,50. Accoin sheep , there ider risk .The primary source of adaptation and selection is mutation . All of IKBKG, LGALS, IL1B, CCL2, RANTES, MASP2, HMGB1, and S-LZ genes were measured using real-time PCR in both normal and inflammatory reproductive disorder-affected buffaloes. Our results demonstrated that buffaloes with inflammatory reproductive diseases expressed immune genes at higher levels than healthy ones. In order to ascertain the mRNA levels of these immunological markers linked to the resistance/susceptibility to inflammatory reproductive diseases in Italian buffaloes, real-time PCR was used in our experiment for the first time. Our investigation utilized qualitative SNP genetic markers, and quantitative gene expression overcame the drawbacks of past studies. As a result, in both healthy and affected by inflammatory reproductive illness buffaloes, the analyzed gene regulation’s mechanisms have been well-established.In the current work, we proposed that the course of inflammatory reproductive disorders may be influenced by genetic variation in an individual’s transcriptional response to those conditions. Bubalus bubalis associated with endometritis infection revealed that IL-1 and IL-6 gene expression was upregulated by 1.3, 1.7, and 5-fold, respectively, in endometritis-infected buffaloes compared to control animals. In contrast, when compared to uninfected animals, the expressions of IL-10 and TNF-α revealed lowered mRNA of 0.2 and 0.4-fold, respectively [Buffaloes suffering from reproductive problems have a sparsely developed gene expression profile of immunological markers. The findings of a study on the expression of cytokine genes in the uteri of ectively .CCL5, CXCL8, IL6, and IL1B) genes than cows without inflammation [TLR4, MyD88, IL-6, and IL-10 were upregulated at the transcript level, while CD14, TNF-, MD-2, NF-B, and IL-12 were noticeably downregulated [TLR4, TLR7, TNF, NCF4, and LITAF expression. Meanwhile, the expression of the IL10 gene was much decreased [SELL, SLC11A1, and FEZL gene expression were significantly higher in mastitic Holstein and Brown Swiss dairy cows than in tolerant animals [C3, C2, LTF, PF4, and TRAPPC13 are present in the blood and endometrium of dairy calves with subclinical endometritis. Additionally, in contrast to cows with sub-clinical endometritis, the gene expression profiles of C3, CXCL8, LTF, TLR2, and TRAPPC13 were temporally changed in healthy cows’ circulating WBC during the postpartum period [CCL5 gene expression did not significantly alter between the three different time periods during the transition phase [LTF gene for mastitis resistance was discovered to have an up-regulated expression profile in Damascus goats [Staphylococcus aureus mastitis were found to have a considerable increase in pro-inflammatory cytokines, chemokines, and their receptors compared to tolerant goats [TLR4, cytokines, and beta-defensin dramatically increased in mRNA expression. Regarding the sheep’s gene expression profile for immunological markers, according to Darwish et al. [IL5, IL6, IL1-ß, TNF alpha, TLR4, and Tollip than tolerant ewes. In dairy cows, the bovine endometrium of cows with inflammation had greater transcript levels of immunological  gene encodes the regulatory subunit of the inhibitor of kappaB kinase (IKK) complex. When NF-kappaB is activated, genes implicated in cell survival and immunity are also activated [The inhibitor of nuclear factor kappa B kinase regulatory subunit gamma , which encodes two tiny proteins that bind to the CCR2 receptors present on the surface of monocytes and neutrophils, respectively, acts as a potent chemokine for these cells [2+ to encourage chemotaxis towards inflammatory stimuli [CCL2 gene and production and health was investigated using Canadian Holstein cattle [C-C Motif Chemokine Ligand 2  serves as the primary protease in the complement system . AutoimmSP2 gene . NumerouSP2 gene . It was n cattle . High-mon cattle . In addin cattle . An antin cattle . Lysozymn cattle ,78.IKBKG, LGALS, IL1B, CCL2, RANTES, MASP2, HMGB1, and S-LZ) markers. This could be explained by cytotoxic radicals and proinflammatory cytokines generated by the phagocytic cells, both of which cause serious inflammation that harms the damaged tissue [Italian buffaloes with inflammatory reproductive disorders exhibit a marked upregulation in the expression of immune  indicators were discovered using PCR-DNA sequencing in healthy and sick Italian buffaloes with inflammatory reproductive diseases. Furthermore, healthy and affected buffaloes with inflammatory reproductive disorders have different levels of these indicators’ mRNA. By employing genetic markers along with normal welfare during buffalo selection, a promising opportunity exists to lessen the occurrence of inflammatory reproductive disorders thanks to these novel functional variations. The gene domains identified here may facilitate future methods for treating inflammatory reproductive diseases.Single nucleotide variations (SNPs) in the genes for immunological (


LOC_Os04g53310 , LOC_Os05g02070 , LOC_Os06g04200 , LOC_Os06g12450 , and LOC_Os08g09230 , are predicted to be involved in the process of rice grain starch synthesis and to influence grain expansion after cooking. Our findings provide valuable insights and will facilitate genetic research and improvement of CCRGE.Cooking-caused rice grain expansion (CCRGE) is a critical trait for evaluating the cooking quality of rice. Previous quantitative trait locus (QTL) mapping studies on CCRGE have been limited to bi-parental populations, which restrict the exploration of natural variation and mapping resolution. To comprehensively and precisely dissect the genetic basis of CCRGE, we performed a genome-wide association study (GWAS) on three related indices: grain breadth expansion index (GBEI), grain length expansion index (GLEI), and grain length-breadth ratio expansion index (GREI), using 345 rice accessions grown in two years (environments) and 193,582 SNP markers. By analyzing each environment separately using seven different methods , we identified a total of 32, 19 and 27 reliable quantitative trait nucleotides (QTNs) associated with GBEI, GLEI and GREI, respectively. Furthermore, by jointly analyzing the two environments using 3VmrMLM, we discovered 19, 22 and 25 QTNs, as well as 9, 5 and 7 QTN-by-environment interaction (QEIs) associated with GBEI, GLEI and GREI, respectively. Notably, 12, 9 and 15 QTNs for GBEI, GLEI and GREI were found within the intervals of previously reported QTLs. In the vicinity of these QTNs or QEIs, based on analyses of mutation type, gene ontology classification, haplotype, and expression pattern, we identified five candidate genes that are related to starch synthesis and endosperm development. The five candidate genes, namely,  It is the only cereal crop that is primarily consumed as whole grains, which underscores its significance in the field of rice breeding  underlying CCRGE. To date, 47 QTLs for grain length expansion , 10 QTLs2  as an effective method for identifying natural genomic variations associated with quantitative traits . Unlike In this study, we performed GWAS on three traits of CCRGE based on two replicated experiments conducted in two different years (environments) and using seven different methods to analyze the data. We detected 165 related quantitative trait nucleotides (QTNs), including some exhibiting only the effect of QTN-by-environment interaction (QEI). Based on the detected QTNs, we identified five candidate genes through gene ontology (GO), haplotype, and expression pattern analyses. Our findings will facilitate further genetic research and the genetic improvement of CCRGE.22.1A set of 345 rice accessions among the list of the 2.20 and B0) and 15 unbroken and straight cooked grains (L1 and B1) were measured. Subsequently, the grain breadth expansion index (GBEI), grain length expansion index (GLEI) and grain length-breadth relative expansion index (GREI) of each accession were calculated according to the formulae described by 1/L0, GBEI = B1/B0, and GREI = (L1/B1)/(L0/B0) = (L1/L0)/(B1/B0) = GLEI/GBEI.The procedure for quantifying the characteristics of cooking-caused rice grain expansion was performed according to 2.3http://iric.irri.org/resources/3000-genomes-project). The core genome set of 404K SNPs  was employed for the analysis. A stringent quality control process was performed, which involved removal of the SNPs that had more than 20% missing calls and a minor allele frequency (MAF) smaller than 5%. As a result, a total of 193,582 SNPs were retained for subsequent analysis.The SNP data of the 345 rice accessions were obtained from the 3K Rice Genomes Project (2.42) using the plink software  plots and the Admixture program as described by software .2.5https://github.com/YuanmingZhang65/IIIVmrMLM) and mrMLM v4.0.2  single-environment analysis, namely, analyzing each environment separately; and (2) two-environment analysis, namely, analyzing the two environments jointly. For single-environment analysis, we employed two R packages: 3VmrMLM  the SNP effect prediction software snpEff.v1.9 (https://www.ricedata.cn/ontology/), and all genes with these GO classifications were retrieved; and (3) genes that meet both steps 1 and 2 were screened out and then subjected to haplotype analysis, where different haplotypes exhibiting t-test significance were considered as candidate genes.Based on the distinct LD decay in each rice chromosome, the left and right REff.v1.9  was empl2.7http://rice.uga.edu), including shoots (library name in NCBI: SRR042529), leaves-20 days (OSN_AA and OSN_CA), pre-emergence inflorescence (OSN_AC), post-emergence inflorescence (OSN_AB), anther (OSN_AD), pistil (OSN_AE), seed-5 DAP , seed-10 DAP (OSN_AK), embryo-25 DAP (OSN_AG) and endosperm-25 DAP (OSN_AH and OSN_BH). A heatmap was generated to visualize the gene expression patterns across the different tissues.The expression profiles of the candidate genes in various tissues were obtained from the Rice Genome Annotation Project database  decreased with the increase of physical distance on every chromosome as well as in the whole genome  were found to be stable because they were detected in the two environments simultaneously (qGLEI-3.5s and qGREI-3.6s) and 6:25062099 (qGLEI-6.4s and qGREI-6.4s), both of which were associated with GLEI and GREI; and 5:5369111 (qGBEI-5.3s and qGREI-5.2s), which was associated with GBEI and GREI  and 8, 8 and 6 suggested QTNs (P > 2.58E-07 but LOD ≥ 3.0) associated with GBEI, GLEI and GREI, respectively  and qGLEI-5.6i (SNP 5:25726382) were also detected as QTN qGREI-2.2s and qGREI-5.8s in the single-environment analysis, respectively  and GREI (as qGREI-8.5i) simultaneously , 119 genes were found, of which 26 were located within the R2 half-decay distance around the detected QTNs/QEIs  classifications on the China Rice Data Center’s website (TNs/QEIs Table S5LOC_Os04g53310 (OsSSIIIb), LOC_Os06g04200 (wx) and LOC_Os08g09230 (OsSSIIIa) exhibited significant haplotype differences for GBEI; LOC_Os04g53310, LOC_Os05g02070 (OsMT2b) and LOC_Os06g12450 (OsSSIIa) displayed significant haplotype differences for GLEI; and all of the genes except for LOC_Os06g12450 showed significant haplotype differences for GREI  was expressed mainly in leaf and pre-emergence inflorescence but not in seed or endosperm; LOC_Os05g02070 (OsMT2b) was expressed mainly in post- and pre-emergence inflorescence and in embryo of 25 DAP (days after pollination), but not in endosperm. This suggests that these two genes maybe not closely or indirectly associated with endosperm development. In contrast, LOC_Os06g04200 (wx), LOC_Os06g12450 (OsSSIIa) and LOC_Os08g09230 (OsSSIIIa) exhibited high expression in 10 DAP seed, and the highest expression in 25 DAP endosperm, but no expression in embryo, indicating their potential involvement in starch synthesis or endosperm development.To further verify the potential impact of these candidate genes on the regulation of starch synthesis and endosperm development, we analyzed the expression patterns of the five candidate genes in various tissues based on data from the Rice Genome Annotation Project database Figure 84qGREI-5.2s and qGREI-5.6s, were commonly detected in two environments. This may be due to changes in the relative effects of different genes for these traits in different environments, indicating that the genes controlling these traits interacted with the environments. Joint analysis of the two environmental datasets using the 3VmrMLM method revealed 21 QEIs for three traits, also indicating the interaction between QTN and environment. Actually, ANOVA results showed significant genotype-by-environment interaction in the three traits  or after cooking (L1 and B1), and is directly proportional to GLEI while inversely proportional to GBEI. Evidently, genes governing GBEI and GLEI may also impact GREI in principle. In other words, the QTLs for GREI may exhibit pleiotropic effects on its component traits or correlated traits. In this study, we did identify 4 QTNs that simultaneously influence GREI and GLEI, and 2 QTNs that simultaneously affect GREI and GBEI  identified in this study were found to be in close proximity to four of the aforementioned QTLs).As mentioned above in the introduction, there were 10, 47 and 15 reported QTLs controlling length, width and length-width expansion caused by cooking in rice grain. Upon comparing these QTLs with the QTNs mapped in this study, we observed that 12, 9, and 15 QTNs for GBEI, GLEI, and GREI detected in this study were located within the intervals of one or more previously reported QTLs Table S6OsMT2b encodes a metallothionein that binds to metal ions and scavenges reactive oxygen species (ROS). OsMT2b (LOC_Os05g02070) transcription level and inhibit 26S proteasome-mediated OsMT2b protein degradation, thereby facilitating ROS clearance, delaying programmed cell death (PCD) of endosperm cells, and ultimately increasing the accumulation of storage substances, and reducing chalkiness rate. In this study, a SNP site is present in the 5’-UTR region of OsMT2b near qGLEI-5.1s , OsSSIIa (LOC_Os06g12450), and OsSSIIIa (LOC_Os08g09230) are crucial genes involved in the biosynthesis of starch in rice grains. wx gene encodes granule-bound starch synthase (GBSS), a major enzyme responsible for amylose synthesis  is a gene that encodes soluble starch synthase in rice. Its expression level and activity directly impact the synthesis and quality of starch in rice endosperm. OsSSIIIb can interact coordinately with OsSSIIIa, and loss of function of both genes leads to an increase in resistant starch content in cooked rice , LOC_Os05g02070 (OsMT2b), LOC_Os06g04200 (wx), LOC_Os06g12450 (OsSSIIa), and LOC_Os08g09230 (OsSSIIIa). These findings can be instrumental in identifying genes and conducting in-depth genetic research on CCRGE.In this study, data of GBEI, GLEI and GREI, three traits related to rice grain cooked expansion, were collected from 345 rice accessions in two distinct environments. Utilizing 193,582 SNP markers, seven methods were employed to identify QTNs based on single-environment data, while the 3VmrMLM method was utilized to identify QTNs and QEIs based on two-environment data. A total of 165 reliable QTNs/QEIs were detected, with 60, 46 and 59 of them being associated with GLEI, GBEI and GREI, respectively. Additionally, 26 genes related to starch synthesis or endosperm development were found to be located around these QTNs/QEIs. Further haplotype and expression pattern analyses led to the identification of five candidate genes, namely The original contributions presented in the study are included in the article/YZ and WW conceived and designed the experiment. KT, LL, EK, EN, ML, WN and SA measured the phenotypes of the traits. YZ, KT and XX analyzed the data. YZ and KT wrote the draft. WW revised the manuscript. All authors contributed to the article and approved the submitted version.


Lr genes) were exposed to Kazakhstani Puccinia triticina (Pt) races at the seedling stage. Field tests were performed to characterize leaf rust responses at the adult plant growth stage in the 2020–2021 and 2021–2022 cropping seasons. The wheat collection showed phenotypic diversity when tested with two virulent races of Pt. Thirteen wheat genotypes (18.6%) showed high resistance at both seedling and adult plant stages. In most cases, breeding material originating from international nurseries showed higher resistance to LR. Nine Lr genes, viz. Lr9, Lr10, Lr19, Lr26, Lr28, Lr34, Lr37, Lr46, and Lr68, either singly or in combination, were identified in 47 genotypes. Known Lr genes were not detected in the remaining 23 genotypes. The most commonly identified resistance genes were Lr37 (17 cultivars), Lr34 (16 cultivars), and Lr46 (10 cultivars), while Lr19, Lr68, Lr26, and Lr28 were the least frequent. Four Lr genes were identified in Keremet and Hisorok, followed by three Lr genes in Aliya, Rasad, Reke, Mataj, Egana and Almaly/Obri. The molecular screening revealed twenty-nine carriers of a single Lr gene, ten carriers of two genes, six carriers of three genes, and two carriers of four genes. Most of these accessions showed a high and moderate level of APR (Adult plant resistance) and may be utilized for the incorporation of Lr genes in well-adapted wheat cultivars. The most effective combination was Lr37, Lr34, and Lr68, the carriers of which were characterized by a low disease susceptibility index. The obtained results will facilitate breeding programs for wheat resistance in Kazakhstan.Leaf rust (LR) is the most widespread disease of common wheat worldwide. In order to evaluate leaf rust resistance, 70 uncharacterized wheat cultivars and promising lines with unknown leaf rust resistance genes ( Triticum aestivum L.) is one of the world’s key grain crops and contributes significantly to food security. With over 781.38 million tons of annual production, wheat has become one of the most prevalent and important crops on the planet [stat.gov.kz (accessed on 20 April 2023)) [Puccinia triticina [P. triticina on crops causes serious disease, resulting in yield losses reaching over 50% [P. triticina is observed on the territory of Southern Kazakhstan [Wheat  to the most common races of the fungus in the region [Avr genes, somatic hybridization, and geographical migration, new, more virulent races appear [Wheat resistance to the leaf rust pathogen is associated with the presence of appropriate resistance genes. Each or gene” . The hige region . As a res appear ,30. In as appear .Lr genes are already known to be distributed on all 21 wheat chromosomes; most of them come from wild relatives  [Lr9 gene was translocated into wheat from Aegilops umbellulata [Lr28—from Aegilops speltoides [Lr1, Lr10, and Lr21. These genes cause a low level of infection, manifested by the appearance of hypersensitivity patches or small uredinia encircled by chlorosis or necrosis [According to the Catalogue of Gene Symbols, more than 80 species) . For exaellulata , and Lr2eltoides . Seedlineltoides . ASR geneltoides . This tyeltoides . Two geneltoides . Examplenecrosis . The lonnecrosis .P. triticina. The single APR genes are not able to completely prevent the formation of urediniums and provide an immune response [Lr34, which is present in wheat germplasm all over the world [Lr34 gene encodes an ATP-binding cassette transporter protein (ABC transporters) and might be involved in the secretion of antifungal molecules. The proteins of this family have a similar basic structure, consisting of two cytosolic nucleotide-binding domains and two hydrophobic transmembrane domains. Identical homologous proteins were found in rice (OsPDR23) and Arabidopsis [Sr), yellow rust (Yr), and powdery mildew (Pm)—Lr34/Sr57/Yr18/Pm38 [Lr37/Yr17/Sr38 [Lr67/Sr55/Yr46/Pm46 [Adult plant resistance (APR) genes are expressed at post-emergence stages . The keyresponse . APR genhe world , is the he world . The Lr3bidopsis . There ar18/Pm38 , Lr37/Yrr17/Sr38 , and Lr6r46/Pm46 . The useLr34, Lr46, and Lr68 have shown long-term resistance. It was also shown that it is still effective to use varieties and lines with the genes Lr9, Lr10, Lr19, Lr26, Lr28, Lr34, Lr37, and Lr68 to control the Kazakhstan leaf rust populations [Lr genes in the diverse wheat germplasm from Kazakhstan as well as advanced lines originating from the breeding program IWWIP  developed by CIMMYT-ICARDA.Gene pyramiding can be carried out both by traditional breeding methods and by indirect selection using DNA markers associated with resistance genes . Traditiulations ,50. The Lr genes.The purpose of this study was to assess a collection of winter wheat for LR seedling and adult plant resistance (APR) and to investigate the potential for resistance in wheat germplasm using molecular markers linked to p < 0.001) for wheat genotypes, while the race effect was found to be significant (p < 0.001)  .Pt races (TJTTR and MKTTQ) for resistant and susceptible genotypes based on mean values is shown in A wide variation was observed in leaf rust disease severity based on infection types (IT) 0–4, ranging from very resistant to very susceptible in the wheat collection. Further, the frequency distribution of the infection types (ITs) produced by the two different Pt race MKTKQ (IT—0–2). Five cultivars  showed immune (IT = 0) reactions, and nine cultivars  showed resistance (IT = 1) reactions (b2) for leaf rust resistance was revealed (0.90).Thirty-six accessions (51.4%) demonstrated resistant or moderately resistant reactions to eactions . A highep < 0.001). Correlation analysis between the response of seedlings to race TJTTR and the average coefficient of infection (ACI) showed a significant positive correlation in 2021  and 2022 . This analysis revealed a positive relationship between the leaf rust average coefficient of infection (ACI) and responses to the race MKTKQ in 2021  and in 2022 .Reactions of wheat seedlings to race TJTTR were strongly associated with resistance to race MKTKQ  analysis. The high level of heritability (hb2—0.79) of the disease susceptibility index among wheat genotypes was shown  and susceptible (MS-S) accessions . The disas shown .To identify the most promising wheat varieties, an analysis of productivity was carried out, which made it possible to evaluate the wheat collection based on the plant height , the days to heading (DH), the spike lengths , the mean number of spikelets/spike (SS), the number of grains per spike (GS), the weight of grain per spike , and the thousand kernel weight  . The larp < 0.001), as well as between AUDPC and WGS  and AUDPC and GS . Analysis between NDVI and DH has shown significant positive correlations , as well as between NDVI and PH  (p < 0.001), GS , and TKW . A positive correlation was noted between NDVI and DH .There was a significant negative correlation in 2021 between AUDPC and TKW  . In 2022In order to examine the association between traits, a principal component analysis (PCA) was performed and visualized as separate biplots for 2021 and 2022 . PCA wasBiplot analysis, based on the reaction of wheat accessions to the leaf rust pathogen and productivity traits, showed that the samples Rasad, Hisorok, Koksu, Gozgon, Kokbidaj, and Alihan showed the most resistant reaction to the pathogen and high productivity.Lr genes were individually identified in each corresponding NIL , as well as in cv Pavon 76 (Lr46) and in cv. Parula (Lr68). Molecular screening results for the presence of relevant Lr genes are presented in Lr9 gene. The marker is closely linked to the gene, as evidenced by the absence of recombination between them [Lr9 was found in seven of the fifty-five genotypes, including Mataj, Raminal, Yzhnaya 12, Hisorok, Egana, Anar, and Almaly/Obri.In order to test 70 varieties and lines of winter wheat, nine closely linked specific markers for nine investigated een them . Of the Lr10 gene was carried out using the specific STS marker FI.2245lr10-6/r2 [Lr10 was found in six wheat accessions: Aliya, Hazarti Bashir, Hisorok, Layagatlii 80, Egana, and Yr2/Octyabrina.The search for the r10-6/r2 . The marAgropiron elongatum (Host) Beauvois with the gene Lr19 located on chromosome 7DL. This translocation is also associated with the yellow coloration of the endosperm, which limits its use in bread wheat breeding. Zhang and Dubcovsky developed a set of markers for alleles of the phytoene synthase 1-Psy-B1 gene, which also allows the detection of the presence of Lr19 [Lr19/Sr25 gene complex was detected in three wheat accessions , as evidenced by the presence of a 191 bp PCR product.Common wheat received a translocation from  of Lr19 . The marLr26), stem (Sr31), and yellow (Yr9) rust are located on the short arm of chromosome 1 of rye (1RS) and have been transferred to wheat through translocations [Lr26 [Lr26 gene was confirmed by amplification of the 1100 bp product in eight genotypes: Keremet, Bunyodkor, Shafag 2, Layagatlii 80, Naz/GF55-3, 416-SP-2, 9-CP and 14-CP.The genes for resistance to leaf .The SSR marker WMC 313 linked to his gene . The Lr2osome 4A . A 320-bLr34 gene is linked to the R genes for yellow rust, Yr18; stem rust, Sr57; and powdery mildew, Pm38. The presence of the APR gene Lr34/Yr18 was assessed using the codominant STS marker csLV34, linked at a distance of 0.4 cm from the Lr34 locus [Lr34 and was found in 16 wheat accessions, including Almaly, Karaspan, Karlygash, Raminal, Rasad, Egana, Naz/GF55-2, Almaly/Obri, 416-SP-2, 2-CP, 4-CP, 5-CP, 15-CP, 16-CP, 17-CP, and 19-CP.The 34 locus . The 150Lr37/Sr38/Yr17 gene complex localized in the short arm of the 2NS chromosome of Triticum ventricosum (Tausch) was translocated to the short arm of the common wheat chromosome 2AS. The CAPS marker Uric-Ln2 was used to identify wheat genotypes carrying the 2NS translocation [Lr37 gene was found in 17 wheat cultivars, including Aliya, Matai, Kyzylbidai, Keremet, Kokbidai, Rasad, Reke, Hisorok, Alikhan, Krasnovodopadskaya 210, 3-CP, 7-CP, 9-CP, 11-CP, 14-CP, 15-CP, and 17-CP.The location . The Lr3Lr46 gene, which is a complex locus providing multifactorial resistance to yellow rust (Yr29), stem rust (Sr58), and powdery mildew (Pm39) [Lr46 [Lr46, including cvs Alatau, Azharly, Akbidaj, Batyr, Farabi, Mataj, Keremet, Koksu, Rasad, and Reke.The m rust Sr, and pow9) [Lr46 . ScreeniLr68 was found in three wheat genotypes: Naz/GF55-3, Alihan, and Almaly/Obri. Carriers of the resistance gene were identified using the dominant STS marker csGS, mapped at a distance of 1.2 cM proximal to Lr68 [The marker linked to  to Lr68 .Lr19 and Lr24 genes [Lr9, Lr19, Lr24, Lr25, and Lr28, and that of the North Kazakhstani population to Lr19, Lr24, Lr25, Lr28, Lr36 and Lr45 [Lr9 and Lr19 was confirmed for both races of the pathogen on Thatcher differential lines, which indicates the effectiveness of these genes in providing seedling resistance.Rust diseases were and still are one of the key reasons for the decline in yields and deterioration in the quality of wheat grain both in Kazakhstan  and arou24 genes ,65. Earland Lr45 . In our P. triticina that causes leaf rust was assessed in this study during the seedling and adult plant stages. One of the primary objectives of breeding programs is the identification of resistant genotypes [The resistance of 70 wheat genotypes to the pathogen enotypes ,67. AccoLr genes and gene complexes. A total of 47 carriers of resistance genes were identified. In varieties Egemen 20, Kazakhstanskaya 10, Progress, Prezident, Rausin, Zhalyn, Daulet, 428/MK-122A-1, Faravon, Steklovidnaya 24, Sultan 2, Naz/Immun 78, Naz/GF55-5, Derbes, APK/Progress, Gozgon, 415-SP-2, 6-CP, 8-CP, 10-CP, 12-CP, 13-CP, and 18-CP, none of the tested Lr genes were identified. The Gozgon (5R), 13-CP (IT-0), 17-CP (20MR), and 415-SP-2 (10MR) genotypes demonstrated a high level of resistance to P. triticina, indicating that extra Lr genes are giving resistance in these genotypes.Molecular screening of seventy wheat accessions was conducted with linked nine markers to identify both carriers of single resistance Lr resistance genes in wheat breeding material were discovered in several earlier studies [Lr genes. Twenty-nine cultivars with one Lr gene were identified. Ten accessions of wheat were carriers of two Lr genes. Among the 70 accessions produced in Kazakhstan, three leaf rust resistance genes  were demonstrated to occur at high frequency: 24.3%, 22.8% and 14.3%, respectively.The origins of  studies ,17,50,68Lr9, six (8.6%) carried the gene Lr10, and three accessions (4.3%) had Lr19 and Lr68 each; Lr26 and Lr28 were found in eight (11.4%) and five (7.1%) cultivars, respectively. These leaf rust resistance genes showed evidence of providing adequate protection in the investigated genotypes. Two genes (Lr9 and Lr34) were identified in Raminal; Lr10 and Lr26 were found in Layagatlii 80; Lr37 and Lr19—in Kyzylbidaj; Lr26 and Lr68—in Naz/GF55-3; Lr68 and Lr37—in Alihan; Lr26 and Lr34—in 416-SP-2; Lr26 and Lr37 in 9-CP and 14-CP; Lr34 and Lr37—in 15-CP and 17-CP.Seven (10%) carried the leaf rust resistance gene Lr37 and Lr34 genes differed in the highest frequency of occurrence . The Lr37 gene still provides a sufficient level of resistance, which indicates the need for its introduction into breeding programs [Lr46 gene provides “slow-rusting”, although it is a less effective gene compared to the Lr34 gene. When combined with Lr34 and/or Lr68, it provides an almost immune response to the leaf rust pathogen [Lr34 is the first cloned slow-rusting gene that has been stable for over 50 years [Lr19 gene against leaf rust races and the Sr25 gene against stem rust was previously confirmed in an extensive collection of wheat germplasm [Lr9, Lr19, and Lr28 genes are still effective in China [Lr9 and Lr19 in Bulgaria [Lr28 in the US of America [Lr26 was overcome in Chinese and Indian leaf rust populations [The programs ,71,72,73pathogen . Race nopathogen . Lr34 is50 years . The effermplasm . Lr9, Lrin China , India, in China , Slovakiin China , Iran [8in China , France in China , Egypt [in China ,83, and in China , but patBulgaria , and Lr2 America . Also, rulations ,79.Lr37, Lr34, and Lr68, the carriers of which were characterized by a low disease susceptibility index (φ—0). In six varieties, three resistance genes were found: Lr10, Lr28, Lr37—in Aliya; Lr34, Lr37, Lr46—in Rasad; Lr28, Lr37, Lr46—in Reke; Lr9, Lr37, Lr46—in Mataj; Lr9, Lr10, Lr34—in Egana; Lr9, Lr34, Lr68—in Almaly/Obri. Two cultivars were carriers of four resistance genes: Lr37, Lr26, Lr46 and Lr19 (Keremet); Lr9, Lr10, Lr37 and Lr19 (Hisorok). These cultivars showed a high level of resistance at the stage of an adult plant for the entire growing season (φ—0). Different responses of cultivars carrying the same resistance genes to P. triticina may be associated with the cumulative effect of genes, the presence of unidentified genes, different expression levels of resistance genes, and other biotic and abiotic factors [Lr genes.In our study, the most effective combination was the presence of  factors ,88. FutuTriticum aestivum L.), including 42 cultivars grown and/or produced in Kazakhstan, 8 cultivars/advanced lines originating from the breeding program IWWIP  developed by CIMMYT-ICARDA, and 20 advanced lines selected from the Kazakhstan-CIMMYT breeding program , Transfer/6*TC (Lr9), TC*6/Exchange (Lr10), TC*7/Tr (Lr19), TC*6/ST-1-25 (Lr26), CS2D-2M (Lr28), TC*6/PI58548 (Lr34), and TC*6/VPM (Lr37) were used as positive controls in Lr gene detection.The collection of 70 winter wheat genotypes  [3 spores). Pustules of leaf rust appeared on the leaves 8–10 days after inoculation, from which inoculum was collected on the 12th day using a mechanical cyclone collector in a zero-size capsule. Spores were dried up to 20–30% relative humidity and then sealed in glass vials. The spores were preserved in an ultra-low refrigerator at −70 °C until further use. Urediniospores were heat treated in a water bath at 40 °C for 5–7 min to break cold-induced dormancy upon removal from storage [Spore collection, storage and reproduction were performed under controlled conditions at the Institute of Plant Biology and Biotechnology, Almaty, Kazakhstan. Leaves bearing the uredinia of leaf rust, 40 mg/L) . Leaf se storage ,91.P. triticina were differentiated using the three-letter nomenclature of Long and Kolmer (1989), which served as the basis for the virulence codes for the isolates [Lr1 (RL6003), Lr2a (RL6000), Lr2c (RL6047), and Lr3 (RL6002); set 2: Lr9 (RL6010), Lr16 (RL6005), Lr24 (RL 6064), and Lr26 (6078); set 3: Lr3ka (RL6007), Lr11 (RL6053), Lr17 (RL6008), and Lr30 (RL6049); set 4: Lr2b (RL6019), Lr3bg (RL6042), Lr14a (RL6013), and Lr14b (RL6006); set 5: Lr15 (RL60052), Lr18 (RL6009), Lr19 (RL6040), and Lr20 (RL6092). For all seedling tests, seeds were sown in 12-cm pots and placed at 18 °C until germination. Using a spray gun, seedlings at the two-leaf stage were inoculated with urediniospores from individual rust samples suspended in light mineral oil at a concentration of 2–3 mg/mL (5–10 × 103 spores). The inoculated seedlings were kept for a day in a climatic chamber at 70% humidity and a temperature of 18°, followed by 20 °C with 18 h of light [Puccinia triticina [Races of isolates . Virulenisolates . Set 1: of light . The seeof light . Infectiriticina . CultivaThe TJTTR race was characterized by high virulence (85%) and was more aggressive than the MKTKQ race (70%).P. triticina isolates, TJTTR and MKTKQ. Under laboratory conditions, the plants were grown in plastic containers (5–8 grains of each cultivar). At the first leaf phase (10–12 days), plants were sprayed with each race’s spore suspension at a concentration of 2–3 mg/mL (5–10 × 103 spores). The incubation of infected plants was carried out according to the parameters described above for race identification. The seedling resistance of wheat collected to the isolates of leaf rust was assessed 10–12 days after inoculation according to the Mains and Jackson scale [A collection of 70 varieties and lines of winter wheat was screened at the seedling stage for two leaf rust races. Plant reactions to leaf rust at the seedling stage were evaluated for the two on scale .2. Treatments and management techniques for fertilizers matched those often advised for the area [3/ha and kept free from weeds.The experimental material was phenotyped during the 2021 and 2022 growing seasons at the Kazakh Research Institute of Agriculture and Plant Growing (KRIAPG), Almalybak , Almaty region. Three replicates were used in a totally randomized design for the experiment. The individual plot size was 1 mthe area . Fertilihttp://weatherarchive.ru (accessed on 15 April 2023)). In May, the amount of precipitation exceeded the norm, which led to an increase in environmental humidity and contributed to the effective infection of plants with spores of Puccinia triticina  were used to inoculate field plots in both test years (2021 and 2022). The ratio of the urediospores of the five selected races forming each year’s inoculation was determined according to their frequencies in the previous years (2020). The percentages of the five races used to make the urediniospore mixtures for the 2021–2022 test were TJTTR (30%), MKTKQ (30%), TDTTR (15%), TFTTQ (15%) and MFTTQ (10%). The susceptible cultivar Morocco was used to multiply the inoculum. Plants were inoculated with a mixture of spores and talc (1:100) at a rate of 20 mg spores per 1 m2 (5–10 × 104 spores) at the boot stage [Mixed races of ot stage ,96. Afteot stage . The spoot stage ,97, whicProductivity was characterized by the major components, namely plant height , days to heading (DH), spike lengths , the mean number of spikelets/spike (SS), grains/spike (GS), the weight of grain/spike , and thousand kernel weights . The weight of a thousand kernels was estimated in grams with the measurement of the mass of seeds after adjusting the moisture content to 12% . Given tAccording to Saari and Wilcoxson (1974), the average Coefficient of Infection (ACI) was determined by multiplying the severity values by the constants for infection types: R (resistant) = 0.2; MR (moderately resistant) = 0.4; MS (moderately susceptible) = 0.8; and S (susceptible) = 1 . The foliy—an assessment of disease at the ith observation;it—time (in days) at the ith observation;n—the total number of observations.The susceptibility index (φ) is calculated from the ratio of the AUDPC of the sample to the AUDPC of the susceptible control.bh2 = SSg/SSt, where SSg is the sum of squares for genotype and SSt is the total sum of squares.In order to determine genotypic and year variances among genotypes for traits of productivity and leaf rust resistance, analysis of variance (ANOVA) was performed using R-studio software, and coefficients of Pearson correlation were calculated using the mean values of the characters assessed. Principal component analysis was performed, and biplots were prepared using R-studio software in R version 3.5.3 . The broLr genes were employed according to certain approved protocols. The polymerase chain reaction (PCR) was conducted using the primers and annealing temperature settings that were specified for each Lr gene in the references (2 (25 mM), 0.2 µL Taq polymerase (5 units µL) , 2.5 µL 10×PCR buffer and 12.8 µL ddH20. TBE buffer  was used to separate the amplification products, and ethidium bromide was added [Each genotype’s genomic DNA was extracted using the CTAB method from the fresh leaves of individual plants at the two-leaf seedling stage . The conferences . A Bio-Ras added . A 100-bPuccinia triticina were tested. The results indicated a significant positive correlation between seedling resistance and adult plant resistance for 2021 and 2022. A highly significant negative correlation was found between the AUDP and the weight of a thousand kernels in susceptible accessions. Twelve wheat accessions that were resistant both at the seedling and adult plant stages were selected, and they can be used directly in breeding programs to improve the leaf rust resistance of wheat. The molecular screening revealed twenty-seven carriers of a single effective Lr resistance gene, ten carriers of two Lr genes, six carriers of three Lr genes, and two carriers of four Lr genes. Large-scale single-gene variety cultivation places pathogens under intense selection pressure, which may eventually cause the establishment of an epiphytotic disease [Lr genes that were discovered in this study.In this study, a collection of 70 winter wheat genotypes showed phenotypic diversity in leaf rust resistance. Two virulent races of  disease ,106. In


N-acyl homoserine lactones (AHLs) as the autoinducer were detected in bacteria isolated from hydrothermal vents in the Okinawa Trough. A total of 18 and 108 isolates possessed AHL-producing and AHL-degrading abilities, respectively. Bacteria mainly affiliated with Rhodobacterales, Hyphomicrobiales, Enterobacterales and Sphingomonadales showed QS activities; QQ was mainly associated with Bacillales, Rhodospirillales and Sphingomonadales. The results showed that the bacterial QS and QQ processes are prevalent in hydrothermal environments in the Okinawa Trough. Furthermore, QS significantly affected the activities of extracellular enzymes represented by β-glucosidase, aminopeptidase and phosphatase in the four isolates with higher QS activities. Our results increase the current knowledge of the diversity of QS and QQ bacteria in extreme marine environments and shed light on the interspecific relationships to better investigate their dynamics and ecological roles in biogeochemical cycling.Quorum sensing (QS) is a chemical communication system by which bacteria coordinate gene expression and social behaviors. Quorum quenching (QQ) refers to processes of inhibiting the QS pathway. Deep-sea hydrothermal vents are extreme marine environments, where abundant and diverse microbial communities live. However, the nature of chemical communication in bacteria inhabiting the hydrothermal vent is poorly understood. In this study, the QS and QQ activities with  N-acyl homoserine lactones (AHLs) has been widely studied in a diverse range of Gram-negative bacteria. Moreover, the AHL-producing genes have been identified in many bacteria, including luxI which leads to the formation of 3OC6-HSL in Vibrio spp., and lasI which is involved in 3OC10-HSL in Pseudomonas aeruginosa [Quorum sensing (QS) is a bacterial communication system that enables bacterial populations to regulate their group behaviors according to density . QS reliruginosa .Trichoderma colonies [Alphaproteobacteria and Gammaproteobacteria [Actinobacteria [Firmicutes [luxI genes in the global ocean metagenomes, indicating that AHL-QS may be of great significance for communication among microorganisms, and the regulation of biological behaviors in marine habitats. Additionally, AHL-QS was associated with large-scale bioluminescence events caused by algal blooms [Marine bacteria with AHL-mediated QS (AHL-QS) activities have been isolated from multiple marine habitats, such as marine snow , corals colonies  and sea colonies . These bbacteria . In addibacteria ,16, Firmrmicutes , archaearmicutes  and cyanrmicutes  could prrmicutes  identifil blooms  and was l blooms .Ruegeria mobilis Rm01, which was isolated from marine particles, could be regulated by QS [Trichodesmium consortia could double the activity of phosphatase in the epibiotic bacteria, thereby promoting phosphorus circulation [AHL-QS exerts an important role in regulating the elements cycling in the global ocean by affecting the activity of extracellular hydrolase and biofilm formation. For example, Jatt et al.  determined by QS . In addiculation .Quorum quenching (QQ) inhibits QS pathways by interfering with the production, release, recognition, and degradation of signaling molecules, thereby affecting various physiological functions dependent on bacterial QS ,24. Inde2, CH4, NH4+, iodine and potassium [Sulfurimonas and Sulfurovum), sulfate-reducing bacteria (Thermosulfidibacter and Desulfothermus) and methanogens  living in the hydrothermal vents [In deep-sea hydrothermal vents, the dramatic changes in temperature (approximately 4–301 °C) and pH (ca. 5.0–7.6) vary in different marine environments . Compareotassium . In addial vents ,34,35,36In this study, we explored the diversity of QS and QQ isolates, and the regulatory function of QS on extracellular enzymatic activities in bacteria isolated from hydrothermal fields of the Okinawa Trough. The AHL-based QS and QQ activities of culturable bacteria were screened, and the biofilm formation of QS and partial-QQ strains was detected to reveal the relationship between QS/QQ and biofilm production. In addition, the effects of QS on the extracellular enzymatic activities of QS bacteria were explored.Bacteria used in this study were isolated from seawater, sediment and macroorganisms  from the hydrothermal fields of the Okinawa Trough during the HOBAB2 voyage in 2014 and the HOBAB4 voyage in 2016 and 2018. The samples were collected by a TV-grab sampler or the remotely operated vehicle (ROV) Faxian during the cruises conducted by the R/V Kexue. The information about samples, including longitude and latitude and depth, were listed in −1 to 10−3, and the sediment samples were soaked in 0.85 % (w/v) saline serially diluted from 10−1 to 10−3, and 100 μL of each dilution was spread onto the surface of marine agar 2216E (MA) plates [https://www.ezbiocloud.net). The phylogenetic tree of 16S rRNA genes from QS strains, their closest type strains and the reported AHL-producing strains was conducted using MEGA v11 based on the neighbor-joining algorithm [The seawater samples were serially diluted from 10) plates  and R2A ) plates . The maclgorithm . The phylgorithm .Agrobacterium tumefaciens (pCF218) (pCF372) A136 [A. tumefaciens A136 was added into the LB medium with 1% agar and X-gal (250 μg/L), and then the Oxford cups were placed on the plate to form wells. Meanwhile, fresh cultures were added into the wells, and the C6-HSL and MB were used as positive and negative controls, respectively. After co-incubation at 28 °C for 24 h, positivity was indicated by the presence of indigo spots. In addition, the high-throughput method described by Su et al. [6-HSL and MB were used as positive and negative controls, respectively. All the experiments were performed in quadruplicate.Screening for AHL-producing and AHL-degrading bacteria was conducted using the bacterial biosensor 72) A136 ,41,42. A72) A136 . Brieflyu et al.  and mixe6-HSL (representative for AHLs with short acyl chains) and C12-HSL (representative for AHLs with long acyl chains) were used as the substrates. Briefly, C6-HSL or C12-HSL was mixed with the bacterial culture and MB medium (negative control), and maintained at 28 °C for 24 h. After incubation, the supernatant was obtained after centrifugation at 4 °C and 6000 rpm for 10 min, and filtered through a 0.22 μm filter (JINTENG® PES syringe filter). The supernatant was then mixed with an A136 X-gal assay solution in 96-well microtiter plates, and incubated at 28 °C for another 24 h. All the experiments were performed in triplicate. Positive results were indicated by reduced indigo color compared with the negative control.AHL-degrading activity was detected using the high-throughput method described by Tang et al. . C6-HSL 6-HSL or C12-HSL, HEPES buffer, and A136 X-gal assay solution were mixed and added into a 96-well microtiter plate, and incubated at 28 °C for 24 h. All the experiments were performed in triplicate. Positive results were indicated by a reduced indigo color compared with the negative control.The QQ activities of QS strains were further identified using the high-throughput method. In short, QS strains were inoculated into MB with shaking (170 rpm) at 28 °C for 24 h. The cells were harvested after centrifugation at 4 °C and 6000 rpm for 10 min and re-suspended in HEPES buffer  for sonication. Additional centrifugation for the lysed cells was conducted at 4 °C and 12,000 rpm to obtain the crude enzyme supernatant. The crude enzyme supernatant, C6- and C12-HSL-degrading abilities were quantified using a crystal violet (CV) assay, as described previously [w/v) CV. Then, the CV in the biofilm was dissolved with 150 μL of 95% acetic acid, and the resulting OD was measured at 570 nm (OD570). MB was used as a negative control, and all the experiments were performed in triplicate. The significant difference between the experimental groups and the controls was calculated by the t-test .Biofilm production of all the bacteria with QS activities, and 23 isolates with Ceviously  with sli−1) [t-test .The extracellular enzyme (EE) activities were quantified following the methods described by Hmelo . Five EE−1)  or inactProteobacteria, Bacteroidetes, Actinobacteria and Firmicutes , Hyphomicrobiales (3), Enterobacterales (3), Sphingomonadales (2), Pseudomonadales (1), Cellulomonadales (1), Dermabacterales (1), Cytophagales (1), Flavobacteriales (1) and Bacillales (1)  . Four isdae SCR2 .6-HSL and C12-HSL were used as the representative AHLs with short and long acyl chains, respectively. Among them, 60 (38.37%) and 89 cultures (55.98%) showed C6-HSL degradation and C12-HSL degradation, respectively. A total of 41 (25.78%) isolates were capable of degrading both C6-HSL and C12-HSL , Gammaproteobacteria (26), Betaprotebacteria (3), Firmicutes (32), Actinobacteria (8) and Bacteroidetes (4). At the order level, it was shown that the QQ strains were mainly affiliated with Bacillales (33), Rhodospirillales (19), and Sphingomonadales (9). Furthermore, bacteria showing both C6-HSL- and C12-HSL-degrading activities were mainly affiliated with Bacillales (10) and Rhodobacterales (9)  .6- and C12-HSL-degrading activities, respectively. Among the isolates from sediment samples, 23 (41.07%) and 33 (58.93%) showed C6- and C12-HSL-degrading activities, respectively. As for cultures from macroorganism, 17 (35.41%) and 33 (56.25%) of the isolates were capable of degrading C6- and C12-HSL, respectively. The QQ cultures isolated from macroorganism sand seawater samples belonged mainly to Alphaproteobacteria and Gammaproteobacteria. QQ strains isolated from sediment were affiliated mainly with Firmicutes. It is noteworthy that more bacteria from sediment samples were found to exhibit QQ activities  and 29 (52.73%) cultures exhibited the Ctivities .6-HSL-degrading activities, including Klebsiella michiganensis BODM11, Cellulomonas taurus BOS2, N. indicus LLJ939, Sphingobium yanoikuyae RASR5, Thalassococcus profundi RWAS1 and Yoonia rosea YESM7. Meanwhile, five cultures exhibited C12-HSL-degrading activities, including Arenibacter palladensis CCM2, Roseovarius indicus CCR3, Cyclobacterium marinum CCS19, N. indicus LLJ939 and S. yanoikuyae RASR5. QS strains N. indicus LLJ939 and S. yanoikuyae RASR5 exhibited both C6-HSL- and C12-HSL-degrading activities , 3 (14%), 2 (7%) belonging to Rhodobacterales, Rhizobiales and Sphingomonadales, respectively. In addition, Su et al. [luxI belonging to Alphaproteobacteria existed in particulate organic matter (POM) samples collected from coastal water and sediment in the Yellow Sea of China. In this study, 9 out of 55 isolates belonging to Alphaproteobacteria produced AHLs, with affiliation to Rhodobacterales (4), Hyphomicrobiales (3) and Sphingomonadales (2)  production [R. indicus CCR3, Y. rosea YESM7, P. rhizosphaerae CJG 283 and T. profundi WRAS1 were affiliated with the Roseobacter Clade and showed QS activities  ; they wearticles . For exaoduction ,51. In otivities . Indeed,articles  have beeng study .Gammaproteobacteria are among the most common AHL producers isolated from marine habitats [Gammaproteobacteria isolates were AHL producers. They were affiliated with Enterobacterales (3) and Pseudomonadales (1). Enterobacterales are usually found in marine aquatic environments and have emerged as major players in antimicrobial resistance worldwide. Jatt et al. [Pantoea ananatis B9, which belongs to Enterobacterales and was isolated from natural marine snow particles, could produce six AHLs, and its extracellular alkaline phosphatase activity was enhanced when adding exogenous AHLs. In our study, three isolates belonging to Enterobacterales showed QS activities, and they were all isolated from marine organisms inhabiting hydrothermal fields , Gammaproteobacteria (63.41%) and Firmicutes (69.56%) showed a relatively higher proportion of QQ activities. Sequentially, Su et al. [R. mobilis Rm01 exhibited the degrading activities of C10-HSL, C12-HSL and C14-HSL because of AHL lactonase. Additionally, we found that both Ruegeria atlantica SIR11 and Ruegeria arenilitorisi CCM11, isolated from shrimp gut and crab gill, respectively, degraded C12-HSL, but only R. atlantica SIR11 attacked C6-HSL , Hyphomicrobiales (3), Enterobacterales (3) and Sphingomonadales (2), and QQ strains mainly belong to Bacillales (33), Rhodospirillales (19) and Sphingomonadales (9). It implied the abundant QS and QQ microbial resources in the extreme marine environment. In addition, the diversity of QQ strains was higher than that of QS strains, and they were more likely to degrade long-chain AHLs compared with short-chain AHLs. The biofilm formation might help the QS and QQ strains survive in the hydrothermal environments. Moreover, the β-glucosidase, aminopeptidase and phosphatase activities in bacteria isolated from hydrothermal environments could be regulated by QS. In conclusion, our study revealed the diversity of culturable QS and QQ bacteria and explored the QS regulation of extracellular enzyme in deep-sea hydrothermal environments. In the future, the QS mechanisms and QQ enzymes will be further studied to explore the important roles of QS and QQ in regulating biogeochemical cycles in hydrothermal vents.This study revealed the diversity of QS and QQ strains isolated from hydrothermal vents in the Okinawa Trough. The QS and QQ bacteria belong mainly to