Shading stress is a common abiotic stress faced by crops. Rice (Oryza sativa) is a light-loving crop. To clarify the characteristics and physiological mechanisms underlying the response of rice varieties with different shade tolerances to shade stress, 'Minghui 63' (a relatively shade-tolerant restorer line) and 'Fuxiangzhan' (a high-quality conventional rice variety with poor shade tolerance) were used as materials in this study. Shade stress treatments were applied at the seedling stage and from the heading to maturity stage, respectively. Differences in photosynthetic traits, biomass, yield traits, and the expression of genes related to photosynthesis and the light reaction pathway were compared between plants subjected to shade stress and those grown under normal light conditions. After shade stress at the seedling stage, the photosynthetic rate, SPAD value (relative chlorophyll content), soluble sugar content, root-shoot ratio, and biomass of both varieties were decreased. The reductions in photosynthetic rate, SPAD value, and soluble sugar content of 'Fuxiangzhan' were greater than those of 'Minghui 63', while the reduction in biomass was smaller than that of 'Minghui 63'. After shade stress from the heading to maturity stage, the growth periods of 'Minghui 63' and 'Fuxiangzhan' extended by 3 and 4 d, respectively. The contents of chlorophyll A and carotenoids in both varieties increased, while the photosynthetic rate decreased, and the number of sterile pollen grains increased. Fifteen genes related to photosynthesis and the light reaction were differentially expressed in at least 1 variety, but the variation trends differed between the 2 varieties. For 'Minghui 63', the biomass decreased by 22.37%, 1000-grain weight by 5.94%, seed setting rate by 10.69%, and yield by 15.45%. For 'Fuxiangzhan', the biomass increased by 19.75%, the number of grains per panicle decreased by 18.04%, the seed setting rate decreased by 14.80%, and the yield decreased by 26.68%. The yield reduction of 'Minghui 63' under shade stress was smaller than that of 'Fuxiangzhan'. The results of this study clarified the responses of 'Minghui 63' and 'Fuxiangzhan' to shade stress, which is beneficial to the research on the shade tolerance mechanism of rice and can serve the innovation of shade-tolerant rice germplasm and variety breeding.

The genetic improvement of wheat (Triticum aestivum) processing quality is a crucial objective in breeding programs, while the composition of high molecular weight glutenin subunits (HMW-GS) and the presence of the 1BL/1RS translocation are 2 significant genetic factors that affecting the processing quality of wheat. In this study SDS-PAGE and acid-polyacrylamide gel electrophoresis (A-PAGE) were employed to systematically characterize the HMW-GS composition and 1BL/1RS translocation background in 151 new wheat varieties (lines) from the 2023 Henan provincial wheat regional trials. The results revealed 12 distinct HMW-GS subunit types, with 3 variants at the Glu-A1 locus, 5 at Glu-B1, and 4 at Glu-D1. The frequency of the high-quality subunit 5+10 was relatively low, accounting for only 19.21%. A total of 22 types of HMW-GS subunit combination forms were detected. The average quality score was 6.83 points. The subunit combination with the highest distribution frequency was 1/7+9/2+12 (accounting for 27.81%), while the high-quality subunit combination 1/7+8/5+10 only accounted for 8.61%. Additionally, 62 wheat varieties (lines) were identified as carrying the 1BL/1RS translocation chromosome, accounting for 41.06% of the tested materials. 14 wheat varieties (lines) have been identified, which possess HMW-GS with a score of 10 and were non-1BL/1RS translocation lines. These varieties (lines) hold potential value for breeding strong-gluten wheat. Therefore, the proportion of new wheat varieties (lines) containing high-quality HMW-GS combinations was not high, and the average subunit score was relatively low. However, the distribution frequency of the 1BL/1RS translocation lines had decreased. This study provides crucial genetic background information for wheat breeders in Henan province. The selected non-1BL/1RS translocation lines carrying high-quality subunit combinations could serve as important resources for developing high-yield and high-quality wheat varieties, offering significant guiding value for improving wheat processing quality.

The COBL (COBRA-like) gene family constitutes a significant gene family, which is involved in the regulation of cell wall amplification. To investigate the function of COBL genes in peanut (Arachis hypogaea), diploid species peanuts Arachis duranensis and Arachis ipaensis , this study mined and identified the peanut COBL gene family and analyzed them for their physicochemical properties, conserved structures, phylogenetic trees and tissue expression patterns. The results showed that there were a total of 51 COBL gene family members in cultivated peanut, diploid peanuts Arachis duranensis and Arachis ipaensis. The peanut COBL gene family members were analogous to those of Arabidopsis thaliana and soybean (Glycine max), which were divided into 2 subfamilies. Each subfamily contained genes that were homologous to those in A. thaliana and soybean. These homologous genes exhibited a high degree of sequence consistency, which indicated the conservativeness of the COBL family genes. The COBL family was shown to to play a pivotal role in the growth and development of the peanut at various stages of its lifecycle. This finding suggested that COBL genes played important regulatory roles in the development of multiple tissues. In the context of salt and drought stress, the expression levels of certain genes in the roots were found to be elevated, suggesting that the activation of these genes in the roots played a crucial role in the perception and response to stress signals when plants were subjected to adverse environmental conditions. This study provides a reference for subsequent functional studies of peanut COBL genes.

Glutamate cysteine ligase (GCS) is a key rate-limiting enzyme in the biosynthesis of glutathione (GSH), a crucial thiol compound that plays a vital role in plant response to abiotic stresses such as cadmium (Cd). To investigate the biological function of the GCS gene in rapeseed (Brassica napus) under Cd stress, the Cd hyperaccumulator variety 'Nanyou 868' was used in this study. The complete coding sequence (CDS) of the GCS gene was cloned by RT-PCR and named as BnaGCS (GenBank No. PV694275). Sequence analysis revealed that the ORF of BnaGCS was 1 536 bp in length, encoding 511 amino acids. The pBWA(V)HS-BnaGCS-1209-osgfp overexpression vector was constructed and introduced into Nicotiana benthamiana via Agrobacterium tumefaciens-mediated transformation. Molecularly confirmed transgenic plants were subjected to Cd stress, and gene expression was analyzed, and both cadmium content and GSH content were measured. qRT-PCR results showed that under Cd stress, the transcription level of BnaGCS in both roots and shoots of transgenic tobacco was significantly up-regulated (P<0.05), with a higher up-regulation amplitude in roots than in shoots. Cd and GSH content measurements revealed that Cd stress significantly increased Cd content in both roots and shoots of wild-type (WT) and transgenic plants (P<0.05). The Cd content in roots of transgenic tobacco was significantly lower than that in WT, while the Cd content in shoots was significantly higher than that in WT (P<0.05). Under Cd stress, the GSH content in both wild-type and transgenic tobacco significantly increased, while the GSH content in transgenic tobacco was significantly lower than that in WT (P<0.05). These findings suggested that BnaGCS might be involved in the response of transgenic tobacco to Cd stress. This study provides a theoretical foundation for elucidating the function of BnaGCS and the mechanism of glutathione biosynthesis.

Pyroxsulam as an acetolactate synthase (ALS) inhibitor for controlling weeds in highland barley (Hordeum vulgare var. nudum) fields, ALS plays an important role in herbicide resistance in highland barley through the ALS gene. To investigate the role of the HvnALS gene in conferring pyroxsulam resistance in highland barley, this study utilized a pyroxsulam resistant variety 'Qing 0306' (R) and a susceptible variety 'Qing 0160' (S). Primers were designed based on transcriptome sequences to clone the HvnALS gene, followed by corresponding bioinformatic analysis. The results of ALS activity assay showed that the relative ALS activity of the resistant variety 'Qing 0306' (R) was extremely significantly higher than that of the control from 1~4 d after pyroxasulfone treatment. qPCR was employed to analyze the expression levels of the HvnALS gene in different resistant naked barley varieties under pyroxsulam treatment. The results showed that, the full length of the HvnALS gene (GenBank No. KAE8811959.1) was 1 941 bp, encoding 646 amino acids. Compared to known ALS gene amino acid sequences, no amino acid mutation sites were found in either variety. Multiple protein sequence alignment and phylogenetic tree analysis indicated that the HvnALS protein possesses the PLN02470 domain and exhibits the closest genetic relationship to the HvALS protein of barley (Hordeum vulgare). qPCR analysis revealed that the expression of this gene rapidly increased after pyroxsulam stress. At 1 d after application, its expression in the resistant material 'Qing 0306' (R) was ectremely significantly higher than that in the susceptible variety (P<0.01). Analysis of HvnALS gene expression following pyroxsulam stress suggested that the high-level resistance in 'Qing 0306' (R) might be associated with the overexpression of the target enzyme, rather than mutations in the target enzyme gene locus. This overexpression was likely the primary mechanism leading to pyroxsulam cross-resistance in highland barley.This study provides a theoretical reference for the genetic improvement of pyroxsulam-tolerant highland barley varieties.

The respiratory burst oxidase homolog (RBOH) gene family plays a crucial role in plant development and stress responses, but its members and functions in blueberry (Vaccinium spp.) remain unclear. This study aimed to identify and analyze the expression patterns of VcRBOH gene family, providing insights into its stress resistance mechanisms and molecular breeding. Based on whole-genome data of blueberry, VcRBOH gene family was identified using bioinformatics methods. The physicochemical properties of encoded proteins, gene structures, phylogenetic relationships, cis-acting elements in the promoters, and expression patterns were systematically analyzed. A total of 16 VcRBOH genes were identified on 13 chromosomes in blueberry, containing conserved NADPH (nicotinamide adenine dinucleotide phosphate) oxidase domains. Phylogenetic tree analysis classified these proteins into 3 subgroups. The results of collinearity analysis indicated that the VcRBOH gene was closely related to Arabidopsis thaliana, and there are 18 pairs of collinearity relationships. Promoter analysis demonstrated that the VcRBOH gene family promoters harbor abundant hormone-, stress-, growth and development-related regulatory elements. The transcriptome data and qRT-PCR results indicated tissue-specific expression patterns of VcRBOH genes among different cultivars. The qRT-PCR results showed that VcRBOHA, VcRBOHB, VcRBOHC, VcRBOHD, VcRBOHJ, VcRBOHK and VcRBOHL were expressed across various organs and were induced by pathogenic stresses such as anthracnose (Colletotrichum spp.) and gray mold (Botrytis cinerea). This study lays a theoretical foundation for further functional studies of disease resistance genes and resistance breeding in blueberry.

Hypoxia at high altitude can cause irreversible damage to brain tissue. By comparing and analyzing the differential expressions of cluster of differentiation 146 (CD146) and hypoxia-related factors—hypoxia inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), Beclin-1 and cysteine aspartate specific protease 3 (Caspase-3), in different brain tissues between yak (Bos grunniens) and yellow cattle (Bos taurus), this study explored their correlation with brain hypoxia adaptability of yak. Brain tissues (cerebrum, cerebellum, corpora quadrigemina and medulla) of adult yaks and cattle were collected for H&E, PAS, Nissl and Masson staining and immunohistochemical analysis. Based on the results of staining, various parameters were measured, including the proportion of pyramidal cells, Purkinje cell area, the distribution of glycogen accumulation, neuronal distribution and nerve fiber contents. The results showed that the differences in brain tissue between yaks and yellow cattle were mainly concentrated in the cerebellum and medulla, with all parameters in the cerebellum being significantly higher than those in the medulla (P<0.05). For interspecific differences, the average optical density (AOD) in PAS staining and the positive rate in Nissl staining were significantly higher in yak brain tissue than that in yellow cattle (P<0.05), and the proportion of nerve fibers in Masson staining was significantly higher in the yak cerebrum and cerebellum than that in yellow cattle (P<0.05), but significantly lower in the yak medulla and corpora quadrigemina (P<0.05).Immunohistochemical results showed that the differential expressions of CD146 and hypoxia-related factors were mainly in cerebellum and medulla. For inter-regional differences, the expressions of CD146 and hypoxia-related factors in the yak cerebellum were significantly higher than those in the medulla (P<0.05), whereas in yellow cattle, their expressions in the cerebellum were lower than those in the medulla. For interspecific differences, the expressions of CD146, HIF-1α, and VEGF in the yak cerebellum were higher than those in yellow cattle, while the expression trend of Beclin-1 was the opposite, the expressions of CD146 and hypoxia-related factors in the yak medulla were significantly lower than those in yellow cattle (P<0.05). Combined with the biological functions of CD146 and hypoxia-related factors, it was speculated that CD146 might play a role in the adaptation of yak brain to hypoxic environment. This study provides basic data for further exploring the specificity of hypoxia adaptation of yak brain tissue and the regulation mechanism of CD146 and hypoxia-related factors under hypoxia.

Prostaglandin F2α (PGF2α) plays a critical role in luteolysis by mediating apoptosis of luteal cells through its receptor. However, PGF2α does not exhibit pro-apoptotic effects on early-stage corpus luteum (CL) cells. Ferroptosis, a recently discovered form of programmed cell death driven by lipid peroxidation and dependent on iron ions, remains unclear in its role during PGF2α-mediated apoptosis of luteal cells at different developmental stages. This study investigated the impacts of PGF2α on iron content, ferroptosis, and autophagy-related gene expression in ovine CL during early, mid, and late luteal phases. Forty-eight synchronized healthy ewes (Ovis aries) were randomly allocated into 6 groups (experimental vs. control for each phase). Experimental groups received 0.15 mg PGF2α on day 5 (early), 9 (mid), and 13 (late luteal phase), while controls received saline at corresponding time points. CL tissues were collected 3 h post-injection for analyses. Results revealed that PGF2α significantly increased iron content throughout the luteal phase (P<0.05). Phase-specific responses were observed: Early-phase PGF2α upregulated anti-ferroptosis genes (SLC7A11, NRF2, FTH1), pro-ferroptosis gene ACSL4, and ferroptosis-related proteins (Ferritin, GPX4, SLC7A11, LC3B, NCOA4), while downregulating GPX4 and HSPA8 (P<0.05). Mid-phase treatment enhanced SLC7A11, ACSL4, ATG5, TPD52, HSPA5, HSPA8, and corresponding proteins, but suppressed NRF2 (P<0.05). Late-phase administration upregulated P53, ACSL4, GPX4, ATG5, HSPA5, HSPA8 genes and GPX4/SLC7A11/LC3B proteins, while downregulating NRF2, FTH1, and Ferritin (P<0.05). Mechanistic analysis revealed temporal regulation patterns: Early-phase PGF2α enhanced antioxidant defenses via SLC7A11/NRF2/FTH1 axis and iron sequestration to resist ferroptosis; mid-phase activated ACSL4-mediated pro-oxidative environment and ATG5-dependent autophagy, accelerating lipid peroxidation; late-phase triggered P53 activation, NRF2 suppression, and ACSL4 amplification, culminating in iron overload and irreversible ferroptosis-driven luteolysis. These findings demonstrate that PGF2α orchestrates sequential regulation of ferroptosis and autophagy pathways to drive CL transition from early antioxidant protection to terminal regression. This study provides theoretical basis for interventions in luteal dysfunction.

Enhancing the immunity and antioxidant capacity is a critical issue that urgently needs to be addressed in modern broiler production. Medicinal plants and probiotics exhibit anti-inflammatory, antioxidant, and immunomodulatory properties. This experiment aimed to investigate the effect of dietary supplementation with Glycyrrhiza uralensis extract (GUE), Lactobacillus acidophilus (Lac) and their combination on immune and antioxidant functions of broilers (Gallus gallus). A total of 420 one-day-old male Liangfenghua broilers were randomly assigned to 4 groups each with 7 replicates, and 15 broilers per replicate. The broilers were fed a basal diet (Con group), a basal diet with 0.1% GUE supplementation (GUE group), basal diet with 1.5% Lac supplementation (Lac group), and basal diet with 0.1% GUE and 1.5% Lac supplementation (GUE+Lac group), respectively. The experiment lasted for 84 d. The results were as follows: 1) Compared with the Con group, the activities of serum, jejunum and liver superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) of broilers in the supplementation groups were significantly increased (P<0.05), while the malondialdehyde (MDA) content was decreased (P<0.05); 2) The serum immunoglobulin A (IgA) and immunoglobulin G (IgG), jejunum villus height, villus height/crypt depth (VH/CD) ratio and secretory IgA (sIgA) content of broilers in the supplementation groups were significantly increased (P<0.05); 3) The mRNA expression of jejunal mucosal immune-related genes mucin 2 (MUC2), cluster of differentiation (CD) 4 positive T cells CD⁴⁺ and CD⁸⁺, interleukin-4 (IL-4) and IL-10 at 56 and 84 d, and the liver nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were significantly increased (P<0.05), while inducible nitric oxide synthase (iNOS), IL-6, nuclear factor kappa-B (NF-κB) and toll-like receptor 4 (TLR4) in jejunal mucosal, cysteine protease-1 (Caspase-1), Caspase-3 and NOD-like receptor protein 3 (NLRP3) in liver was significantly reduced (P<0.05). In summary, dietary supplementation with GUE and Lac can improve the intestinal morphology, immune and antioxidant functions, inhibit inflammatory response and apoptosis in broilers, and the combined supplementation has a synergistic effect. This study provides a basis for the development and utilization of G. uralensis extract and L. acidophilus synbiotic.

The OCA2 (oculocutaneous albinism Ⅱ) gene is an important gene in melanin biosynthesis, and involved in pigmentation in the eye, skin, and hair colors of animals. Mutations in this gene could result in oculocutaneous albinism type Ⅱ in human. The black patchs on the body surface are key phenotypic characteristics in Oujiang color common carp (Cyprinus carpio var. color), however, the genetic basis and underlying mechanisms of their formation remain unclear. In this study, The high-generation selected strains of Oujiang color common carp was used to perform the cloning, expression and promoter transcriptional activity studies of the OCA2 gene, and gene knockout experiment was carried out by the CRISPR/Cas9 technology. The results revealed that the OCA2 gene consisted of 23 exons and 22 introns, encoding 834 amino acids with over 97% homology with the other fish species in Cyprinid family, which indicated highly structural and functional conservation. The qPCR analysis of the OCA2 gene showed that its expression levels were significantly higher in the eye and skin tissues in the whole white with black patch than those in the whole white color types in Oujiang color common carp. The dual-luciferase reporter assays indicated a complex regulatory mechanism in the promoter region of the OCA2 gene, and a key regulatory element during the 600~1 200 bp fragment before the start codon. CRISPR/Cas9-mediated knockout of exon and promoter regions in the OCA2 gene resulted in the numbers reduction by 70% and the areas decreasing by 40% of melanocytes from surface skin and eye tissues in the mutant. This study revealed that the OCA2 gene plays an important role in pigmentation in Oujiang color common carp.

Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disease characterized by the production of large amounts of lipid deposits in the liver, which poses a serious health risk to humans. In recent years, there has been a gradual increase in the incidence of fatty liver disease in pets and farm animals, which has resulted in serious economic losses to the farming industry. Angiotensin-converting enzyme 2 (ACE2) has been shown to positively regulate anti-inflammatory, anti-injury and anti-fibrotic effects in a variety of diseases, including fatty liver. The previous study of our group found that ACE2 might exert some anti-injury effects on NAFLD induced by exogenous stimuli, but its exact protective mechanism is not clear. In this study, normal rat (Rattus norvegicus) hepatic stromal BRL-3A cells were induced with 0.025~0.200 mmol/L oleic acid (OA) for 24 h to establish a non-alcoholic fatty liver disease (NAFLD) cell model, and hepatocellular injury markers were measured to determine the optimal induction conditions. Subsequently, the cells were treated with either exogenous ACE2 active protein or siRNA targeting ACE2, after which lipid deposition was observed through staining, and changes in parameters including total anti-oxidative capacity (T-AOC), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and inflammatory factors were detected. Western blot detected the protein expression of cellular ACE2, and mas receptor (MasR); ELISA detected the content of Ang Ⅱ and Ang 1-7 in the supernatant of the cells, and clarified the effect of ACE2 on NAFLD cellular damage. Results showed that: 1) NAFLD cell model was successfully established, analyzed by oil red staining to detect the changes of triglyceride (TG) content and reactive oxygen species (ROS) in hepatocytes, and the induction condition of NAFLD cell model was selected to be 0.100 mmol/L oleic acid for 24 h, and 3 different concentrations of OA (low, medium, and high (0.025, 0.100, and 0.200 mmol/L)) were determined for the follow-up test. 2) Compared with the control group, ACE2 protein expression was significantly down-regulated in the high concentration OA-treated group (P<0.05), and there was no significant difference in ACE2 expression in the low- and medium-concentration groups, the expression levels of ACE2 and MasR in the group treated with the addition of ACE2-activated proteins were significantly elevated (P<0.05), and after the transfection of siRNA-ACE2, the expression level of ACE2 was significantly reduced (P<0.05), and MasR expression level was reduced but the difference was not significant; 3) In medium concentration OA treatment, ACE2 and MasR expression was up-regulated, Ang 1-7 and Ang Ⅱ levels were increased, lipid accumulation was increased, oxidative stress was aggravated, antioxidant capacity was weakened, inflammatory factors and NO content were extremely significantly elevated (P<0.01), iNOS activity was extremely significantly enhanced (P<0.01), and anti-inflammatory factor content was extremely significantly reduced (P<0.01); 4) Compared with the OA-treated group, the ACE2 active protein-treated group cellular ACE2, MasR, and Ang 1-7 levels were significantly or extremely significantly up-regulated (P<0.05 or P<0.01) and Ang Ⅱ levels were down-regulated. The relative intracellular lipid droplet area and TG content were significantly reduced (P<0.05), while ALT, AST viability and T-AOC were enhanced to different degrees, and oxidative stress and inflammatory response were reduced.Transfection with siRNA-ACE2 treatment reversed the above changes. The experimental results suggested that treatment with ACE2-activated protein could alleviate cellular lipid deposition, attenuated cellular oxidative stress and inflammatory response, and had a certain alleviating effect on OA-induced NAFLD in rat hepatocytes. The mechanism was mainly realized by degrading Ang Ⅱ, activating Ang 1-7/Mas, and acting on the NO/iNOS signaling pathway. Thus, this study revealed a mechanism by which ACE2 alleviates NAFLD at the cellular level and provide an important theoretical foundation for developing ACE2 as a potential therapeutic target for NAFLD.

Continuous cropping barriers of Pinellia ternata have become an important factor limiting its yield and quality. To clarify the types of pathogenic fungi in the soil of multi-year continuous cropping of P. ternata,fungi were isolated and purified from the soil after 7 years of continuous cropping in this study. Pathogenic fungi were screened and identified via in vitro inoculation methods, and their pathogenicity was evaluated. To further understand the occurrence pattern of P. ternata root rot, the biological characteristics of the pathogenic fungi were investigated, including their growth conditions under different temperatures, light conditions, pH levels, carbon sources, and nitrogen sources were further investigated, along with the lethal temperature for the fungi. Experimental results showed that 22 fungal strains were isolated from the soil of 7 years of continuous cropping of P. ternata, among which 4 strains exhibited pathogenic ability: Fusarium falciforme E, F. solani N, F. keratoplasticum H, and F. oxysporum O. All the above pathogenic bacteria could utilize various carbon sources, grow rapidly at a temperature of 25~30 ℃, pH 9~10 and under light conditions, and lose their growth activity when treated at 56~61 ℃. This study provides an experimental basis for the identification and control of pathogenic fungi in the soil of continuous cropping of P. ternata.

Genome editing technology refers to a biotechnology enabling site-specific and heritable modifications of target genes within the genome. Among these technologies, CRISPR/Cas9 has emerged as the most widely adopted genome editing technologies due to its simplicity, high efficiency, and multiplex editing capability. It has been extensively applied in the genetic improvement of major oil crops including soybean (Glycine max), rapeseed (Brassica napus), camelina (Camelina sativa), and peanut (Arachis hypogaea). In this article, comprehensive overview of CRISPR/Cas genome editing technologies and their molecular mechanisms were provided, the metabolic pathway of fatty acid biosynthesis in plants was summarized, and recent advancements in genome editing for enhancing oil yield and improving fatty acid profiles were highlighted. The review provides technical support and guidance for creation of excellent germplasm and the cultivation of new varieties of oil crops through genome editing technologies.

For patients with end-stage organ failure, organ transplantation is often the only viable treatment. However, there is an extreme shortage of organs available for transplantation globally. Regenerative medicine offers alternative ways to generate donor organs, with major strategies including xenotransplantation, tissue engineering, bioprinting, and organoid technology. However, these methods have their own limitations and drawbacks, restricting their clinical applications. In recent years, several clinical trials of xenotransplantation have been conducted, involving the transplantation of gene-edited pig organs into live human patients, but current reports indicate that these patients did not survive for long periods. Blastocyst complementation to create human-animal chimeras represents another potential pathway to obtain organs for human transplantation. This involves generating animal embryos with cell-lineage or organ deficiencies through gene editing to create a "vacant" microenvironment, which is then populated with human pluripotent stem cells (hPSC). Despite many obstacles and challenges, scientists are continuously refining and attempting this approach in hopes of clinical application. This review summarized the current research on interspecies chimeras and examined the existing challenges and potential strategies in this field, thereby providing a theoretical reference for future studies.

Rice (Oryza sativa) blast is one of the most important diseases that threatens rice yield stability. In recent years, the incidence and severity of rice blast have been increasing in Northeast China. The inactivation of the function of Bsr-d1 (broad spectrum resistance digul) can weaken the degradation of hydrogen peroxide and improve rice resistance to Magnaporthe oryzae; Bsr-k1 (broad-spectrum resistance Kitaake-1) encodes a TPR protein, and its functional inactivation mutant bsr-k1 leads to the enrichment of OsPAL (phenylalanine ammonia-lyase) family gene mRNA and increased lignin synthesis, thereby enhancing the immune response and giving rice broad-spectrum resistance. The study designed targets for the first exon of the 2 broad-spectrum disease resistance genes Bsr-dl and the third exon of Bsr-kl and constructed gene editing vectors. Agrobacterium tumefaciens-mediated rice transformation technology was used to introduce the gene editing vector into the recipient rice variety 'TH899', and T0 generation plants with various different variants of the Bsr-dl and Bsr-kl genes were obtained. Analysis of the T₂ generation gene-edited materials that did not contain transgenic components showed that both single and simultanecous mutations of the Bsr-dl and Bsr-kl genes showed enhanced resistance to rice blast, among which the selected double-gene-edited rice materials had significantly better disease resistance than single-gene-edited materials. In addition, the single-plant yield and seed length-to-width ratio of some gene-edited lines also changed. In summary, this study used gene editing technology to simultaneously edit the Bst-dl and Bsr-kl genes, which was able to significantly enhance the resistance of 'T1899' to rice blast more than single-gene editing, and the yield traits of some mutants were also changed. The results of this study showed that combinatorial editing of rice blast resistance genes regulating different physiological pathways could effectively improve rice disease resistance. This study demonstrates the great potential of multi-gene editing in improving rice disease resistance and provides new material resources for breeding rice blast-resistant japonica rice.

In the research and standardization of detection methods for genetically modified (GM) products, qualitative and quantitative detection, implementation of labeling regulations, and safety supervision, certified reference materials (CRMs) for GM detection are indispensable. GM soybean (Glycine max) 'DBN9004', a herbicide-tolerant soybean line independently developed in China, obtained a biosafety certificate for production and application in 2020. There is an urgent need to develop certified reference materials for its safety supervision and detection. In this study, using homozygous GM soybean 'DBN9004' and non-transgenic receptor soybean 'Jack' as raw materials, the leaves were frozen, ground, and genomic DNA was extracted. The reference materials DBN9004a and DBN9004c were prepared by weighing and mixing to achieve DBN9004/Lectin copy number ratios of 100% and 3%, respectively. Each concentration of the standard substances was aliquoted into 500 tubes. Using the droplet digital PCR method, according to General Principles and Statistical Principles for Certification of Reference Materials (JJF 1343), the homogeneity evaluation, stability evaluation, processing of joint certification data, and uncertainty assessment of reference materials with 2 concentrations were carried out. The results showed that, the homogeneity test results for both materials were below the critical value F_{0.05 (14,30)}(2.04), indicating good intra- and inter-unit homogeneity. The reference materials can be stored and transported stably for 10 d at temperatures below 37 ℃, and long-term stability at -20 ℃ was confirmed for up to 9 months. After 10 freeze-thaw cycles. The collaborative study involving 9 laboratories showed no outliers in the copy number concentration and ratio measurements, with normal distribution and equal precision. The copy number concentrations and expanded uncertainties for DBN9004a and DBN9004c were (4.43±0.38)×10⁴ and (6.32±0.65)×10² copies/μL, respectively. The DBN9004/Lectin copy number ratios and expanded uncertainties were (98.1±9.8)% and (3.08±0.32)%, respectively. The developed gradient genomic DNA reference materials for GM soybean DBN9004 exhibit good homogeneity and stability during transport and storage. They meet the requirements for qualitative and quantitative detection of GM soybean 'DBN9004' and provide reliable certified reference materials for the safety supervision and implementation of quantitative labeling regulations for GM soybean 'DBN9004'.

CRISPR/Cas9 gene editing technology has been successfully applied to various crops, and targeted mutagenesis of target genes as well as targeted improvement of target traits have been achieved. However, no mature and efficient gene editing system has been established in peanut (Arachis hypogaea). In this study, 2 U6 promoters, namely AhA3U6 and AhB9U6, were isolated from the peanut variety 'Huayu 23'. Subsequently, these promoters were employed to substitute the GmU6 promoter in the original vector pBGK041-GmU6. The homologous genes of the peanut AhPEPC1 (PEPC: phosphoenolpyruvate carboxylase) gene, namely Aradu.A52DW and Araip.RUX3H, were selected as the target genes in this study, and the CRISPR-Cas9 recombinant vectors pBGK041-AhA3U6-AhPEPC1 and pBGK041-AhB9U6-AhPEPC1 were successfully constructed. Subsequently, all 3 vectors were introduced into Agrobacterium rhizogenes for the induction of hairy root formation. The transformation positive rates were determined to be 78.4%, 83.5%, and 83.0%, respectively. The results indicated that the gene editing efficiency of pBGK041-GmU6-AhPEPC1 was merely 1.7%, whereas that of pBGK041-AhA3U6-AhPEPC1 and pBGK041-AhB9U6-AhPEPC1 reached 4.2% and 2.6%, respectively. Furthermore, the 3 vectors were introduced into peanut plants via the pollen tube pathway, with conversion efficiencies of 30.0%, 33.7%, and 29.2% being achieved. The sequencing results of transgenic seeds showed that no gene editing was detected in the pBGK041-GmU6-AhPEPC1 vector transgenic seeds, while the editing efficiencies of the pBGK041-AhA3U6-AhPEPC1 and pBGK041-AhB9U6-AhPEPC1 vector transgenic seeds were 2.2% and 1.3%, respectively. The oil content of the transgenic kernels was 56.88% and 57.49%. Replacement of the original GmU6 promoter with endogenous peanut promoters AhA3U6 and AhB9U6 in the pBGK041-GmU6 vector induced detectable mutations in both transgenic peanut hairy roots and kernels. This promoter substitution enhanced editing efficiency and elevates kernel oil content by approximately 4%. This system provides technical support for the implementation of peanut gene editing and new germplasm resources for the breeding of high-oil varieties.

As an important immunomolecule molecule, lymphocyte activation gene 3 (LAG3) is a potential molecule connecting immunosuppression and neuroregulation, and its functional study is of great significance for revealing the mechanism of stress-induced neuro-immune imbalance. This study predicted the physicochemical properties, structural domains, and modification sites of bovine (Bos taurus) LAG3 protein. The coding sequence of the LAG3 extracellular domain was amplified from bovine peripheral blood lymphocyte cDNA by gene cloning technology. The prokaryotic expression vector pET-28a-LAG3 was constructed and transformed into Escherichia coli BL21 (DE3) to induce the expression of recombinant protein. The inclusion bodu-form recombinant protein purified by nickel column affinity chromatography was used as the immunogen to immunizing BALB/c mice (Mus musculus) to obtain polyclonal antibodies. Indirect ELISA was emplyed to detect antibody titers, Western blot was used to analyze antibody specificity. The results showed that the coding sequence of bovine LAG3 gene was 1 551 bp in length, encoding 516 amino acids. The protein molecular formula was C₂₅₅₉H₃₉₇₄N₇₂₈O₇₁₂S₁₁, with a relative molecular mass of 56.68 kD. It was a hydrophilic, non-lipid -soluble, unstable alkaline protein containing 1 signal peptide, 1 transmenbrance region, 48 potential posphorylation sites, 3 N-glycosylation sites and 19 antigenic epitopes. The concentration was 1.03 mg/mL after purification, and the prepared polyclonal antibody could bind to the total protein of bovine peripheral blood lymphocytes, showing a single band with a size of 45 kD. The highest antibody titer reached 1∶1 048 576. In conclusion, this study characterized the structural features of bovine LAG3 and successfully prepared a high-titer, high-specificity mouse anti-bovine LAG3 polyclonal antibody. These findings provide key tools for in-depth research on expression patterns, subcellular localization, and immunoregulatory mechanisms of LAG3 in bovine stress responses.

Melon (Cucumis melo) is an annual trailing herbaceous plant that plays a significant role in the agricultural industry of the Taigu district in Shanxi province. In recent years, viral diseases have severely impacted melon production in this region. To identify the pathogens responsible for these viral diseases and to establish rapid detection techniques, this study collected symptomatic melon samples from Taigu district that exhibited spots, mosaic patterns, shrinkage, and necrosis. These samples were analyzed comprehensively using RT-PCR combined with bioinformatics. The results indicated that the presence of quadruple viral pathogens in melo specimens, specifically for: Cucumber mosaic virus (CMV), Cucumber green mottle mosaic virus (CGMMV), Cucurbit aphid-borne yellows virus (CABYV), and Watermelon mosaic virus (WMV). Mixed infections were found to be prevalent. Based on homology comparisons and phylogenetic analyses of the coat protein (CP) sequences of the viruses mentioned above, the results indicated that the amplified isolates obtained in this study CABYV isolate Fenghuang, CMV isolate Fengyun, CGMMV isolate Longquan, and WMV isolate Zhouquan exhibited sequence identities of over 99.5%, 98.0%, 99.6%, and 97.0%, respectively, when compared with corresponding virus sequences deposited in the GenBank database. Each isolate had a relatively close genetic relationship with the corresponding isolate of the same virus. In response to the phenomenon of complex infection of sweet melon virus diseases in the local area, a multiplex RT-PCR detection system was established to simultaneously detect CMV, CGMMV, CABYV and WMV. Multiple RT-PCR amplification was performed on materials with complex infection of 4 viruses, resulting in 4 specific bands of 1 106, 690, 521, and 400 bp, respectively. The multiplex RT-PCR detection system developed in this study allows for rapid identification of melon viral diseases and provides technical support for the monitoring and prevention of melon virus diseases.