Pathogenesis-related protein 1 (PR1) is an important defense protein in plants, its production and accumulation in plants is one of the important responses to biotic and abiotic stresses. In previous study, TaPR1 was found to be induced by Puccinia triticina (Pt) and participate in wheat defense response to leaf rust. In the early stage of this study, the TaPR1 promoter sequence was successfully obtained by gene cloning and named pTaPR1, among which the 900 bp sequence (pTaPR1₉₀₀) had strong activity. This study further analyzed the sequence composition and possible regulatory factors of pTaPR1. Preliminary analysis based on the PLACE and PlantCARE databases showed that pTaPR1 contained the core elements including TATA-box, CAAT-box and a large number of light-responsive elements, as well as the ABA-responsive element and the MYB-binding site. Yeast one-hybrid (Y1H) technique was used to screen the transcription factors that might bind to pTaPR1₉₀₀ involved in regulating TaPR1 gene expression, 12 candidates were obtained. The interaction between 4 candidates including U-box E3 ubiquitin ligase, pollen protein, glycine A3 protein and heavy metal-associated isoprenoid phycobiliprotein with pTaPR1₉₀₀ were initially verified. Taken together, this study provides theoretical reference for exploring the transcriptional regulatory mechanism of TaPR1.

Breeding salt tolerance and late blight-resistant potatoes (Solanum tuberosum) is an important requirement for the development of the potato industry. In this study, key genes responding to salt stress and the invasion of the late blight pathogen Phytophthora infestans were screened by transcriptome sequencing. The results showed that the infection area of P. infestans on the leaves significantly increased after being treated with 100 mmol/L NaCl for 1 week, namely, salt stress significantly weakened the resistance of tissue-cultured potato to P. infestans. Transcriptome sequencing revealed that the expression of 3 780 potato genes changed significantly under salt stress, in which the cystatin 6 (CYS6) gene significantly downregulated. qRT-PCR results further confirmed that StCYS6 gene expression was downregulated in potato leaves after salt stress, but upregulated after P. infestans invasion. Transient expression of the candidate gene StCYS6 in Nicotiana benthamiana leaves significantly enhanced resistance to P. infestans. Inactivation mutations showed that StCYS6 protein promoted plant resistance to P. infestans independent of its cysteine protease inhibitor activity. The analysis of salt tolerance and pathogen resistance of StCYS6 transgenic potato plants showed that the degree of wilting of potato leaves was significantly reduced, indicating that StCYS6 could significantly improve the salt tolerance of potatoes; However, StCYS6 transgenic potato plants did not significantly improve their resistance to P. infestans. This study obtained a candidate gene StCYS6 that could enhance the salt tolerance of potatoes, providing basic data for cultivating new varieties of stress resistant potatoes.

Fruit coloration is mainly determined by flavonoids such as anthocyanins, and MYB transcription factors play an important role in the regulation of anthocyanins. In order to explore the effect of strawberry (Fragaria vesca) MYB gene family on strawberry fruit coloring, in this study, 114 MYB gene family members were identified by homology alignment of the MYB gene family sequences in the Arabidopsis thaliana and strawberry gene databases. The chromosomal positioning, physicochemical properties, subcellular localization and secondary structure analysis of the strawberry MYB gene family were analyzed by bioinformatics analysis software to determine the naming of the genes and the location of the main expression. The gene family was classified and its functions were predicted based on collinearity, gene structure, and promoter cis-acting element prediction. The results showed that MYB family members were predominantly expressed in the nucleus, and all but FvMYB7 were hydrophilic proteins. Among the 114 members, 107 proteins exhibited interactions with each other. A relative synonymous codon usage value was greater than 2.0. This indicated that there were highly preferred codons in strawberry MYB gene. 114 members could be divided into 3 subgroups according to the phylogenetic tree. Except FvMYB72 which didn't contain any action elements, all other members contained light, abiotic stress, hormone and meristem response elements. qRT-PCR analysis indicated that most members were in the S3 (50% coloring period) with high relative expression levels, however, the expressions of FvMYB33, FvMYB57, and FvMYB75 were higher in the S4 (full coloring stage). The tissue-specific expression profile results showed that only the FvMYB33 was specifically expressed in the fruit peel, and the results of anthocyanin content showed the highest content was in S4 period. It was speculated that FvMYB33 was the main gene regulating strawberry fruit coloring. This study provides a reference to further explore the mechanism of MYB gene family in strawberry fruit coloring.

The main pigment responsible for the coloration of plant petals is anthocyanin. Based on previous transcriptome data from petunias, the highly expressed 4-coumarate CoA ligase 13 (Ph4CL13) gene was identified during the bud coloring stage, suggesting its involvement in anthocyanin synthesis. Anthocyanin primarily determines petal color in plants. To investigate the role of Ph4CL13 in anthocyanin synthesis, this study used Petunia hybrida 'Ultra' as the material. The 1 653 bp cDNA fragment of Ph4CL13 (Peaxi162Scf00089g00045.1) was isolated from the petunia genome. Phylogenetic analysis revealed a close relationship between Ph4CL13 and Fa4CL-1 and Fa4CL-2. qPCR analysis revealed that Ph4CL13 was highly expressed during the coloring stages, suggested its involvement in anthocyanin synthesis and impact on petunia flower coloration. Silencing Ph4CL13 via virus-induced gene silencing (VIGS), the results showed that the corollas of petunia turned white, and the level of anthocyanin significantly decreased in Ph4CL13-silenced plant compared to control. qPCR analyses demonstrated that Ph4CL13 silencing transcriptionally repressed several anthocyanin biosynthesis genes, including chalconesynthase (CHS), chalcone isomerase (CHI), flavanone-3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), flavonoid synthase (FLS), suggested that Ph4CL13 played a critical role in the anthocyanin synthesis stage of flower coloring process, that might be a potential complex feedback mechanism among these genes related to anthocyanin synthesis. This study preliminarily revealed the important role of Ph4CL13 in anthocyanin synthesis, providing a reference for further exploring its molecular mechanisms in flower color development.

Rhododendron species have ornamental, economic and medicinal value, while most of the excellent varieties of Rhododendron cultivated at present have poor heat tolerance, and the increase in temperature will seriously affect the normal growth and development of Rhododendron. Heat stress transcription factors (HSFs) plays a key role in the regulation of plant heat shock response or heat tolerance. In this study, phytoene desaturase (RpPDS, OQ628048) and RpHSFC1a (OQ628046) genes of Rhododendron×pulchrum were cloned, and the virus induced gene silencing (VIGS) system was constructed with RpPDS as the reporter gene and Tobacco rattle virus (TRV) as the vector. The bioinformatics of the RpHSFC1a gene and its function in response to high temperature stress were analyzed. Using phenotypic observation, PCR and qPCR methods, TRV could infect and replicate and metastasize in the leaves of Rhododendron×pulchrum, and yellowing will occurred in leaves when the expression of RpPDS gene is inhibited, the result showed that TRV virus-mediated gene silencing system could be applied in Rhododendron×pulchrum. The CDS of the RpHSFC1a gene length totally 891 bp and encodes 296 amino acids, and RpHSFC1a contained HSF_DNA-bind superfamily conserved domain. RpHSFC1a was predicted to be a hydrophilic and transmembrane protein. Phylogenetic analysis showed that RpHSFC1a protein had the highest homology rate with HSFC1 of Diospyros lotus, with a homology of 75.8%. Under heat stress, RpHSFC1a gene was significantly up-expression, the chlorophyll parameters F_v/F_m, PI_ABS and ET_o/RC in the leaves of the strain of RpHSFC1a gene silenced were significantly lower than those of the control group. The DI_o/RC was significantly higher than that of the control group. It was shown that the low expression of RpHSFC1a gene could reduce the ability of Rhododendron×pulchrum to resist high temperature stress, and it is speculated that the RpHSFC1a gene participates in the regulatory mechanism of Rhododendron×pulchrum in response to high temperature stress. This study provides a theoretical reference for the study of heat tolerance, and genetic resources for improving the high temperature stress tolerance of Rhododendron.

The long cultivation cycle and poor quality of lily bulbs by produced in China is a important factors that lead to long-term dependence on imports. The plant-specific transcription factor NAC plays an important role in regulating plant growth and development, resistance establishment, etc., and mining, identifying and studying related genes in lily is a strategy to solve the above problems. In this study, one gene transcript carrying NAC domain was screened in the lily transcriptome database, and the gene was cloned and identified as LoNAC48 gene (GenBank No. PP078617) based on PCR. Bioinformatics analysis showed that LoNAC48 gene contained a complete open reading frame with a length of 777 bp encoding 258 amino acids, which was an unstable hydrophobic protein, and phylogenetic tree analysis showed that it was highly homologous to Lilium regale LrNAC48 protein. The results of subcellular localization showed that LoNAC48 was a nuclear transcription factor, and tissue expression analysis showed that LoNAC48 gene was present in bulbs, roots, flowers and other tissues and organs expression, among which the expression level was the highest in leaves; The results of different stress responses showed that the expression of LoNAC48 gene was susceptible to low temperature, high temperature, drought, salt and hydrogen peroxide (H₂O₂) stresses, Under low temperature, salt, and hydrogen peroxide stress, the expression level was significantly upregulated at 12 hours compared to the control, and significantly decreased at 24 hours of drought stress, and the expression pattern of LoNAC48 was almost the same under salt stress and hydrogen peroxide stress. Phenotypic analysis of transgenic Arabidopsis thaliana lines showed that overexpression of LoNAC48 gene could significantly promote the growth of Arabidopsis thaliana and increase the leaf area. These results indicated that LoNAC48 gene had the dual functions of stress resistance and regulation of plant growth and development. This study provides genetic resources for molecular-assisted breeding of lily and improvement of lily traits.

Lavender (Lavandula) is an important aromatic crop, and the differences in the quality of its essential oil are mainly caused by variations in the content of terpenoids, such as linalool and linalyl acetate. Studying the relevant genes that may be involved in regulating the biosynthesis of essential oils in lavender can provide genetic resources for breeding excellent lavender germplasm. The basic Helix-Loop-Helix (bHLH) transcription factor family plays a crucial role in plant growth, development, and secondary metabolism. In this study, the lavender LibHLH75 gene (GenBank No. OR019688) was cloned from the 'Zahua' of lavender (Lavandula×intermedia）, with an open reading frame of 831 bp, encoding 276 amino acids, which was closest to the bHLH75-like protein from Salvia splendens and bHLH75 protein from S. hispanica. qRT-PCR analysis showed that the LibHLH75 gene was expressed in different tissues of lavender and at different flowering stages of the corolla, with the highest expression level at the calyx and bud stages. The results of subcellular localization and transcriptional activation activity identification indicated that LibHLH75 encoded protein was localized in the nucleus, and had transcriptional activation activity. This study preliminary analyzed the function of LibHLH75, which provides a theoretical basis for further revealing the molecular regulatory mechanism of bHLH transcription factors in the biosynthesis of lavender terpenoids.

RING finger proteins, which usually have E3 ubiquitin ligase activity, are important in regulating plant growth and development and in response to biotic and abiotic stresses. However, studies on the RING finger gene family of Brachypodium distachyon are limited. In this study, the RING finger gene family of B. distachyon was systematically analyzed using bioinformatics methods, and its expression under drought, salt, and abscisic acid (ABA) stress treatments was preliminarily analyzed by qPCR. The results showed that a total of 666 RING finger genes were identified in the B. distachyon genome, which were unevenly distributed on 5 chromosomes, and 39 intergenic duplications occurred. The phylogenetic analysis divided the RING finger gene family into 9 subclusters, and the members of each subcluster were highly homologous and contained similar conserved structural domains and motifs. The covariance analysis among species showed that the RING finger gene family originated from genome duplication, and that gene fragment duplications and tandem duplications played a key role in its expansion and evolution. There were 303 RING finger collinearity gene pairs in rice (Oryza sativa) and B. distachyon, whereas there were only 48 pairs in Arabidopsis thaliana and B. distachyon, and a total of 39 RING finger gene pairs in B. distachyon genomes. Tissue expression profiles showed that 547 RING finger genes were expressed in different tissues of B. distachyon, and 39.8% of the genes were highly expressed in male flowers of B. distachyon. Differential expression analysis showed that 29 RING finger genes might be involved in the response to drought, salt and abscisic acid (ABA) stress. qPCR results showed that 26 RING finger genes were significantly up-regulated in drought stress (P<0.05), 3 RING finger genes were down-regulated, 27 RING finger genes were up-regulated and 2 were down-regulated in salt stress; 15 RING finger genes were up-regulated, 9 RING finger genes were down-regulated, and 5 RING finger genes were at the same level as the control in ABA stress. The present study provides a theoretical basis for revealing the function and evolution of the RING finger gene family and its role in the regulation of drought stress.

As an endogenous timekeeping system, the circadian clock system plays an important role in regulating behavioral activities and physiological functions in mammals. To investigate the role of the core clock gene brain and muscle arnt-like protein 1 (BMAL1) in apoptosis of goat endometrial epithelial cells (GEECs), the CDS of BMAL1 gene was cloned from 24-month-old goat (Capra hircus) uterine tissues, and the physicochemical properties of the gene and its coding protein were analyzed and predicted using the bioinformatics software; the expression patterns of BMAL1 were detected using qPCR in different tissues of goats as well as GEECs after synchronization. The effect of BMAL1 gene overexpression on GEECs apoptosis was detected by flow cytometry, qPCR, and Western blot (WB). The results showed that the expression of BMAL1 gene showed significant diurnal differences in various tissues and at different time points. The total length of CDS fragment of goat BMAL1 gene was 1 881 bp, which was highly conserved in mammals. And protein encoded by BMAL1 could participate in multiple physiological processes by interacting with proteins such as circadian locomotor output cycles kaput (CLOCK), nuclear receptor subfamily 1, group D, member 1 (NR1D1), and casein kinase 1δ (CSNK1D). Flow cytometry results showed that BMAL1 overexpression significantly reduced the apoptosis rate in GEECs (P<0.05). qPCR and Western blot results showed that BMAL1 overexpression led to a significant decrease in the expression levels of pro-apoptotic genes BCL2-associated X protein (BAX) and tumor suppressor gene P53 in both mRNA and protein, and the expression of apoptosis suppressor gene B cell lymphoma-2 (BCL-2) significantly elevated (P<0.05). This study provides a preliminary basis for analyzing the molecular regulatory mechanism of the ruminant circadian clock system and further exploring the biological functions of the goat BMAL1 gene.

Yes-associated protein (YAP) and TEA domain transcription factor 1 (TEAD-1) are the core factors of the Hippo signaling pathway and play an important role in regulating cell proliferation and organ growth. In order to detect the expression of YAP and TEAD-1 in different developmental stages of yak (Bos grunniens) placenta, and to explore the physiological role of Hippo signaling pathway in regulating placental cell proliferation and organ growth regulation. In this study, placental tissue samples of pregnant yaks were collected and divided into different developmental stages (<2 months old,3 months old,4 months old,>5 months old) based on fetal head-to-hip distance. The expression of YAP and TEAD-1 was detected and analyzed by qRT-PCR, Western blot (WB) and Immunohistochemistry (IHC) technology were used to detect the expression of these genes. The results showed that YAP and TEAD-1 were mainly expressed in fetal chorionic trophoblast giant cells, columnar trophoblast cells and maternal placental crypt epithelial cells. With the progress of pregnancy, the expression of YAP and TEAD-1 mRNA and protein in yak placenta tissue change significantly, and the change trend of YAP and TEAD-1 was basically the same. The co-expression characteristics and the correlation between the expression level and the placental development stage suggested that YAP and TEAD-1 might be involved in the regulation of yak placental development through the Hippo signaling pathway, and play an important role in the physiological function of placental tissue and pregnancy maintenance. The results provides a theoretical basis for elucidating the mechanism of yak placental development and pregnancy regulation.

The major cat (Felis catus) allergen, Fel d1 is composed of 2 heterodimers: chain 1 (CH1) and chain 2 (CH2). Fel d1 could induce allergic reactions to humans. In this study, the genome sequences of CH1 and CH2 of 38 domestic cats of different breeds were analyzed by bioinformatics, and 2 single-guide RNAs (sgRNAs) were designed at the exon 2 of CH2, then the constructed target vector of CRISPR/Cas9 was transfected into the cat fetal fibroblasts cell. The mutation efficiency of the CH2 was verified by PCR and Sanger sequencing. The results showed that the CH1 and CH2 sequences contained 12 and 51 polymorphic loci, and most of these loci were located on the GC-rich of intron 2, and some of them were located on exon 2, intron 3 and exon 3. In the overall evolution, CH1 was more conservative than CH2. The gene editing efficiency of CH2 was totally 40%. There were 2 types of mutation in CH2, which eliminated potential antigen sites. The gene editing efficiency of type 1 with deletion of 45 bases was 35%, and the gene editing efficiency of the type 2 with deletion of 44 bases was 5%. This study provides a way for further use of the target vector to obtain Fel d1 gene mutant cell lines for somatic cell nuclear transfer or embryo injection to obtain hypoallergenic cats.

Vascular endothelial growth factor B (VEGFB) is an important regulator of glucose uptake in skeletal muscle. Knockout of VEGFB can promote glucose uptake in skeletal muscle tissue of diabetes mice (Mus musculus), and improve glucose intolerance and insulin resistance in diabetes mice. However, there is no research to clarify the specific regulation of VEGFB on glucose uptake of skeletal muscle cell. In order to explore the regulatory effect of VEGFB on glucose uptake by skeletal muscle cell, this study used mouse myoblast C2C12 as a model, and used qPCR to detect the gene expression of VEGF signaling related genes and glucose transporter 4 gene (GLUT4) during the differentiation process of C2C12. The effects of VEGFB, VEGF receptor (VEGFR) inhibitor Axitinib and PI3K inhibitor Wortmanin on glucose consumption and uptake during C2C12 differentiation were detected by glucose oxidase method and 2-(N-(7-nitrobenz-2-oxa-13-diazol-4-yl) amino)-2-deoxy-D-glucose (2-NBDG) method. Western blot was used to detect the expression of GLUT4 on the cell membrane and in the cytoplasm and the activation of PI3K/AKT. The results of qPCR showed that the gene expression of VEGFB (P<0.01), VEGFR2 (P<0.01) and GLUT4 (P<0.05) increased significantly after differentiation of C2C12 compared with that before differentiation. Exogenous addition of VEGFB significantly increased the glucose uptake of C2C12 during differentiation (P<0.05), and significantly promoted the expression of GLUT4 on the cell membrane (P<0.05), and activated the PI3K/AKT pathway. But the VEGF receptor inhibitor Axitinib could reverse the promotion of VEGFB on glucose uptake, GLUT4 membrane localization and PI3K/AKT activation. In addition, PI3K inhibitor Wotmanin treatment could also reverse the effect of VEGFB on glucose uptake and GLUT4 cell membrane expression. The results of this study indicate that VEGFB could promote GLUT4 translocation to C2C12 cell membranes through the VEGFR-PI3K/AKT signaling pathway, thereby promoting glucose uptake by C2C12 during differentiation. This study reveals the regulatory effect of VEGFB on glucose uptake during the differentiation of myoblast C2C12, providing a new scientific understanding of the regulation of skeletal muscle glucose metabolism, and a basic reference for in-depth research on the regulation mechanism of skeletal muscle glucose metabolism.

The growth hormone (GH)-insulin-like growth factor (IGF) axis is the main endocrine axis that regulates fish growth. In order to explore the regulatory mechanism of growth hormone receptor (GHR) in the growth difference between female and male blotched snakehead (Channa maculata), the ghr gene in C. maculata was cloned, gene structure was analyzed, and the expression patterns of ghr in different adult tissues and liver at different developmental stages were clarified, meanwhile, its response to exogenous sex steroid hormones were also analyzed. The results showed that 2 602 bp cDNA sequence of the ghr gene in C. maculata was obtained, including 100 bp 5'-UTR, 1 872 bp ORF, encoding 623 amino acids, and 630 bp 3'-UTR. The genome sequence of ghr gene in C. maculata was 18 118 bp long, including 9 exons and 8 introns. 899 bp 5'-flanking sequence was identified, which contained androgen receptor (AR), bromomain-containing protein 4 (BRD4) and suppressor element-1-silencing transcription factor (REST) and other transcription factor binding sites. ghr was highly expressed in the liver of adult C. maculata, and its expression in males was significantly higher than that in females (P<0.01). The expression analysis of ghr gene at different developmental stages displayed that the highest expression of ghr gene was found in the liver of 365 d male C. maculata, which was significantly higher than that in females at the same period (P<0.01). Hormone induction experiments showed that long-term injection of 17α-ethynylestradiol (EE2) inhibited ghr expression in male and female C. maculata, long-term injection of 17α-methyltestosterone (MT) inhibited ghr expression in female C. maculata, but promoted ghr expression in males within a certain period of time. The results demonstrate that GHR might play a key role in the growth difference between females and males and it is also regulated by sex steroid hormones in C. maculata. This study provides the reference for further exploring the molecular mechanism of growth regulation in blotched snakehead.

Flo8 (flocculation 8) gene, a key transcription regulatory factor downstream of the cAMP pathway, plays an important role in the regulation of fungal growth and development. In previous study, the involvement of StFlo8 in regulating the appressorium development and the formation of infection filaments in the early stage of Setosphaeria turcica was identified. In the present study, StFlo8 fused with glutathione S-transferase (GST) tag were heterologously expressed via the Escherichia coli expression system. Further, the GST pull down technology was used to screen the proteins that may interact with StFlo8 from the mycelial proteins of S. turcica, and the yeast two hybrid technology was used to verify the interaction of some of the proteins. The results showed that the target product with a size of 116 kD obtained by prokaryotic expression was consistent with the size of GST-StFlo8, 41 proteins that may interact with StFlo8 were obtained from the successfully purified expression product through GST pull down. After validating a subset of proteins using the yeast two-hybrid technique, it was revealed that Flo8 interacts with FAD-dependent oxidoreductase (GenBank No.165285). This study preliminarily clarified the interaction protein of StFlo8, providing informative references for further elucidating the regulatory molecular mechanism of the activity of StFlo8 in S. turcica.

The N⁶-methyladenosine (m⁶A) is the most common RNA modification in eukaryotes. Proteins containing the YT521-B (YTH) domain recognize the m⁶A site and mediate RNA metabolism. However, the function of m⁶A recognition proteins in plant pathogenic fungi has not been reported yet. This study systematically identified and analyzed the m⁶A recognition proteins in 127 plant pathogenic fungi using bioinformatics method. Furthermore, the expression patterns of the genes encoding m⁶A recognition proteins during the development of Setosphaeria turcica were analyzed. The results showed that a total of 225 m⁶A recognition proteins were identified in the tested 127 fungi, and 93% of the fungi in the Basidiomycota phylum had 2 m⁶A recognition proteins. Physicochemical analysis showed that 67.56% of the m⁶A recognized proteins identified were alkaline. Notably, significant differences in the acidity and alkalinity of these proteins were observed within different phyla and fungal lifestyles. Phylogenetic analysis revealed that 225 m⁶A recognition proteins could be classified into 5 groups, 2 of which shared a closer evolutionary relationship with humans (Homo sapiens). Additionally, m⁶A recognition proteins from fungi with the same phylum and lifestyle tended to cluster together on evolutionary branches. The conserved site analysis uncovered the existence of several conserved-type sites in the sequences of the YTH domain across the 225 m⁶A recognized proteins. A gene that encodes the m⁶A recognition protein, namely StYTH1, was identified in S. turcica. Expression analysis revealed that StYTH1 had the highest expression level in the developmental stage of appressorium, suggesting that StYTH1 might be involved in regulating the development of appressorium. The present study provides basic materials for an insightful revelation of the functions of the m⁶A recognition proteins.

Mating system is an important trait in oomycetes, effector PITG_02860 was segregated and linked with the mating type of A1 of Phytophthora infestans. In order to investigate the function of the P. infestans PITG_02860 gene, protoplasts transformation protocol was optimized under the conditions of the mycelium cultured with soybean juice for 72 h, and then lysed by the complex enzyme (10 mg/mL lysing enzyme, 5 mg/mL cellulase) at 26 ℃ for 45 min, resulted in the highest rate of preparation of protoplasts, which was up to (31.5±2.36) protoplasts/μL; and the regeneration of the protoplasts in the medium of rye could reach to (3.34±0.20)%. Based on this transformation system, the PITG_02860 gene silencing vector was constructed with the vector pTOR-mRFP as the backbone and transformed into strain 88069. Functional verification of the 2 silenced strains revealed that, compared to the wild-type, the number of oospores produced during sexual reproduction significantly decreased and the rate of oospore deformity significantly increased (P<0.05), the growth of hyphae was hindered, the number of zoospores significantly decreased (P<0.05), and the pathogenicity to potato (Solanum tuberosum) leaves was significantly reduced (P<0.05). The above results indicated that PITG_02860 was not only a key gene for the normal development of sexual reproduction of P. infestans, but also an important virulence function gene when infecting potatoes. The study provides new ideas for sexual reproduction occurrence and virulence mechanism of P. infestans.

CRISPR/Cas9 technology, as the third generation of gene editing technology with simple design, convenient and quick operation, and low cost of use, has set off a technological revolution in the field of life sciences research. Currently, CRISPR/Cas9 has been successfully applied to gene editing in eukaryotic and prokaryotic organisms such as animals, plants and microorganisms. In recent years, CRISPR/Cas9 has also been used in the study of insects, and has shown great promise in the exploration of new technologies for pest control. This paper mainly provides an overview of the working principle of the CRISPR/Cas9 system and summarizes the progress of this technology in pest gene function and pest control. This review provides a reference for the use of CRISPR/Cas9 to carry out broader research in insect molecular biology and pest management strategies.

As a mitochondria-targeted antioxidant, mitoquinone mesylate (MitoQ) has been proved to relieve oxidative stress damage of mitochondria, target mitochondria to resist apoptosis, and maintain body health. Existing research has proven that MitoQ plays an important role in cell damage, organ damage repair and the treatment of various diseases. This paper reviews the biological functions of MitoQ such as antioxidant, resistance to cell inflammation and apoptosis, and its mitochondrial targeting, and systematically summarizes the research progress of MitoQ in the treatment of neurodegenerative diseases, cardiovascular injury, metabolic organ injury, and diabetes. This review provides reference for the use of MitoQ in cell biology and clinical disease treatment.

Fluoroquinolones (FQs) are one of the most commonly used antibiotics for human medicine and animal breeding. Due to their large dosage, FQs are widely retained in the environment, posing potential threat to human and animal health. Microbial degradation is an ideal way to eliminate FQs in the environment at this stage, which has the characteristics of high efficiency, green and low cost. At present, the research on microbial degradation of FQs mainly focuses on the screening of degradable microorganisms and the identification of degradation products. This paper reviewed the functional characteristics of FQs degradation of bacteria, fungi, microalgae and microorganisms, and summarized the common degradation pathways and key enzymes. This review provides a theoretical basis for alleviating the environmental residue problem of FQs.

The retinal pigment epithelium 65 (RPE65) gene plays an important role in the visual cycle; when the RPE65 gene is mutated, there is a loss of cone and rod cell function, and loss of vision in severe patients. In order to construct an animal disease model with RPE65 gene mutation; this study utilized transcription activator-like effector nucleases (TALEN) gene editing technology to construct expression vectors for Exon3 and Exon7 of the RPE65 gene in Beagle dogs (Canis lupus familiaris), and combined with microscopic manipulation to create RPE65 gene-edited dogs. The dogs' visual function was tested by clinical observation, routine ophthalmologic examination, fundus photography (FP), optical coherence tomography (OCT), and full-field electroretinography (ERG). T7E1 digestion and gene sequencing results showed that 2 RPE65 gene-edited male dogs were finally obtained. Ophthalmic phenotypic analysis showed that the fundus blood vessels of RPE65 gene-edited dogs were sparse and slender. The ERG results showed that the amplitude of the a-wave and b-wave potentials in photoreceptor cells decreased, and the peak time prolonged, indicating impaired retinal function. In this study, RPE65 gene-edited dogs were constructed by using TALEN gene editing technology with visual dysfunction, which may provide a reference for studies related to retinal diseases induced by mutations in the RPE65 gene, and also provide a good animal disease model for the development of therapeutic treatments and the screening of clinical drugs.

Porcine parvovirus type 6 (PPV6) is a new swine (Sus scrofa) Porcine parvovirus, which can cause symptoms of reproductive dysfunction and seriously threaten the healthy development of pig industry. In order to develop an indirect ELISA (iELISA) method for rapid detection of PPV6 antibody, the highly conserved encoded gene virion protein (VP1)(348~675 aa) of the popular PPV6 strain was used as the target fragment, and the gene was amplified from the DNA of PPV6 positive sample by PCR. The recombined plasmid pET30a-PPV6-VP1 was expressed in Escherichia coli BL21 (DE3) and was induced by IPTG. The recombinant VP1 protein with a relative molecular weight of 40 kD was purified by Ni-NTA. The iELISA was developed by using the recombinant VP1 protein as coating antigen, and studied the specificity, sensitivity, repeatability and coincidence rate of the method. The coating concentration of VP1 protein was 1 ng/μL, and the detection positive threshold was OD₄₅₀>0.62. The dilution concentration of serum and HRP-rabbit anti-pig IgG was 1∶100 and 1∶60 000, respectively. The iELISA was not cross reaction with positive sera of Porcine reproductive and respiratory syndrome virus (PRRSV), Classical swine fever virus (CSFV), Pseudorabies virus (PRV), Japanese encephalitis virus (JEV), Porcine circovirus type 2 (PCV2), PPV1. The sensitivity of iELISA was 1∶3 200. The method was of high duplicability with less than 10% variation of intra-and inter-batch coefficients. The result of iELISA was positive correlation with virus neutralization test, and the positive coincidence rate with Western blot method was 96.6%. The above results showed that the established iELISA method had good specificity, sensitivity, and repeatability. The positive rate of PPV6 antibody was 5.31% in 452 pig serum samples collected in some provinces in the western of China from 2018 to 2022, which preliminarily proved that the disease had been infected in the western of China. This study successfully truncated expression VP1 (348~675 aa) protein and established iELISA of PPV6 with better specificity, sensitivity, repeatability and coincidence rate, which provides material for serological investigation and test kit development of PPV6.