Adenosine 5'-monophosphate-activated protein kinase (AMPK), as a key enzyme in mammalian cell metabolism, plays an important role in the maturation of oocytes. However, the role of AMPKα1, the catalytic subunit of AMPK, remains unclear in the maturation of mouse (Mus musculus) oocytes. In this study, the different functional sites of AMPKα1 were mutated by splicing by overlap extension PCR (SOE-PCR) to obtain AMPKα1-D157A, AMPKα1-T172D, AMPKα1-S485A and AMPKα1-S485A-S173A mutants, respectively. After verifying their functions in Hela cell lines, cRNAs were transcribed in vitro, and the AMPKα1 genes containing various mutation sites were overexpressed in mouse oocytes by microinjection technology. The results showed that AMPKα1 and its mutants were widely distributed in the cytoplasm of Hela cells, and cells overexpressing AMPKα1-S485A mutants showed increased proliferation and activity. Meanwhile, overexpression of AMPKα1 and its mutants in mouse oocytes showed aggregative distribution, and it was seemed to be associated with chromatin separation during meiosis. Compared with the control group, the overexpression of each group except the AMPKα1-S485A overexpression group accelerated the maturation process of mouse oocytes, and the germinal vesicle breakdown (GVBD) rate of the oocytes overexpressing AMPKα1-S485A mutant was the same as that of the control group. In addition, the overexpression of AMPKα1-S485A could increase the polar body excretion rate of oocytes and reduce the mortality rate of oocytes. These results showed that AMPKα1 played an important role in the maturation of mouse oocytes, and this study provides a reference for the subsequent study of AMPKα1 in mouse oocytes.

Phytophthora root and stem rot (PRR) represents one of the destructive diseases in soybean (Glycine max) production, generally causing 10%~50% yield loss every year. Transgenic biotechnology provides an effective strategy for preventing and controlling the occurrence of PRR. The small G protein, known as the 'molecular switch', is not only involved in the process of regulating the adaptation of plants to external environmental stimuli during plant growth and development, but also participates in the establishment of plant defense responses. In this study, Gossypium barbadense Rac family small G protein 1 gene (GbRac1) was introduced into the soybean cultivar 'Williams 82'. Integration and expression of the foreign gene in transgenic soybean plants were confirmed by Southern blot and reverse transcription-PCR (RT-PCR) analysis. The PRR tolerance of the T₃ and T₄ generations of transgenic soybean lines was evaluated using a hypocotyl inoculation method. At 7 d after inoculation with Phytophthora sojae race 1, typical PRR symptoms such as wilting leaves and plant death were observed in the non-transformed plants. In contrast, most of transgenic plants showed no visible symptoms and developed normally with the survival rate of 70.68%~90.63%, higher than that of the control plants (40.74%~50.85%). However, no significant differences in agronomic traits were observed between the transgenic and non-transformed plants in the field conditions when no P. sojae inoculation involved. Taken together, these results demonstrated that over-expression of GbRac1 significantly enhanced the tolerance of the transgenic soybean to PRR without negative consequences on the agronomic performance. This results also pave the way for broadening disease resistance in soybean and development of the PRR-resistant soybean cultivars.

The GSTU7 gene encodes glutathione transferase. In the previous research, this group found out that GbGSTU7 (GenBank No. GB_A03G0575) gene responds to the process of resistance to Fusarium wilt. In order to verify the GbGSTU7 gene in the resistance of Gossypium barbadense to Fusarium wilt, the Tobacco rattle virus-mediated virus induced gene silencing system was used to silence the GbGSTU7 gene in the disease-resistant material of Gossypium barbadense '06-146', and qPCR technology was used to analyze the expression of GbGSTU7 gene. The physiological race 7 of Fusarium wilt was used to infect the the GbGSTU7 gene silenced plants to study their disease resistance and measure the glutathione peroxidase activity of Gossypium barbadense. The results showed that the silencing efficiency of GbGSTU7 gene was 74%, and the disease index for cotton Fusarium wilt increased 31.4, the soluble protein content increased by 3 times, and the activity of glutathione peroxidase decreased by 2 times, indicating that the GbGSTU7 gene played an important role in the anti-fusarium wilt reaction of Gossypium barbadense. The silencing of GbGSTU7 affected protein transport and cooperated with glutathione peroxidase to resist infection by pathogens. This study provides a reference for the in-depth study of the mechanism of the GbGSTU7 gene involved in the resistance to Fusarium wilt of Gossypium barbadense.

Calcium-dependent protein kinase (CDPK) is a specific type of Ser/Thr protein kinase to plants, which plays an important role in plant growth, development and stress signal transduction. In order to explore the function of CDPK genes in the growth and development of pecan (Carya illinoinensis) and Chinese hickory (Carya cathayensis), bioinformatics methods were applied to identify the members of CDPK gene family from pecan and Chinese hickory genome. MEGA7.0 software was used to carry on the multiple sequence alignment, classification and phylogenetic tree construction. The gene structures and conserved motifs of CDPK family were also analyzed. The results showed that 28 (CILCDPK1~28) and 25 (CCACDPK1~25) CDPK members were successfully identified in pecan and Chinese hickory, respectively. In order to explore the homologous evolutionary relationship between the two plants and other species, a phylogenetic tree was constructed using CDPKs in pecan, Chinese hickory, Arabidopsis and rice (Oryza sativa). The phylogenetic tree showed that all CDPK proteins could be divided into 4 subgroups. Cis-acting element analysis showed that all CDPK genes contained hormone-response elements, and abscisic acid-response elements were the most abundant. Transcriptome data analysis of pecan and Chinese hickory showed that 3 CILCDPKs and 2 CCACDPKs might play an important role in the 3 key stages of embryonic development. In addition, 3 CDPK genes showed high expression during pecan grafting and healing, which might play important roles in callus and vascular tissue formation of pecan. qPCR analysis showed that CILCDPK1/3/6/11/13/19/25 were up-regulated under drought stress. The present study provides a reference basis for exploring the mechanism of CDPK gene family in plant growth and development and drought stress response.

R2R3-MYB transcription factor is one of the largest transcription factor family in plants, and it is widely involved in the regulation of plant secondary metabolism. Based on the sequencing data of the full-length transcriptome of Crocus sativus, explored the temporal and spatial expression characteristics of R2R3-MYB transcription factors and screend the CsR2R3-MYB transcription factors related to the secondary metabolism of C. sativus, bioinformatics methods were used to identify and systematically analyzed the CsR2R3-MYB transcription factor, and the temporal and spatial expression pattern of the CsR2R3-MYB transcription factor was explored by qRT-PCR. Totally 42 R2R3-MYB transcription factors were screened from the C. sativus transcriptome. Physical and chemical properties analysis of the proteins showed that the CsR2R3-MYB proteins were all unstable hydrophilic proteins. Subcellular localization prediction showed that most of the CsR2R3-MYB were present in the nucleus. The phylogenetic tree was constructed together with Arabidopsis thaliana R2R3-MYB, and the results showed that the CsR2R3-MYB transcription factors could be divided into 26 subclasses. According to the function conserved properties of homologous proteins, CsMYB9, CsMYB16, CsMYB17, CsMYB18, CsMYB20, CsMYB21, which were homologous to the S7, S19, and S20 subclasses, were predicted to participate in the secondary metabolism regulation of active substances in C. sativus by responding to jasmonic acid (JA) signals. The results of temporal and spatial expression analysis showed that CsMYB5, CsMYB6, CsMYB7, CsMYB9, CsMYB10, CsMYB13, CsMYB17, CsMYB19, CsMYB21, CsMYB22, CsMYB23, CsMYB26, CsMYB28, CsMYB30, CsMYB31 and CsMYB32 showed relatively high expression levels in the stigma tissues and responds to JA signals. The previous studies of this research group showed that JA could promote flavonoids biosynthesis. According to the prediction results of the phylogenetic tree and the temporal and spatial expression characteristics, screened CsMYB9, CsMYB17, and CsMYB21 regulate the secondary metabolism of C. sativus, by responding to JA signals. This research provides a reference for the further exploration of the functions of CsR2R3-MYB. And it also provides ideas for the study of transcription regulation of C. sativus secondary metabolism.

Prostate cancer is a heterogeneous disease with a high degree of clinical and genetic heterogeneity, thus providing multiple therapeutic targets.To investigate the expression profiles of 14-3-3 protein family isoforms in different cell lines of prostate cancer and to search for specific drug targets for the treatment of prostate cancer.The expression patterns of each isoform of the 14-3-3 protein family in different prostate cancer cell lines were analyzed by qPCR (real-time fluorescence quantitative PCR) in combination with the UALCAN-TCGA database. The effects of different expression levels on the proliferation and migration ability of 22RV1 cells were demonstrated by up- and down-regulating the expression of tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ) gene using CCK-8 and cell scratch assay. It was found that the expression of YWHAZ in cancer tissues was lower than that in normal tissues when GLEASON SCORE was lower than 8, while the relative expression of YWHAZ in prostate cancer cell lines showed a significant upward trend in the middle and late stages as Gleason score (GS) was elevated. The results of CCK-8 assay and cell scratch assay revealed that the interference and overexpression of 14-3-3ζ expression levels in 22RV1 cells severely affected the proliferation and migration ability of the cells YWHAZ. The above results implied that YWHAZ may play a role as a proto-oncogene in the middle and late stages of prostate cancer development. The related results provide a theoretical basis and experimental ideas for the subsequent study of drugs targeting this gene.

Tibetan pig (Sus scrofa) is a representative non model animal in plateau habitat. The study of its plateau response mechanism can provide a reference basis for the exploration of hypoxia adaptation mechanism of other animals. In order to improve and optimize the annotation information of Tibetan pig genome, analyze the characteristics of Tibetan pig lung tissue response to hypoxic environment at transcription level, this study performed RNA-seq sequencing of lung tissues in Tibetan pigs at different altitudes to construct cDNA library, and the reassembled sequencing data was compared and analyzed with the reference genome of Tibetan pigs. The results showed that the 5' end and 3' end of 334 genes were extended, and the 5' end of 1 313 genes and the 3' end of 2 734 genes were extended respectively; 809 new transcripts were excavated and 95 new transcripts at different altitudes were differentially expressed (P<0.05). GO analysis results showed that the differential new transcripts were significantly enriched in the process of cell metabolism, enzyme catalytic activity and cellular components (P<0.05); KEGG pathway enrichment results found that the differential new transcripts were significantly enriched (P<0.05) in the pathways such as programmed cell necrosis and nuclear factor-κB (NF-κB) signaling pathway. Nine new transcripts with significant differences were randomly selected for qPCR verification, and the results were consistent with that of RNA-seq. This study provides a useful supplement and reference for improving the annotation information of Tibetan pig genome and a reference basis for further studying the genetic mechanism of Tibetan pig adapting to different altitude environment.

The myogenic factor 5 (MYF5) gene is considered to be an important gene affecting muscle growth and development. The aim of this study was to investigate the effect of genetic variation of MYF5 gene on growth traits in sheep (Ovis aries), in order to provide effective molecular genetic markers for the breeding of new breeds (lines) of high quality meat sheep. Three SNP loci (124510044, 124509770 and 124507708) with polymorphism and significant interbreed differences were screened. The selected loci were examined by flight mass spectrometry in 383 sheep from the 3 crosses and correlated with growth traits at birth and 3 months of age. The results showed that locus 12451044 detected 3 genotypes, which were moderately polymorphic (0.25 ≤PIC< 0.50) in all populations; locus 124509770 detected 3 genotypes, which were low polymorphic (PIC< 0.25) in the H₁ generation population and moderately polymorphic (0.25 ≤PIC< 0.50) in the other populations. Only one genotype deletion was detected at locus 124507708. In the H₂ generation population, individuals with CT genotype at locus 124510044 had significantly higher body height at 3 months of age than TT genotype (P<0.05), and individuals with CG genotype at locus 124509770 had significantly higher body height at 3 months of age than GG genotype (P<0.05); individuals with GG genotype had significantly higher chest circumference at 3 months of age than CC genotype, regardless of generation (P<0.05 ). 124510044 and 124509770 loci were complete linkage disequilibrium, producing 4 haplotypes and 5 diploids; individuals with diploid H4H4 had significantly higher chest circumference at birth and at 3 months of age than H1H1 (P<0.05). The results of this study showed that genetic variation in the MYF5 gene has an effect on growth traits in sheep, and the loci 124510044 and 124509770 can be used as potential candidate genetic markers for growth and development and meat production performance in meat sheep.This study provides an important reference for the MYF5 gene in the selection and breeding of meat sheep, and provides a theoretical basis for molecular marker-assisted breeding in meat sheep.

Ciliary neurotrophic factor (CNTF) is a cytokine with multiple biological functions in vivo. It is an important candidate gene affecting muscle development. The aim of this study was to investigate the tissue expression pattern of CNTF gene in sheep (Ovis aries), scan its SNPs and analyze its association with growth traits in a large population. In this study, 1 399 Hu sheep with clear family tree and accurate phenotype records were raised in a specialized performance measurement house. Before morning feeding at 80, 100, 120, 140, 160 and 180 d, respectively, the weight of the sheep was weighed, and body length, body height, chest circumference and cannon circumference were measured at the same time. At the end of the assay period, whole blood samples of each sheep were collected by jugular vein for genomic DNA extraction. They were slaughtered immediately and tissue samples were taken. According to design the CNTF mRNA sequence specific primers, RNA from the collected tissue samples was extracted and reverse transcribed to cDNA, using qPCR technology to detect the CNTF gene expression in 10 tissues of Hu sheep. Genomic DNA was extracted from whole blood samples of the experimental population using genomic DNA extraction kit according to the instructions. Mixed DNA was used as the template to amplify the designed PCR primers. SNPs of the gene was scanned by mixed pool sequencing method, and the SPSS 23.0 software was used for correlation analysis with sheep growth traits. This results found a high positive association between growth traits, with a higher correlation between weight and body size indicators such as body length, body height, chest circumferences and cannon circumferences (P<0.01). The ovine CNTF gene was ubiquitously expressed, with expression observed in 10 tissues including heart, liver, spleen, lung, and kidney, with higher expression in liver and rumen. The sheep CNTF gene fragment was amplified and sequenced, a g.2576 C>G polymorphism locus was detected in intron 1 of the sheep CNTF gene. The SNP sites were genotyped by KASPar technique, and 3 genotypes were obtained, namely CC, CG and GG. In the experimental population, the genotype frequencies of CC, CG and GG genotypes were 0.13, 0.12 and 0.75, respectively, and the frequencies of alleles C and G were 0.19 and 0.81, respectively. The polymorphism information content of this locus was 0.26, which showed moderate polymorphism, and its heterzygosity (He), homozygosity (Ho) and effective number of alleles (Ne) were 0.31, 0.69 and 1.45, respectively. The results of trait association analysis showed that the polymorphism locus was significantly correlated with the growth traits such as body weight, body height, body length, chest circumference and cannon circumference of Hu sheep from 80 to 180 d (P<0.05). The body weight at 80, 100, 120, 140 and 160 d, body height at 80, 100 and 120 d and chest circumference at 160 d of individuals with GG genotype were significantly higher than those with CG genotype (P<0.05). The body height at 140, 160, 180 d, chest circumference at 80, 100, 140 and 180 d and cannon circumference at each stage were significantly higher in individuals with GG genotype than those with CG genotype and CC genotype (P<0.05). These results indicated that the CNTF gene g.2576 C>G locus could be used as a candidate molecular marker for breeding of growth traits in Hu sheep.

Rumen is an important digestive organ of ruminants, early supplementation with starter can meet the nutritional needs of rapid growth of lambs (Ovis aries) and promote the early development of rumen. This study was conducted to investigate the effects of supplementation with starter on gene expression in rumen tissues of lambs to clarify the possible mechanism of starter on rumen development. Twenty-two healthy and disease-free male Hu lambs (birth weight=(3.65±0.49) kg) were randomly divided into 2 groups, control group and supplementary feeding group. Control group was fed with milk replacer, while supplementary feeding group was fed with milk replacer and starter. At the age of 28 days, rumen pouch tissues were collected for transcriptome sequencing analysis. The result showed that 61.70 and 58.95 Gb of clean reads were obtained in control group and supplementary feeding group, respectively, and the comparison efficiency with the reference genome was 87.85%~89.45%. Compared with the control group, 868 differentially expressed genes were screened after supplementary feeding, among which 464 genes were up-regulated and 404 genes were down-regulated. And it was mainly concentrated in GO functions such as enzyme activity, ion channel and membrane transport and KEGG signaling pathways such as steroid hormone biosynthesis, arachidonic acid metabolism, peroxisome proliferator-activated receptor (PPAR) signaling pathway, regulation of lipolysis in adipocytes and cyclic adenosine monophosphate (cAMP) signaling pathway. Correlation analysis showed that recombinant IQ motif containing GTPase activating protein 2 (IQGAP2), recombinant solute carrier family 16, member 1 (SLC16A1), ankyrin repeat domain-containing protein 45 (ANKRD45), 3-hydroxy-3-methylglutaryl-CoA, synthase 2 (HMGCS2), phosophoenolpyruvate carboxykinase 2 (PCK2) were significantly positively correlated with the length of rumen papilla, the concentration of acetate, propionate, butyrate and total volatile fatty acid (TVFA)(P<0.05), while ceramide synthase 3 (CERS3), recombinant keratin 4 (KRT4), solute carrier family 22A17 (SLC22A1), recombinant insulin like growth factor binding protein 6 (IGFBP6) were significantly negatively correlated (P<0.05). Supplementation with starter may promote rumen development by regulating the expression of genes related to rumen epithelial proliferation and nutrient generation. This study provides a basis for further exploring the mechanism of supplemental starter on rumen development.

Heat shock proteins (HSPs) are closely related to fish response to temperature stress. To investigate the response characteristics of the heat shock proteins family genes, hsp90b1 and hspb1, in Larimichthys polyactis subjected to temperature stress, In this study, full-length CDS of hsp90b1 and hspb1 in L. polyactis was cloned, and detected their spatiotemporal expression characteristics and expression patterns after acute and long-term temperature stress. The full-length CDS sequence of hsp90b1 (GenBank No. OK340652) was 2 406 bp, encoding a predicted 801 amino acid. While the full-length CDS sequence of hspb1 (GenBank No. OK340653) was 609 bp encoding 202 amino acid. Tissue expression analysis results showed that the hsp90b1 gene expression is the highest in the skin, while hspb1 gene expression was the highest in the muscle. As the natural water temperature was 20 ℃ (control group), L. polyactis was treated with acute high (32 ℃) and low (6 ℃) temperature stress. The results showed that the acute temperature stress could induce significant high expression of hsp90b1 in the liver. Differently, hspb1 expressed higher with high temperature and expressed lower with low temperature (P<0.05). During long-term temperature treatments (with 13 ℃ as the control, fish was treated at 6, 8, 16 and 20 ℃, respectively and maintained for 7 d). It was found that the expression of hsp90b1 in the liver of the 4 treatment groups was significantly higher than that in the control group, which temperature was 13 ℃ (P<0.05), while the expression level of the high-temperature group (16 ℃ and 20 ℃) was higher than that in the low-temperature group (6 ℃ and 8 ℃); Unlike hsp90b1, hspb1 still showed the characteristics of high expression at high temperature and low expression at low temperature (P<0.05). The present study indicated that hsp90b1 and hspb1 were significantly differentially expressed in L. polyactis during temperature stress, suggested that hsp90b1 and hspb1 played an important role in the response to temperature stress. The results of this study provide an important foundation for exploring the molecular mechanism of the L. polyactis in response to temperature stress.

The circadian clock genes are the key gene to control the circadian rhythm. Light is the environmental variable that has the greatest impact on the circadian clock, changes in the photoperiod lead to changes in the circadian expression mode of the circadian clock genes, which further affect the various physiological activities such as biological living habits, development and reproduction. In order to investigate the effects of different photoperiods on the circadian rhythm of Nile tilapia (Oreochromis niloticus), the circadian expression of biological core circadian clock genes bmal1 (brain and muscle arnt-like 1), clocka (circadian locomotor output cycles kaput a), cry5 (cryptochrome 5) and per1b (period 1b) in brain, liver and eye of Nile tilapias was analyzed under 3 different photoperiods, the light (L)/dark (D) ratios were 8 h light and 16 h dark (8L∶16D), 12 h light and 12 h dark (12L∶12D), 16 h light and 8 h dark (16L∶8D), light intensity was 1 000 lx, the light start time is set to the zeitgeber time (ZT) 0 h. The result showed that bmal1, clocka, cry5, per1b displayed a circadian rhythm in brain, liver and eye under normal photoperiod condition (12L∶12D); Under 12L∶12D, the acrophase of positive regulatory genes bmal1 and clocka in liver (ZT 13.98 h, ZT 12.02 h) was earlier than that in brain (ZT 17.31 h, ZT 14.37 h), and that of negative regulatory genes cry5 and per1b in liver (ZT 7.28 h, ZT 0.92 h) was later than that in brain (ZT 4.66 h, ZT 0.76 h); Long light (16L∶8D) led to the delayed acrophase of clocka in brain, the increased amplitude of cry5 in liver; Short light (8L∶16D) caused the increased amplitude of per1b in brain, bmal1 in liver and cry5 in eye. It indicated that Nile tilapia had a biological clock system similar to other known fish. The biological rhythms of different tissues were not exactly the same. Environmental factors such as photoperiod would affect the expression of clock genes and the response of clock genes in different tissues was different. This study provides a reference basis for further elucidating the regulation mechanism of circadian clock system in Nile tilapia.

The properties of farmland soil polluted by acid mine drainage (AMD) change greatly, which is potentially harmful to microbial growth. In order to study the impact of arsenic containing AMD on the microbial community structure in paddy field, soil samples were collected from contaminated and clean fields near one arsenic rich coal mine of Xingren city, Guizhou province. The physical and chemical properties of overlying water and soil were determined. The high-throughput sequencing technology was used to analyze the microbial community composition and diversity changes in contaminated and clean areas. The results showed that the soil in contaminated area was acidified, the pH ranged from 3.60 to 4.40, and the average contents of iron and arsenic were 114.26 g/kg and 98.70 mg/kg respectively. While the average soil pH in clean area was 5.93, and the average contents of the two elements were 106.25 g/kg and 21.63 mg/kg, respectively. Proteobacteria was the main category of microorganisms in the two areas, with an average relative abundance of 29.96%. The diversity of soil microbial community in S1 and S2 of contaminated area was significantly different from that in clean area (P<0.05). Compared with the clean area, the relative abundances of acidophilic genera Gp1, Gp2 and Gp13 increased by 3.23%, 2.88% and 1.38%, respectively. Desulfosporosinus and Desulfitobacterium were only detected in the soil of contaminated area. The abundances of Geobacter and Anaeromyxobacter in iron reducing bacteria group were relatively high. Their average proportions in contaminated area were 1.58% and 1.60%, respectively, and 1.62% and 3.02% respectively in clean area. Redundancy analysis and Spearman correlation analysis showed that soil acidification and arsenic contamination were the main environmental pressures on soil microbial community structure. The study can help to further reveal the relationship between the variation of soil environmental conditions contaminated by arsenic containing AMD and the change of microbial community structure, and provides a theoretical basis for the ecological restoration of contaminated field.

Pseudomonas aeruginosa has multiple drug resistance, and the formation of biofilm is one of its important drug resistance mechanisms. Glycyrrhizin (GLY) has antibacterial effect. In order to investigate the effects of antibacterial activity and biofilm formation of GLY on clinical isolates of multidrug resistant P. aeruginosa, G1 strain (GenBank No. MZ683158) and G2 strain (GenBank No. MZ683159) were isolated from the eye secretions of rabbits suffering from keratitis (Zhejiang Shengzhou wool rabbit 'White King', Leporidae angora). Further, the minimum inhibitory concentration (MIC) of GLY on G1 and G2 strains was determined by microdilution method, crystal violet staining was used to quantify biofilm, the effect of GLY on biofilm morphology was observed by scanning electron microscope (SEM), and the expression of pelA, algA, rhlI, rhlR and pslA was analyzed by qPCR. The results showed that MICs of GLY against G1 and G2 strain were 40 and 20 mg/mL, respectively; both G1 and G2 strains could form biofilm, and G1 strain had stronger ability. The biofilm formation of G1 and G2 strains decreased significantly with the treatment of 20 mg/mL GLY (P<0.01). SEM observation showed that 5 mg/mL GLY significantly reduced the biofilm formation; under the treatment of 10 mg/mL GLY, the bacteria were dispersed and no obvious biofilm structure was found. The expression of pelA increased after 5 mg/mL GLY treatment without signifcant difference, while those of algA, rhlI, pslA and rhlR were significantly decreased (P<0.01). After 10 mg/mL GLY treatment, the relative expression of pelA, algA, rhlI, rhlR and pslA were significantly decreased (P<0.01) compared with the control. In conclusion, GLY had a inhibitory regulatory effect on the synthesis of P. aeruginosa biofilm in a dose-dependent manner, which might be due to the down regulating of related genes. This study provides a reference for in-depth study of the effect of GLY on P. aeruginosa biofilm and the treatment of P. aeruginosa keratitis in rabbits.

Sugarcane (Saccharum spp. complex) is a major species of the gramineous family and an economically important crop, which plays an important role in the development of world agricultural economy and the guarantee of sugar supply. One hand, sugarcane is known as a pioneer crop, as it has good resistance or tolerance to abiotic adversity. However, its production and productivity are constrained by various abiotic and biotic stresses, resulting in a serious loss in cane yield and sugar content. On the other hand, sugarcane flowering and fruiting requires strict light and temperature conditions, and hybridization is generally adopted by selecting suitable sites to induce the parents to flower in all countries, together with a photoperiod chamber. Normally, many commercial cultivated varieties generally do not blossom in the process of cultivation, and there is no seed even though they flower in a certain location in some years. In addition, sugarcane is an industrial raw material crop, and its product sucrose is carbohydrate and does not contain protein. Therefore, the safety risk level of transgenic sugarcane is low, second only to non-food crops, such as cotton. Furthermore, sugarcane is an asexually propagated crop, and once excellent transgenic individual plants are obtained, its population can be rapidly expanded by tissue culture. Genetic engineering has become an important means for sugarcane to make up for the defects of traits derived from traditional crossbreeding and to accelerate the process of genetic improvement. Regarding why sugarcane is a suitable species for transgenic improvement, this paper highlighted the achievements of improvement of insect pest resistance and disease resistance, briefly reviewed the improvement of other traits and the work of sugarcane as a bioreactor. Then, we focus on the factors affecting the efficiency of genetic transformation and the expression level of exogenous target genes in sugarcane. Additionally, the progress of gene-editing in sugarcane was reviewed. Finally, the future of transgenic sugarcane was prospected. The present review aims to promote the application of genetic engineering in sugarcane improvement.

Dehydration proteins (dehydrins, DHN) are group 2 members of the late embryogenesis abundant proteins (LEA). DHN is usually produced and accumulated when plants are subjected to adversity stress to enhance plant tolerance to adversity, and therefore plays an important role in plant resistance to adversity. This paper reviews the protein structure and functional characteristics of dehydrin, as well as its role under abiotic stresses such as low temperature, drought and salt stress, with a view to providing a reference for in-depth studies of dehydrin. The application prospect of using modern biotechnology to use dehydrin for improving plant tolerance to adversity is foreseen. This review summarizes the function of dehydrin, explores the action mechanism of dehydrins and analyzes the molecular mechanism of dehydrins in the review, so as to breed new varieties and maintain high and stable yields and sustainable development.

Food-borne microorganisms are important factors affecting food quality and safety. Droplet digital PCR (ddPCR), which can be used for the absolute quantitative detection of nucleic acids, plays an increasingly important role in the detection of food-borne pathogens. In order to quantify the genome copy numbers of food-borne pathogens, a strategy for simultaneous detection of the DNA copy numbers of Salmonella typhi, Staphylococcus aureus and Listeria monocytogenes by multiplex droplet digital PCR (ddPCR) was developed in this study. Primers and probes targeting ttrA/ttrC、FMN-binding glutamate synthase、invasion associated endopeptidase gene were selected to evaluate the feasibility and applicability of the strategy by specific verification using TaqMan probe qPCR. The probes for Salmonella typhi and Staphylococcus aureus were labeled with 6-carboxy-fluorescein (FAM) fluorophore, and the probe for L. monocytogenes was labeled with hexachloro fluorescein (HEX) fluorophore. The detection systems of simplex ddPCR for 3 target strains were constructed. On this basis, through the optimization of primer and probe concentration and annealing temperature, a multiplex ddPCR detection system for simultaneous, rapid and absolute quantitative detection of genomic copy numbers of Salmonella typhi, Staphylococcus aureus and L. monocytogenes in the same reaction system was constructed. The multiplex ddPCR detection system could obtain fluorescent amplification with 8 (2³) distinct clusters under the optimal condition obtained by means of annealing temperature optimization, and the quantitative linear range of these 3 pathogens were 235~0.23、284~0.18、380~0.42 copies/µL, which were almost the same as simplex ddPCR for single strain, and the linear correlation coefficient R² in the quantitative detection range were all greater than 0.999 with excellent repeatability. The constructed multiplex ddPCR detection system had strong specificity, good stability and wide quantitative range, which could provide technical support for absolute quantitative detection of food-borne pathogens without cultivation.