β-N-oxalyl-L-α,β-diaminopropionic acid (β-ODAP) is a neuro-excitatory amino acid found in Lathyrus sativus and also named as dencichine in Panaxnoto ginseng known for its hemorrhage stopping property. The biosynthesis of β-ODAP was proven to be related to several basic metabolisms especially sulfur and cysteine (Cys) metabolism. In this report, cysteine synthase (CS) genes were cloned from L. sativus based on the previous transcriptomics data. Firstly, multiple sequence alignment and phylogenetic tree analysis were conducted to detect the sequence characteristic and classification of LsCS genes. To confirm the property of LsCS gene expression product, functional complementation to Escherichia coli Cys-auxotrophic strain NK3, bacterial expression, recombinant protein purification and enzymatic activity assay were conducted. And then the key site for CS activity was detected via point mutants of LsCS. At last, gene expression level of LsCS in developing seeds was analyzed via qRT-PCR. The results suggested eight LsCS genes including LsCS1~LsCS7 and LsCAS separately in L. sativus. The LsCS genes were found homologous to those in soybean and other species and belong to β-substituted alanine synthase gene family. Expression of LsCS genes in E. coli Cys-auxotrophic strain NK3 resulted in bacterial growth and enzymatic activity assay suggesting a high cysteine synthase activity of LsCS, which were verified to be PLP-dependent. Among LsCS genes, LsCS1、LsCS2、LsCS4、LsCS6 and LsCS7 expressed with high level during seed development. These results will help to reveal gene functions of LsCS in β-ODAP biosynthesis of L. sativus.

Melanophilin (Mlph) gene plays fundamental roles in regulating the transportation of mature melanosomes in vertebrates, and its mutations can lead to hair or feather color fading in animals. However, the molecular function of Mlpha gene in aquatic animals need to be explored. In this study, spatial and temporal expression of Mlpha gene were investigated during different developmental stages in zebrafish (Danio rerio), and Mlpha gene was also knocked-out by using the CRISPR/Cas9 technology to exam body color changes in the F₂ homozygous mutants in zebrafish. The results of in situ hybridization and real-time quantitative PCR (qRT-PCR) expression indicated that Mlpha gene expressed at all the collected developmental stages, including 6, 12, 24, 30, 36, 48, 60 hpf (hours post fetilization) and 1, 2, 3, 4, 5,10, 15 dph (days post hatching) and the expression level increased with the increase of dendritic melanocytes in zebrafish. Meanwhile, the highest expression level was identified on the abdomen of zebrafish at 5 dph when obvious melanin band appeared. Similarly, the in situ hybridization results also showed the detectable signals and the strongest signal at 5 dph. After Mlpha gene was knocked-out by using CRISPR/Cas9 technology, the larval individuals of F₁ mosaic mutant zebrafish displayed slow expansion of melanin in juveniles and fragmented black stripe in adults. Under observation using the steroscope, the numbers of melanocytes and their coloration were not significantly reduced at the stripe fragments, but their morphology was reduced and dendritic structure was degenerated. Furthermore, the development speed of Mlpha knockout zebrafish was slower than that of wild-type zebrafish, and the diffusion speed of melanin was also significantly slower than that of wild-type zebrafish, indicated that melanosome dispersion was affected if Mlpha gene was knocked-out. In the F₂ homozygous mutant zebrafish, the Mlpha protein could not be completely translated owing to a terminator codon production in the second exon of this gene, and the adults appeared small melanocyte in skin. Moreover, in situ hybridization and qRT-PCR expression analysis revealed that the expression levels of Rab27a and myosin Va (MyoVa) which interacted with Mlpha were significantly down-regulated in the F₂ homozygous mutants (P<0.05). It is speculated that the mutation of Mlpha gene may lead to its inability to bind to MyoVa, resulting in the slow aggregation and transportation of melanosomes. It may also be that the mutant can't bind Rab27a, which makes melanosomes unable to transport along actin filament. In addition, there was no dimming of melanin pigments in adult zebrafish, indicating that the mutation of Mlpha gene did not affect the synthesis process of melanin, but may affect the transport process of melanin. Our study indicated that Mlpha gene played an essential role in regulating the transportation process of melanin, and its mutation could affect black stripe formation in zebrafish. The results of this study provide a reliable reference for studying the genetic basis and mechanism of melanoma pigmentation or formation in fish. However, the mechanism of Mlpha gene regulating body color in fish needs to be further studied.

The leaf rust (Puccinia triticina) in Triticum aestivum secretes various special effectors, which invade host cells and play a toxic role to destroy the plant defense response. It is of great significance to analyze the pathogenic mechanism of leaf rust for the development of disease control technologies and strategies. In previous study, leaf rust physiological race PHNT were mutagenized with ethyl methanesulfonate (EMS) and virulent mutants of leaf rust resistance gene Lr19 were isolated successfully. Then RNA sequencing was performed on samples collected from Lr19-virulent mutants and wild-type strains. In this study, Signal P, TMHMM, Big-PI Predictor, Target P and Effector P online prediction software was used to predict effector candidates. The results showed that a total of 301 effector candidates were screened out of 10 870 protein sequences of leaf rust, and the length of signal peptides ranged from 19 to 23 aa. The amino acids with the highest appearance frequency in signal peptides were leucine and alanine. The prediction results of subcellular localization showed that effector candidates were mainly located extracellular. Three new motifs were obtained by MEME prediction analysis. In order to determine the expression pattern of effector candidates in different time course of wheat infected by leaf rust, 7 candidate effector proteins rich in cysteine, glycine or serine were randomly selected from 301 candidate effector proteins, and their gene expression was analyzed by qRT-PCR. The results showed that 7 genes were up-regulated in the early stage of leaf rust infection, which might play a role in the process of leaf rust infection. In order to verify the toxic function of the effectors, 7 genes were constructed to pEarleyGate103 with GFP label by Gateway cloning. The toxicity function of effector candidates was verified by Agrobacterium tumefaciens-mediated transient transformation. The results showed that PTTG_27401 could inhibit BCL2-associated X (BAX)-induced cell necrosis which meant potential toxic function. The present study provides basic information for further identification of PHNT effectors and analysis of pathogenic mechanism.

Maize is the main grain-forage crop in China, and the kernel test weight (KTW) is an index to evaluate the quality grade of maize. With the breeding and popularization of high-density-tolerance maize varieties increasing, the effect of planting density on maize quality was began to study. In this study, 248 maize inbred lines with rich genetic diversity were used as association population, genome-wide association studies (GWAS) was used to explore KTW trait in multiple environments with different planting densities. Using 830 57 SNP markers distributed in the whole genome, the effect of planting density on KTW trait was explored and find out the related loci and candidate genes for controlling KTW trait. The results showed that planting density had a significant effect on KTW trait. The inbred lines 'XF134', '4676A', '811A', 'Ji 4112', etc. were less affected by the density and could maintain higher KTW, which was suitable for planting in high density environment. This study identified 14 significant SNPs, which were located on chromosomes 1, 3, 4, 5, 6, 7 and 10, respectively. And the phenotypic variation explained (PVE) by single locus ranged from 1.84% to 30.91%. There were 72 candidate genes found by searching the candidate genes in the range of 120 kb upstream and downstream of SNP. The results of enrichment analysis of candidate genes were mainly involved in 7 biological processes, 8 cellular components and 6 molecular functions. Seven candidate genes GRMZM2G079263, GRMZM2G079617, GRMZM2G403609, GRMZM2G180659, GRMZM2G160840, GRMZM2G104254 and GRMZM2G057281 may be related to KTW. These genes were involved in the regulation of endosperm development, photosynthesis and resistance to adversity. This study provides help for further explore the genetic principle and mechanism of KTW and molecular assisted selective breeding.

Cadmium is a heavy metal pollutant with high toxicity to many organisms, which enters into the environment along with industrial and agricultural activities. Vacuolar sequestration is an important mechanism to coping with cadmium stress in plants. Vacuolar H⁺-pyrophosphatase (V-H⁺-PPase), a proton pump in vacuolar membrane of plant cells, plays important roles in the transmembrane transport of inorganic ions. In this study, a V-H⁺-PPase gene, BnVP1 (GenBank No. MW029619) from ramie (Boehmeria nivea) was isolated by the rapid amplification of cDNA ends (RACE) method based on the previous transcriptome analysis of ramie under cadmium stress. The open reading frame sequence of BnVP1 was 2 292 bp and encoded 763 amino acids. BnVP1 was a transmembrane protein belonging to H_PPase superfamily, which contained 15 transmembrane domains. BnVP1 shared high amino acid sequence similarity (90.04%~92.27%) with typeⅠ V-H⁺-PPase proteins from Prunus persica (XP_007208066.1), Malus domestica (XP_008359855.1), Theobroma cacao (XP_017977384.1) and Gossypium raimondii (XP_012476518.1). The promoter sequence of BnVP1 contained several stress-related cis-acting elements. qRT-PCR showed that the expression of BnVP1 had no tissue specificity. The expression of BnVP1 in the root and leaves of ramie increased rapidly to maximum under cadmium stress, which indicated that the expression of BnVP1 was significantly induced by cadmium. The study enriches the research content of the molecular mechanism of cadmium tolerance in ramie, and provides a reference for the study of cadmium tolerance mechanism of Bnvp1 gene.

Jasmonates play a key role in plant response to biological stresses, and jasmonate ZIM-domain (JAZ) is a critical factor in jasmonic acid (JA) signal pathway. The present study screened out 24 members of JAZ family in pigeonpea (Cajanus cajan) genome, and explored phylogenetic analysis, gene chromosome mapping and protein conserved domain analysis. According to previous transcriptome data, the expression patterns of JAZs in roots and leaves were analyzed, and the expression of JAZs in leaves in response to pathogenic fungus Cc1-1 was detected by reverse transcription-PCR (RT-PCR). The results showed that there were 129~376 amino acid residues in JAZ family, and the isoelectric points were 5.94~9.37. JAZ family members could be divided into 5 subfamilies by MEGA software, and they were unevenly distributed on 6 chromosomes. The subcellular localization prediction showed that CcJAZ12 and CcJAZ18 were located in the chloroplast and CcJAZ19 in the cytoskeletal, and all the other 21 JAZs were located in the nucleus. The prediction results of cis-acting elements in the promoters showed that JAZ family genes had certain amount of JA responsive elements, and a large number of light responsive elements and a small number of abscisic acid, salicylic acid and other hormone regulatory elements. Transcriptome analysis showed that the expression of JAZs in leaves was higher than that in roots, and the expression of CcJAZ19 was the highest, which might play a more important role in the response of leaves to biological stresses. After infection of pigeonpea leaves by pathogenic fungus Cc1-1, the gene expression of JAZs in the leaves increased gradually with the infection time, and the expression of CcJAZ8, CcJAZ19, and CcJAZ23 was significantly up-regulated. The expression level of CcJAZ19 was up-regulated to 2.5 times (P<0.05) at 6 h after infection, indicating that it might participate in the defense response of JA signal to fungi. The present study provides basic data for the follow-up study on molecular regulation mechanism of JA signal.

Sweet osmanthus (Osmanthus fragrans) is traditional famous flowers in China, and its floral bud differentiation process is affected by ambient temperature. Plant specific transcription factors TCP (teosinte branched 1/cycloidea/proliferating cell) involved in regulation of plant cell, organ and tissue formation or other physiological processes, and plays a significant role during floral bud development in plants. In this study, 12 OfTCPs cDNA sequence were cloned based on the transcriptional data of O. fragrans 'Yanhonggui'. Among them, 7 OfTCPs had complete ORF lengths ranging from 807 to 1 365 bp. The length of the proteins encoded by TCP ranged from 268 to 454 aa with molecular weights from 28.24 to 49.49 kD and the isoelectric points were from 6.57 to 9.08. The protein domain analysis showed that OfTCP contained basic helix-loop-helix (bHLH) domain; The phylogenetic tree analysis showed that twelve OfTCPs could be divide into the ClassⅠand ClassⅡ, and the same subgroup proteins contained similar conservative element. The qRT-PCR results showed that three genes OfTCP5, OfTCP9 and OfTCP12 might be the key genes to promote flower bud differentiation in response to low temperature response after the different temperature treatment (19 and 25 ℃), however, the response of OfTCP9 to low temperature was relatively slow and play an important role in the late stage of flower bud differentiation. Meanwhile, OfTCP5 and OfTCP12 had no self-activating activity and toxicity. This study lay the foundation for further research on the molecular mechanism of the TCP transcription factor in response to environmental temperature changes in the regulation of flower bud differentiation of Osmanthus fragrans.

Previous study in this group discovered that the expression level of insulin-like growth factor Ⅰ (IGF1) receptor (IGF1R) gene in liver tissue of Chinese Holstein cows (Bos taurus) during the dry period was significantly higher than that in early and peak lactation periods through transcriptome sequencing. IGF1R is a receptor tyrosine kinase that plays a critical role in the signaling of the IGF family. With directly sequencing the encoding regions and 2 000 bp of upstream and downstream flanking regions of IGF1R gene using the pooled DNA from 40 sires to verify the IGF1R gene genetic effects on lactation traits in dairy cows. The identified SNPs were genotyped in 947 Chinese Holstein cows individually by target sequencing technology. Association analysis was conducted between the SNPs and 5 milk traits using the animal model. As a result, a total of 7 SNPs were detected including 1 SNP located in exon, 5 SNPs located in intron region and 1 SNP located in 3' flanking region. Of them, single-marker association analysis results showed that 6 SNPs were found associated with milk yield, fat yield, protein yield and protein percentage significantly (P=0.0146~<0.0001). The dominant effects or allelic substitution effects of 7 SNP sites reached significantly difference (P<0.05; P<0.01). Through linkage disequilibrium (LD) analysis, 7 SNPs were linked (r²=0.85~0.99) and formed 3 blocks. Haplotype-based association analysis showed that block 1 was not significantly associated with all the 5 milk traits (P>0.05). While block 2 and block 3 were associated with milk yield, fat yield and protein yield traits (P=0.0003~<0.0001), in which H1 and H2 were dominant haplotypes that increased milk yield, fat yield and protein yield, respectively. In conclusion, IGF1R gene had significant impacts on milk yield, fat yield and protein yield traits in dairy cow, implying it could be used as a molecular marker in genomic selection.

Breast carcinoma amplified sequence 2 (BCAS2) can participate in DNA damage responses, protein stability changes and pre-mRNA splicing processes in a variety of cells. There are many studies on the role of BCAS2 gene in germ cell formation, early embryonic development, and ovarian granulosa cells in Mus musculus, but whether this gene is involved in the regulation of granulosa cell proliferation or apoptosis in livestock follicles has not been reported. In this study, follicle stimulating hormone (FSH) was added to porcine granulosa cells cultured in vitro to observe whether it affected the expression of BCAS2; then BCAS2 was knocked down using RNAi technology to detect cell apoptosis and the expression levels of related genes. The results showed that both BCAS2 mRNA and protein expression were significantly increased after 24 h of treatment with 100 ng/mL FSH (P<0.05). After BCAS2 knockdown, the apoptosis rate of granulosa cells was significantly increased (P<0.05), the expression of B-cell lymphoma-2 gene (Bcl-2) was significantly decreased (P<0.05), the expression of proliferating cell nuclear antigen gene (PCNA) was extremely significantly decreased (P<0.01), and the expression level of BCL2-associated X protein gene (Bax) was significantly increased (P<0.05). In addition, the mRNA expression of tubulin β gene (TUBB), TUBA1A and estrogen receptor 2 gene (ESR2) were extremely significantly decreased (P<0.01), ESR1 expression was significantly decreased (P<0.05), and tumor protein p53 gene (p53) expression was significantly increased (P<0.05). It showed that BCAS2 gene knockdown can induce granulosa cells apoptosis in Sus scrofa, and the regulatory mechanism may be involved in the possible pre-mRNA splicing, estrogen receptor (ER) transcriptional regulation, or p53 pathways. This study provides a theoretical basis for further revealing the potential regulatory mechanism of follicular development in domestic animals.

Tryptophan is the second or third limiting amino acid for weaning pigs (Sus scrofa), which plays an important role in the regulation of feeding intake, growth, immunity and intestinal development. The 4 different ratio of tryptophan to lysine was conducted to investigate the effects of dietary tryptophan level on serum biochemical index, organ IDO and intestinal tight junction protein gene expression in weaning pigs. A total of 240 weaning pigs (Large White×Landrace×Duroc) with an initial body weight of (6.5±0.2) kg were randomly divided into 4 treatments with 6 repetitions and 10 pigs each repetition and the experiment lasted for 28 d. The dietary ratios of tryptophan to lysine were 0.15, 0.18, 0.21, and 0.24. The result showed that compared with the group with tryptophan to lysine ratio of 0.15, when the ratio was increased to 0.18, 0.21 or 0.24, the expression of IDO gene in the spleen, kidney, stomach and ileum of weaning pigs was significantly reduced (P<0.05), and the expression of jejunum IDO mRNA in the 0.24 group was significantly increased (P<0.05). When the dietary ratios of tryptophan to lysine increased from 0.15 to 0.24, the expression of occludin and ZO-1 in the duodenum and ileum were significantly reduced (P<0.05), the jejunum expression levels of ZO-1 mRNA in the 0.18 and 0.24 groups were significantly decreased (P<0.05), while the jejunum occludin mRNA expression levels in the 0.21 group were significantly increased (P<0.05). When the ratio of dietary tryptophan to lysine increased from 0.15 to 0.24, the level of serum urea nitrogen was influenced in weaning pigs (P<0.05). It was found that when the ratio of tryptophan to lysine was adjusted from 0.15 to 0.24, the gene expression of IDO and small intestinal tight junction protein in the organism could be significantly regulated, and nitrogen metabolism of the organism could be also affected in this study. In conclusion, the optimal ratio of tryptophan to lysine for weaning pigs was 0.18. The results provides a theoretical basis for the application of tryptophan in the diet of weaning pigs.

Quaking (QKI) is a member of RNA-binding protein family, and involved in the regulation of cell proliferation, differentiation and apoptosis. In order to explore the biological functions of QKI family genes in goat (Capra hircus) muscle development, Jianzhou big-eared goat was used as the research object in present study. The coding sequences of goat QKI family genes were obtained by TA cloning and analyzed by bioinformatics. qRT-PCR and Western blot were used to detect the expression of QKI family genes in different tissues (brain, heart, liver, spleen, lung, kidney, longissimus dorsi, semimembranosus, semitendinosus, gastrocnemius, psoas major, and adductor) of goats (3 d after birth) and developmental stages (embryonic 90 d, 105 d, 135 d; and 3 d, 150 d after birth) of the semimembranosus muscle, and their expression during the proliferation and differentiation of goat skeletal muscle satellite cells (SMSCs). The results showed that the complete coding sequences of goat QKI-5 (GenBank No. MW651981), QKI-6 (GenBank No. MW651982), and QKI-7 (GenBank No. MW651983) genes were obtained by amplification, with the length of 1 026, 978, and 960 bp, which encoding 341, 325, and 319 amino acids, respectively. Nucleotide sequence alignment revealed that QKI-5, QKI-6, and QKI-7 were highly conserved among species, and had the closest phylogenetic relationship with sheep (Ovis aries) and cattle (Bos taurus). The secondary and tertiary structures of goat QKI-5, QKI-6, and QKI-7 protein were mainly composed of α-helix and random coil, and there was a common conserved functional domain, KH (K homology) domain. The results of qRT-PCR showed that the expression of QKI-5, QKI-6, and QKI-7 genes in goat skeletal muscle tissues was significantly higher than that in other tissues (P<0.05); In the semimembranosus muscle of different developmental stages, QKI-5, QKI-6, and QKI-7 showed a trend of increasing first and then decreasing, with the highest expression at 3 d after birth (P<0.05). Western blot results showed that the expression of QKI-5 protein was only detected in goat skeletal muscle tissue, and mainly expressed after birth. Furthermore, QKI-5, QKI-6, and QKI-7 genes showed increasing expression tendency during the proliferation and differentiation of goat SMSCs, and reached the peak at 7 d after differentiation (P<0.05). The present study cloned and obtained the complete coding sequences of goat QKI-5, QKI-6, and QKI-7 genes, and revealed their high expression in goat skeletal muscle tissues and increasing expression patterns during proliferation and differentiation of goat SMSCs, indicating that QKI family genes might play an important role in goat skeletal muscle development. The present study provides basic data for further investigation of molecular mechanism on goat muscle development regulated by QKI.

Cryopreservation of poultry sperm has great potential in artificial insemination, breeding selection and disease prevention. Although cryopreservation measures have been continuously optimized, the fertilization ability of spermatozoa after cryopreservation is still lower than that of fresh sperm. The purpose of this study was to investigate the difference of gene expression between fresh and thawed Gallus sperm. The gene expression levels in fresh and frozen sperm were detected by Affymetrix chicken genome-wide expression profile chip. The signal pathways and candidate genes related to freezing injury of chicken sperm were screened, and some differentially expressed genes (DEGs) were verified by qRT-PCR. The results showed that 687 DEGs were between the 2 groups (|Log₂(FC)|>1; q-value<0.05). Among these genes, 615 were significantly upregulated and 72 were significantly downregulated in the fresh sperm group. Gene Ontology (GO) analysis showed that 196 DEGs of all significantly regulated genes were involved in mitogen-activated protein kinase (MAPK) signal pathway, ATP activation, cell cycle and other biological processes. KEGG analysis showed that DEGs were significantly (P<0.01) enriched on carbohydrate metabolism, signal transduction, cell growth and death. Four DEGs (cold-inducible RNA-binding protein (CIRBP), 90 kD heat shock protein (HSP90), HSP70 and Ras homolog family member A (RHOA)) were selected and verified by qRT-PCR,and the results were in accordance with the above expression profile. Therefore, MAPK pathway, cell cycle regulation pathway and CIRBP, HSP90, HSP70 and RHOA genes may be important regulatory pathways and candidate genes leading to significant differences in cryopreservation. This study would provide a theoretical reference for understanding the potential molecular mechanism of frozen-thawed chicken sperm damage and how to improve the fertility of frozen-thawed chicken sperm.

Egg production trait is one of the economically important traits that have long attracted attentions from duck (Anas platyrhynchos) breeders, and screening genetic markers associated with egg trait is of great significance for molecular marker-assisted breeding in duck. To explore the association of retinol-binding protein 4 (RBP4) gene polymorphisms with duck egg trait and identify the potential molecular markers, in this study, Nonghua White ducks were used as the experimental animals to detect single nucleotide polymorphisms (SNPs) in exons 1~6 and introns 1~3 of the RBP4 gene using PCR amplification combined with direct sequencing technologies, and their correlations with laying performance and egg quality traits were also analyzed. The results showed that one SNP namely g.266T>G was identified at the 266^th nucleotide within intron 1 of the RBP4 gene in Nonghua White duck, and the resulting 3 genotypes (TT, TG and GG) were present in this population, with 'T' being the dominant allele. Population genetics analysis showed that the heterozygosity (He), homozygosity (Ho), effective number of alleles (Ne) and polymorphic information content (PIC) at this mutation site was 0.408 7, 0.407 9, 1.697 7 and 0.277, respectively, being moderately polymorphic (0.25<P<0.50). Results from χ² test showed that this SNP site was not in Hardy-Weinberg equilibrium (P<0.05). Correlation analysis showed that genotypes at g.266T>G site were extremely significantly correlated with egg-white height (P<0.01), as manifested by higher egg-white height observed in individuals with TT genotype compared to those with TG and GG genotypes (P<0.01). Besides, this SNP could have the potential to influence egg production number, eggshell strength and egg weight, because individuals with GG genotype had higher egg production number and eggshell strength but lower egg weight compared to those with TG and TT genotypes. In summary, the g.266T>G SNP in intron 1 of the RBP4 gene has a significant impact on egg quality traits in Nonghua White duck and can be exploited as a candidate molecular marker for selection of duck egg production trait. This study provides a theoretical basis for future molecular marker-assisted breeding in Nonghua White duck.

Synaptonemal complex 3 (SCP3) and DNA meiotic recombinase 1 (DMC1) are key factors in meiosis, which have not been reported in Silurus meridionalis. In present study, PCR, fluorescence immunohistochemistry and Western blot were used to clone and analyze the sub-cellular distribution and tissue expression of scp3 and dmc1; GST pull-down was used to verify in vitro protein-protein interaction. The results showed that the CDS of scp3 (GenBank No. MW075231) and dmc1 (GenBank No. MF770581) were 723 and 1 029 bp, encoding 240 and 342 amino acids, respectively. Homology analysis showed that the homology of S. meridionalis SCP3 and DMC1 to Tachysurus fulvidraco were as high as 89.17% and 97%, respectively. In phylogenetic analysis, S. meridionalis SCP3 and DMC1 were clustered with fish, and then clustered with tetrapod. Reverse transcription-PCR (RT-PCR) results showed that scp3 and dmc1 were only specifically expressed in ovary. The results of qRT-PCR showed that relative expression of scp3 and dmc1 had the highest level in 55 dah (days after hatching), which were significantly higher than that of 30 and 90 dah. The results of fluorescence immunohistochemistry showed strong positive signals of SCP3 in nucleus and cytoplasm of oocytes, and strong positive signal of DMC1 in the nucleus of oocytes. GST pull-down assay showed that SCP3 interacted with DMC1 in vitro. This study could provide basic data for further study on the function of SCP3 and DMC1 during meiosis, and reference for artificial breeding of S. meridionalis to obtain high-quality eggs.

Heat shock proteins (HSPs) are highly conserved among different species, which are involved in protein folding, stabilization of biological matrix, assembly of macromolecules, degradation of polypeptides and transcriptional regulation. To study the expression profiles of HSP70-4 in different tissues or under the stress of fasting and bacteria invasion, The sequence of HSP70-4 gene were identified based on full-length transcriptome sequencing of Acipenser dabryanus. The full-length cDNA sequence of HSP70-4 was 4 029 bp, which included 234 bp of the 5'-UTR, 1 245 bp of the 3'-UTR and 2 550 bp of the ORF (GenBank No. MZ285757) that encoded 849 amino acids. Phylogenetic analysis showed that the A. dabryanus HSP70-4 gene clustered mostly close to the A. ruthenus HSP70-4, supported with a significant bootstrap value (100). HSP70-4 was universally expressed in all 11 of the tested tissues including liver, gonad, muscle, brain, eye, skin, head kidney, heart, intestine, spleen, and gill. The expression of HSP70-4 was the highest in the eye and was also detected at relatively high levels in the brain and muscle, and relatively weakly expressed in the liver, head kidney, skin, and intestine. Under starvation stress, the expression of HSP70-4 was upregulated, and significantly upregulated at 14 d post-fasting in intestine and muscle. HSP70-4 in the skin, spleen and head kidney was significantly upregulated (P<0.05) at 12 h post-challenge towards Edwardsiella tarda (1×10⁷ CFU/mL) infection and the upregulation of HSP70-4 in the gill started at 24 h post-challenge. The results showed that short-term bacteria infection resulted in significantly upregulated expression of HSP70-4 gene in the gill, skin, spleen and kidney of A. dabryanus. This study demonstrated that HSP70-4 was involved in the biological process of fighting hunger and infection by pathogenic bacteria in A. dabryanus. This study demonstrated that HSP70-4 was involved in the biological process of fighting hunger and infection by pathogenic bacteria and provided a theoretical basis for the further study on the mechanism of induced differential expression of HSP70-4 gene and the stress resistance mechanism of A. dabryanus.

Glutamate dehydrogenase (GDH) is an enzyme widely found in organisms related to nitrogen metabolism, and GDH genes of lower organisms are often used for the improvement of nitrogen assimilation efficiency in crops. In order to explore the high quality GDH genes for genetic improvement of crops, FgGDH gene was cloned from Fusarium graminearum, the protein FgGDH was prokaryotic expressed and purified, and its enzymatic kinetics characteristics were determined in vitro in this study. The results showed that the aminating activity of FgGDH was significantly higher than that of deaminating activity, indicating that FgGDH is more inclined to catalyze NH₄⁺ and α-ketoglutaric acid (2-OG) to generate glutamic acid and promotes the assimilation of NH₄⁺. In addition, the Km values of FgGDH for NH₄⁺ and 2-OG were (3.71±0.21) mmol/L and (6.44±0.32) mmol/L, respectively, which were significantly lower than those of OsGDH4 in rice. These results indicate that the affinity of FgGDH for NH₄⁺ and 2-OG is marked higher than those of endogenous OsGDH4 in rice, and FgGDH has a higher ammonia assimilation efficiency. This study provides a theoretical basis for the genetic improvement of nitrogen assimilation efficiency in rice and other crops by using FgGDH gene.

The general secretion pathway (Sec) is an essential pathway to regulate protein transport and widely exists in various types of cells. The ATPase can stimulate the Sec pathway, which promotes the transmembrane transport of enzyme proteins, and increases the secretion of trypsin in ruminant's pancreas. Based on the signal network, this review describes the regulation factors of functional amino acid and the mechanism of protein transport through the Sec pathway, which provides a theoretical basis for the regulation principle of secretory cells and the improvement of animal digestion by nutritional means.

Mutton has low cholesterol content, high nutritional value, delicious and juicy taste and other excellent characteristics, while its taint smell reduces the acceptance of some consumers. The flavor of mutton is produced by the Maillard reaction, lipid reaction, thiamine thermal degradation of the flavor precursors during cooking and heating. The article summarized the flavor precursors that form the flavor of mutton, the production pathways of flavor substances, the volatile flavor compounds, the research techniques of flavor precursors and the influence of nutritional factors and genetic factors on the flavor of mutton, which provides a certain theoretical basis for the study of mutton flavor.

Listeria monocytogenes is a Gram-positive facultative intracellular bacterial pathogen that can infect humans and animals, causing listeriosis in neonates, pregnant women, elderly persons, and individuals whose immune system is compromised. Live L. monocytogenes is adept at generating strong antigen-specific T-cell responses due to its unique features during host infection, making it an attractive vector for cancer immunotherapy. Increasing studies have shown that the attenuated Listeria is becoming an efficient tool for cancer immunotherapy. Several Listeria-based tumor vaccines undergo being developed, such as the cervical cancer and canine osteosarcoma. Clinical trials have exhibited impressive therapeutic efficacy in these cancer models. This review will summarize the recent studies and progress about Listeria-based cancer immunotherapy, which will help readers understand the current technology and development of tumor immunotherapy vaccines. This review could provide a reference for developing new strategies and technologies for tumor prevention and treatment.

Starch is one of the important quality characters of potato (Solanum tuberosum), and the development of linkage molecular markers could promote the breeding of high starch content varieties. In this study, 106 F₁ individuals and 240 F₂ individuals and their parents tetraploid potato wild species (S. demissum) 'YSP-4' (female parent) and low starch line 'MIN-021' (male parent) were used as materials, a candidate interval associated with starch content was excavated based on the previous results of specific-locus amplified fragment sequencing (SLAF-seq) combined bulked segregant analysis (BSA) strategy, two SSR markers chr2-SSR14 and chr2-SSR22 linked to starch content were developed in this interval, respectively, and verified by F₂ segregation population. The results showed that the correlation between the detection results of chr2-SSR14 and chr2-SSR22 markers and phenotypic identification in F₂ segregation population was 89.3% and 92.9%, respectively. After correlation analysis by SPSS software, the correlation coefficients (r) of the 2 markers were 0.68 and 0.769, respectively, which were significantly correlated at the level of 0.01 (P<0.01). The chr2-SSR14 and chr2-SSR22 markers developed in this study could be used for marker-assisted selection of high starch content trait in potato, which is of great significance for the breeding of new varieties of tetraploid potato with high starch content.

Herbicide-tolerant transgenic soybean (Glycine max) ZH10-6 is a new herbicide-tolerant genetically modified soybean line with glyphosate degradation gene (glyphosate acetyltransferase, GAT) and glyphosate resistance gene (pseudomonas fluorescens G2 strain 5-enolpyruvyl shikimate-3-phosphate synthase, G2-EPSPS) studied by the Institute of Crop Science, Chinese Academy of Agricultural Sciences. In order to establish anevent-specific detection method with it, multiplex PCR (MPCR) reaction with 4 sets of PCR primers covering different regions of the insertion and border sequence and 1 primer set for the soybean internal standard gene Lectin were evaluated.. Through primer-specific testing, optimization of PCR reaction system and procedure, assessment of reaction sensitivity and stability for the different generations of the transgenic soybean, a multiplex PCR detection method for the transgenic soybean ZH10-6 was established. The results showed that the method had great specificity. The optimal primer concentration (μmol/L) ratio of the 5 targets in the MPCR detection system (border A, border B, border C, border D, endogenous gene Lectin): 0.8∶0.3∶0.1∶0.2∶0.05. The detection sensitivity of each target could reach 1%, which could be stably inherited among 3 generations, and was suitable for rapid and efficient detection of ZH10-6 insertion sites. The detection method established in this study improved the detection efficiency and reduced the detection cost. It provides a new detection technique for identification of the new variety of transgenic herbicide-tolerant soybean ZH10-6 transformant and its derivatives,which is technical support for its biosafety assessment, administrative supervision and intellectual property protection.