Leaf shape is one of the important agronomic traits of maize. Moderate leaf shape is of great significance to increase maize planting density and yield. Leaf shape is regulated by multiple genes and genetic loci. However, the molecular mechanism of leaf morphological development in maize is still unclear. In this study, wrinkled leaf 1 (wl1) which was a leaf shrinkage mutant, was obtained by treating pollen of maize inbred line Mo17 with ethyl methanesulfonate. Genetic analysis showed wl1 was a recessive mutant controlled by a single gene. wl1 showed that small ridges and trichomes on the leaf surface, and the number of stomata decreased compared with wildtype. Paraffin section showed that the epidermal cell layers on the wl1 leaf was doubled. Besides, maize plant height, length and width of ear leaf, the first and second leaf above ear leaf were decreased compared with wildtype (P<0.01). Transcriptome sequencing analysis indicated the expression of several genes involved in photosynthesis relative biological processes and cellular component were decreased in wl1. Meanwhile, photosynthetic measurement showed the net assimilation rate, stomatal conductance and transpiration rate of wl1 were significantly lower than wild type (P<0.01). wl1 was located on chromosome 1 within the interval of physical distance of 1.7 Mb by using map-based cloning. According to analyzing the expression level of candidate genes in interval, genome resequencing and sequence alignment with seven different maize inbred lines, the candidate gene of wl1 was preliminarily identified as Zm00001d027266, which encoded a DeSI (desumoylating isopeptidase)-like protein. This study lays the foundation for the cloning of wl1 gene and the molecular mechanism analysis of maize leaf shape development.

Cluster of differentiation 302 (CD302), also known as DEC-205-associated C-type lectin-1 (DCL-1), is a type I transmembrane C-type lectin receptor. It participates in the processes of cell adhesion, migration, endocytosis and phagocytosis, and plays an important role in resisting pathogenic infection. In this study, the LjCD302 cDNA sequence (GenBank No. MT468568) was obtained from Japanese sea bass (Lateolabrax Japonicus) tissue transcriptome. It spanned 2 280 nucleotides (nts) and comprised an open reading frame of 729 nts, which encoded a polypeptide of 242 amino acids (aa) with a molecular weight of 26.9 kD and an isoelectric point of 4.97. LjCD302 composed of a signal peptide sequence (aa 1-27), a C-type lectin-like domain (CTLD) (aa 30-165), a transmembrane domain (aa 179-201) and a cytoplasmic tail (aa 202-242). The aa sequence homology analysis showed that LjCD302 shared the highest homology with that of the barramundi perch (Lates calcarifer) (82.0%). Phylogenetic tree analysis showed that CD302 from fish, amphibians, reptiles and mammals grouped into different clusters, and LjCD302 belonged to the fish cluster and had the closest evolutionary relationship with the barramundi perch CD302. Real-time quantitative PCR analysis showed that LjCD302 mRNA in healthy Japanese sea bass was mainly expressed in the liver, followed by the head kidney, and its expression was significantly up-regulated in head kidney, gill, intestine, spleen, and liver of Japanese sea bass after Vibrio harveyi infection (P<0.05). The CTLD domain of LjCD302 was prokaryotically expressed, and its antiserum was prepared. Western blot analysis showed that LjCD302 expression was significantly up-regulated in the head kidney of V. harveyi-infected Japanese sea bass (P<0.05). In summary, the expression of LjCD302 mRNA and protein was positively correlated with V. harveyi infection, suggesting that it may play a crucial role in the antibacterial immune response in fish. The results provide a basis for further research on the function of fish CLR and its mechanism of action in antibacterial immunity.

As an important factor that affects the spike number per unit area of wheat (Triticum aestivum), spike number per plant (SNPP) is sensitive to nitrogen level. In order to detect loci of SNPP response to nitrogen stress and to assist selecting and improving the nitrogen use efficiency or nitrogen stress sustain ability of wheat varieties, genome wide association study (GWAS) was conducted. The association panel consisted of 132 wheat varieties and their advanced lines derived from the north Yellow and Huai Valley Wheat Region of China. SNPP in 8 environments under 2 different nitrogen levels and tolerant index (TI) of SNPP to low nitrogen stress were investigated. Affymetrix Wheat 55K SNP was used to achieved the genetic data. Finally, 4 loci were identified to associated with SNPP in two or more environments accounting for 10.20%~17.5% of the phenotypic variation. And 3 loci associated with TI of SNPP were detected in 2 environments with the phenotypic variation ranging from 11.4% to 16.3%. Evaluation of the genetic effects and frequency of the alleles in the significantly associated loci suggested that it was important to strengthen the selection of the AX-108733649-2B, AX-108889870-4A, AX-111667100-5B and AX-111039914-7D loci for genetic improvement of wheat varieties with high nitrogen use efficiency.

Thioredoxin (TRX) contains highly conserved typical domains with oxidative active site WCG/PPC, and is a class of small molecular multifunctional proteins widely existing in organisms. Nucleoredoxin (NRX), a member of the TRX superfamily, was contained multiple domains typical and atypical of TRX, and has the potential of redox activity. According to bioinformatics analysis, the Triticum aestivum TaNRX1-D contains 3 TRX domains: domain 1 contained WCPPC, domain 2 contained GYPPV and domain3 contained WCGPC, while the domain 2 did not harbour such an activity site. In order to further explore the different structural domain influence on TaNRX1-D protein activity, this experiment truncated the protein sequence at different lengths according to the predicted physicochemical properties of the TaNRX1-D domain protein. The corresponding protein products were obtained by constructing prokaryotic expression vectors and purified, respectively. TaNRX1-D possesses TRX activity in vitro. Compared with 1.0 µmol/L or 2.5 µmol/L, 5.0 µmol/L TaNRX1-D displayed high insulin reduction activity in a dose-dependent manner. Among the 5 domains of TaNRX1-D, the insulin reduction activity of the 5 domains as follows: NRX-D3 (NRX-domain3)>NRX-D1 (NRX-domain1)>NRX-D2 (NRX-domain2), and each of the 5 domains alone was less active than the NRX-DQ protein. The analysis of three structural domains inside the protein on the reducing activity of TaNRX1-D protein will provide a new reference for further understanding of the function of TaNRX1-D protein.

Stress associated proteins (SAPs) are the A20/AN1 zinc-finger proteins which widely involved in plant response to abiotic stress. The present study investigated the correlation between TaSAP2-6A gene marker and agronomic traits in a variety of environments to provide a basis for the exploration and utilization of excellent allelic variation in wheat (Triticum aestivum). In this study, TaSAP2-6A gene (GenBank No. JQ768347.1) was isolated from wheat, with the full-length genome sequence of 3 409 bp including the upstream sequence (2 714 bp), 3' untranslated region (167 bp) and coding region (528 bp). The coding region encoded 175 amino acids consisting of an A20 domain and an AN1 domain. Using a set of 'Chinese Spring' nullisomic-tetrasomic lines, TaSAP2-6A gene was located on chromosome 6A. It was consistent with the predictions made from URGI (Unité de Recherche Génomique Info) website. The polymorphism of TaSAP2-6A gene sequence was detected by 32 wheat accessions with high diversity, and no nucleotide variation was detected in the coding region, but one InDel (insert-deletion, T/-) site was identified at 1 527 bp site of the upstream sequence. A dCAPS (derived cleaved amplified polymorphic sequence) marker was developed based on the InDel, named as InDel-1527. The genotypes of a natural population consisted of 323 accessions were detected by the marker InDel-1527 of TaSAP2-6A. Association analysis revealed that this marker was significantly associated with plant height, grain number per spike and grain yield per plant in ten environments (year×site×treatment) with well-watered, drought-stressed, heat-stressed, and drought-heat-stressed treatments. The InDel (T) was considered as the superior allelic variation to decrease plant height, and increase grain number per spike and grain yield per plant. The results provide a gene resource and its functional marker for marker-assisted selection in wheat.

Ginger (Zingiber officinale) has high economic value as medicinal and food resources. 6-gingerol is high important bioactive component in the pharmacological action of ginger. In the present study, a local ginger cultivar of Chongqing was used as the material. Liquid chromatograph-mass spectrometer (LC-MS) technology was used to detect the main components of pungent principle. Four cDNA sequences of 6-gingerol biosynthetic relative enzyme genes including phenylalanine ammonia lyase (PAL), cinnamate 4-hydroxylase (C4H), 4-coumarate:CoA ligase (4CL) and caffeoyl-CoA-o-methyltransferase (CCoAOMT/CCOMT) (GenBank No. MN887496, MN887498, MN887497 and MN887499) were cloned and qRT-PCR was used to detect the expression of the genes in ginger rhizome at different developmental stages. Correlation analysis was performed between the expression of each gene and the 6-gingerol increase.The results showed that 6-gingerol accumulated very quickly in the 1-month ginger material after sowing, and increased slightly in the subsequent stages. PAL, C4H, 4CL and CCoAOMT genes were all highly expressed in 1~2 months ginger after sowing, suggesting a correlation between each gene and 6-gingerol synthesis. Correlation analysis showed that the expression of CCoAOMT rather than the others was significantly positively correlated with 6-gingerol synthesis (P<0.05). These results indicated that the biosynthesis of 6-gingerol existed stage specificity, and there was a positive regulatory relationship between CCoAOMT gene and 6-gingerol biosynthesis. The results of this study will lay the foundation for the further study of the function of 6-gingerol synthesis-related enzyme genes in Z. officinale.

Calcineurin B-Like proteins (CBLs) are a kind of important protein which can involve in decoding calcium signals in plants. They play important role in plant response to various stresses. The cDNA of SsCBL3 (GenBank. MN850491) cloned from sugarcane (Saccharum officinarum) cultivar 'ROC22' under low potassium stress by reverse transcription-PCR (RT-PCR). SsCBL3 contained a complete open reading frame of 678 bp encoding a protein with 225 amino acids. The sequence features of SsCBL3 analyzed by bioinformatics. The molecular weight of SsCBL3 was 25.767 47 KD and the isoelectric point was 4.82, which was an acidic protein. The structure of SsCBL3 protein was analyzed showed that it contained 3 EF-hand functional domains and 1 FPSF motif which interacted with CIPK kinases. Evolutionary analysis further revealed that SsCBL3 has high homology to the CBL3 from gramineous crop, and had the closest relationship with Zea may. The result of qRT-PCR showed that the expression of SsCBL3 was changed under low nitrogen, low phosphorus, low potassium, drought, salt and ABA stresses. Moreover, the different stress times also change expression of SsCBL3. The expression of SsCBL3 strongly induced by drought or salt stresses. The results of qRT-PCR indicated that SsCBL3 gene may play an important regulatory role in sugarcane under stress. The results of research provides a theoretical basis for cultivating sugarcane varieties with stress-resistant.

TCP (teosinte branched 1, cycloidea, proliferating cell factors) transcription factors are widely involved in plants, which play an important role in regulating plant growth and development, morphogenesis, abiotic stress and other physiological processes as a kind of plant-specific transcription factor. They can regulate the expression of downstream target genes to transform the cell division process, and play a role in a variety of hormone regulatory pathways. Lagerstroemia indica, as an excellent kind of summer flowering woody plant, is commonly used in gardens. Because of its rich germplasm resources and diverse plant architecture, it is a good material for investigating woody plant architecture. In order to clarify the role of the TCP gene family in Lagerstroemia, a total of 44 TCP genes were identified in Lagerstroemia based on transcriptome data. The bioinformatics and gene expression analyses were used to analyze the characters and expression patterns of LfiTCPs. The results showed that the proteins encoded by the LfiTCPs were uniform and stable in physical and chemical properties, and were mostly localized in the nucleus. Phylogenetic analysis showed that the LfiTCPs were divided into 2 branches, and the number and proportion of which were similar to other species. LfiTCPs had a relatively conservative structure, which could maintain the TCP domain intact, and those LfiTCPs in 2 branches had obvious structural differences. Most of the LfiTCPs were similarly expressed in standard and dwarf crapemyrtles,14 of them were initially screened out based on the differential expression results. Among which, LfiTCP12;2 and LfiTCP15;2 showed significant differences between non-dwarf and dwarf progenies. The result of qRT-PCR confirmed that they were highly expressed and lowly expressed in dwarf progenies, respectively. This study provides the information for further research on the plant architecture of Lagerstroemia indica, potentially promoting the molecular breeding process of woody plant architecture.

A high frequency induced transformation system of hairy roots by Agrobacterium rhizogenes, that is an effective method to verify the gene function. Here, a genetic transformation system of Betula luminifera by A. rhizogenes was established, and had primarily optimized the culture condition of genetic transformation hairy root. First, 7 A. rhizogenes were used to infect B. luminifera leaves, and ArQual was obtained with the highest hairy root induction rate (69%). The optimal system for hairy root induction of B. luminifera was as follows: Leaves were pre-cultured in 1/2 MS medium for 2 days, infected for 20 minutes, and co-cultivation with acetosyringone (the concentration of 400 μmol/L) and cefotaxime (400 mg/L). In addition, the effect of different foreign gene vectors on hairy root induction was not significant. Based on the above system, pCAMBIA13011 vector containing β-glucosidase gene (GUS) gene and pGWB5 vector containing green fluorescent protein (GFP) gene were electroporated into ArQual, and transgenic hairy roots of B. luminifera were successfully induced with a conversion rate of 36.4%. In order to expand the application range of above genetic transformation system, the hypocotyls of B. luminifera seedlings (short cycle culture) were transformed by soaking and the stem segment of B. luminifera (long period culture) were punctured. After 30 days, the hairy roots were detected by GUS staining, fluorescence microscopy and PCR, and the results showed that GUS and GFP genes were successfully transformed and expressed, illustrated the different expression vector could express in new hairy roots of B. luminifera. In general, a high-frequency induction system for hairy roots of B. luminifera was established, which could be used to quickly verify gene function and expression in woody plants.

Moso bamboo (Phyllostachys edulis) is an important economical bamboo species in China. Shoot bud germination and development is one of the main factors limiting its productivity. The phosphatidyl ethanolamine-binding protein (PEBP) family contains FLOWERING LOCUS T (FT) subfamily, TERMINAL FLOWER 1 (TFL1) subfamily and MOTHER OF FT AND TFL1 (MFT) subfamily. The FT subfamily plays an important role in the growth and development of plants. In order to explore the function of FT gene in the process of shoot bud development, 2 FT homologous genes, PheFT6 and PheFT17, were cloned in Ph. edulis, and their expression patterns were analyzed by qRT-PCR, and their interaction proteins were explored by yeast two-hybrid. The results showed that the lengths of the coding regions of PheFT6 (GenBank No. MT976159) and PheFT17 (GenBank No. MT976160) were 525 and 528 bp, encoding 174 and 175 amino acids, respectively, which contained the PEBP domain. The PheFT6 gene typically contains 4 exons and 3 introns, while the PheFT17 gene has additional 2 exons inserted, resulting in alternative splicing that produce at least 4 transcripts. Phylogenetic tree analysis showed that PheFT6 and PheFT17 were far from rice Heading date 3a (Hd3a) and Arabidopsis thaliana FT. qRT-PCR analysis showed that the expression of PheFT6 and PheFT17 genes showed obvious circadian rhythm, with low expression level in the daytime and high expression level at night. The expression of PheFT6 gene was induced by low temperature (12 ℃), while the expression of PheFT17 gene was induced by warm temperature (28 ℃). Compared with the seedlings cultured at 12 ℃, the seedlings treated at 28 ℃ had higher tillering ability. The expression of PheFT6 and PheFT17 genes was strongly inhibited by drought, and the inhibition increased with the increasing of drought degree. The yeast two-hybrid results showed that both PheFT6 and PheFT17 did not interact with PheGF14b and PheGF14c in 14-3-3 (G-box factor 14-3-3 protein), but interacted with PheTB1L-1 and PheTB1L-2, homologues of Hub protein BRANCHED1(BRC1)/TEOSINTE BRANCHED1 (TB1), during branching, which suggested that PheFT6 and PheFT17 might be involved in branching and tillering in Ph. edulis. This study provides a reference for further analysis of the biological functions of PheFT6 and PheFT17, and could provide basic data for the study on molecular mechanism of shoot development.

Neonatal Fc receptor (FcRn) can specifically recognize and transport IgG across the mucosal barrier. In order to determine the interaction between porcine (Sus scrofa) FcRn and IgG Fc segment CH2 domain, the porcine IgG CH2 gene was cloned by the reverse transcription (RT-PCR) and further subcloned into prokaryotic expression vector PEGX-4T-1. The fusion protein was induced by isopropyl β-D-thiogalactoside (IPTG) and purified by glutathione S-transferase (GST) Affinity Column. In addition, IgG CH2 gene was subcloned to eukaryotic expression vector pEGFP-C1 and Co-transfection was carried out to determine whether the IgG CH2 and FcRn co-located in African green monkey (Chlorocebus sabaeus) kidney cells (COS-7). The interaction of IgG CH2 and FcRn was verified by coimmunoprecipitation. Furthermore, the binding ability of IgG CH2 and FcRn under different pH condition was detected by ELISA. The results showed the amplified porcine IgG CH2 gene was 330 bp, encoding 110 amino acids. The relative molecular weight of fusion protein GST-CH2 was 38.4 kD. The GST-CH2 in the induced bacteria existed in the form of soluble protein and inclusion body. The soluble GST-CH2 protein will possess the better ability of combining the FcRn. Laser confocal microscope observation showed that the IgG CH2 and FcRn were located in the cytoplasm and had an aggregation phenomenon. The coimmunoprecipitation showed that the corresponding target bands could be detected in all the coprecipitation strips through labeled protein GFP and Flag, which indicated IgG CH2 could bind FcRn. By ELISA, it was found that the IgG CH2 and FcRn had the binding ability when pH value was 6.0, but they were not combined at pH 7.4, suggesting that the combination of IgG CH2 and FcRn was dependent on pH. In conclusion, the results showed that FcRn and CH2 domain of IgG Fc segment had a co-localization and interaction, their combination was pH-dependent, which provides materials for the construction of Fc-based small-size antibody fusion protein.

The growth and development of the gastrointestinal tract of suckling piglets directly affect the survival rate of piglets, and the survival rate of piglets is the main factor that determines the production efficiency of the pig industry. To explore the development of intestinal microbial community in Bamei piglets (Sus scrofa) at different ages, fecal samples of Bamei suckling piglets were collected in 1~3 d (colostrum, COL) and 8~10 d (ordinary milk, ORD), and the piglets grew up and were healthy under the same conditions. Microbial data were analyzed using the 16S rRNA V3+V4 hypervariable region second generation Illumina HiSeq2500 sequencing method. Relative abundance of Lactobacillus and Fusobacterium in 1~3 d were significantly higher than those in 8~10 d (P<0.05). The milk intake in 8~10 d was significantly higher than that in 1~3 d (P<0.05), and the milk intake was strongly negatively correlated with Lactobacillus and Fusobacterium (r<-0.6, P<0.01). KEGG function predicted that the environment adaptation pathway, excretion system pathway, immune system pathway, and the amino acid metabolism pathway were significantly enriched in 8~10 d (P<0.05). Understanding the changes of intestinal flora of piglets in the early stage can provide a reference for piglet feeding and management during lactation.

Increasing litter size is the primary aim of breeding in Mongolia sheep (Ovis aries). Growth and differentiation factor 9 (GDF9) is an oocyte-derived growth factors that play an important role in ovarian folliculogenesis in mammals. Many studies have shown that the mutations in GDF9 were associated with litter size and ovulation rate in several sheep breeds. The aims of this study were to detect the association between the g.46547645T>G SNP of the GDF9 gene and litter size in Mongolia sheep, and detect the genetic diversity in Mongolia, Ujimqin, Hulunbuir (big and short tail types), Small-tailed Han and Hu sheep breeds, as well as to validate the effect of the g.46547645T>G SNP on the promoter activity of GDF9. A total of 416 sheep were used in this study including 260 Mongolia sheep, 36 Ujimqin sheep, 30 Hulunbuir sheep (big tail type), 30 Hulunbuir sheep (short tail type), 30 Small-tailed Han sheep and 30 Hu sheep. The g.46547645T>G in GDF9 were genotyped using the PCR-restriction fragment length polymorphism (PCR-RFLP) method, and then performed the association and genetic diversity analyses. The effect of g.46547645T>G on the promoter activity of the GDF9 was detected by the luciferase reporter assay. The χ² test indicated that the g.46547645T>G SNP is under Hardy-Weinberg equilibrium in the 6 sheep breeds. There were 3 genotypes TT, TG and GG of the g.46547645T>G SNP in Mongolia sheep, 2 genotypes TT and TG in Ujimqin sheep, Hulunbuir sheep (big tail type), Hulunbuir sheep (short tail type), Small-tailed Han sheep and Hu sheep populations. The values of polymorphism information content (PIC) of the g.46547645T>G SNP presented with related low polymorphism in the Mongolia sheep, Ujimqin sheep, Small-tailed Han sheep and Hu sheep breeds, and presented with moderate polymorphism in the Hulunbuir sheep (big tail type), Hulunbuir sheep (short tail type) breeds. The result of association analysis showed that the individuals with TG genotype of g.46547645T>G SNP had extremely significantly more lambs than the individuals with TT genotype in Mongolia sheep (P<0.01). In addition, GDF9 promoter activity analysis showed that the G allele at g.46547645T>G could significantly decrease luciferase activity compared with that of the control (P<0.05), the T allele at g.46547645T>G could significantly increase luciferase activity compared with that of the control (P<0.05), and the G allele at g.46547645T>G could extremely significantly decrease luciferase activity compared with that of the T allele (P<0.01). This study may facilitate effective marker-assisted selection to increase litter size in Mongolia sheep populations, as well as bring new insights into GDF9 expression.

Chemokine-like factor (CKLF) superfamily member 2 (CKLFSF2), named as CKLF-like MARVEL transmembrane domain-containing 2 (CMTM2), plays an important role in spermatogenesis, bone formation and body development. Therefore, it is speculated that the CMTM2 gene might also play an important role in normal growth and development of animals. This study explored CMTM2 genetic polymorphisms and its correlation with growth traits in Inner Mongolia cashmere goats (Capra hircus), and obtained the growing-related genetic markers, hoping to accelerate the breeding process of Inner Mongolia cashmere goat and superfine cashmere goats. In this study, Inner Mongolia cashmere goat (including 460 yearling sheep and 215 adult sheep) were selected as the research objects, and genome DNA was extracted from ear tissues to perform PCR amplification for the detection of insertion/deletion (InDel) mutation of CMTM2 gene, and the correlation between the variations and growth traits was analyzed. The results showed that there was a 14-bp InDel mutation in the CMTM2 gene promoter region of Inner Mongolian cashmere yearling and adult goats, there were 2 types of alleles I and D, and the allele frequency of I was higher than that of D in both yearlinging and adult goats. Homozygous insertion type (I), heterozygous type (ID) and homozygous deletion type (DD) were produced in the mutation. The analysis of polymorphic information content (PIC) indicated that the site was moderately polymorphic in both yearling and adult goats (0.25<PIC<0.50), and this variation did not conform to Hardy-Weinberg equilibrium in the yearling goat population (P<0.05). Further correlation analysis showed that in the yearling goats (n=460), the mutation was significantly correlated with height at hip cross (P<0.05). In the adult goat population (n=215), the mutation site was significantly correlated with body height (P<0.05), heart girth (P<0.05), height at hip cross (P<0.05), chest depth (P<0.05), and fluff fineness (P<0.05). In all of the goat population (n=675), the mutation site was significantly correlated with fluff fineness as well (P<0.05). The above results suggested that the 14-bp InDel within CMTM2 gene promoter region had significant or extremely significant correlation with some growth traits of Inner Mongolia cashmere goats, and different analysis methods showed that the mutation was significantly associated with fluff fineness of goats. Therefore, CMTM2 gene could be used as a candidate gene for the selection of growth traits in Inner Mongolia cashmere goats. The present study provides reference for the selection of Inner Mongolia cashmere goats and superfine cashmere goats.

Mandarin fish (Siniperca chuatsi) has low utilization rate of artificial feed and high breeding cost. In present study, in order to improve the palatability and utilization rate of artificial feed for mandarin fish, Bacillus spp. with enzyme producing ability was screened from the intestine of healthy mandarin fish. The intact intestines of 16 healthy mandarin fish were selected aseptically, and the homogenate was diluted in a stepwise manner to screen the strains producing amylase, protease and cellulase by the plate coating method. After comprehensively analyzing the enzyme-producing ability of the strains, a strain GY65 with strong enzyme-producing ability was selected. The strain GY65 was determined to be Bacillus velezensis according to morphological observation, physiological and biochemical characteristics analysis and genes evolution analysis of 16S rRNA and gyrA. The growth characteristics showed that the optimal pH of strain GY65 was 7, and the growth of strain GY65 was promoted by the low concentration of Mg²⁺, Ca²⁺, Mn²⁺ and Fe³⁺. Biosafety test showed that the bacteria was sensitive to most antibiotics; and non-pathogenic to mandarin fish, snakehead (Channa argus), nile tilapia (Oreochromis niloticus) and zebra fish (Danio rerio). In conclusion, the strain GY65 has good biological safety and the potential as a feed additive for mandarin fish. This study provides a reference for improving the utilization rate of artificial feed for mandarin fish.

As a new breeding method, genomic-wide selection has revolutionized the dairy industry and played an important role in the genetic breeding of other animals. With the improvement of livestock reference genome, the development of genotyping chip technology and sequencing technology, genomic selection will have a greater impact on animal breeding. In this paper, the principles and methods, advantages and disadvantages and influencing factors of whole-genome selection, as well as its application in ruminants such as cattle and sheep were briefly reviewed. The problems and challenges of whole-genome selection were discussed, and the development prospects of whole-genome selection were prospected, which provides a new idea for breeding improvement of ruminants such as cattle and sheep.

With the development of industry, the content of heavy metals in the earth-surface is increasing, which seriously threatens human health. Among the technologies for treatment of heavy metal pollution, microbial remediation technology has attracted much attention due to its advantages of low cost, simple operation and environmental friendliness. However, the low efficiency of microbial remediation technology limits its application. Synthetic biology combines biotechnology and engineering theory. Through the de novo design of microbial gene network, microorganisms can complete various tasks of artificial design. Synthetic biology has many applications in improving the efficiency of microbial remediation of heavy metals. Under the guidance of synthetic biology, a large number of microbial biosensors for heavy metal detection, microbial functional strains for heavy metal adsorption, and microbial factories for the production of heavy metal nano-particles have been developed. This review provides a new systemic and complete solution for the comprehensive treatment of heavy metals.

With the continuous expansion of genetically modified soybean market, Chinese citizens have become more and more disputable about the food safety and environmental risks of genetically modified soybean (Glycine max). More and more countries require labeling of genetically modified food, and some countries also have requirements on the content of genetically modified ingredients. Therefore, in order to establish a rapid and effective detection method for Bar-transgenic soybean quantitative detection, 18 strains of phosphinothricin acetyltransferase (PAT) monoclonal antibody (MAb) with good specificity were prepared in mice (Mus musculus) immunized with purified PAT protein, and a panel of monoclonal antibodies (9F5 and 3G12) that can be used for sandwich enzyme linked immunosorbent assay (ELISA) were screened using the segmented expression peptides of PAT expressed in Escherichia coli, and a rapid quantitative double antibody sandwich ELSIA (DAS-ELISA) for PAT was established based on this panel of monoclonal antibodies. ELISA plate coated with the capture antibody 9F5, and co-incubated with detector antibody 3G12 (2 μg/mL) labeled with HRP for 30 minutes at 37 ℃, color rendering at room temperature and away from light for 15~20 minutes. The detection limit was 1.69 ng/mL and detection range of PAT protein was 0.5~8.0 μg/mL. The coefficient of variation in the plate and between plates was less than 10%. With high sensitivity and stability, the detection could be completed in 50 minutes. This method provides technical support for detection of Bar-transgenic soybean.

The medicinal components of Uncaria rhynchophylla, rhynchophylline and isorhynchophylline, are regulated by various enzymatic reaction genes. Screening the best internal reference genes in the qRT-PCR system of U. rhynchophylla is of great significance for the study of gene expression of medicinal components in U. rhynchophylla. The present study aimed to select reference genes stably expressed in different tissues, different shading treatments and different ethylene treatments in U. rhynchophylla. U. rhynchophylla was selected as the material, and the expression of 8 reference genes of β-actin (β-act), α-tubulin (α-tub), 18S ribosomal RNA (18S rRNA), translation elongation factor (ef-1), glyceraldehyde-3-phosphate dehydrogenase (gapdh), β-tubulin (β-tub), ribosomal protein (rpl) and ubiquitin-ligase enzyme (ubc) in different tissues and different treatments were analyzed by qRT-PCR. The GeNorm program was used to determine the number of reference genes, and then NormFinder and BestKeeper programs were combined to evaluate the stability of the reference genes. Finally, the geometric mean method was used to integrate the results obtained from different software and obtained comprehensive ranking. The results showed that single reference gene could be used in different tissues and treatments. In different tissues, ef-1 should be selected as the reference gene; under shading treatments, gapdh should be selected as the reference gene; under ethylene treatments, both ubc and β-act could be used as the reference gene. If a relatively stable reference gene is used in multiple expression studies, ubc should be selected. This study provides correction and standardization genes for the expression analysis of related genes in different experimental systems of U. rhynchophylla, and is helpful to improve the accuracy and reliability of the experiments.

In wild animals, horns are used to fight for survival and mates. However, in the intensive farming system, polledness is a favorable trait because helps avoid the cost and protect animal welfare. Polledness is an autosomal dominant trait with multiple variants/alleles. Three types of polled variants have been reported on chromosome 1 of taurine cattle (Bos taurus) - the Friesian variant that occurred in dairy cattle breeds, the Celtic variant that was found in beef and dual-purposed breeds, and the Mongolian variant that was identified in Mongolian cattle. In this study, based on a genotyping platform that combines KASP (kompetitive allele-specific PCR) assay and microfluidic chip, a molecular diagnostic method was developed to detect the three polled variants simultaneously. The new KASP assay was further validated by using the conventional molecular experiments including PCR amplification, gel electrophoresis and Sanger sequencing, demonstrating that the new method was accurate and reliable and could be applied in large-scale testing. Interestingly, the Mongolian polled variant was detected in a bull of Sanhe cattle, an indigenous Chinese cattle breed. In addition, a previously unpublished deletion of 2 bp was identified in the Friesian variant that could be used as a marker associated with polledness. These results provide a technical basis for molecular breeding for polled cattle.