S-phase kinase-associated protein 2A (SKP2A) is a F-box protein, which is involved in regulating cell division and improving tolerance to osmotic stress in Arabidopsis thaliana. In order to study the role of SKP2A in the process of wheat (Triticum aestivum) response to abiotic stresses and fungi pathogen infection, the present study further analyzed TaSKP2A gene and its encoded protein based on the full length ORF of wheat TaSKP2A. The results showed that TaSKP2A encoded a polypeptide of 381 amino acids, predicted isoelectric point of TaSKP2A was 6.83 and the molecular weight was 40.87 kD. TaSKP2A was non-secreted protein according to the signal peptide prediction, and its subcellular localization was nuclear. TaSKP2A belonged to F-box/LRR protein based on the C-terminal domain. qRT-PCR results showed that TaSKP2A was significantly higher expressed in pistil and flag leave, while lower expressed in young stem and stamen of wheat. After treating with salicylic acid (SA) and NaCl, the expression of TaSPK2A was up-regulated at 24 and 48 h, respectively. When treating with H₂O₂, TaSPK2A showed gradually down-regulation. The relative expression of TaSPK2A in resistance combination at all inoculation timepoints were higher than that in susceptible one, and the highest expression (8.1 times higher when comparing to sample at 0 h) occurred in resistance combination at 12 h after inoculation with leaf rust pathogen (Puccinia triticina). Yeast (Saccharomyces cerevisiae) two hybrid was used to screen the target protein interacting with TaSPK2A, the results showed that chitinase (CHI) and S-phase kinase-associated protein 1 (SKP1), a scaffolded protein of Skp1-Cullins-F-box (SCF) complex, could interact with TaSPK2A. These results suggested that TaSKP2A might play an important role in response to salt stress and leaf rust pathogen infection, the TaSPK2A may be a member of SCF complex. The results lay basic foundation for analyzing the functional knowledge of SKP2A and further exploration of the regulatory network and mechanism of SKP2A.

Productive tiller number (PTN) is one of the important factors for the yield performance of wheat (Triticum aestivum) and a complex quantitative trait controlled by polygenes. In this case, it is critical to explore the main-effect quantitative trait loci (QTL) and its closely linked molecular markers by the strategies of QTL mapping and meta-analysis in molecular genetic improvement in wheat breeding. In this study, a set of recombinant inbred lines (RIL) of wheat under different water conditions was used to map QTLs for the PTN trait. These identified QTLs were further integrated with reported QTLs for meta-analysis. The results showed that phenotypic values of PTN in the RIL varied widely with significantly transgressive segregation which were sensitive to water stress and thus indicated a low heritability. Under different water conditions, a total of 22 additive QTLs were identified and distributed on all chromosomes except chromosome 4A, accounting for 5.86%~13.84% of phenotypic variations. A total of 97 QTLs governing the PTN from 15 mapping populations were used to construct QTL consensus map in wheat. Consequently, 33 of meta QTLs and their closely linked molecular markers were available and confirmed. The minimum interval of meta QTL was 1.94 cM. The study would be useful to fine QTL mapping and molecular marker-assisted selection breeding for the productive tiller number in wheat.

Restorer lines is one of the important parents of three-line hybrid rice (Oryza sativa). Excavating its high-yielding genes is great important for improving the yield of hybrid rice. A recombinant inbred lines (RIL) population derived from a cross-combination of 'Luhui8258' and 'Yanghui34'. The RIL and 2 parents were grown under 3 different environments including Deyang, Suining and Luzhou of Sichuan province, and 8 yield traits including biological yield per plant (BYP), harvest index (HI), yield per plant (YP), effective panicles per plant (EP), spikelet per panicle (SP), full grain per panicle (FGP), seed setting rate (SSR, as a percentage), 1000-grain weight (TGW) were investigated. On the basis of constructing genetic linkage map consisted 184 DNA markers, a total of 62 QTLs for yield-related traits distributing on all of the 12 chromosomes were identified. Among them, 2 QTLs synergistic alleles were derived from 'Luhui8258' were detected repeatedly in 3 environments, namely, it were qTGW3 and qFGP6-1, with the contribution rate of 5.92% (Deyang), 7.79% (Suining), 19.04% (Luzhou) and 8.80% (Deyang), 10.90% (Suining), 4.96% (Luzhou), respectively. In addition, 17 yield-related QTL detected in two of these environments, included qBYP6-2, qBYP9 (QTL for BYP), qHI3-1, qHI3-2, qHI12 (for HI), qYP3-2, qYP12 (for YP), qEP2-1, qEP9-2 (for EP), qSP6-1、qSP12 (for SP), qFGP2-1, qFGP2-2, qFGP6-2, qFGP12-2 (for FGP) , qSSR2, qSSR6-1 (for SSR). Among them, 11 QTL synergistic alleles were derived from 'Luhui8258', including qHI3-1, qHI3-2, qBYP6-2, qBYP9, qYP3-2, qEP2-1, qEP9-2, qSSR2, qSSR6-1, qSP6-1, qFGP6-2, the other 6 QTL synergistic alleles were derived from 'Yanghui 34'. This research provides the basis and technical support for fine mapping, cloning and molecular marker-assisted selection of rice yield-related traits in near future breeding program.

Lateral branch length, one of the most important architectural traits, acts as an effective breakthrough point for architecture improvement of vine plants, such as cucumber (Cucumis sativas), melon (C. melo), and watermelon (Citrullus lanatus). In this study, the short-lateral-branching material D581 (♀) and long-lateral-branching material H465 (♂) were used as parents to establish F₂ and F_2∶3 populations, which were further used to study the variation pattern of lateral branch length and then map the QTLs controlling the lateral branch length as well as predict the candidate genes related to this trait. The results showed that the variation of lateral branch length revealed a typical normal distribution in F₂ and F_2∶3 populations, and the estimated broad sense heritability of the 6 statistic indices of lateral branch traits exceeded 60%. A total of 253 SSR and InDel markers were adopted to construct a genetic map containing 12 linkage groups, covering 1 433.3 cM distance of the melon genome with an average distance of 5.67 cM between the markers. Totally, 31 QTLs were detected in spring and autumn with the phenotypic variance explained (PVE) for a single QTL varying from 6.39% to 16.46%. Seven QTLs detected by various statistic indices or in various environments were co-located in SSR029716-ECM88 region of LG8, with a cumulative PVE of 75.66%. Subsequently, by means of deep resequencing of the parents (>30×) and development of new InDels in the targeted region, this region was further delimited to a 185 kb region which contained 31 annotated genes. Of these genes, three genes were previously reported to participate in plant hormone signal transduction and predicted as the putative candidate genes controlling lateral branch development in melon. The QTLs and candidate genes identified in this study acted as helpful directions for the current gene discovery of lateral branch traits of melon, offering a theoretical basis for molecular improvement of plant architecture in melon.

Celery (Apium graveolens) is one of the most economically important leafy vegetables worldwide. Current celery genetic research requires some valuable molecular markers. A total of 4 034 SSR loci were identified in 39 619 unigenes assembled from 3 sets of high quality reads of celery transcriptome data in GenBank. Of these, di-nucleotide repeat motifs were found to be the most abundant, which accounted for 43.9% (1772), followed by mono- (26.7%, 1078) and tri- (21.0%, 846) nucleotide repeat motifs. Tetra-, penta-, and hexa- nucleotide repeats accounted for 1.3% (52) and the other was compound type (7.1%, 286). A set of 39 pairs of primers from the designed 375 SSR primers were successfully validated to yield clear and stable high quality specific PCR products in 5 selected representative celery accessions from different sources. Using fluorenscently labelled PCR and multiplex capillary electrophoresis, all the 39 markers detected 110 alleles in 76 celery accessions, ranging from 2~4 with an average number of 2.821 alleles and 2~9 genotypes with an average of 4.308 per locus. Average polymorphism information content (PIC) and discrimination power (PD) were 0.256 (0.013~0.581) and 0.379 (0.026~0.764) for per locus, respectively. Genetic diversity analysis of the 76 celery accessions indicated that for local celery and celery, the average numbers of alleles were 2.641 and 2.436, the average Nei's indices were 0.347 and 0.194 and the average Shannon's indices were 0.589 and 0.351, suggesting greater genetic diversity in local celery than that in celery. Analysis of molecular variance (AMOVA) of A. graveolens showed that 61.72% of genetic variation was within celery individuals, 22.07% among individuals within local celery or celery population, and 16.22% between local celery and celery populations. The cluster results of PCA (principal component analysis) and UPGMA (unweighted pair-group method with arithmetic means) dendrogram separated the 76 celery cultivars into 2 groups mainly corresponding to celery and local celery, respectively. However, compared to celery, local celery formed a more discrete cluster, which indicated local celery probably had more complex genetic background. The study could enrich SSR markers and provide multiplex capillary electrophoresis detection system based on fluorescence labeling for genetic diversity analysis and identification of a large number of celery germplasm resources.

WRKY transcription factor is a zinc finger protein unique to plants. Based on the transcriptome data of tartary buckwheat, a WRKY transcription factor gene FtWRKY10 was cloned in this study. Sequence analysis results showed that FtWRKY10 was located on chromosome 5 of tartary buckwheat genome, its DNA length was 4 303 bp, which contained 6 exons and 5 introns, and its ORF was 1 647 bp. The deduced FtWRKY10 protein was a classic WRKY transcription factor of type I, and it had the closest phylogenetic relationship with AtWRKY20 of Arabidopsis thaliana. qRT-PCR showed that its expression was induced by drought stress in tartary buckwheat. Molecular identification results showed that FtWRKY10 protein showed transcriptional activation activity in yeast and located in the nucleus of transgenic Arabidopsis thaliana cells. The transcription factor could specifically bind with its target DNA motif of W-box in yeast and plant cells and initiate the expression of the reporter genes, respectively. In Arabidopsis, overexpression of FtWRKY10 could enhance the tolerance of Arabidopsis germination to drought and ABA treatments. Under drought stress, the overexpression of FtWRKY10 could increase the content of chlorophyll and proline, the activity of SOD (Superoxide dismutase), POD (Peroxidase) and CAT (Catalase), and also up regulate the expression of stress resistance related genes of responsive to dehydration protein genes (RD29A, RD29B, and RD22), responsive to ABA protein gene RAB18, and serine/threonine protein kinase gene KIN1. For conclusion, the FtWRKY10 encodes a type I WRKY transcription factor related to drought tolerance. This study could provide strategies for exploiting and utilizing the excellent stress resistance traits from tartary buckwheat, and supply basic materials for further enriching the functions of WRKY family.

The zinc finger-homeodomain (ZF-HD) is a family of homologous heterologous box proteins, a plant-specific transcription factor that plays an important role in plant growth and development. In order to explore the function of ZF-HD gene family in the growth and development of Phyllostachys edulis, the family's physicochemical properties, phylogenetic relationship, gene structures and expression patterns were analyzed in detail by bioinformatics based on Phyllostachys edulis genome and related transcriptome data in this study. The results showed that 23 ZF-HD members in the genome with fewer introns (1~2) of P. edulis, and their gene structures and motifs were relatively conservative. The amino acid sequences of ZF-HD gene family had different domains, including zinc-finger (ZF) and homeodomain (HD), which could bind to DNA. Phylogenetic tree showed that ZF-HD gene family could be divided into 6 subfamilies. Combined with the results of multiple sequence alignment and homologous modeling analysis, ZF-HD belonged to the C2H2-type Zn-finger domain super-gene family. The upstream promoter sequences contained multiple cis-acting elements related to hormone response. The collinearity relationship with rice (Oryza sativa) was higher than that with Arabidopsis thaliana, and there was a doubling event of gene duplication. In combination with Gene Ontology (GO) enrichment analysis, RNA-seq data of ZF-HD genes were analyzed under different hormone treatment conditions and in different stages of P. bud development. It was found that ZF-HD could respond to the regulation of exogenous gibberellin (GA) and naphthalene acetic acid (NAA). The high expression of some gene family members in P. edulis shoots indicated that they might be involved in the rapid growth and development of P. edulis shoots. The results of this study will help to reveal the biological functions of the ZF-HD gene family in the rapid growth and development of P. edulis.

BBX (B-box) protein is a subfamily of zinc finger transcription factor protein family, which is widely involved in plant growth and development. And, they also play an important role in abiotic stress response. Eucalyptus is an economic tree species in southern China. However, most cultivated Eucalyptus species are sensitive to abiotic stresses, which made the extension of Eucalyptus cultivation range and the improvement of cultivation benefit limited. In order to enrich gene resources of abiotic stress resistance and provide the basis for molecular assisted breeding for Eucalyptus, total of 21 EgrBBX gene family members were identified from the Eucalyptus grandis genome. Chromosomes distribution, protein sequence structure, and the classes and distribution of cis-elements in promoters were characterized with software of mg2c, Protparam, MEGA and PlantCARE. And, tissue specific expression of EgrBBX family genes and their expression profiling under low temperature, drought and salinity were also analyzed with qRT-PCR. The results showed that EgrBBX genes, named EgrBBX1~21, were distributed at 9 chromosomes. The encoded protein sequences contained classic B1, B2 and CCT domains and could be classified into 5 classes: B1+B2+CCT type ⅠandⅡ, B1+B2, B1+CCT and B1. The results of promoter element analysis indicated that a large number of light-responsive elements and abiotic stress response elements such as ABRE (ABA-responsive element), MBS (MYB binding site), LTR (low-temperature responsiveness), and HSE (heat-stress responsive element) existed in promoter sequences of EgrBBX family gene. qRT-PCR results showed that EgrBBX1, EgrBBX7, EgrBBX9, EgrBBX13 and EgrBBX18 were mainly expressed in leaves, and EgrBBX3, EgrBBX6 and EgrBBX15 had relatively high expression levels in stems. Under the treatments of different time at 4 °C, drought and high- salinity (200 mmol/L NaCl), most EgrBBX gene expression were changed. During the time of low temperature treatment, expression of EgrBBX4, EgrBBX7, EgrBBX8, EgrBBX14, EgrBBX16 and EgrBBX17 showed strong inhibition. EgrBBX8, EgrBBX11, EgrBBX12 and EgrBBX18 showed significant up-regulated expression at 1 d after drought treatment, and the induction effect weakened or disappeared after 2 d. Under high salinity treatment, 13 of 21 EgrBBX genes reached expression peak after 12 h treatment and then gradually decreased. The results in this study provided a basis for further revealing the functions of EgrBBX family in abiotic stress response, and could facilitate the screening of EgrBBX genes which are stress-resistant in Eucalyptus.

Luteinizing hormone receptor (LHR) and prolactin receptor (PRLR) genes play essential roles in mammalian reproductive development. But still no any available research regarding these genes in yak (Bos grunniens). Therefore this study was planned to explore the role of LHR and PRLR genes in different tissues of Yak during different developmental ages (6, 24, 36 and 72 months). The mRNA expression levels of LHR and PRLR in the testis tissues of yak at different developmental stages and organs of yak were detected by qRT-PCR, while the protein expression of LHR and PRLR were evaluated by immunohistochemistry.The results showed that mRNA expression levels of LHR and PRLR genes in yak testis increased significantly from infancy to sexual maturity (P<0.01), but there was no significant difference between adult and sexual maturity. The expression level of LHR gene mRNA in different organs of adult yak was higher in testis than in kidney, skeletal muscle and lung (P<0.05). While the expression level of PRLR mRNA was higher in kidney, followed by skeletal muscle, testis and spleen (P<0.05). Additionally, immunohistochemistry findings revealed that LHR and PRLR proteins were located in testicular supporting cells, mesenchymal cells, primary spermatocytes and secondary spermatocytes, and they were increased gradually from infancy to sexual maturity stage and tended to be stable in adulthood. These findings displayed that LHR and PRLR may be vital for sperm maturation, in the biological functions of interstitial cells, and in regulating the development of other non-gonadal tissues. These results provide an important theoretical basis for revealing the molecular mechanism of testis development and reproductive regulation in yaks.

Caveolin-1 (Caveolin-1, CAV1) is a membrane protein associated with endocytosis, extracellular matrix organization, cholesterol distribution, cell migration and signaling.The aim of the present study was to explore the expression and biological function of CAV1 in the reproductive system of female yak (Bos grunniens) under normal physiological conditions. In this study, the uterus, ovaries and fallopian tubes of female yaks at different reproductive stages (follicula, luteal and gestation) were used as materials. The mRNA and protein expression of CAV1 were analyzed by gene cloning, qRT-PCR, Western blot and immunohistochemistry (IHC). The results showed that the CAV1 gene (GenBank No. MN127963) was highly conserved. The CAVI gene sequence of yak was different from ordinary cattle (Bos taurus), but the encoded amino acid was constant. The CAVI gene of yak had the closest relatives to Bos taurus and Ovis aries, and the farthest relatives to Felis catus and Equus saddle. and the encoded protein was a stable hydrophobic membrane protein. CAV1 was expressed in the main organs of the female yak reproductive system, while the expression level of CAV1 protein in the oviduct, ovary and uterus in the follicular phase was significantly higher than that in the luteal phase and gestational period (P<0.05). The expression level of CAV1 protein in the fallopian tube of the gestational period was significantly higher than that in the luteal phase (P<0.05). The expression level of CAV1 protein in the ovary and uterus of the luteal phase was significantly higher than that in the gestation phase(P<0.05). Immunohistochemistry results showed that CAV1 is expressed in the oviduct mucosa epithelium, serous gland, ovarian follicle membrane, follicular granule layer, luteal cells, reproductive epithelium, endometrium and uterine gland. This study provides a theoretical basis for further exploration of CAV1 involved in mammalian reproductive physiology regulation, which is helpful to the study of the breeding rules of mammals in the plateau.

The content and distribution of intramuscular fat (IMF) is the material basis for the formation of Bos taurus marbled meat. There is a difference in fat deposition capacity between Simmental cattle (B. taurus) and Angus cattle (B. taurus). The IMF content in Angus is significantly higher than that in Simmental cattle. In this study, high-throughput sequencing technology was used to screen the genes related to backfat deposition, in order to provide reference for the research on the mechanism of fat deposition in Simmental and Angus cattles. Three 15-month-old bulls from each breed were fattened and slaughtered to determine the meat quality of the carcass. Transcriptome sequencing and bioinformatics analysis were performed on the backfat fat tissues of the 2 breeds. The results showed that there were no significant differences in crude fat, water loss, ribeye area and pH between the 2 breeds, while the shear force of Simmental cattle was significantly higher than that of Angus cattle (P<0.01). Analysis of transcriptome data showed that there were 1 296 differentially expressed genes (DEGs) in the backfat tissue of the 2 breeds, which mainly enriched in metabolic process, growth and development etc. of 55 function items and lipid metabolism, carbohydrate metabolism, energy metabolism etc. of 44 metabolic pathways. According to the pathway and function analysis, 3 genes including pyruvate dehydrogenase kinase 4 (PDK4), nuclear receptor subfamily 4 group A member 1 (NR4A1) and prospero related homeobox (PROX1) were selected for gene expression analysis by quantitative real-time PCR. Quantitative results showed that the relative expression of PROX1 and PDK4 in the backfat of Angus cattle was 47% and 14% higher than that in the Simmental cattle (P<0.01), and the relative expression of NR4A1 in the Simmental cattle backfat was 30% higher than that in the Angus cattle (P<0.05). The differentially expressed genes obtained in this study could be used as candidate genes for the improvement of beef quality traits and provide basic information to further explore the genetic mechanism of backfat growth and development.

In order to explore the genetic effects of the untranslated region polymorphism of bone morphogenetic protein receptor IB (BMPR-IB) gene on the reproductive performance of sheep (Ovis aries), Multiple Hu Sheep (HM), Singleton Mongolian Sheep (MS), Twins Mongolian Sheep (MT), Singletons Oula Sheep (ZS), Twins Oula Sheep (ZT) and Singletons Australian Sheep (AS) was selected in this study to detect SNPs in the 3'-UTR of BMPR-IB gene by DNA sequencing, the correlation between polymorphism loci and litter size was also studied. Two mutation sites of 1339 (G/A) and 1354 (A/G) were detected at 1 145~1 500 bp in the 3'-UTR region of BMPR-IB gene, and further analysis was performed on the 1354 locus (corresponding to the 3019 locus in the mRNA sequence of BMPR-IB gene NM_001009431.1). The results showed that wild genotype AA, heterozygous mutant genotype AG and homozygous mutant genotype GG were detected in MT and ZT, the dominant allele was A; wild genotype AA and heterozygous mutation genotype AG were detected in MS, ZS and AS, the dominant allele was A; heterozygous mutation genotype AG and homozygous mutation genotype GG were detected in HM, the dominant allele was G. The result of polymorphism information content (PIC) analysis showed that HM, MT, MS, ZS and AS were in low polymorphism (PIC<0.25); and ZT was in moderate polymorphism (0.25<PIC<0.5). There was a significant difference in the 1354 locus of 3'-UTR region of BMPR-IB gene among the multiple-lamb, twins-lamb and singletons-lamb populations (P<0.05). The genotype of AG and GG had significant effects on the fecundity in HM, MT and ZT. These results suggested that the 1354 locus in the 3'-UTR region of BMPR-IB gene might be regarded as a molecular genetic marker locus for multiple-lamb and twins-lamb in sheep. This study provides a theoretical scientific basis for breeding sheep with high fecundity.

The SLC26A7 (solute carrier family 26 member 7) gene in the members of the sulfate/anion transporter gene family (SLC26) can be used as a selective chloride ion channel to regulate renal electrolyte balance and mediate thyroid hormone biosynthesis. Its mutation causes thyroid insufficiency and affects individual growth and development. In this study, comparative genomics and bioinformatics methods were used to analyze the transposon insertion of the pig (Sus scrofa domesticus) SLC26A7 gene, and the polymorphisms in different breeds and their effects on the growth characteristics of Yorkshire were detected. The results showed that SLC26A7 gene had a 280 bp ERV insertion in intron 1, and the frequency of introduction of ERV⁺ is higher than that of Chinese native pig breeds. The age of 100 kg body weight of ERV^+/+ individuals in the Yorkshire were significantly higher than that of ERV^-/- individuals (P<0.05), and the ERV^+/+ individuals is significantly lower than the ERV^-/-(P<0.05). Insertion of ERV in the first intron of SLC26A7 gene inhibited its promoter activity. This study confirmed that the ERV insertion in pig SLC26A7 intron 1 had a significant effect on the age of 100 kg body weight and pre-slaughter weight (P<0.05). The results provided a reference for the application of this molecular marker in pig molecular breeding.

Inositol-1, 4, 5-triphosphate receptor type Ⅱ (ITPR2) is a key regulatory factor of calcium ion transmembrane transport activity, which affects eggshell quality. In this study, the tissue expression of ITPR2 gene was detected by qRT-PCR, and the SNP sites of protein domain were identified by PCR purification and direct sequencing combined with DNAStar screening. The genetic effect of SNP sites on eggshell quality was analyzed by general linear model. The results showed that ITPR2 gene was expressed in 11 tissues, and the expression of ITPR2 gene was in the order of pancreas>eggshell gland>glandular stomach>kidney>pectoral muscle>spleen>small intestine>lung>liver>heart>muscle>stomach, and the expression of ITPR2 gene in pancreas and eggshell gland was significantly higher than that in other tissues (P<0.01). Three moderate polymorphisms (0.25<PIC<0.50) were detected in exon 16, intron 14 and intron 37 of ITPR2 gene in Sansui ducks (Anas platyrhyncha domestica), among which there was a strong linkage disequilibrium between g.128767757 A>G and g.128659402 G>T, g.128662092 C>T; g.128659402 G>T and g.128767757 A>G deviated from Hardy-Weinberg equilibrium (P<0.05). Correlation analysis showed that the mutation site g.128662092 C>T in exon 16 affected the eggshell quality, and TT genotype significantly increased the eggshell strength (P<0.05); 3 SNPs produced 4 haplotypes and 10 diplotypes, H4 was the dominant haplotype, H2H4 was the dominant diplotype; combination genotype had different effects on eggshell quality, H1H1 and H1H2 significantly increased the eggshell quality strength (P<0.05). The prediction results of ITPR2 protein function showed that the protein encoded by this gene contained 2 647 amino acids, which belonged to non-soluble and unstable protein; there were 6 N-glycosylation sites, and the second and third structures were mainly α-helix and irregular curl. The results of this study suggested that the CT genotype of ITPR2 gene (g.128662092 C>T) in Sansui duck was beneficial to the enhancement of eggshell strength, and the combination of three SNP loci could significantly improve eggshell quality, which could be used as a genetic marker for eggshell quality selection.

Traditional methods for identification and classification of plant nematodes are mainly based on morphological characters and morphometrics. These methods are subjective and cumbersome, and are difficult to make reliably taxonomy and identification. The molecular method for taxonomy and identification based on DNA not only provides objective information, but also can reveal the evolution of different groups of nematodes. In this study, 18S rRNA gene fragments of 94 plant parasitic nematode populations, which belonging to Rhabditida, Dorylaimida and Triplonchida, were amplified with primer sets NEMF1/S3 and A/S3, respectively. The sequences were compared and edited by DNAstar software, and were used to construct the phylogenetic trees by Mega 6.0 software. The results showed that the phylogenetic tree constructed based on the sequences, which PCR amplified with primers A/S3, can reflect the phylogenetic relations of nematode species within different groups and the same group. The results provide reference to study phylogeny of plant nematodes in the future.

Gene splitting technology can effectively avoid the escape of target traits in genetically modified crops to the environment. In order to cultivate a new glyphosate-resistant aroA_A1501 transgenic rice (Oryza sativa) using gene resolution technology, 13 possible resolution sites were screened out based on the advanced structure of aroA_A1501 protein in this study. aroA_A1501 was split into N-terminal (An) and C-terminal (Ac) by PCR, and is combined with N-terminal (In) and C-terminal (Ic) of SspDnaE intein to form fusion genes An-In and Ic-Ac. Based on pETDuet-1, a total of 40 prokaryotic expression vectors containing An-In, Ic-Ac and An-In / Ic-Ac were constructed. The constructed prokaryotic expression vector was introduced into the auxotrophic Escherichia coli strain ER2799. It was proved by functional complementation tests that aroA_A1501 was split at the positions 141/142, 224/225, and 230/231. The protein was reassembled into a functional intact protein mediated by the protein intein. The glyphosate tolerance test proved that 230/231 was the most suitable resolving site. The reassembled protein after resolving was 3 times more tolerant to glyphosate than the complete aroA_A1501 protein. This study provides basic data for the later cultivation of glyphosate-resistant transgenic rice with aroA_A1501 gene.

Novel duck reovirus (NDRV) disease is a serious infectious disease for poultry in China. Chlorogenic acid (CHA) is a main component of many traditional Chinese medicines, and plays an important role in antiviral research. Exploring the antiviral effect of CHA on NDRV which might provide a basis for the clinical treatment of NDRV to some extent. Firstly, based on S3 gene sequence (GenBank No. KJ879932) of NDRV, the method of real-time fluorescent quantitative PCR (qPCR) was established. Secondly, maximum safe concentration of CHA on baby hamster (Mesocricetus auratus) kidney cell line BHK-21 cells was determined by CCK-8 kit. Finally, the anti-virus test was performed. The experiment was divided into 2 groups. GroupⅠ: NDRV were inoculated in BHK-21 cells before different concentrations of CHA were added. Then the freeze-thaw solution of cells was collected periodically. GroupⅡ: The method of virus adsorption was the same as that of groupⅠ. Before treatment, the virus were proliferated on BHK-21 cells for 10 h, and the viral load after 10 h proliferation was quantitatively detected as the base value. After 10 h of virus proliferation, different concentrations of CHA were added, and the follow-up procedure was the same as that of Group Ⅰ. Negative and positive control were set in both groups. And the remaining experiment procedures were the same as those in groupⅠ. The cytopathic effects of each group were observed regularly, and the collected samples were quantitatively detected by qPCR. The results showed the lowest detectable viral nucleic acid of the method was 92.5 copies/μL, and the maximum safe concentration of CHA on BHK-21 cells was 128 μg/mL. The effect of anti-NDRV of CHA indicated that different concentrations of CHA could inhibit the proliferation of NDRV in vitro, and this inhibitory effect had an obvious dose-effect relationship. This study could provide basic data for the study of NDRV mechanism and its practical application.