Plant height (PH) is an important agronomic trait of wheat (Triticum aestivum), which is closely related to high and stable yield. Nitrogen is an essential mineral nutrient for plant growth and development, critically affects PH and yield of wheat. The sensitivity to nitrogen varies among different cultivars. In order to explore genetic loci influencing PH and PH tolerance to low nitrogen stress, and to provide excellent genetic resources of dwarf and high nitrogen efficiency, in this study, 132 elite wheat varieties (lines) mainly from the north Yellow and Huai Valley Wheat Region of China in recent 30 years were selected to carry out genome-wide association study (GWAS). PH and tolerant index (TI) of PH to low nitrogen stress under 8 environments with different nitrogen fertilizer conditions were identified. The genetic data were acquired by Affymetrix Wheat 55K SNP chip. Finally, 13 SNPs distributed on 1B, 1D, 2A, 2B, 2D, 3B, 4D, 5A, 6B and 7D were identified to be significantly associated with PH in 3 or more environments, which could explain 9.6%~26.6% of the phenotypic variation. Among them 4 significant association SNPs on 1B (2), 3B and 7D could be identified in both low nitrogen environment (LN) and high nitrogen environment (HN). AX-108824683-7D was significantly associated with PH in 7 environments, accounting for 16.8%~26.6% of the phenotypic variation. This locus has not been reported in previous studies, suggesting that it is a new dwarf gene locus. 7 SNPs were identified to be significantly associated with tolerant index of PH to low nitrogen stress, among which AX-111004994-4B was stably identified in 2 environments, explained 12.4%~13.1% of the phenotypic variation. The results provide references for dwarfing and high nitrogen efficiency in wheat molecular breeding.

Stem length and stem diameter, as two important traits of maize (Zea mays) stem, are closely related to maize lodging resistance, also directly or indirectly affect maize yield and plant architecture, and they are important target traits in maize breeding. This study performed a high-resolution QTL mapping for stem length and stem diameter using the multiple QTL model implemented in R/qtl, together with 19 838 SNP markers. The introgression line population of 866 lines was derived from a cross between a typical maize inbred line W22 and a typical accession of teosinte. The results showed that there were extensive genetic variations in stem length and stem diameter, which were typical quantitative traits controlled by multiple micro-effect genes. A total of 2 QTLs controlling stem length was located on chromosome 1 and 4 respectively, with the phenotypic contribution rate of 4.51% and 2.68%, and the additive effect of 0.41 and 0.39 cm, respectively. In addition, 5 QTLs controlling stem diameter were located on chromosome 1, 2, 5, 6 and 7, respectively, and the phenotypic contribution rate of each QTL varied from 2.06% to 8.32%, and the additive effect varied from 0.03 to 0.09 cm. Further analysis of candidate genes for the largest effect QTL (qSD5-1) of stem diameter, 13 possible candidate genes were screened out. This study might provide a theoretical basis for elucidating the genetic basis of stem length and stem diameter, the cloning of related genes and molecular marker-assisted selection breeding in maize.

Homeobox (HB) genes play an important role in diverse physiological and developmental processes of plants. The HB genes of many plants have been identified. However, the complete identification and classification of potato HB genes is still lacking. In this study, a total of 35 HB genes in the potato (Solanum tuberosum) were identified. The physicochemical properties, phylogenetic relationships, chromosome locations, gene and protein structure, and potential functions were analyzed in detail by bioinformatics. The tissue-specific expression patterns of potato HB genes were also analyzed with qRT-PCR. According to the Neighbor-joining phylogenetic tree and domain architecture, 35 potato HB genes were classified into 6 subfamilies. An analysis of the exon-intron structures and conserved motifs provided further insight into the evolutionary relationships between these genes. Synteny analysis of potato, Arabidopsis and tomato genomes provided a reference for the potential functional relevance of these HB genes. All of the potato HB genes were distributed on 12 chromosomes. Through the RNA sequencing (RNA-Seq) and qRT-PCR analyses, many potato HB genes including StBEL35 (S. tuberosum bell-type homeobox 35), StKNOX1 (S. tuberosum KNOTTED1-like homeobox 1) and StKNOX9 showed a relatively higher expression levels on storage organs (including stolon and tuber). Most of these genes showed higher expression levels in mature tubers, indicating that they may play a role in the later stages of tuber development. Combined with previous studies, these genes may be involved in gibberellins (GA) biosynthesis, thereby affecting tuber formation. This study will facilitate the functional analysis of these genes and provide comprehensive information for further analyses of the molecular functions of the potato HB gene family, and provide a theoretical basis for further molecular breeding in potato.

In production practice, the identification of Chinese cabbage (Brassica rapa ssp. pekinensis) hybrids mostly rely on field planting and phenotypic identification, which is inefficient and inaccurate. Under the condition of more and more mature molecular marker research technology, it is necessary to establish a rapid, efficient and accurate identification technology of Chinese cabbage hybrids based on molecular marker technology. Considering this, a total of 86 pairs of insertion/deletion (InDel) primers were used to identify 40 Chinese cabbage hybrids by PCR amplification and polymorphism detection. Powermarker V3.0 software were used to analyze the polymorphic results. MEGA 7.0 software were performed for cluster analysis of hybrids. In order to screen core primers suitable for DNA fingerprint construction of different kind of Chinese cabbage hybrids, the percentage of heterozygous state for each pair of primers were compared in every group. The results showed that, 189 polymorphic loci were obtained, with an average of 2.2 alleles for each primer combination. The heterozygosity index ranged from 0 to 0.648 2 and averaged at 0.304 2. The major allele frequency ranged from 0.391 9 to 1.000 0 and averaged at 0.701 1. The number of alleles detected by each pair of primers ranged from 1 to 4, of which 74 pairs of primers could only detect 2 alleles. Then, cluster analysis revealed that, based on producing area, the 40 hybrids were clearly divided into imported and domestic varieties, among which the domestic varieties could be further divided into folded-, cylindric-, and fluffy-type varieties.The number of loci with heterozygous state ≥50% and their distribution in genome was significantly different in the 4 groups. Finally, 5 pairs of primers with heterozygous state ≥50% and allele number equal to 2 were selected from each group for construction of core primers. Other 32 Chinese cabbage hybrid varieties which were sold in market in recent years were detected with core primers. The detection results of pure loci were assigned a binary numeral 0 or 1 according to the mobility ratio of electrophoresis from small to large, while heterozygous loci were assigned letter H. All primers were ordered according to chromosome number they located from A01 to A10 and primers on the same chromosome were ordered according to their physical position from small to large. DNA fingerprint of 72 hybrids were constructed. The selected core primers will be universal and compatible with DNA fingerprint construction of new hybrid cultivars created in future. The results could also provide reference for gemplasm screening and hybrids identification of Chinese cabbage.

Drought is one of the abiotic limiting factors that seriously affect the growth of crops. NAC (NAM, ATAF and CUC) transcription factor is one type of transcription factors in plants, it play an important role in regulating plant growth, development and response to abiotic stress. In order to study the regulation function of NAC transcription factor in drought stress of kenaf (Hibiscus cannabinus), a NAC like gene was isolated from kenaf transcriptome database, named HcNAC1 gene (GenBank No. MT799841). The coding sequence of HcNAC1 contained 1 044 bp, encoding 347 amino acids, protein molecular weight 38.77 kD, isoelectric point was 8.78. The HcNAC1 genes with NAM/NAC family conservative area, containing about 160 amino acid conservative in the protein N-terminal structural domain, with 2 nuclear localization signal (NLS) sequence. Phylogenetic tree analysis showed that kenaf HcNAC1 had the closest evolutionary relationship with Hibiscus syriacus. The expression level in different tissues and within 24 h of drought stress was detected by reverse transcription PCR (RT-PCR) and qRT-PCR, research results showed that HcNAC1 gene was expressed in root, stem, leaf, flower and fruit, HcNAC1 gene was expressed under PEG6000 stress and had most expression level at 12 h, and then the expression level gradually decreased. HcNAC1 played an important regulatory role in response to drought stress. The study explored the correlation between HcNAC1 expression and drought stress. and could provide a reference for further research on the molecular mechanism of HcNAC1 transcription factors involved in drought stress of kenaf.

'Xuehua' is one of main cultivars of pears (Pyrus bretschneideri) in China, but the appearance quality is seriously affected by its rough surface, large fruit dot and other peel characters. In this study, according to peel characters, the young fruit were sprayed with a concentration of 200 mg/L chlorogenic acid (CGA) every week from 4 weeks after full bloom. The results showed that, compared with water spray (control), the fruit surface became relatively smooth and the fruit dot became significantly smaller after sprayed with CGA. When the fruit was mature, the firmness and index of absorbance difference (I_AD) of pericarp chlorophyll became significantly lowered, but the single fruit weight, soluble solid content (SSC), titratable acid (TA) and peel color values of L, a, b were significantly increased. At the same time, compared with the control, spraying CGA could significantly reduce the lignin content and increase the total CGA content in the peel, and reduce the activity of phenylalanine ammonialyase (PAL), peroxidase (POD) and polyphenol oxidase (PPO) in the peel most of time, but had little effect on the activity of cinnamyl alcohol dehydrogenase (CAD). In addition, the gene expression of PAL1, POD2, PPO1, PPO4, PPO5, PPO6, CAD2 and CAD3 were significantly down-regulated in the early post-spray application period. In a word, the results of this study indicated that exogenous spraying of CGA could inhibit the lignin synthesis in the peel and the increase of fruit dot areas, and improve the skin characteristics of 'Xuehua' pear, which provides a new idea and basis for the effect of exogenous chlorogenic acid on the appearance quality of 'Xuehua' pear fruit.

CBF (CRT/DRE binding factor) gene plays an important role in ennhancing the ability of plants to resist low temperature stress. In order to verify the cold-resistance function of CcCBFc gene from Cinnamomum camphora, the mechanism of the cold-resistant effect of the CcCBFc in Arabidopsis thaliana was revealed, and the biological function of this gene in Arabidopsis against cold stress was presumed. In the early stage of this laboratory, 4 CcCBFs (CcCBFa, CcCBFb, CcCBFc, CcCBFd) were cloned, and the CcCBFc (GenBank No. KP336741) was initially verified to improve the drought resistance, salt resistance and cold resistance of the transgenic tobacco. This study was based on this, which used transgenic CcCBFc Arabidopsis and wild-type as materials to detect antioxidant enzyme activity under low temperature stress (4 ℃). Antifreeze effect of transgenic CcCBFc Arabidopsis was detected by -4 ℃ simulated freezing injury. qRT-PCR was used to detect the expression level of antioxidant gene of transgenic CcCBFc Arabidopsis at 6 d after low temperature stress, and sequenced transgenic CcCBFc Arabidopsis and wild type at transcriptional level under low temperature stress. The results showed that CcCBFc could increase the peroxidase and catalase activities of transgenic Arabidopsis, and had a positive effect on the antifreeze of transgenic Arabidopsis, and could promote the expression of antioxidant genes in transgenic Arabidopsis. Transcriptome sequencing results of the wild-type and transgenic CcCBFc Arabidopsis indicated that the function of the exogenous CcCBFc gene was related to the metabolic process, catalytic activity and binding activity, and was involved in the regulation of the endoplasmic reticulum. The synthesis of proteins and plant hormones enhanced the cold tolerance of Arabidopsis. This study provides a reference for further revealing the function of CcCBFc in the response mechanism of low temperature stress.

Exosomes (EXs) are a kind of small cellular vesicles with a diameter of 30~150 nm, which mediate the material transport between cells and play a significant regulatory role in development and maturation of gamete. Some studies also have shown that seminal plasma EXs (spEXs) played an important role in preservation and fertilization of sperm. However, the research on the expression regulation of spEXs miRNAs have not been reported at present. In this study, the expression of porcine spEXs miRNAs and their potential regulatory effect on gene expression in sperm fertilization were explored. The semen of elite Landrace boar (Sus scrofa domestica) was used to extract the spEXs, which were identified by electron microscopy, analysis of particle size and marker protein expression. Then the spEXs were extracted and used to sequence the miRNAs. Next, the highly expressed miRNAs were analyzed and their target genes were predicted. Finally, enrichment analysis of Gene Ontology (GO) and KEGG pathways were conducted. A total of 617 miRNAs were identified and 110 new miRNAs were discovered. The functional analysis of miRNAs with high expression showed that they had potential regulatory effects on sperm motility, metabolism and capacity. Among these miRNAs, the highly expressed miR-10 family (ssc-miR-10a-5p and ssc-miR-10b) could potentially regulate the expression of phosphodiesterase (Pde), inhibited the degradation of the second messengers (cAMP or cGMP) in sperm. Then the activation of cAMP/(protein kinase A) PKA signaling pathway and the phosphorylation of seminal a kinase anchoring proteins (AKAP) promoted the maintenance of sperm fertility. Sperm mobility inhibitor (SPMI) could inhibit sperm movement by inhibiting the motility of flagellum ATPase, which was a potential target gene of ssc-miR-204. And the high abundant ssc-miR-28-5p could target downstream regulatory N Myc downstream regulator 2 (Ndrg2), which inhibited the production of key enzymes in glycolysis. The enrichment analysis showed that target regulation of ssc-miR-28-5p could promote glycolysis and increase the amount of lactic acid in sperm, which was consistent with the study that sperm contained a lot of lactic acid. The found miRNAs found will provide basic data for preservation of porcine semen and development of semen in this study, which has a significant research value in the study of sperm fertilization mechanism.

Meat quality is the main economic traits affecting the efficiency of sheep breeding. In order to select the high-efficiency commercial herds, transcriptome sequencing was applied for identifying the genes related to sheep growth performance and meat quality from the genetic level. The crossbred commercial generation herd (F₁) of Small-tailed Han sheep ewe (STH)×Suffolk ram (SFK) and STH×Mongolian sheep ram (MG) was selected as the object. The slaughter performance and meat quality of these population were measured, meanwhile, transcriptome sequencing of longissimus dorsi muscle was also carried out for identifying the genes related to sheep growth performance and meat quality. The results showed that the production performance, slaughter performance and meat quality of STH (♀) × SFK (♂) generation was better than that of STH (♀) × MG (♂) generation. Transcriptomic results suggested that a total of 397 differentially expressed genes (DEGs) were identified in two generation. Most of these genes were mainly enrichmented in 46 Gene Ontology (GO) items such as growth and development and 26 pathways such as carbon metabolism, PI3K-Akt (phosphatidylinositide 3-kinases/protein kinase B), and amino acid synthesis. Finally, 3 genes of adiponectin (ADIPOQ), insulin-like growth factor binding protein 7 (IGFBP7) and actin α1 cardiac muscle(ACTC1) were taken as the candidate genes associated with meat quality traits according to results of function and pathway analysis. The results of qRT-PCR revealed that the relative expression of ADIPOQ and IGFBP7 in STH × SFK generation were significantly higher than that of STH (♀)×SFK (♂) generation (P<0.01), however the expression level of ACTC1 was significantly lower than that of STH (♀)×MG (♂) generation (P<0.01). These results indicated that the STH × SFK generation was superior to the STH×MG generation in slaughter performance and meat quality. The differentially expressed genes selected can be taken as candidate genes for improving the sheep meat quality.

Fibroblast growth factor 21 (FGF21) is an important endocrine regulatory protein factor, which plays an important role in ontogenesis and metabolism. In order to analyze the effect of polymorphisms of ovine FGF21 gene on growth traits and carcass traits in lambs (Ovis arise), to filter variation of FGF21 gene. Blood samples of 5 different commodity sheep were collected, including 170 sheep were tested for polymorphism, 756 Romney lambs were analyzed for correlation which relevant production data were collected. DNA were extracted and polymorphisms in Exon-1 and Exon-3 regions of FGF21 gene were detected by PCR-single strand conformation polymorphism (PCR-SSCP), and the relationship between allele of Exon-3 regions and production traits in lambs was investigated using generalized linear models (GLMs). In total, 6 SNPs were detected in Exon-1 and Exon-3 regions of ovine FGF21 gene, the nucleotide substitution c.554 C/T in Exon-3 resulted in amino acid change (p.Pro185Leu). The association analysis results showed that the allele in Exon-3 region of ovine FGF21 gene had different effects on growth traits of male and female lambs. In male lambs, the presence of A₂ was associated with decreased tailing weight and weaning weight (P<0.05), and in female lambs the presence of B₂ was associated with decreased weaning weight and pre-weaning growth rate (P<0.05). No associations were detected between the other allele and growth traits in population. Meanwhile, the results showed that the presence of A₂ was associated with decreased hot-carcass weight (H-W), the presence of B₂ was associated with increased H-W, should yield and total yield (P<0.05). No associations were detected between the other allele and carcass traits in male lambs. Ovine FGF21 gene had abundant polymorphisms. The allele of Exon-3 of ovine FGF21 gene might have different effects on growth traits of male and female lambs, and also might affect some carcass traits, which could improve the growth traits and carcass traits of lambs, and provide basis molecular genetic data for improving important economic traits of sheep, FGF21 gene could be an effective molecular marker for marker-assisted breeding of sheep, these results could provide scientific basis for genetic improvement of sheep.

The Tibetan chicken (Gallus gallus domesticus), a chicken breed endemic to Tibetan Plateau, has adapted to the hypoxic, high-altitude environment over a thousand years. The study on the genetic diversity of the Tibetan chicken is of great significance for genetic conservation and inbreeding programs. In this study, 4 haplotypes were identified based on D-loop sequencing in Tibetan chicken (n=40), and each representative of 4 haplotypes were selected for total mitochondrial genome sequencing and analyzed together with published mitochondrial genome data of red jungle fowl. Four haplotypes belonged to 3 previously published clades, i.e., Clade A, clade B and clade E. Based on D-loop sequencing data, the average haplotype diversity and nucleotide diversity were 0.658±0.065 and 0.004 42±0.000 94, respectively. The mitochondrial genome of Tibetan chicken is 16 785 bp in size, consisting of 22 transfer RNA (tRNA) genes, 2 ribosomal RNA (rRNA) genes, 13 protein-coding genes and 1 non-coding control region. There were 7 overlapping genes and 17 intergenic regions. The phylogenetic tree had different cluster branching based on D-loop sequences and the mitochondrial genome. The results indicated that Tibetan chicken populations in this study have relatively low nucleotide and haplotype diversity and likely share multiple maternal lineages. The D-loop sequence has limited power for the resolution of phylogenetic relationships in comparison with the complete mitochondrial genome. The results of the current study can be used as baseline genetic information for genetic conservation program and in breeding.

Plant endophytic bacteria are important microbial resource. The universal primer pairs for bacteria 16S rDNA gene amplification show similarity with plant DNA sequence from organelles such as chloroplast and Mitochondria, which brings great challenge to the high-throughput sequencing of endophytic bacteria. The aim of present study is to eliminate the interference of non-target DNA to high throughput sequencing of rice(Oryza sativa) endophytic bacteria, screen out of the optimal primer pair for analysis of the endophytic bacteria in rice seeds and describe their community structure. Seeds of japonica rice variety Nipponbare were used as material. Seven universal primer pairs targeted bacterial 16S rDNA gene for high throughput sequencing were selected as candidate primers. The results of bioinformatics analysis revealed that the primer pairs dP1, dP3 and dP7 showed low homology with plant organelle DNA sequences. They were less likely to amplify the gene fragments of rice chromosomes and organelles.Thus, they were selected as the initial screening primers and tested by 16S rDNA gene high throughput sequencing using Illumina MiSeq pyrosequencing. The three selected primers produced effective DNA fragments, except that the dP7 primer pairs need to use high fidelity Taq enzyme. After quality control (QC), decontamination treatment and organelle_filtered, the number of residual good reads from dP7 products were 11.7 and 5.3 times of that from dP1 and dP3, which could reduce the interference of rice organelle gene sequences to a certain extent, and produced the most OTUs (632). So dP7 was selected as the optimal primer pair. Furthermore, rice seeds diverse bacterial endophytic which were divided into 5 Phylum, 8 class, 13 order, 32 family, and 51 genera. Among them, the first dominant bacterium genera were Bacillus and Lactococcus, and their relative abundance were 64.46% and 15.07%, respectively. The second sub-dominant bacterium genera were Pantoea and Exiguobacterium, and their relative abundance were 4.27% and 3.61%. In conclusion, different primers have great influence on the results of high throughput sequencing of plant-associated bacterial communities. Among all the tested primer pairs, dP7 was the optimal primer pair for the analysis, with which more diversity endophytic bacterial communities were obtained in rice seeds. The study provides basic data for the future research on the microbial resources and the interaction mechanism between endophytic bacteria and rice host.

Porcine epidemic diarrhea virus (PEDV) is the pathogen that causes porcine (Sus scrofa domesticus) epidemic diarrhea. PEDV infection mainly causes severe diarrhea, vomiting and dehydration in pigs, resulting in extremely high mortality of pigs. The mortality rate of piglets within 6 days of age after infection with PEDV is even as high as 100%, which has caused great economic losses to the pig industry in the world, which seriously affected the healthy development of the pig industry. In order to study the pathogenic mechanism of PEDV, PEDV was inoculated into Vero cells cultured in vitro, and total RNA from non-infected PEDV control group, PEDV 12 h group, and PEDV 24 h group Vero cells was extracted for Illumina high-throughput sequencing. 56789250, 48406834, 52574920, 51348394, 62905540, 54244594, 50051472, 67574592, and 57516788 sequencing raw data were obtained, respectively. The raw data was filtered, and 56126966, 47748166, 51932440, 50756174, 62190894, 53605948, 49571034, 66728750, and 56851386 filtered data (clean reads) were obtained. The correlation coefficients between samples were all greater than 0.98, indicating that the correlation between samples was qualified. Bioinformatics analysis of the sequenced data revealed that compared with the group uninfected with PEDV, there were 4 differentially expressed genes in the group infected with PEDV 12 h (P<0.05), and 1 498 differentially expressed genes in the group infected with PEDV for 24 h (P<0.05), respectively; compared with the group infected with PEDV for 12 h, there were 1 643 differentially expressed genes in the group infected with PEDV for 24 h (P<0.05). Five differentially expressed genes were randomly selected for qRT-PCR verification analysis, and the verification results were consistent with the transcriptome sequencing results, indicated that the transcriptome sequencing results were relatively reliable and can be used for further bioinformatics analysis. The Gene Ontology (GO) function annotation analysis was performed on the differentially expressed genes screened above. The GO annotations of differentially expressed genes were classified into three aspects of biological process, cell composition, and molecular function, respectively, and involved cell process, biological regulation, metabolic processes, cell composition, organelle composition, and catalytic activity. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was performed on the above-mentioned differentially expressed genes, and the results showed that when Vero cells were infected with PEDV for 12 h, 4 differentially expressed genes in the African green monkey (Chlorocebus sabaeus) genome gene annotation were enriched in the NOD-like receptor signaling pathway and microRNAs in cancer signaling pathway; For 24 h post-infection, some cells exhibited severe cytopathic effects, while most of the cells were not detached from the well bottom of culture plates, the differentially expressed genes in the African green monkey genome gene annotation were mainly concentrated in TNF (tumor necrosis factor) signal pathway, P53 (tumor protein 53) signal pathway, Jak-STAT (Janus kinase/signal transducer and activator of tran-ions) signal pathway, MAPK (mitogen-activated protein kinase) signal pathway and immune inflammation, respectively. According to the signal pathway activated by PEDV infection, it can be judged that when cells were infected with PEDV, they can affect the virus's ability to invade and replicate by regulating autoimmunity and inducing apoptosis. The differentially expressed genes found in this study provide data references for further research on the pathogenic mechanism of PEDV.

Baculoviruses are a family of enveloped, rod-shaped, double-stranded DNA viruses which are specifically pathogenic to arthropods, mainly belonging to the orders Lepidoptera, Hymenoptera, and Diptera. ac78 of the archetype of the Baculoviridae, Autographa californica multiple nucleopolyhedrovirus (AcMNPV) was one of the 38 core genes of baculoviruses. Previous studies showed that Ac78 played an important role in the baculovirus life cycle, and Ac78 conserved amino acid (aa) 2~25 and 64~88 were essential for Ac78 functions during baculovirus life cycle, but the mechanism by which Ac78 employs that affected the budded virion (BV) production and multiple nucleocapsid-enveloped occlusion-derived virion (M-ODV) formation was still unknown. An aa alignment of the Ac78 homologs showed that Ac78 N-terminal aa 9 (R9) and 22 (K22) were highly conserved in baculoviruses, suggesting that R9 and K22 might be important in the functions of Ac78. In the present study, the ac78 with a HA tag prior to the stop codon, in which the codon of R9 or K22 was mutated into the codon of alanine (A), together with the enhanced green fluorescence protein gene (egfp; referred to as gfp in the present study) and the AcMNPV polyhedrin (polh) gene, was inserted into the polh locus of the ac78-knockout AcMNPV bacmid (bAc78KO) to construct the recombinant viruses, vAc78R9A:HA or vAc78K22A:HA. All recombinant viruses were confirmed by PCR analysis and further DNA sequencing. The ac78-knockout virus (vAc78KO), ac78-repaired virus (vAc78:HA), vAc78R9A:HA, or vAc78K22A:HA bacmid DNA was transfected into Sf9 (Spodoptera frugiperda IPLB-Sf21-AE clonal isolate 9) cells, viral replication and infection were monitored by a fluorescence microscope. At 24 h post transfection (h p.t.), no obvious differences in the amounts of GFP-positive cells were observed among all 4 groups, indicating comparable transfection efficiencies. At 72 h p.t., almost all Sf9 cells transfected with vAc78R9A:HA or vAc78:HA bacmid DNA showed GFP fluorescence, the number of fluorescent cells increased slightly onwards in the vAc78KO- or vAc78K22A: HA-transfected cells, but the amount of GFP-positive cells in Sf9 cells transfected with vAc78K22A:HA was a little more than that in Sf9 cells transfected with vAc78KO. Using virus supernatants harvested from vAc78KO-, vAc78:HA-, vAc78R9A:HA-, or vAc78K22A:HA-transfected Sf9 cells, viral growth curve analysis was performed by fifty percent tissue culture infective dose endpoint dilution assay to further evaluate the ability of vAc78R9A:HA and vAc78K22A:HA to produce infectious BVs. There was no significant difference between vAc78R9A:HA and vAc78:HA in capabilities to generate infectious BVs, while the titer of vAc78K22A:HA was approximately 100-fold lower than that of vAc78:HA at 120 h p.t. The results showed that the R9 mutation had no effect on infectious BV production, but the K22 mutation significantly reduced the production of the infectious BVs. Thin sections of vAc78KO-, vAc78:HA-, vAc78R9A:HA-, or vAc78K22A:HA-transfected Sf9 cells at 72 h p.t. were observed using an electron microscope. In vAc78:HA- or vAc78R9A:HA-transfected Sf9 cells, numerous M-ODVs and occlusion bodies (OBs) embedding the numerous M-ODVs emerged in the ring zone, but in the vAc78KO- or vAc78K22A:HA-transfected cells, the M-ODVs in the ring zone and OBs were significantly fewer than those in the vAc78:HA-transfected Sf9 cells, showing that the R9 mutation did not affect the M-ODV formation, or the embedding of M-ODVs into the OBs, while the K22 was important in the formation of M-ODVs and the embedding of M-ODVs into the OBs. Taken together, the results demonstrated that Ac78 R9 was not essential for the infectious BV production and M-ODV formation, but K22 played an important role in the Ac78 functions. This study enriches the information of Ac78, and would lay a foundation for uncovering the mechanism employed by Ac78.

At present, bacteria are the main microorganisms to inhibit tobacco bacterial wilt, and Actinobacteria are few. In this study, Actinobacteria, which can be used to develop and produce bio organic fertilizer for controlling tobacco bacterial wilt, were screened to determine their taxonomic status. Firstly, antagonistic Actinobacteria against tobacco bacterial wilt were screened from tobacco (Nicotiana tabacum) rhizosphere soil by the method of inhibition zone assessment. The taxonomic status of antagonistic Actinobacteria was determined by polyphase analysis of phenotypic and physio biochemical characterizations, in combination with genetic characteristics. The results showed that 3 Actinobacterial strains with good antagonistic effect against tobacco bacterial wilt were obtained, and the 3 Actinobacterial strains could inhibit the pathogen of tobacco brown spot disease (Alternaria alternata) and other bacterial wilt pathogens of Solanaceae plants such as capsicum (Capsicum annuum), tomato (Solanum lycopersicum) and potato (Solanum tuberosum). In the 3 strains, SC-105-B-10 (GenBank No. KT633863) belonged to Streptomyces microflavus, L2-003-A-5 (GenBank No. KT119348) belonged to Streptomyces albidoflavus, and SC-168-A-2 belonged to Streptomyces pratensis (GenBank No. KR140205). This study obtained 3 Actinobacterial strains, which have potential for the development of bio-organic fertilizer for controlling tobacco bacterial wilt.

R2R3-MYB transcription factors play a crucial regulatory role in a series of plant biological processes, and is involved in the growth and development, synthesis of primary and secondary metabolites, and hormone signaling of fruit and vegetable plants. R2R3-MYB transcription factors are targeted by researchers, which can promote the creation of new consumer products and the development of the fruit and vegetable industry. This review summarized the gene structure characteristics of R2R3-MYB transcription factor family members, and discussed their regulatory roles in the formation of fruit and vegetable qualities, and their molecular regulatory mechanisms were described, in which the directions to future research were suggested.

Salmonellosis is a common disease in poultry, which is mainly caused by multiple Salmonella serotypes. It can be transmitted to human through fresh or undercooked meat, eggs and milk. The immune response and genetic resistance of chicken (Gallus gallus) to Salmonella has been studied through different animal infection models. High-throughput approaches will further accelerate the study of mechanism of genetic resistance to Salmonella. Here, the factors affecting the animal infection model, the basic research methods and the mechanism of chicken genetic resistance to Salmonella infection were reviewed. With the appearance of more resistance genes and finer genome map, it is expected to select out resistant breeds, and reduce the spreading of Salmonella in poultry and human.

The specific sites of physical association between endoplasmic reticulum (ER) and mitochondria are known as mitochondria-associated endoplasmic reticulum membrane (MAM), which plays an important role in the regulation of mitochondrial dynamics, calcium signaling, lipid processing and transportation, mitochondrial autophagy and endoplasmic reticulum stress. These physiological functions of MAM are very important for oocyte maturation. It has recently become clear that many proteins in MAM are crucial for oocyte maturation. This article reviews the research progress of MAM involved in oocyte maturation in recent years in order to provide a new direction for understanding the regulation mechanism of oocyte maturation, oocyte quality identification and improving the success rate of assisted reproduction.

Interspecies distant hybridization is one of the methods to create new germplasm of fruit trees, but how to detect the true interspecies hybrids has always lacked efficient and reliable technical means. This research was based on genomic research and the comparative analysis of the whole genome sequence of apple(Malus) and pear (Pyrus). Based on the indels from the 2 genomes, the InDel specific molecular marker was designed. This study combined PCR detection, and screened out 6 pairs of specific molecular markers with high specificity, good repeatability and clear amplification bands. Among them, 3 pairs of specific sequences from pear genome and 3 pairs of specific sequences from apple genome could effectively detect the differences between them. The above 6 pairs of specific molecular markers were used to identify 311 individuals from the progeny of the cross between the 3 groups of pear and apple. The results showed that the progeny of 7 strains could simultaneously amplify specific bands of apple and pear, which were identified as inter-genus hybrids that accounting for 2.3% of the total hybrid progeny. Among them, 1 was Malus domestica cv. 'Starking'×'Comice', 3 were 'Golden Delicious'×'Dangshansuli', 3 were 'Fuji''×'Comice'. Therefore, 6 pairs of inter-generic specific primers for pear and apple can be used for identification of distant hybrids at seedling stage. The technical method developed in this research to use gene-specific molecular markers to identify the intergeneric hybridization of 'Rosaceae' Pyrus and Malus would not only provide efficient and accurate detection markers for the distant hybridization of apples and pears, but also provides a valuable example for identification of inter-genus hybrids of other fruit trees.

Wild rice stem is the expanded edible fleshy stem of Zizania latifolia, a Gamineous plant, when it is invaded by Ustilago esculenta. The selection of appropriate internal reference genes is important for analyzing the related gene expression profile in the study of the interaction between Zizania latifolia and Ustilago esculenta. In this study, the expression profiles of 8 candidate genes in U. esculenta were evaluated by qRT-PCR under different environmental conditions including 4 temperatures, 3 carbon source media, 5 developmental stages in vitro, and 5 time points during infection process. Data were analyzed using geNorm, NormFinder and BestKeeper software. In order to test the stability of the selected genes, the endoglucanase 1 gene (UeEgl1) was used as the test gene. The results showed that glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH) and α-tubulin expressed stably in all samples and were suitable for expression pattern analysis of U. esculenta. The results of this study could provide an effective correction tool for gene differential expression analysis of U. latifolia, ensure the accuracy and reliability of gene expression results, and provide reference for further analysis of the function of related genes in the process of interaction between U. latifolia and Z. latifolia.