Required for Mla12-required resistance (RAR1) as an important element for plant disease resistance, is widely involved in plant resistance response. To study the structure and expression characteristics of SiRAR1 in foxtail millet (Setaria italica), the CDS sequence and promoter sequence of SiRAR1 gene were separately cloned with the leaf cDNA and genomic DNA from resistance material 'Shilixiang' as template by PCR, and the biological characteristics of SiRAR1 were predicted by bioinformatics analysis. Then the expression patterns of SiRAR1 in different tissues and the Uromyces setariae-italicae-induced stress response were surveyed by qRT-PCR, respectively. Lastly the prokaryotic expression characteristics for the gene were detected by SDS-PAGE. The results showed that the ORF of SiRAR1 gene contained 765 bp and encoded 254 amino acids (GenBank No. MK814879). The predicted protein molecular weight was 28.04 kD and the theoretical isoelectric point (pI) was 8.13. The largest secondary structure element of SiRAR1 protein was random coil, and the smallest was beta turn. Phylogenetic analysis showed that SiRAR1 protein had the closest evolutionary relationship with plants in the same family, and had the highest genetic homology (92.44%) with the hypothetical protein C2845_PM12G15490 (RLM79685.1), a gene from chromosome 12 of Panicum miliaceum. The qRT-PCR analysis showed that SiRAR1 gene was expressed in roots, stems, leaves and panicles in foxtail millet. There was the highest expression level in roots and the lowest in stems, and the expression levels of SiRAR1 were similar in panicles at booting and heading stages. The qRT-PCR analysis also showed that under the infection of Uromyces setariae-italicae, the expression level of SiRAR1 gene in the resistant material 'Shilixiang' was up-regulated from 12 to 36 h and peaked at 36 h, and up-regulated at 96 h in the susceptible material 'Yugu1'. The peak expression of SiRAR1 gene in 'Shilixiang' was about 3 times higher than that in 'Yugu1' (P<0.01), and its peak expression in 'Yugu1' was about 1.6 times higher than that in 'Shilixiang' (P<0.05). The results showed that the expression of SiRAR1 gene had earlier up-regulated time, longer duration and stronger up-regulated range in resistant plants than that in susceptible plants. The prokaryotic expression vector pET30a-SiRAR1 was induced to express SiRAR1 fusion protein with 33 kD by 0.1 and 0.4 mmol/L isopropyl β-D-thiogalactoside (IPTG), respectively. The results provide a theoretical foundation for further study on the SiRAR1 function and its disease resistance mechanism in foxtail millet.

Due to environmental pollution and human activities, the amount of loach resources in China has been sharply reduced, and germplasm resources have been degraded. In order to explore the genetic diversity and population genetic structure of different geographical populations of loach (Misgurnus anguillicaudatus) in China, 10 microsatellite markers were used to analyze the genetic diversity of 7 populations in Poyang Lake region and Pearl River Basin of China (Poyang Lake, Yingde, Qingyuan, Liannan, Nanxiong, Meizhou, Liuzhou). The results showed that the average number of allele (Na) and effective allele (Ne) per locus were 7 and 2.5, and the average observed heterozygosity (Ho) and expected heterozygosity (He) were 0.327 4 and 0.396 0. The average population inbreeding coefficient (Fis), genetic differentiation coefficient (Fst) and number of migrants per generation (Nm) of the 7 loach populations were 0.093 0, 0.372 0 and 0.422 1, indicating that the level of genetic exchange among the 7 populations was low and there was a high degree of genetic differentiation. A total of 79 alleles were amplified in 10 pairs of microsatellite primers, the Na, Ne, Ho, He and polymorphic information content (PIC) ranged from 3.0 to 7.5, 1.7 to 5.5, 0.307 4 to 0.534 7, 0.329 0 to 0.861 5, and 0.316 8 to 0.529 4, respectively. The genetic distance between the 7 loach populations was 0.045 3~1.602 3. The genetic relationship of Liuzhou and Nanxiong population showed the closest with lowest distance of 0.045 3, while the genetic relationship of Qingyuan and Meizhou population showed the farthest with highest distance of 1.602 3. The clustering analysis based on genetic distance showed that the Liuzhou, Nanxiong, Liannan and Meizhou populations were clustered into one branch, while the Qingyuan, Yingde and Poyang Lake populations were clustered into another branch. Above results demonstrated that the level of genetic diversity in loach in China was moderate and the genetic differentiation among loach populations was affected by factors such as self-migration ability and geographical isolation. This study provides a theoretical basis for further selection of the basic group of loach fine breeds and the protection of loach germplasm resources.

Carotenoids, as an important nutrient, are very common in carrot (Daucus carota). Lycopene, a kind of carotenoids, plays important roles in growth, development, and response to abiotic stress in higher plant. Phytoene synthase (PSY) is one of the key enzymes of lycopene biosynthesis. In this study, DcPSY2 (GenBank No. NM_001329167.1) was cloned from carrot cv. 'Junchuanhong' by RT-PCR (reverse transcription PCR). It contained an ORF with the length of 1 317 bp and encoded 438 amino acids. The amino acid sequences of DcPSY2 and the PSY proteins of 17 other plants were compared, the results showed that the overall similarity of PSY proteins was 82.22 %, which indicated that PSY proteins were conserved. Evolutionary relationship exhibited that DcPSY2 protein had the closest evolutionary relationship with the PSY protein in mahogany (Bixa orellana). DcPSY2 protein was a hydrophilic and non-secretory protein without signal peptide. It belonged to isoprenoid biosyn C1 superfamily and was distributed in various organelles. Secondary structure prediction demonstrated that DcPSY2 contained 53.65 % α-helix, 5.48 % β-fold, 12.79 % extended main chain and 28.08 % random curl. The results of qRT-PCR showed that DcPSY2 gene expression changed significantly in response to 4 kinds of abiotic stresses, including high temperature (38 ℃), low temperature (4 ℃), drought (200 g/L PEG6000) and high salt (200 mmol/L NaCl). Under high temperature or high salt treatments, the relative expression level of DcPSY2 reached the peak after 2 h. Whereas under low temperature or drought treatments, the expression level of DcPSY2 increased significantly at 1 h (P<0.05). The results provide a basic material for the functional study of DcPSY2 gene in carrot growth and its application study in stress resistance breeding.

Chinese cabbage (Brassica rapa spp. pekinensis) originated in China, and is an important vegetable in China and even in East Asia. Virus disease is one of the most important diseases for Chinese cabbage production, in which Turnip mosaic virus (TuMV) is the main pathogen. The inheritance pattern for TuMV resistance is very complicated in Chinese cabbage. In order to better understand the similarities and differences of resistance sites between different TuMV resistant materials, the high-generation inbred line TuMV resistant material '73' and National TuMV source resistant material '8407' were used as core parents to study the similarities and differences of TuMV resistant sites in 9 resistant materials ('73', '8407', 'Zao 219', '07-372', C07-14', 'Henan 304', '322', '826' and '07-55'). By crossing and inbreeding, F₁ and F₂ populations of 'Zao219'×'8407', '07-372'×'8407', '07-14'×'8407', 'Henan304'×'8407', '322'×'8407', '826'×'8407', '07-55'×'8407', '73'×'8407', 'Zao219'×'73', '07-372'×'73', 'Henan304'×'73', '322'×'73', '07-55'×'73', '07-372'×'322', 'Henan304'×'322', '07-372'×'Henan304', 'Zao219'×'322', 'Zao219'×'Henan304' and 'Zao219'×'07-372' were constructed. The C4 strains of TuMV was inoculated on the above F₁, F₂ and parent materials. The similarities and differences of resistance sites among different combinations were judged according to the resistance performance of F₁ and the resistance isolation of F₂. The results showed, in combinations '73'×'8407' and '322'×'8407', both F₁ were resistant, individual susceptible plants were found in both F₂ populations, indicating that the resistance sites in '73' and '322' were linked to that in '8407' but not allelic. In combination '07-55'×'8407', F₁ was resistant, no susceptible plants were found in F₂, but 3 grade 1 and 6 grade 3 plants, indicated that the resistance site in '07-55' and '8407' were incomplete allelic, should be linked. In combinations '322'×'73' and '07-55'×'73', F₁ and all of F₂ plants were resistant, no susceptible plants were found in F₂, indicating that the resistance sites of '322', '07-55' and '8407' were allelic, and their resistance was controlled by the same gene. In combinations 'Zao219'×'8407', '07-372'×'8407', '07-14'×'8407', and '826'×'8407', all of the F₁ and F₂ plants were resistant, no susceptible plants were found in F₂, indicating that the resistance sites of 'Zao219', '07-372', '07-14', '826' and '8407' were allelic, and their resistance were controlled by the same gene. In combination 'Henan304'×'8407', F₁ was susceptible, both resistant and susceptible plants were found in F₂, and the segregation ratio of resistant and susceptible plants was 9∶7, indicating that the resistance sites of 'Henan304' and '8407' were not allelic, TuMV resistant genes in 'Henan304' and '8407' were complementary. In combination 'Henan304'×'322', F₁ was resistant, both resistant and susceptible plants were found in F₂, and the segregation ratio of resistant and susceptible plants was 15∶1, indicated that the resistance sites of 'Henan304' and '322' were not allelic, TuMV resistant genes in 'Henan304' and '322' were inherited independently. This result provides a basis for the next step of targeted selection of disease resistant materials and research on the molecular markers of resistance to TuMV in Chinese cabbage. It also broadens the germplasm resources of resistance of Chinese cabbage for TuMV resistant breeding, and provides theoretical guidance for the selection of parents and offspring of resistance breeding of Chinese cabbage to TuMV.

Ethylene responsive factors (ERFs) are plant-specific transcription factors that play important roles in plant developmental process and in response to abiotic stress. In order to explore the function of OsERF103 (GenBank No. XM_015768788) in rice (Oryza sativa) , bioinformatics methods were used to analyze the sequence characteristic of OsERF103. For subcellular localization analysis, the fusion expression vector of pBWA(V)HS-OsERF103-Glosgfp was constructed and transformed into rice protoplast, then laser scanning confocal microscopy was used to observe the subcellular localization of OsERF103 fusion protein in rice protoplast; Expression analysis of OsERF103 in different tissues and under different abiotic stress conditions was conducted by qRT-PCR. The results showed that OsERF103 was a hydrophilic and unstable protein without signal peptide and transmembrane structure, and contained an AP2 (APETALA2) domain. Phylogenetic analysis revealed the OsERF103 was evolutionarily closest to a abscisic acid repressor 1-like (ABR1-like) protein of Oryza brachyantha. The results of subcellular localization showed that OsERF103 was located in the nucleus. OsERF103 transcripts were detected in all sampled tissues at booting stage, with the highest level detected in root and weak level detected in leaf and young spike. OsERF103 transcripts were induced by high temperature, low temperature, PEG6000, high salt and abscisic acid (ABA), and differential expression patterns of OsERF103 were showed under the different treatment conditions. This study provides basic data for further exploring the biological function of OsERF103.

The 'Tonami' large-fruit mutant derived from the natural mutant of 'Tonami' apple. To analysis the reason of large-fruit mutant of 'Tonami' apple (Malus domestica 'Tonami'), fruit size, cell ploidy, cellular structure and transcriptome in the early development of fruit were studied in 'Tonami' apple and its large-fruit mutant. Results showed that weight of average single fruit of 'Large-fruit Tonami' apple was significantly higher than the contrast (P<0.05), was 1.5 times of the contrast. There was no significant difference in the cell size of the fruits, but the number of cells in the 'Large-fruit Tonami' at 28 d after full bloom and at the ripeness was significantly higher than that of 'Tonami' (P<0.05). Cell ploidy were the same of 'Large-fruit Tonami' apple and contrast. Transcriptome of fruit after full bloom 14 and 28 d were compared with 'Tonami' and 'Large-fruit Tonami' apple. Fifty-one differential expressed genes (DEGs) were identified. By KEGG analysis, these unigenes were found to be implicated in plant hormone signal transduction pathway. Five genes that might be related to large-fruit mutant were selected from that pathway, these genes were mainly involwed in cytokinin signaling transduction pathway and ethylene signaling transduction. The study showed that large-fruit mutant of 'Tonami' apple might be related to the changes of plant hormones in fruit development. This study would provide basic information for further studying the cytological and molecular basis on apple large-fruit mutant.

SnRK2 (sucrose non-fermenting-1-related protein kinase) is a kind of plant-specific Ser/Thr protein kinase, which plays an important role in plant stress resistance. In order to explore the number, structure and expression difference of SnRK2 gene family of strawberry (Fragaria vesca) in response to abiotic stress, based on the conserved sequences of SnRK2 genes in Arabidopsis thaliala and rice (Oryza sativa).Using bioinformatic tools, the homologous comparison, gene structure, systematic evolution, conserved motif, physical and chemical properties of protein, protein secondary structure and sequential action elements of strawberry SnRK2 gene family were analyzed, and qRT-PCR was used to analyze the gene expression under simulated stress. Ten members of SnRK2 gene family were identified from strawberry genome database, divided into 3 subfamilies, the number of coding amino acids is 258~366, and the molecular weight was 29 469.75~41 481.68 D, the theoretical isoelectric points were between 4.68 and 9.10, which were distributed on 4 chromosomes of strawberry. Gene structure and motif analysis showed that most genes had 9 exons, and the distribution of motif in the same subfamily was basically similar. The results of subcellular localization prediction showed that FvSnRK2 gene family was mainly expressed in cytoplasm. The secondary structures of protein were mainly α-helix and irregular curl. Analysis of the upstream 2 kb cis-acting element found that all members of the FvSnRK2 gene family contain MYB and MYC transcription factor response elements. The results of qRT-PCR analysis showed that the relative expression of 5 genes in FvSnRK2 gene family reached the peaks at 12 h after 10 % PEG6000 processing, and the response of FvSnRK2.4 was the most obvious, which was 15 times higher than that at 0 h. After 100 μmol/L ABA (abscisic acid) treatment, the relative expression of 6 genes was the highest at 24 h, and 2 genes had the highest relative expression at 12 h, FvSnRK2.5 and FvSnRK2.9 responsed strongly, which were 16.2 times and 42.4 times higher than that at 0 h, respectively. The relative expression of 7 genes was highest at 2 h after 200 mmol/L NaCl treatment, the response of FvSnRK2.10 was 65.2 times higher than that at 0 h, and FvSnRK2.5 was the highest at 12 h, which was 94.7 times higher than that at 0 h. FvSnRK2.5 and FvSnRK2.10 had strong response after ABA and NaCl treatment, and played important roles in stress resistance signal regulation in strawberry. The present research provides theoretical reference for functional verification and breeding of strawberry SnRK2 gene.

Lycoris sprengeri is perennial herbaceous plants of Amaryllidaceae, Lycoris with the red and blue multicolor flower, which is a good material for cut flower, potted flower and landscape gardening, so L. sprengeri has the very high commercial value. To enrich the germplasm resources and cultivate varieties in L. sprengeri, a R2R3-MYB transcription factor gene LsMYB5 cDNA sequence (GenBank No. MK779710) was successfully screened by using reverse transcription PCR (RT-PCR) and rapid amplification of cDNA ends (RACE). The LsMYB5 gene length was 871 bp, and the open reading frame (ORF) was 681 bp, which encoded 226 amino acids. The c-terminal of LsMYB5 protein contained conservative EAR inhibition domain (pdLNLD/ELxiG/s) and zinc finger domain (CX-(1-2), CX-(7-12), CX-(1-2)C), which may have transcriptional inhibition function. LsMYB5 gene mainly expressed in leaves and the fading period of petals, and there were significant differences in the expression of LsMYB5 in clones with different colors (P<0.05). To further analyze LsMYB5 gene function, prokaryotic expression vector of pET-30(a)-LsMYB5 was constructed and transformed into Escherichia coli BL21 (DE3), the expression of recombinant proteins was induced under the induction of isopropyl glucosinolates galactose glucoside (IPTG), and the His label protein was purified. The results will provide the theory basis for screening structural genes regulated by LsMYB5 transcription factor. which related to flower color formation in Lycoris sprengeri.

Crowtoe (Lotus corniculatus) is a legume forage with broad application prospects, but the efficiency of promotion and utilization is seriously affected by drought. In order to solve this problem, after many years of research, the high drought tolerance crowtoe germplasm B08 was selected by this team, finally. In order to further study and explore the drought-resistance mechanism of germplasm B08, suppression subtractive hybridization (SSH) technology was used to construct the positive and negative SSH cDNA library of B08 under drought stress. 600 positive clones were randomly selected from the forward and reverse libraries for sequencing analysis. The results showed that: (1) 574 and 595 high quality sequences were obtained in the forward and reverse libraries, with an average length of 207.5 bp and 242.7 bp, respectively. Finally, 632 unigenes sequences were obtained by assembling high quality sequence data, of which 178 were forward library and 454 reverse library. (2) The 632 unigenes were compared with public databases by BLAST, and 147 genes were annotated successfully. Among them, 80 were known genes, 67 were unknown genes, and 485 new genes were found. (3) The results of NCBI Non-Redundant Protein Sequence Database (nr) annotation showed that there were high homology with Populus trichocarpa, Malus domestica, Spinacia oleracea, Hordeum vulgare, Brassica napus, Cajanus cajan, Cicer arietinum, Coffea canephora and so on. The forward and reverse libraries were classified using Gene Ontology (GO) program. The genes involved in cell, cell part, organcell, binding, metabolic process, and catalytic activity, cellular process accounted for a large proportion. (4) The obtained unigenes involved the synthesis of plant photosynthesis, energy metabolism, hormone synthesis, protein metabolism, nucleic acid metabolism, ion transport and signal transduction. (5) Substances closely related to drought resistance of plants had been found, such as serine/threonine-protein phosphatase, calmodulin-like protein (CML), indole-3-acetic acid-induced protein ARG7, mitogen-activated protein kinase (MAPK) et al. The results of this study provides information for further cloning and verifying the drought-resistant genes, digging new drought-related genes, understanding the molecular mechanism of drought-resistance, and breeding new varieties with high drought-resistance.

Major genes related to growth traits of goat (Capra hircus) were selected as marker assisted selection which can improve the accuracy of selection and accelerate breeding progress. The current study aimed to analyze gene expression characteristics of insulin-like growth factor-1 (IGF-1) gene in skeletal muscle, SNPs distribution and its association with growth traits of goats. qRT-PCR was used to analyze IGF-1 gene expression level in masseter, trapezius, semispinalis, arm triceps, extensor carpi radialis longus, longissimus dorsi, psoas, quadriceps femoris, biceps femoris of Fuqing goat and Nubia black goat. PCR sequencing and high-resolution melting (HRM) technologies were carried out to screen SNPs and analyzed their association with growth trait in Fuqing goat (n=124), Nubia black goat (n=152), Jianyang big ear goat (n=121), Daiyun goat (n=20). The results revealed that IGF-1 gene was expressed in 9 skeletal muscle tissues of Fuqing goat and Nubia black goat, and the expression level was different in skeletal muscle tissues and breeds. The expression level of IGF-1 gene in 9 skeletal muscle tissues of Fuqing goat was lower than that of Nubia black goat. The expression level of IGF-1 gene in masseter and quadriceps femoris was extremely significantly different between the 2 breeds (P<0.01), and the expression level of IGF-1 gene in arm triceps and trapezius and longissimus dorsi was significantly different between the 2 breeds (P<0.05). The exon sequence of IGF-1 gene was highly conserved, and no mutation was found in Fuqing goat and Nubia black goat. Most of the 3 SNPs of the 4 breeds showed moderate polymorphism distribution, PIC was 0.18~0.38; the 3 SNPs had obvious variety specificity on the growth traits of goats, and had significant influence on some growth traits of Jianyang big ear goat and Nubia black goat, but had little influence on the growth traits of Fuqing goat. The results of this study reveals the difference of IGF-1 gene expression in the skeletal muscles of Fuqing goat and Nubia black goat, and provides basic data for further exploring the mechanism of goat growth and development. The research results unveile that 3 SNPs within IGF-1 have strong effects on many growth traits of goats, which could be used as candidate markers for marker assisted selection of greater growth performance.

Bian chicken (Gallus gallus) is a valuable genetic resource as a Chinese native chicken. It is of great significance to explore the genetic diversity and molecular genetic evolution of the population at the molecular level. In this study, RAD-seq simplified genome sequencing technology was employed to identify SNP markers in Bian chickens and compare the genetic differences between the Bian chicken population and the other breeds population by calculating the genetic statistics. Functional annotation was conducted for the selected genes during evolution. The results showed that 319 727 SNP markers were identified in Bian chickens. The average observed heterozygosity (0.193) and nucleotide diversity (0.231) were in the medium levels, but inbreeding coefficient(0.167) was relatively high.The linkage disequilibrium analysis showed that there was no strong linkage disequilibrium between the alleles of Bian chickens population. The clustering results of the structure model showed that Bian chicken and Dagu chicken, Anyi gray chicken, Langshan chicken had a close relationship and belonged to the same type. 200 selected genes were identified through the Fst and θπ testing. Gene Ontology (GO) and KEGG analysis showed that these selected genes were mainly enriched in metabolism, neuroactive ligand-receptor interaction, mitogen-activated protein kinase (MAPK), vascular smooth muscle contraction signal pathways. The function analysis of the genes including nitric oxide synthase (NOS2), heterochromatin protein 1 binding protein 3 (HP1BP3), aprataxin (APTX), prostaglandin endoperoxide synthase 2 (PTGS2) and phospholipase A2 group 4 type alpha (PLA2G4A) was helpful to understand the molecular mechanism of special traits such as cold resistance, hypoxia, fertility, nervous system development and stress resistance in Bian chicken. The results of the study provide molecular basis for the genetic evolution and evaluation and conservation of local chicken breeds.

Bombyx mori nucleopolyhedrovirus (BmNPV) is a highly pathogenic virus in the sericultural industry and often results in severe economic losses. Some proteins are found to show antiviral activity against BmNPV in previous studies, but little is known about the correlations between expression levels of these antiviral related genes and resistance abilities of different genetic materials against BmNPV. In this study, to explore the correlations, the highly resistant parent (R) was crossed with the highly susceptible parent (S) to generate different types of genetic materials such as F₁, F₂, and backcrosses. The resistance to BmNPV and the corresponding relative expression levels of 5 antiviral related genes including Bmlipase-1, serine protease Ⅱgene (BmSP-2), ribosomal protein S3A gene (BmS3A), suppressor of profilin 2 gene (BmSop2) and nicotinamide adenine dinucleotide phosphate oxidoreductase gene (BmNOX) in different genetic materials were detected by BmNPV attack test and fluorescent quantitative PCR (qRT-PCR) technology.The results showed that BmSP-2 and Bmlipase-1 specifically expressed in midgut, other genes showed no tissue specificity. The relative expression levels of 4 genes (Bmlipase-1, BmSP-2, BmNOX and BmSop2) were higher in midguts of different types of genetic materials with stronger viral resistance, which had significant differences in different types of genetic materials, being consistent with their resistance abilities. However, there was no correlation among the expression levels of BmS3A. The highest level of consistency between expression levels of these antiviral related genes and resistance abilities of different types of genetic materials against BmNPV had been observed in Bmlipase-1. It was concluded that relative expression levels of Bmlipase-1 could be one of the basis in selecting anti -BmNPV materials in silkworm breeding.

Entomopathogenic nematodes (EPNs) are a class of highly effective and safety biocides whose capacity of reproduction is an important factor which affect the yield and quality of these EPNs as products potential. According to the differentially expression genes (DEGs) profile of the rabditid-EPN Heterorhabditidoides chongmingensis constructed in previous study, a RNA-binding protein lin-41 coding gene (lin-41) involved in regulating the development of nematodes was selected. The role of lin-41 in the growth and development of H. chongmingensis was revealed by identifying the biological functions of this gene in the nematode. Conservative analysis results showed that the conserved region of lin-41 of H. chongmingensis contained the non Hodgkin's lymphoma (NHL) superfamily domain and multiple NHL repeats. The amino acid sequences of lin-41 of H. chongmingensis, Ancylostoma duodenale, Dictyocaulus viviparus, Caenorhabditis elegans, C. brenneri and other related nematodes were highly conserved. The conserved NHL superfamily domain of the functional region fragment of lin-41 was selected as interfering fragment, and then the double-stranded RNA (dsRNA) expression vector of the fragment was constructed by L4440 plasmid vector and transformed into Escherichia?coli HT115 (DE3). The expression of the target gene was disturbed in the H. chongmingensis by feeding with the transformed E. coli HT115 (DE3). The effects of RNAi on the expression level of lin-41 and the reproductive related G protein α subunit gene (goa-1) were detected by qRT-PCR. The biological characteristics changes of the H. chongmingensis nematodes after lin-41 RNAi were recorded. The relative expression of lin-41 in H. chongmingensis after fed with the transformed E. coli HT115 (DE3) inducted by isopropyl-beta-D-thiogalactopyranoside (IPTG) with 0.1 mmoL/L for 4 h was reduced by 72.08% than that of the L4440 group. The number of eggs laid by the lin-41 RANi group was significantly less than that of the L4440 group, with an average of 45 eggs reduced (P<0.05). However, there was no significant change in the hatching rate of eggs between the 2 groups. These results indicated that the decreased in the expression of lin-41 neither affected the normal development and the hatch of eggs nor the quality of eggs. There were no significant change in body length between the 2 groups, although the female rate of the lin-41 RNAi group was slightly higher than that of the control group. These results indicated that the reducing expression level of lin-41 in H. chongmingensi neither affected the growth nor the sex differentiation of the nematode. In addition, RANi of lin-41 significantly reduced the expression level of goa-1 (P<0.05), which indicated that lin-41 not only participated in the oogenesis of nematodes but also affected the spawning behavior of nematodes in the complex reproductive system of H. chongmingensis by influencing the expression level of other gene related to reproduction in nematodes. This result also suggested that the regulatory network of egg laying behaviors of H. chongmingensi including both lin-41 and goa-1. The biological function of lin-41 in H. chongmingensis was explored by RNAi method in present study, which involved in regulating the oogenesis of nematodes to affect the egg production of H. chongmingensis and in the egg laying behaviors of the nematode by regulating expression level of goa-1. This study provides important theoretical basis and technical support for the subsequent expansion, development and utilization of H. chongmingensis nematodes in agriculture.

Shengzhou Nane (Prunus salicina var. taoxingli) usually ripe in mid-July, high temperatures and frequent rains accelerate water loss, softening, microbial attack and decay during storage. The aim of this study was to investigate and identify pathogens, reduce occurrence of diseases, and isolate and identify pathogenic bacteria as well as to detect the possible host. Initially, effects of light, temperature, pH and different C and N sources on colony growth was explored, and NaHCO₃ and chitosan oligosaccharide (COS) were used for antibacterial test. The results showed that the pathogen was Byssochlamys sp. (B-TXL-02), which can cause disease in a variety of fruits, including pear (Pyrus spp.), apple (Malus domestica), peach (Prunus persica), cherry (Prunus avium), apricot (Prunus armeniaca), grape (Vitis vinifera), mango (Mangifera indica), banana (Musa spp.) and others fruits, indicating that the host had broad spectrum. Continuous illumination inhibited colony growth. Optimal factors for Byssochlamys sp. growth were temperature 30 ℃, pH 5, D-fructose and glucose for C source, beef extract and yeast extract for N source. When added into the potato dextrose agar, NaHCO₃ and COS could significantly inhibit colony growth in the first 6 d and the first 12 d, respectively, by inhibiting spore germination and inhibited mycelial growth. Treated with 0.3% NaHCO₃+1.6% chitosan oligosaccharide, the postharvest rot rate of fruit was significantly reduced. This study provides a practical method for prevention and control of Byssochlamys sp. for Shengzhou Nane during postharvest storage.

Fusarium wilt of watermelon (Citrullus lanatus) is caused by Fusarium oxysporum f.sp. niveum (FON) which is considered as the most important soil-borne pathogens. Some safe antagonistic strain can control FON growth and reduce Fusarium wilt of watermelon. In order to screen an antagonistic strain against FON, the rhizosphere soil from vegetable crops on Qinxin farm in Qiqihar Heilongjiang province was collected and used in this study. The antagonistic strain was screened out using plate confrontation method, and identified by morphological, physiological, and biochemical and bio-control enzyme analyses, as well as 16S rDNA sequence analysis. Antifungal activities of sterile fermentation filtrate from different growth stage of antagonistic strain and the stability of sterile fermentation filtrate under different conditions were determined using growth rate method. FON spore morphology and cell membrane integrity were analyzed by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The results showed that the antagonistic strain was classified as Bacillus methylotrophicus, named as WC. The WC strain could product amylase, glucanase, protease and siderophores. The sterile fermentation filtrate from the stationary phase had stronger antifungal activity against FON compared to those from the exponential stage and death phase, and the inhibition rate of FON colony growth was 61.57%. The stability tests showed that the sterile fermentation filtrate had stronger antifungal activity against FON under 10 ℃ for 30 min compared to other testing temperature, with inhibition rate of 62.78%. The antifungal activity from WC fermentation filtrate reached to 74.54% in pH 7, the stabilities of fermentation filtrate were poor in acid and base conditions. The filtrate could be stored with stable antifungal activity at 4 ℃ for 30 d and was insensitive to UV-irradiation. The analyses from SEM demonstrated that treatment with fermentation filtrate from WC led to wrinkling and depressed changes on the surface of FON spores. The CLSM revealed that the fermentation filtrate from WC damaged membrane integrity. The above results indicate that WC is a high-efficiency antagonistic strain against FON, and could be used to control Fusarium wilt of watermelon in future.

The definite taxonomic classification and fermentation conditions of the functional bacterial could be more efficient for biofertilizer development. Plant growth-promoting rhizobacteria (PGPR) strain MT-002-B-12 is promising isolate for tobacco biofertilizer production as a potential supplementary microbe. The aim of this study is to identify its taxonomic classification, and further optimize the conditions in fermentation. The phylogeny of PGPR strain MT-002-B-12 was analyzed by phenotypic features, physiological-biochemical characteristics and 16S rDNA gene sequence alignments. The fermentation conditions of MT-002-B-12 to produce indole-3-acetic acid (IAA) were optimized by combining the single factor condition analysis and response surface analysis method. The result of classification showed that MT-002-B-12 was Klebsiella variicola. The fermentation experiments showed that the optimum fermentation conditions for MT-002-B-12 was culture volume 30% (V/V), initial pH 6.0, temperature 25 ℃, shaker speed 150 r/min, culture time 34 h; and the optimum content of nutrients in the medium was 1.11% sucrose, 1.36% peptone, 0.15‰ MgSO₄·7H₂O, 0.34‰ CaCl₂·2H₂O. The strain MT-002-B-12 enriched the species of growth promoting bacteria, and could be used as a strain resource for the development of tobacco biofertilizer.

Nonalcoholic fatty liver disease (NAFLD) is a clinical pathological syndrome characterized by hepatic steatosis and lipid storage. It is caused by fatty acid synthesis, oxidation, transport, insulin resistance, oxidative stress, gluconeogenesis and other aspects. It was found that the protein menin encoded by the tumor suppressor gene MEN1 can be widely involved in pathways that regulating the liver metabolism. MEN1/menin was an important regulator of liver metabolism and assumed to be closely associated with the pathogenesis of fatty liver disease. This article generally elucidate the research progress of menin in regulating the lipid metabolism, glucose metabolism, insulin resistance and also inflammation.

The transgenic technology has been concerned greatly by people all over the world nowadays. Genetically modified organisms (GMOs),especially the GM crops have caused several controversial issues, including health problems, ecological environmental risks and even ethical concerns. From the perspective of 'Functional Nucleic Acid' and 'Biosensors', this review summarized molecular amplification techniques, different ways of signal output and nanomaterials -based functional nucleic acid biosensors for detection of genetically modified ingredients. Finally, the challenges and trends in the future development of genetically modified components' detection are prospected. This review will be helpful for promoting the development of test techniques for GMOs and the progress of functional nucleic acid-based biosening disciplines.

Myostatin (MSTN) is an important negative regulator of muscle development. Mutations of MSTN gene can lead to muscle hypertrophy in many natural species. However, there was not been reported significant muscle phenotype in pigs (Sus scrofa) in association with MSTN mutation. Researchers from Institute of Animal Sciences, Chinese Academy of Agricultural Sciences used zinc-finger nucleases (ZFN) editing technique in combination with somatic cell nuclear transfer and embryo transfer techniques to knock out the MSTN gene of Meishan pig's fetal fibroblast, and obtained the first MSTN gene editing Meishan pigs in the world. The homozygous MSTN mutation (MSTN^-/-) Meishan pigs had a distinct double-muscle phenotype with broad back, full hips and well-developed front shoulders. The lean-meat rate could reach more than 60%, which was nearly 20% higher than wild type. It had reached the same level as exotic pig breeds, effectively solving the technical bottleneck that restricted the development of local black pigs in China. The gene editing pig had not any marker gene or any integrated carrier fragment in the genome, and there was no off-target phenomenon. It was more reliable in animal health and product safety. Additionally, it had been approved by the Ministry of Agriculture to enter the stage of production test safety evaluation. The patent for the preparation of the related achievements and the patent for the protection of MSTN mutant sequences had been granted by the National Invention Patents. At present, this achievement has good industrialization prospects in terms of safety and breeding value, which could bring enormous economic benefits to the society.