For screening of high efficiency and safe nematicidal resource, a Bacillus strain YBf-10 which exhibits extreme toxicity to root-knot nematode was isolated from marine sediment. The 16S rRAN gene of this strain was amplified by PCR, then sequenced and analysed with Blast in NCBI. The result indicated that it showed 99% identity with the 16S rDNA of Bacillus firmus strain Z1-7. It means the isolated Bacillus strain is B. firmus. The YBf-10 strain was cultured in LB medium until spores were mature, the supernatant was harvested by centrifuged. The supernatant was diluted by 10-folds, and bioassay to 2nd stage juveniles (J2) of Meloidogyne hapla was conducted. The results showed that YBf-10 exhibited extreme toxicity, and the adjusted mortality reached to 50% at 24 h, and even up to 100% at 72 h. The supernatant could obviously suppress hatching of eggs of M. hapla to 80% at 48 h. For ascertaining the synthetic phase of the nematicidal substance, samples were collected at interval of culturing times, the supernatant of the samples was prepared by centrifugalization, and assayed toxicity to J2 of M. hapla. The result indicated that, from onset of the stationary phase, it exhibited obvious toxicity. The toxicity enhanced persistently during all the stationary phase, this meant that the nematicidal substance was synthesized at stationary phase of YBf-10 strain. The nematicidal activity of supernatant treated at 80℃ for 30 min showed no obvious change. The protein in supernatant was prepared by ammonium sulfate precipitation with 30% relative saturation, and exhibited no toxicity to J2 of M. hapla. Whereas, the supernatant removed protein exhibited the same nematicidal activity with the raw supernatant. The result indicated that the nematicidal substance produced by YBf-10 was micromolecular compound, not protein. On the basis of above research, we conclude that the Bacillus strain which exhibits toxic to nematode is characterized as Bacillus firmus, and it synthesizes micromolecular nematicdidal compound at stationary stage. The compound exhibits extreme virulence to root knot nematode, and supplies nematicidal resource for exploring biological control nematicide with the YBf-10 strain

The aim of this study was to prepare monoclonal antibody(McAb)against Pasteurella multocida. The DNA fragment encoding the mature domain of P. multocida outer membrane protein A(OmpA) was amplified from the genomic DNA and sub-cloned into pET28a(+) expression vector, 37.6 kD rOmpA fusion protein was expressed mainly as an insoluble protein, optimal solubilization of the recombinant protein was obtained using 8 mol/L urea in lysis buffer. BALB/c mice(Mus musculus) were subcutaneously injected with 100 μg of P. multocida OmpA emulsified by equivolumminal freund's complete adjuvant at the age of 6~8 weeks. Thereafter they were boosted two times with 200 μg of P. multocida OmpA emulsified by Freund's incomplete adjuvant at intervals of three weeks. The spleen cells of BALB/ c mice immunized with recombinant Pm OmpA were collected and infused with SP2/ 0 cell. Sebsequently four hybridoma cell strains were obtained by indirect enzyme linked immunosorbent assay (ELISA). The ELISA titers of antibodies in culture supernatant were 1∶128, 1∶128, 1∶256 and 1∶128, respectively, and ascites titers were 1∶6 400, 1∶6 400, 1∶12 800 and 1∶6 400, respectively. The McAbs did not cross-react with other gram-negative and gram-positive bacterial pathogens, including E. coli, Bordetella bronchiseptica, Pseudomonas aeruginosa and staphylococcus. High titer McAbs were secreted from the hybridoma cells after repeat freezing. The result of Western blotting assay showed that the four Mabs could react with Pm OmpA protein specifically. ELISA test revealed that the 2A2 McAb belonged to the subtype of IgG2b, with a concentration was 130 μg/mL after protein A affinity purification. The purified 2A2 McAb was selected by Western blot and IFA assays. The result indicated that the McAb could react with the Pm isolate strain. The success of this study has built up a solid base for developing a novel diagnostic methodology to the Pasteurella multocida infection in rabbits.

Xinjiang hand stretched noodle (XHSN) is one of the most popular noodles in China, and evaluation for high quality XHSN is essential to success of breeding program. Based on past research achievement of the XHSN and relevant molecular markers for the genes coding high quality XHSN, three types of multiplex PCR systems were developed and validated with 13 Xinjiang winter wheat cultivars and advanced lines with known genes, and used to evaluate the genes associated with XHSN quality in 46 Xinjiang winter wheat(Triticum aestivum L.) cultivars and advanced lines. The first multiplex PCR was used to simultaneously detect genes Ax2*, Pinb-D1b and ω-secalin, the second one was to detect the genes Psy-A1a, Psy-A1b, Psy-A1c, Ppo-D1a and Ppo-D1b, and the third one was to detect Glu-B3a, Wx-B1a and Wx-B1b. The test results showed that the frequencies of genes Ax2*, Pinb-D1b, without ω-secalin, Psy-A1a, Psy-A1b, Psy-A1c, Ppo-D1a, Ppo-D1b, Glu-B3a, Wx-B1a and Wx-B1b in the 46 winter wheat cultivars and advanced lines were 30.4%, 45.7%, 78.3%, 94.5%, 4.3%, 2.2%, 91.3%, 8.7%, 21.7%, 91.3% and 8.7%, respectively. Five (Ppo-D1a, without ω-secalin, Pinb-D1b, Glu-B3a and Ax2*) of those genes related with the high quality for the XHSN in Xinjiang wheats had higher frequencies than those in wheats from other Chinese regions, but the high quality genes Psy-A1b and Wx-B1b had lower frequencies. Meanwhile, 19 combinations of the high quality genes appeared in the tested cultivars and advanced lines, the combinations with one, two, three, four or five high quality genes accounted for 13.0%, 32.6%, 26.1%, 23.9% and 4.3%, respectively. The cultivar with six or seven high quality genes was not found. In all combinations of the high quality genes, Ppo-D1a/without ω-secalin, Ppo-D1a/Glu-B3a/without ω-secalin and Ppo-D1a/Ax2*/Pinb-D1b/without ω-secalin had the highest frequencies. Those results showed that the three systems developed in this study were effective and stable to amplify target bands for these genes and could be used to evaluate the XHSN quality quickly. In order to improve the quality of the XHSN in the future, those high quality genes and their combinations should be maintained for the XHSN. Developing cultivars with lower in both yellow pigment (Psy-A1b) and amylose (Wx-B1b) contents will be important objectives in the XHSN wheat breeding programs in the future. Developed three multiplex PCR systems and identified wheat genetic resources with the high quality genes in this study can be provide valuable information for estimation and breeding of the XHSN cultivars in the future.

Many flowering plants possess a self-incompatibility system to prevent inbreeding. In Brassica oleracea, self-incompatibility is genetically controlled by S-locus cysteine rich protein (SCR) and S-locus receptor kinase (SRK). The SCR is the determinant of pollen S-haplotype specificity. In order to compare the structure of the gene and molecular characterization of the protein among the allelic SCRs, the nested PCR primers were designed on the basis of the conserved amino acids in the signal peptide's cleavage site and the ploy A of mRNA. Here we cloned partial cDNA sequence of SCR from six Brassica oleracea L.. Sequence analysis showed that the cDNA sequence of SCR in D3, E1, 240, A1, N1 and G1 were 319, 311, 290, 288, 385 and 377 bp, respectively, which all encompassed 3'UTR. Their coding regions predicted a protein of 58, 58, 58, 58, 58 and 55 amino acids, respectively. The protein sequences were identical between SCR-D3 and SCR3. SCR-E1, SCR-240, SCR-A1 and SCR-E1 also had the same sequences, and they were all identical to the SCR7. The SCR of G1 was a new S haplotype gene. Although SCR-E1, SCR-240, SCR-A1 and SCR-E1 were the same S haplotype, their 3'UTR were different. For example, the length, the polyadenylation signal and the adenine nucleotide's content were different among them. Sequencing and bioinformatic analysis indicated that there were some differences in the secondary structure and the 3-dimentional structure of the six SCRs, suggesting that the interactions of SCR with SRK required strict complementary space. All SCRs had potential phosphorylation sites, but no glycosylation sites. It showed that the phosphorylation of SCR might play roles in signal transduction of self-incompatibility. Furthermore, the amino acid residues interacting with SRK were situated on the surface of the SCR molecule, and most of these amino acid residues were basic amino acid. So, we suggested that the process of SCR interacting with SRK required the participation of the static charge. Hereby, we provided theoretically a new way for improving self-seed set by chemical reagent, such as neutral salts. Bioinformatic analysis also showed that the differences in the strength of self-incompatibility among different S haploids might be caused by their binding capacity of the SCR and the SRK. The differences in the strength of self-incompatibility among the same S haploids might be caused by the numbers of the SCR and the SRK. Moreover, the SCR were regulated at the mRNA level by the 3'UTR. The evolution speed of SCRs was fast, and the evolution speed of the 3'UTR was even faster than that of the coding regions. The results provide a theoretical basis for further research and utilization of self-incompatibility mechanism

Ogura cytoplasmic male sterile(OguCMS) is the most widely used male sterile type in cabbage breeding. MYB transcription factors play a key role in regulation of plant defense response and multiple development processes. In present experiment, a R2R3-MYB transcription factor which down regulated 10.3 times in cabbage(Brassica oleracea var. capitata) OguCMS lines was cloned by SMART RACE strategy.The full-length cDNA of BOMYB1 was 1 141 bp, which contained a 196 bp long 5' untranslated region, a 246 bp long 3' untranslated region and a 699 bp long open reading frame(GenBank accession number: JN703995). It was localized in the nucleus by subcellular localization prediction. It was an anther preferentially expressed gene in cabbage, which reached its expression peak in the late development. It was induced by the regulation of plant hormones salicylic acid(SA) and jasmonate methyl (JA-ME), and consequently regulated the expression of anther development genes. The experimental results suggests that BOMYB1 may be one of the important genes which involved in OguCMS anther development.

As the model plant in the fruit trees, a number of important quality trait locus has been mapped in peach. Fruit maturity date is an important economic trait in peach. In this study, the self-crossed progenies of peach(Prunus persica (L.) Okubo) were used as material to mapping the maturity genes in peach. 165 SSR primer pairs from the genome of Prunus species were screened by bulked segregant analysis (BSA). Results showed that three primer pairs were related with maturity genes: dominant marker UDP96-003, and co-dominant markers BPPCT015, UDP97-402. The electrophoresis results of these three markers in random samples were analyzed by FsLinkageMap 2.0 and variance analysis for single marker, the result indicated that the rank of the three markers was UDP96-003, UDP97-402 and BPPCT015 with the genetic distance of 21.0 and 13.2 cM, respectively, and all of the three markers were linked with maturity gene. The analysis results from FsQtlMap showed that there was a major-QTL loci between BPPCT015 and UDP97-402 with the LOD of 13.35 and the effect value of 0.623, respectively, and the genetic values were qq :84.59 d, Qq:100.08 d and QQ:115.25 d, respectively. The single-gene effect value of the loci approximately 15 d. The results provide experimental evidence to previous inference, and the locus is anchored between the two SSR markers.

With the changes of soil moisture, the PnAG3 expression changed in the several peanuts (anti-Aspergillus flavus species J11, HY22 and susceptible peanut species JH1012) growing in the pot, while their physiological index related to drought resistance was also changed. The results showed that the Superoxide dismutase(SOD), peroxidase(POD) and catalase(CAT) activities of anti-Aspergillus flatus species had increased 3.15, 2.55 and 4.95 times in 40%~55% of field maximum water capacity treatment group, respectively. But the SOD, POD and CAT activity of susceptible species in the three treatment groups had no significant difference. Malondialdehyde(MDA) contents were increased in all treat groups, and it was higher in susceptible peanut species than that of anti-Aspergillus flavus species. PnAG3 expression levels were higher in anti-Aspergillus flavus species than that of susceptible peanut species. But the maximum expression level of anti-Aspergillus flavus species was appeared earlier than that of susceptible species which was similar to the CAT activity changes. Overall, SOD was continued to rise. But the PnAG3 gene expression, and POD, CAT and MDA showed a tendency of "up-down". Combined with the physiological index related to drought resistance and the PnAG3 expression changes, there was a correlation between the variation trend of PnAG3 expression and adversity physiological indexes. And the PnAG3 expression in resistant varieties was higher than that of susceptible varieties. So that, the function of PnAG3 may be relevant to the peanut stress resistance.This study provides a basis for further research on resistance breeding and germplasm screening of peanut.

Considerable research efforts have been expended to study the factors that are associated with fat deposition in poultry production. Dehydroepiandrosterone (DHEA, 3b-hydroxy-5-androsterone-17-one) is a steroidal compound that is secreted by the mammalian adrenal cortex gland. It is known to be a fat-reducing agent by activation of the steroid hormone receptors. In the present study, cultured primary chicken hepatocytes exposed to DHEA (0, 0.01, 0.1, 1.0 10 and 100 μmol/L, respectively) dissolved in medium were left for 20 min for cAMP assay by RIA(radioimmunoassay) kit. We found that the levels of cAMP were significantly higher in 0.1~100 μmol/L DHEA groups(P<0.05), especially a maximum accumulation in 0.1 μmol/L DHEA group that compared with the control group(P<0.01). Chicken hepatocytes pre-incubated with 0.25 mmol/L IBMX(3-isobutyl-1-methylxanthine) or vehicle for 5 min, followed by the addition of 0.1 μmol/L DHEA, 20 μmol/L forskolin or vehicle for 20 min at 37℃ were tested for adenylate cyclase (AC) activity. Cell lysates isolated from hepatocytes exposed to 0.1 μmol/L DHEA, 0.25 mmol/L IBMX or vehicle were tested for phosphodiesterase (PDE) and cAMP-dependent kinase A (PKA) activity. Additionally, chicken hepatocytes cultured in media as described above were exposed to 0.1 μmol/L DHEA, 20 μmol/L forskolin or vehicle for 1 h at 37℃. The cells were also exposed to 10 μmol/L H89(PKA- selective inhibitor), 2 μmol/L PKA inhibitor peptide (PKI) or vehicle for 30 min at 37℃. Then, 0.1 μmol/L DHEA was added, the cells were cultured for another 1 h, and scraped for the measurement of CREB phosphorylation levels. The results showed that, no significant differences were found in AC activity in both the DHEA and control groups(P>0.05). As expected, a full stimulation was achieved at 20 μmol/L forskolin(P<0.05), a powerful agonist of AC activity. In contrast, 0.1 μmol/L DHEA markedly suppressed PDE activity at 20 min as compared with the control group(P<0.05). The inhibitor IBMX was also potent in suppressing PDE activity during the incubation period(P<0.05). Cells incubated with 0.1 μmol/L DHEA for 20 min exhibited a significant increase in PKA activity(P<0.05). 0.1 μmol/L DHEA treatment lead to a significant increase in CREB phosphorylation(P<0.05). No significant differences was observed in hepatocytes pretreated with PKA inhibitors(P>0.05). The results suggest that DHEA can activate the cAMP/PKA signaling pathway and regulate lipid metabolism by enhancing CREB phosphorylation level, and provide a better understanding for improvements proposed for DHEA in decrease of body fat in broiler chickens.

The aim of present research is to determine the localization of Crumbs-3(Crb3) in mouse testis and its distribution rules on spermatogenesis in seminiferous tubules and developmental process of Leydig's cells. We studied the histological structure and distribution of Crb3 at different postnatal development stages of mouse(Mus musculus) testis by common paraffin sectioning method stained with Hematoxilin and Eosin method and immunofluorescence histochemistry. The results showed that the diameter and thickness of seminiferous tubules increased，their lumens experienced a process from small to big, then narrowed before widened lastly as individual development. Spermatogenic cells in seminiferous tubules were in dynamic development changes, in which there were only some spermatogonium and Sertoli cells at 2th week of age, then all spermatogenic cells except spermatozoa could be seen at 4th week of age, a great quantity of spermatozoa appeared at 4th week of age after proliferation of spermatogenic cells. Then the density of spermatogenic cells increased as development. The shape of Sertoli cells became clear and easily to be recognized, and the volume of leydig cells and eosinophilic cytoplasm increased gradually, while nuclear-cytoplasmic ratio decreased gradually as development. Crb3 protein were mainly distributed in membrane of both spermatogenic cells and leydig's cells, and the positive reaction in leydig's cells were obviously stronger than that in spermatogenic cells.The results hint that Crb3 not only relates with the establishment and maintenance of seminiferous epithelium cells, but also probably participates in regulating the synthesis and secretion of steroid hormone in testis

White-rot fungus(Pleurotus ostreatu) has strong degradation ability of lignin. The experiment was conducted to research effect of different Mn2+ levels on lignocellulose enzyme activity of rape(Brassica campestris L.) straw fermented by white- rot fungus and changes of rape straw tissue structure. The experiment, which was separated into control group(SMc, 0 mmol/L),Test One (SM1, 0.6 mmol/L) and Test Two(SM2, 1.2 mmol/L), selected white-rot fungi P. ostreatu-WY01 to ferment rape straw motionlessly fermented of 40 d on constant temperature of 28℃ in nitrogen-limited mediums which were added Mn2+ of different levels in solid state.The result showed that laccase activity and manganese peroxidase(MnPs) activity of Test One, which reached respectively peak of 211.11 U/g at 6 day and 49 U/g at 10 day, very significantly overtoped other groups(P<0.01); lignin peroxidase(Lips), cellulose enzyme(Cels) and hemicellulase enzyme(Hcels) activity respectively were no difference by adding Mn2+ and tissue structure of straw was accordance with related cellulose enzyme activity. At early period of fermentation, experimental groups Lacs activity reached peak at 6 d and fell fastly until tend to smooth at 10 d while SMc had been living in lower level in process. Lips of SM1 reached peak at 12 d while SM2 and SMc reached peak at 10 d.After peaks,they trended to fall gradually until keep lower level after 20 d. MnPs activity kept low level before 6d of fermentation and ascended suddenly followly. Experimental groups MnPs activity which SM1 and SM2 respectively reached peak at 10 d and 16 d, were greater significantly than SMc. During the fermentation, all treatment groups Cels and Hcels activity had a rising trend and experimental groups were higher than control group. Scanning electron microscopy showed that without any treatment, tissue was very compact and tissue arranged orderly. Empty cavity the arrow refered to the figure might be caused by high temperature sterilization leading to inflation. Processed by the white-rot fungus P. ostreatus-WY01, SMc rape straw tight tissue was destroyed and appeared some loose mesh at 20 d;straw waxy skin arrowed to on left and fibrous tissue was broken down on the right at 40 d; SM1 partly appeared collapsed at 20 d and fibrous tissue was completly decomposed and pulverized at 40 d. SM2 tissue was destroyed in different degree at 20 d and fibrous tissue was completed decomposed at 40 d while it not as effective as SM1. The findings suggested that Mn2+ could induce and benefit degradation of rape straw by white-rot fungus(P. ostreatu-WY01), and comprehensive biodegradation effect was SM1＞SM2＞SMc. The study provides us a basis to develop resources of biological feed using rape straw

Actinobacillus pleuropneumoniae (APP) and Haemophilus parasuis (Hps) are the bacteria of causing porcine disease. In this study, We constructed two recombinant vectores pBOSKΔIC-1 and pBOSKΔIC/ompP2 using APP5 (SW1) strain as a template, and the two recombinant vectores were used for plus-minus screening system with the basis of Kanr and sacB. The mutant strain, named SW1 Δ apxIC/ompP2, was obtained, which was deleted toxin I gene C (apxIC) in SW1 and gomphosised outer membrane protein P2 Hps gene (ompP2). The mutant missed a size of 475 bp apxIC gene but was inserted a size of 1107 bp Hps ompP2 gene compared with the parental strain SW1. No reverse mutation of apxIC gene was observed during after 10 generations, while Hps ompP2 gene still had a good genetic stability in vitro, the mutant strain had no significant difference of he basic growth characteristics with the parental strain (SW1). The results confirmed that an ΔapxIC mutant of A. pleuropneumoniae gomphosised ompP2 gene of H. parasuis was successfully constructed. and it offered a foundation for developing a new bivalent vaccine of APP and Hps

Because of the high similarity in the botanical characters of the Eurasian grape varieties, it is difficult to distinguish or identify a large number of varieties using the traditional single morphological methods, especially the strains and elite cultivars derived from a small number of parents that show similar traits. In this study, random primers with the composition of 11 bases were selected for RAPD amplification of the Eurasian grape (Vitis vinifera L.) varieties, and corresponding map relationships were bulit according to the specific bands, thus plant species were identified through drawing the diagram manually (manual cultivar identification diagram, MCID), which distinguished different varieties efficiently and quickly. This method was convenient, fast and accurate, and 191 grape varieties could be distinguished using 7 times PCR at the molecular level. Compared to the clustering tree, the manual CID could be easy to read and have more practical use. In other words, the primers for distinguishing between any two varieties could be found according to the MCID chart. This method can achieve early identification of seedlings, and has a wide range of versatility in the other species. Furthermore, it will have a positive meaning in the identification of grape germplasm resources and promote sustainable development of wine industry

In this review, recent development and future prospects of biotechnologies application on ornamental horticulture in the world are presented. To bloom the floriculture industry, breeding of new cultivars with improved ornamental and agronomic traits are required, relying on total understandings of principles during specific developmental progresses and application of new biotechnologies. At present, new biotechnologies come out every day, providing breeders with new ideas and methods on breeding of new cultivars. Here, we summarized the application of biotechnology on ornamental horticulture in the fields of cellular engineering, molecular markers, and gene modification. The gene modification in specific traits of ornamental plants was emphasized on, including morphology modification, biotic or abiotic stress resistance improvement, and postharvest life elongation. Finally, some prospects for potential biotechnologies application in future horticultural research and breeding were explored

microRNA(miRNA) plays an important role for lipid metabolism. To find the potential miRNAs regulating fatty acid metabolism of mammary gland of goat, Databases and MFOLD were used for predicting miRNA and analyzing ΔG, respectively. And pri-miRNA were cloned for further study. Prediction results showed that 50 miRNAs including 13 identical miRNAs predicted by 3 databases and 37 identical miRNAs predicted by 2 databases were obtained through predicting 30 genes regulating fatty acid metabolism of goat by MicroCosm, TargertScan and PicTar on line. There were 4 and 2 different miRNA predicted targeting FASN(fatty acid synthase) and BTN1A1(goat butyrophilin, subfamily 1, member A1), respectively. And no miRNA binding sites was predicted on 3'UTR of AGPAT6(1-acylaglycerol-3-phosphate O-acyltransferase 6). There were 3 and 2 numbers of different location respectively on 3'UTR of FASN and BTN1A1 targeting by miR-103. The results of ΔG analysis (ΔG ＞ -20 kcal/mol) of flanking regions of 50 miRNAs binding sites showed that 9 miRNA were related to fatty acid metabolism, miR-103/BTN1A1, miR-15/FASN, miR-23/LPL(lipoprotein lipase), miR-27/PPARγ(peroxisome proliferator-activated receptor γ), miR-29/GPR41(orphan g protein-coupled receptor family, member 41), miR-146/BTN1A1, miR-195/FASN, miR-200/SCD(Stearoyl-CoA desaturase) and miR-497/GPR41. And there were highly homology on the binding sites of miR-103/BTN1A1, miR-27/PPARγ, miR-195/FASN among species，which sthrengthened the possibility of the connection between miRNA and their targets. Also, the numbers of one side and two sides of flanking regions of miRNA/mRNA binding sites whose ΔG were smaller than threshold were 14 and 9, respectively. Among all the predicted targets, FASN, ACOX1(acyl-CoA oxidase 1), LPL and PPARγ were targets of miR-27. FASN, BTN1A1 and ACOX1 were both targets of miR-103 and miR-15. 5 pri-miRNA (pri-miR-103-1/23a/27a/146b/200a) were cloned from DNA template from blood of Xinong Saanen dairy goat(Capra hirus). The sequence and secondary structure analysis demonstrated that all the 5 primary miRNA(pri-miRNA) comprising completed pre-miRNA and the stem loop structure could generate functional miRNA in vivo. The results of homology analysis showed that the homology of 4 pre-miRNA of goat were 100 % with cow except for pre-miR-27a whose homology was 98 % (There was a G located in the 11 base from 3' end of pre-miR-27a of goat instead of A of cow). In conclusion, those 9 miRNAs can regulate fatty acid metabolism in goat mammary gland, and cloning of 5 pri-miRNAs provides a basis for their function studying.

In order to investigate the role of granulin(GRN) during the development of adipocyte, we constructed lentivirus interfering vector targeting on porcine GRN, packaged and infected the pig preadipocyte, differentiation state of preadipocyte was detected by Oil Red O staining and Oil Red O extraction assay, the expression levels of adipocyte differentiation marker genes were analysed by Real-time PCR. The results showed that the virus titer of the lentivirus reached 5×107 TU/mL, and the GRN expression level was remarkably decreased after lentivirus infection, of which, GRN mRNA expression was suppressed by approximate 76% by short hairpin RNA(shRNA2), moreover, adipocyte differentiation was promoted, and adipocyte differentiation marker genes, such as peroxisome proliferator activated receptor γ(PPARγ), sterol regulatory element binding protein 1(SREBP-1C), adipocyte-type fatty acid-binding protein(ap2) and lipoprotein lipase(LPL) mRNA expression levels were all increased. Taken together these data suggested that knocking down of GRN promoted adipocyte differentiation, and GRN may play a negative role in pig preadipcoyte differentiation.