Abstract Peroxisome proliferator-activated receptor α(PPARα) gene exon5 polymorphism was detected by PCR-SSCP and DNA sequencing in total 717 individuals, six breeds(Bos taurus) of Qinchuan cattle, Nanyang cattle, Jiaxian Red cattle, Luxi cattle, Xianan cattle and Angus, and its correlation of this SNP with some carcass and meat traits of 108 Qinchuan cattle was analyzed. T→C mutation was detected at 138 bp in PPARα gene exon5. Chi-square test showed that locus was not in Hardy-Weinberg equilibrium(P＜0.05) in Qinchuan cattle and Xianan cattle, but was in Hardy-Weinberg equilibrium(P＞0.05) in other breeds. In the six breeds, the polymorphism information contents(PIC) were moderate polymorphisms(0.25＜PIC＜0.50). Correlation analysis showed that AA genetype was significantly affected by back fat thickness and eye muscle area(P＜0.05); and BB genetype was significantly affected by water holding capacity(WHC )(P<0.05), showing this locus may be a major QTN or tightly linked back fat thickness, eye muscle area and WHC, it can be a candidate molecular marker for the beef cattle breeding.

Abstract In order to reveal the relationship between three key genes related to ubiquitin-26S proteasome pathway (UPP) and physiological male sterility of wheat(Triticum aestivum), the pure lines of Xinong 1376 and Xinong 2611 with different genetic background were employed to develop two pairs of near-isogenic physiological lines by new Chemical hybrid agents (CHA) SQ-1, and RT-PCR technique was performed to obtain the expression patterns of the three key genes in sterile and fertile anthers at different development stages. The results showed that the transcripts abundance of gene Urm1 (ubiquitin-related modifier 1）in sterile anther significantly increased compared to that in fertile anther for both Xinong1376 and Xinong2611 at different developmental stages; the transcripts abundance of ubiquitin fusion gene S27a increased obviously only in sterile anther of Xinong2611, but increased slightly in sterile anther of Xinong 1376, at all development stage in comparison with fertile anther; the F-box protein in sterile anthers of the two varieties was clearly up-regulated expressed, even specially expressed at key abortion periods of pollen, mononucleated anaphase stage to binucleated stage, but that was expressed weakly in both fertile and sterile anther at mononucleated prophase stage and trinucleated stage. Hereby, it is proved that the transcripts of the three genes in the sterile anthers studied in the paper and increased in transcript abundance are closely related to aborted pollen, and the genes are the promoters for pollen abortion of wheat physiological male sterility induced by CHA SQ-1.

Abstract Cypermethrin(CYP) has deleterious effects on male reproductive function. The objective of this study was to identify whether the effects of β-CYP on sertoli cells dedifferentiation were associated with oxidative stress. A dose of β-CYP(20 mg/kg) was administered to male mice(Mus culus) for 35 d with or without vitamin E(20 mg/kg). The expression and distribution of proliferating cell nuclear antigen(PCNA) and connexin 43(CX43) in testes were detected by immunohistochemistry and the testis total antioxidant capacities were detected. β-CYP not only increaed the expression of PCNA in sertoli cells,but also decreased the expression of CX43 in sertoli cells and total antioxidant capacity(TAC)(P<0.05). Vitamin E reversed the effects of β-CYP on he expression of PCNA and CX43 in sertoli cells and total antioxidant capacity(P <0.05). Therefore, β-CYP can cause sertoli cells dedifferentiation by inducing oxidative stress.

Abstract In order to explore the role of vitamin A(VA) on fatty deposition and cholesterol cleaning, the distribution of hepatic lipid droplet in high-fat diet fed mouse(Mus musculus) was observed by paraffin section preparation methods; the relative level of hepatic stearolysis related gene mRNA was determined by real-time RT-PCR and serum cholesterol(CHO) and high density lipoprotein-cholesterol(HDL-C） levels. Results showed that: first three days there was no significant difference between VA treated and control group on the daily gain(P>0.05); on the 4th, 9th and 27th day the daily gain was significantly lower than that of control group (P<0.05); the subcutaneous fat and fat pad under peritoneum were significantly lower in VA treated (0.1890 ±0.0056) g and (0.3862 ±0.0053) g than that of control group (0.2620 ±0.0020) g and (0.4867 ±0.0052) g (P<0.01); the amount of hepatic lipid droplet was fewer in VA treated than that of control group; the expression levels of scavenger receptor-BⅠ(SR-BⅠ) and hormonesensitive lipase(HSL) mRNA were significantly higher in VA treated than that of control group(P<0.01); peroxisome proliferators activated receptor γ(PPARγ) and sterol regulatory element binding protein-1c (SREBP-1c) were significantly lower in VA treated than that of control group(P<0.05); triglyceride hydrolyase(TGH) mRNA was significantly higher in VA treated than that of control group(P<0.05); and the levels of serum CHO and HDL-C were significantly lower in VA treated than that of control group(P<0.01). Our results suggest that VA profits to fat degradation and cholesterol cleaning.

Abstract Full-length cDNA sequence of transcription factor VpRFL1 was cloned from Vitis pseudoreticulata W.T. Wang according to EST sequence and using RACE method. VpRFL1 genomic DNA sequence was cloned from V. pseudoreticulata W.T. Wang using PCR technique. Sequence analysis showed that the full-length of VpRFL1 (GenBank accession No. FJ356672) was 1324 bp, containing a 1053 bp open reading frame, which encoded 350 amino acid residues. The corresponding DNA sequence was 1599 bp, containing 3 exons and 2 introns. Nuclear localization signal of VpRFL1 was located at N-terminal 19~46 amino acid residues, and the 275~316 amino acid residues at C-end were rich in Cys, which was a C4C4 variant of RING finger. Transient expression of recombinant plasmid VpRFL1/ PBI221-GFP in onion(Allium cepa) epidermal cells showed that VpRFL1 was localized in cell nuclei. The results from the sequence feature and localization analysis suggest that VpRFL1 functions in nuclei and was supposed to be involve in biotic stresses pathway.

Abstract We firstly cloned a glyA gene that encoded serine hydroxymethyltransferase (SHMT) by PCR with the genomic DNA of Aeromonas hydrophila as template. The resulting glyA gene was ligated into vector pGEX-6p-1 to generate recombinant plasmid, pGEX-glyA. Then we transformed the recombinant plasmid into Escherichia coli DH5α. Sequence analysis revealed that the full length of glyA gene (GenBank accession: FJ797607) was 1 254 bp, and encoded a protein with a molecular weight of about 45 kD, composed of 417 amino acids. Further, we expressed the recombinant SHMT in E.coli by IPTG induction and purified the target protein using the GST-tag affinity chromatography. Enzymatic assay revealed that the recombinant SHMT displayed maximum activity at pH 8.0 and 20℃. The activity of SHMT was activated by Cu2+ and Mn2+, and inhibited by Zn2+ and SDS. The characterization of SHMT suggestes that SHMT is a cold-active serine hydroxymethyltransferase and has a potential significance for study of cold-active mechanism and industry applications.

Abstract The effects of different microelement concentrations in the culture medium on browning and differentiation of callus derived from mature embryos of three elite indica rice cultivars, Guichao2, Teqing and 9311 were investigated through the callus induction, two-time subculture, pre-differentiation and differentiation in this study. The results suggested that in the culture medium of pre-differentiation and differentiation, callus browning was inhibited and callus greening frequency was improved when Guichao2 callus was cultured with 5×CC microelement content and 9311 callus was cultured with 3×CC microelement content, but no effect for Teqing callus when increasing the microelement content. This study provides a new insight into indica rice genetic improvement and transformation.

Abstract Human β-defensin 3 (hBD3) gene mammary special expressing vector named pEBB containing EGFP report gene was transfected into bovine fetal fibroblasts by Lipofectmine TM2000 and G418 selection (600 μg/mL) was applied to select positive cells. After identified with PCR, these positive cell clones were donated to produce transgenetic cloned embryo with somatic cell nuclear transplantation followed by monitoring the existence of hBD3 gene in embryo genome using PCR. After tranplanting these trangenetic cloned embryos into 64 receptors' uterus, there were 21 cows pregnant 3 months later. This research provides feasible techniques for getting trans human β-defensin 3(hBD3) transgenetic bovine embryos, producing transgenetic cows and proventing cow mastitis.

Abstract Selection of the best stable reference genes is of crucial importance for the accurate normalization of expression in real-time quantitative PCR. Taking the piglet tissues as example, the expressions of nine housekeeping geens beta-actin(ACTB), beta-2-microglobulin(B2M), glyceraldehyde-3-phosphate dehydrogenase(GAPDH), hydroxymethylbilane synthase(HMBS), hypoxanthine phosphoribosyltransferase 1(HPRT1), ribosomal protein L4(RPL4), succinate dehydrogenase complex, subunit A(SDHA), TATA box binding protein1(TBP1) and Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein and zeta polypeptide(YWHAZ) were detected using real-time quantitative PCR. And their expression stability were observed by analysis of geNorm program. Results showed HPRT1 and HMBS > RPL4 > TBP1> B2M > YWHAZ > SDHA > GAPDH > ACTB. As the best choice, HMBS can be used as normalizing expression of target gene in different piglet tissues, while RPL4 is good reference gene for high abundant transcripts.

Abstract A new gene (UL28) from Duck plaque virus (DPV) DNA gene library constructed in Key Laboratory of Animal Disease and Human Health of Sichuan Province was selected and sequenced. A complete open reading frame (ORF2412) was found by ORF Finder and BLAST. It was amplified by PCR and cloned into pMD18-T vector. The positive plasmid was selected and identified by PCR and incision enzyme. Furthermore, it was confirmed by nucleic acid dot-hybridization to make sure the ORF2412 belonged to DPV DNA, and analyzed by bioinformatics tools of NNPP version 2.2，Translate tool，DNAStar and so on. Furthermore, CHIPS and CUSP methods of EMBOSS package were used to analyze the codon usage. The results showed that the typical characteristic of herpesvirus_UL28 was possessed; it contained one structural domain of PRTP and numbers of feature associated with the phosphorylation sites and N-glycosylation sites. Hydrophobic region was greater than hydrophilic region; it was a kind of protein out of membrane. Phylogenetic demonstrated that it was more closely related to poultry herpesvirus (α-herpesvirus). In addition, DPV UL28 gene was shown strong bias in codon usage.

Abstract CCAAT/enhancer-binding protein α（C/EBPα） gene plays a key role in the process of adipocyte differ- entiation, which is also an important candidate gene for growth and body composition traits. In the current study, polymorphisms of 5′region, coding sequence and 3′region of C/EBPα were detected by sequencing the 9th and 10th population of a designed resource population, the Northeast Agricultural University broiler lines (Gallus gallus) divergently selected for abdominal fat percentage (NEAUHLF). A silent mutation (c.*552G>A) was found at base 552 of the coding sequence. Association analysis indicated that the mutation was significantly related to abdominal fat weight, abdominal fat percentage, liver weight, liver weight percentage, metatarsus circle and shank weight (P<0.05). The abdominal fat weight and abdominal fat percentage of birds with AA genotype were significantly higher than those with GG genotype (P<0.05), and the liver weight, liver weight percentage, metatarsus circle and shank weight femur length of birds with GG genotype were significantly higher than those with AA genotype (P<0.05). The conclusion was drawn that C/EBPα gene can be or link to a major gene that affects fat deposition and skeletal development traits in chickens.

Abstract The embryo developmental capacity after in vitro fertilization with the X-sorted frozen and unsorted frozen sperm were studied. The cleavage and blastocyst rate of embryos fertilized with X-sorted frozen sperm, were significantly lower than that of control group fertilized with unsorted frozen sperm (51.4% vs. 67.9%, P< 0.05; 19.7% vs 29.9 %, P<0.05 ). After the two kinds of frozen sperm were treated with the micro epi-floating non centrifugation and the epi-floating , the cleavage rate of the former was better than that of the latter, but there were not statistically significant difference (50.7% and 65.5% vs. 48.3% and 63.8%, P>0.05). Additionally, no statistical differences were observed in morula rate and blastocyst rate of the cleavaged zygotes fertilized with the X-sorted frozen sperm and unsorted frozen sperm. After co-culturing with sertoli cells and granular cells respectively, the cleavage rate (50.0% and 51.4%) of sexed zygotes co-cultured with feeder cells were not statistically significant difference with that of control group (50.3%, P>0.05). And the morular rate(53.3% and 54.6%) and blastular rate (21.1% and 21.5%) of sexed embryo co-cultured with feeder cells were higher than that of control group (52.2% and 20.4%), but there were no statistically difference (P>0.05).

Bt gene either Cry1Ac or Cry1C was individually transferred into broccoli (Brassica oleracea L. ssp. italica) genome via Agrobacterium-mediated transformation method using Green Comet hypocotyls as explants. And thirty five kanamycin-resistant broccoli plants respectively harboring Cry1Ac or Cry1C were obtained. PCR analysis with primers specific to Cry1Ac or Cry1C identified 27 Cry1Ac positive plants and 13 Cry1C positive plants. Southern blotting analysis with Cry1Ac or Cry1C as probes indicated the Bt gene was intergrated into plant genome. Transgenic broccoli lines with single copy transgene were inbreeded to homozygote and crossed to each other to obtain transgenic lines with double Bt genes. RT-PCR analysis showed the various expression levels in different transgenic lines. The Bt endotoxin protein was about 0.12~0.82 μg /g FW by ELISA assays. Both in vivo bioassays in laboratory and field test supported the improved insect resistance and the resistance level correlated with Bt expression level. Homozygotic broccoli lines with resistance to diamondback moth (Plutella xylostella L.) and imported cabbageworm (Peris rapae L.) were obtained trough selection in successive generations. Double Bt genes improved insect resistance in transgenic broccoli in this study.

Global genome transcription profiles of the rice variety Pei'ai 64S (Oryza sativa ssp. indica), which grown under no stress and subjected to cold, heat, or drought stresses, were conducted. Transcription profiles were gained for leaves and panicles at seedling, booting , and heading stages from plants under normal condition, or the above stresses using the Affymetrix 60 K GeneChip Rice Genome Array. We identified a gene OsCrGtl (Oryza sativa cold responsive glycosyltransferase-like gene, GenBank accession No. FJ828672), which was sharply induced by cold in leaves and panicles at all the developmental stages tested. The transcription profile of OsCrGtl obtained by the GeneChip analysis was confirmed by quantitative real-time RT-PCR analysis. To analyze the function of OsCrGtl in cold tolerance, a cDNA of this gene following amplification was cloned by RT-PCR. Sequence analyses indicated that its cDNA encodes an open reading frame (ORF) of 462 amino acid residues with theoretical M.W. =50.327 kD and pI =5.35. In silico analysis suggested that the ORF contained a glycosyltransferase domain and was similar to proteins of the glycosyltransferase family in indica and japonica, Zea mays, and Arabidopsis thaliana. By analysis of the deduced promoter region of OsCrGtl for putative cis-regulatory elements with PlantCARE software, 9 matches to known cis-elements related to stress responses were founded. Upon the above results and analysis, OsCrGtl is a novel candidate gene involved in cold-tolerance in rice.

Sequence-specific amplification polymorphism (SSAP) retrotransposon-based molecular marker for germplasm identification and genetic relationship analysis in the genus Diospyros were investigated, and the performance of each SSAP primer pair when individually used for aforementioned studies was also studied in detail. Twenty five SSAP primer pairs generated a total of 1 300 robust fragments among inter- and intra-species genotypes examined, of which 1 214 (93.38%) were polymorphic with an average of 48 polymorphic bands per primer pair, indicating that SSAP is a high multiplex ratio and high polymorphic marker system. The analysis of genetic relationships of 28 genotypes based on the molecular data from SSAPs revealed that the Chinese genotypes, Japanese genotypes and related species were distinctly separated and some genotypes known genetic relationships were clustered together, which was consistent with previous studies, suggesting that SSAP was feasible and reliable for the future genetic analysis in the genus of Diospyros. Additionally, three SSAP primer pairs were found to be the most informative. Our results suggest that SSAP can be as a potentially powerful tool for characterization of Diospyros germplasm in the future.

Abstract　A 25 kD antifungal protein was extracted from Coix lacryma-jobi by ammonium sulfate precipitation, cation exchange chromatography, affinity chromatography, gel filtration and reverse-phase HPLC, respectively. Its purity was identified to be 98.23% by HPLC. The molecular weight was determined to be 25 kD by SDS-PAGE. It inhibited mycelial growth in a number of fungal species including Alternaria alternate, Trichoderma viride, and Fusarium graminearum, with an IC50 value of 9.15, 4.50 and 12.21 μmol/L, respectively. Transmission electron microscopy of mycelial of Alternaria alternate revealed marked ultrastructural changes, which included cell wall thickening and deformation, plasmolysis, cell wall perforated, cell content exudated and cell organelle severely damaged.

Abstract An 1 755 bp up-stream regulatory sequence of the transcription factor gene BrWRKY33 was cloned from Chinese cabbage (Brassica rapa subp. pekinensis cv. LongbaiⅡ). Two 5' deletion mutants of the sequence were fused with the reporter gene gus, respectively, and transformed into Arabidopsis thaliana. The results indicated that the W-box sequences present in －1 755~－315 bp region of BrWRKY33 was related to the down regulation, and the 0~－315 bp region contained the essential sequence for gene transcription. Transgenic plants challenged with soft rot bacteria (Erwinia carotovora subsp. carotovora) showed an increase in gus activity, indicating that BrWRKY33 gene play a role in the resistance against soft rot disease.

Abstract The wheat accession H9020-20-12-1-8 is a translocation line previously derived from hybridization and backcross between common wheat (Triticum aestivum L.) and Psathyrostachys huashanica Keng. It shows excellent resistance to the dominant races（CYR32） of Puccinia striiform f. sp. tritici prevalent in China at seedlings. The genetic analysis showed that H9020-20-12-1-8 had one dominant gene to CYR32. The resistance gene temporarily was designated as YrHy. A 248 individuals of F2 segregating population derived from H9020-20-12-1-8 and a susceptible wheat line Mingxian 169 was mapped using simple sequence repeat (SSR) markers, three of the 305 SSR markers on 2BS (Xgwm429, Xwgm770 and Xwmc154) were selected and linked with YrHy and with a distance of 5.4，6.4 and 11.3 cM, respectively. YrHy is a novel resistance gene to wheat stripe rust donated from P. huashanica Keng on the basis of pedigree analysis and molecular results.

Abstract In order to identify the availability of subgerminal cavity microinjection of foreign DNA as an effective way in making transgenic chickens and gene function study, foreign plasmid pEGFP-N1 was injected into the subgerminal cavity of the fresh fertilized eggs before incubation, and the embryos of 0.5，1，3，4，5，6 day-old were collected to identify the metabolism and distribution of the exogenous DNA. The success rate of integration of the foreign DNA was then analyzed by restriction enzyme digestion. The results indicated that the microinjected plasmid could be found in the whole embryos after incubation for 0.5, 1, 3, 4, 5 and 6 days. The positive ratio was 100%, 80.00%, 66.57%, 75.00%, 64.29% and 84.21%, respectively. The foreign plasmid was declined along with the embryos development. 75 % of the hatched chicks showed positive results. However, the fragment of foreign DNA did not integrate into the host cell genome. They were in free or polymerization status.

Adipocyte determination and dfferentiation factor-1 gene(ADD1), a gene associated with adipocyte differentiation in the research of human and rat, plays a role during adipocyte differentiation with PPARγ and C/EBP. In this paper, we cloned a part of the gene sequence from pig, screened it with PCR-SSCP (single strand conformation polymorphism), and used sequencing to identify mutation. The result indicated that a SNP (single nucleotide polymorphism) happened in the conservative domain of the ADD1 gene, which led to a different amino-acid sequence and a new site for proteinase. To verity the relationship between this mutation and meat quality, we chose 136 pigs as the experimental population, all of which were butchered to determine meat quality. The result of polymorphism analysis showed that a novel SNP was found in second exon. The polymorphism was disequilibrium in population and AA genotype was not found in it. The relation analysis showed that intramuscular protein, intramuscular fat and meat color had obviously difference among different ADD1 genotypes of pigs. The ADD1 polymorphism can apply to pig breeding for improving meat quality．

Abstract A number of promoter fragments from Panebacillus polymyxa M-1 were cloned using a promoter-trap vector, pECE7. Three promoter fragments of the highest chloramphenicol (Cm) resistance were selected through the concentration gradient of Cm, which were named p5, p8 and p17, respectively. A pair of primer was designed to obtain the gfpmut3a. Then the gfpmut3a and the promoter fragments were inserted into the Escherichia coli-Bacillus shuttle vector pHY300PLK to construct the vectors of pGFP5, pGFP8 and pGFP17. The new gfp vectors were transformed into E.coli DH5α by heat shock, and the bright fluorescence of E.coli-gfp5, E.coli-gfp8 and E.coli-gfp17 was observed by the confocal laser-scanning microscopy. Meanwhile the new vectors were respectively transformed into Bacillus cereus B905 by electroporation and the engineering bacteria with GFP, Bacillus cereus B905（gfp-5, gfp-8 and gfp-17） were obtained. The results clearly showed bright green fluorescence of the transformants, and the fluorescence of the B905-gfp17 was less than that of the others.

Abstract The polymorphisms of pituitary adenylate cyclase-activating polypeptide gene(PACAP) were detected in a 257 Qinchuan cattles(Bos tarurs) population by using PCR-SSCP and the assocation of genotype with partial growth traits was calculated by the least squire method(LSM). Result showed that a G/A mutation was found at the locus of 3909 bp, which result in 3 genotypes named AA,AB and BB respectively. Allele frequencies of A and B were 0.5992 and 0.4008, respectively. χ2 test showed that the locus was in Hardy-Weinberg equlibrium in the Qinchuan cattle population detected. Variance analysis showed: The body length of individuals with AA, AB and BB genetype were extremely significantly difference ( P<0.01); The wither height of individuals with AA and AB genotype was extremely significantly higher than those with BB (P<0.01), and those with AA genotype was significantly higher than those with AB ( P<0.05); The body weight of individuals with AA and AB genotype was extremely significantly higher than those with BB (P<0.01); The heart girth of individuals with AA genotype was extremely significantly higher than those with BB (P<0.01), those with AA genotype was significantly higher than those with AB genotype(P<0.05), and those with AB genotype was significantly higher than those with BB genotype( P<0.05); The heart width of individuals with AB genotype was significantly higher than those with BB ( P< 0.05); There were not significantly differences in the other growth traits among individuals with different genotypes. The locus can be a potential QTN affecting body length, wither height, body weight, heart girth and heart width or tightly linked with it. It will be a candidate moleculer marker for the Qinchuan cattle breeding.

In this study, phytase gene (phyA2) from Aspergullus nige were transformed into genome of maize (Zea mays L.) rataria by particle bombardment. The results of RT-PCR and Western blotting detections showed that phyA2 were inserted into maize genome and expressed. The assay of phytase activity in maize plant root and utilization of phytate showed that phytase activity of plant roots was a significant factor in the utilization of phosphorus from phytate.

To investigate promotion functions and interactions between TATA/CAAT boxes and darkness inducing motif YTCANTYY in pib promoter, a set of recombined binary plasmids of 3'end deleted pib promoter were constructed. Experiment results from analysis of their transgenic rice (Oryza sativa ssp. japonica) plants showed that the deletion of pib promoter fragments of TATA and CAAT boxes in 3' end remained not only their promotion activity but also their high expression in root system. The 3' deleted pib promoter harboring 2 or 4 YYTCANTYY and intact pib promoter harboring 6 YTCANTYY all possessed darkness inducing activity. After 24 h darkness treatment, GUS activities in the transgenic rice plant roots of above three recombined binary plasmids enhanced clearly and the increases of GUS activities in the stem and leaf were not obvious. These were said that the darkness inducing activity in pib promoter is mostly decided by the motif of YTCANTYY and this motif also can compensate for the missing fuction of TATA/CAAT.

The cry1Ah gene is one of the novel insecticidal genes cloned from Bacillus thuringiensis isolate BT8. A plant expression vector pHUAh harboring the cry1Ah gene was constructed and transferred into maize (Zea mays L.) by microprojectile bombardment. Thirteen regenerated plants were PCR positive. Bioassay results showed that events B1-1 and B1-7 had high resistant to the Asian corn borer (Ostrinia furnacalis). These two events were analyzed for 4 years from T1 toT5 generations. The results showed that foreign gene cry1Ah was expressed stably in maize and could be inherited stably over generations. Cry1Ah protein expressed in events B1-1 and B1-7 and their different tissues were analyzed by ELISA. The results exhibited that the contents of Cry1Ah protein were different between the events B1-1 and B1-7, as well as among different tissues. Results of bioassay to T2~T5 transgenic maize plants suggested that the transgenic plants had high resistance to the Asian corn borer and the resistance can be inherited stably. So events B1-1 and B1-7 can be potential candidates for the breeding of Bt insect-resistant transgenic maize.

Expansins are considered to be the key regulators of cell wall extension during plant growth. In the present study, rapid amplification of cDNA(RACE) ends was used to clone the full length cDNA of TaEXPB8 homeologous genes. Four distinct cDNA sequences, with the length of 1180， 1169， 1150 and 1135 bp were identified and named as TaEXPB8-1， TaEXPB8-2， TaEXPB8-3 and TaEXPB8-4, respectively. Interestingly, putative auxin responsive cis-acting regulatory elements were detected in the promoter region of these four homeologous genes. Further analysis revealed that the chromosome location of TaEXPB8-1 was anonymous. but TaEXPB8-4 was located on Bin6DS（0.99~1.00）. Moreover, TaEXPB8-2 and TaEXPB8-3 were located on same chromosome region Bin6AS（0.35~1.00）, and tandem repeat of expansin genes were also detected in the linear genomic region of rice（Oryzasativa L.）, maize（Zea mays L.） and Arabidopsis. Thus, TaEXPB8-2 and TaEXPB8-3 could be originated from one ancestor gene, suggesting the possibility that these two genes were formed in wheat before the divergence of monocots and dicots. It was shown that TaEXPB8 gene was mainly expressed in elongation zone of wheat primary root, and down-regulated by high concentrations of exogenous auxin treatment, indicating that this gene it might play an important role in wheat root development, especially for root elongation.

Abstract Food safety assessment of genetically modified food is an important step of developing genetically modified products, and animal experiment is the only way to assess the genetically modified food safety. Generally animal study is to evaluate effects on health in the overall level. In this article, animals used are classed large livestock and small rodents, different animal studies on evaluating the safety of genetically modified food are summarized and the prospect of animal study is focused on.

Abstract Recombinant plasmids pKtac1-act1 and pKtac1-act2 were integrated into Comamonas testosteroni chromosome by electroporation respectively, and two engineering strains were obtained and identified by the analysis of PCR and Southern blot. Expression and stability of 3α-hydroxysteroid dehydrogenase/carbonyl reductase(3α-HSD/CR) of engineering strains were detected. The results indicated that the expression of 3α-HSD/CR almost 20-folds increased in two engineering strains than that of the wild type without inducing; and the expressions of activator and 3α-HSD/CR were quite high and stable when engineering strains inoculated in LB medium with antibiotic; but unstable when inoculated in LB medium without antibiotic. The preliminary results indicate that the expression of 3α-HSD/CR can be regulated by the activator gene.

Abstract Based on the morphological, physiological and biochemical characteristics and 16S rDNA sequences analysis, strain AiL3 isolated from mangrove(Acanthus ilicifolius) was identified as Bacillus amyloliquefaciens. Antifungal substance from fermentation of strain AiL3 was deposited by the method of organic solvent extraction and ammonium sulfate, exhibited a high antifungal activity of precipitated by 50% ammonium sulphate precipitation. The results implied that antifungal substance of strains AiL3 was a kind of proteins. It was shown on PDA plates that the development of many pathogens such as Colletotrichum gloeosporioides and Fusarium oxysporum f.sp. cucumerinum was inhibited by the antifungal protein effectively. Their EC50 values were 9.32 and 5.81μg/mL on mycelial growth and conidium germination of C. gloeosporioides. Furthermore, the antagonistic crude protein(20.33μg/mL) could degrade the conidial cell wall and restrain completely the formation of spore.

Abstract Purity identification for hybrid seeds is very important in oilseed rape seed production. Analysis of peroxidase (POD) isozyme of Brassica napus hybrid.-Zhongyouza 12 and its parental lines indicated that one specific band with a relative mobility (Rf) of 0.41 presented in hybrid and male parent, but absent from female parent. So this specific band could be used to identify hybrid of seeds produced from sites where the male parent plants were strictly removed after flowering. For more precise identification, 173 pairs of SSR primers were screened for polymorphic amplification between the parental lines of Zhongyouza 12. Among those primers, six pairs amplified clear specific bands to either of the parents, with no difference among individual plants within lines. Four pairs of the primers, P052, P130，P131 and P154, amplified specific bands of both parents in F1 hybrid plants. Primer combination P131+P154 could create additive band profiles in one reaction system as which amplified by P131 and P154, respectively. In case of seeds produced from difficult conditions where the pollen source out of control, the use of P131 + P154 primer combination could improve the detection sensibility of unknown pollen, and more accurately identify the seed purity of Zhongyouza 12 under the condition of using the same workload. One hundred DNA samples from Zhongyouza 12 single plants were analyzed by using primers P131, P154 and P131+P154, respectively. The results of seed purity were fully consistent, and were very close to the results of field identification.

Abstract Using the loop-mediated isothermal amplification(LAMP), a novel method of gene amplification for Mycoplasma hyopneumoniae detection was established. A set of six primers was designed specificity to recognize the 16S rRNA gene, which is a relatively conserved region of M. hyopneumoniae. The LAMP-based assay could be completed within 30 min at 63℃. Results showed that the LAMP-based assay, compared with PCR, had high superiority. The detection limit of the LAMP assay was found to be 10 copies per reaction determined by using a recombined plasmid containing the target sequence, which was 10-fold higher than that of the PCR assay. In addition, both LAMP and PCR were highly specific to M. hyopneumoniae. Furthermore, the LAMP and PCR assay was applied to 127 clinical nasal swab samples collected from pig farms. For the LAMP assay, all the nasal swab specimens tested positive, while for the PCR assay, 116 nasal swab specimens tested positive. The data confirmed that LAMP was more sensitive than PCR for the detection of clinical samples. The LAMP assay is easy to perform, extremely rapid, highly sensitive, specificity and cost-effective, therefore it has the great potentiality suitable for diagnosis of M. hyopneumoniae both in well-equipped laboratories and in field situations.