Freshwater mussels play an important role in many aquatic ecosystems, but little is known about their biodiversity and conservation genetics. In order to provide genetic information of freshwater mussels in China, microsatellite markers were used to investigate genetic diversity of five species of freshwater mussels in family Unionidae, Hyriopsis cumingii, Cristaria plicata, Lamprotula caveata, Schistodesmus lampreyanus , Acuticosta chinensis, which were collected from Qinglan Lake, Jiangxi Province. 8 primers for polymorphic DNA microsatellite loci were sieved from 20 primers which were developed from oyster mussel (Epioblasma capsaeformis) (Bivalvia: Unionidae) in North America and freshwater pearl mussel (Margaritifera margaritifera L.) (Bivalvia: Margaritiferidae) in Central Europe. Allelic richness ranged from 2 to 6 alleles per locus and averaged from 2.8750 to 4.000 alleles for five mussel populations. Heterozygosity levels varied from 0.4417 to 0.6333 for average observed heterozygosity (HO) and 0.4430 to 0.5706 for average expected heterozygosity (HE), average polymorphism information content (PIC) ranged from 0.368 to 0.498, which revealed that five populations showed higher genetic diversity. Many loci in different populations deviated from Hardy Weinberg equilibrium. Genetic distance (D) among five mussel populations ranged from 0.6228 to 1.4724, the maximal D was 1.4724 between H. cumingii and C. plicata, and the minimal D was 0.6228 between S. lampreyanus and L. caveata. The study of genetic diversity for freshwater mussels can provide guidelines for resource conservation and genetic improvement.

Differential Display Reverse Transcript Polymerize Chain Reaction (DDRT—PCR)was applied to study gene expression in heat-tolerant variants N1-2-1 and it’s mother plants N1 under heat stress. The results show that 1419 bands were amplified from 40 pairs of primers in the two clonal lines, 100 of them appear different obviously between the two clonal lines and which accounted for 7.2% , 41 of 100 bands which came from heat-tolerant variants N1-2-1. The northern blot results showed that 6 segments of the 41 were confirmed to express only in the N1-2-1, those fragments were then cloned into vectors for sequencing, sequence analysis through Internet Blast searching showed that 2 of them related to response of rice and potato under heat stress respectively, one of them related to anthocyanin 5-aromatic acyltransferase , and the other 3 fragments（DF2、 DF5、DF6） were novel cDNA fragment.

The development of near-isogenic lines (NILs) is one of the important foundations for the establishment of molecular linkage map, the mapping of quantitative trait loci (QTL) and molecular marker-assisted selecton (MAS). In the present study, NILs for two major QTLs of basal root thickness(BRT) and 1000-grain-weight(TGW) were obtained through the foreground selection for target QTL, the background selection for the genome of the recurrent parent of 3 backcross populations( BC1F1、BC2F1、BC3F1) and phenotypic selection for the population of BC3F2. They were 9 of BRT QTL-NILs with BRT ranges of 1.07~1.16mm, over the recurrent parent by 6.11%~15.18% and an average recovering ratio of genetic background (RRGB)of 97.22%; 11 NILs of 1000-grain-weight (TGW) QTL with ranges of 21.25～26.25g, over the recurrent parent by 7.05％～32.16％ and 95.97% of RRGB. The number of molecular markers for background selection in the development of NILs and QTL cloning were also discussed.

Lycopene is a kind of carotenoid. Because of its important role in preventing some diseases such as the cancer and the heart disease the lycopene is becoming the research hotspot as the function food in recent years in the world. Increasing the express of Psy gene and blocking the express of Lcy gene which is the main gene regulating the transformation of the lycopene is the convenient way to enhance the content of lycopene. The primers were designed according to the gene sequence（NM-121729）、（U46919）and (X86452) in GenBank. The Psy gene was isolated from the cDNA of the Arabidopsis thaliana. The Pds promoter and the DNA segments of the Lcy gene were obtained from the genome DNA of tomato. The 3’end of lcy DNA segment was connected together by an intron to inform the RNA interferential segment then which was inserted in the expression vector with the fruit-specific promoter of phytoene desaturase gene(pds)；The Psy gene was inserted in the same vector with the Pds promoter. Then the expression vector was obtained which can increase the expression of Psy gene and interfere the expression of Lcy gene at the same time by transformant into the plant with the carotenoid.

Polyphenol oxidase pigmentation method was studied using different wheat cultivars and F2 of different cross combinations. The results were as fallows: the suitable condition of seed treatment was that under room temperature 18~25℃, wheat seeds were put in 0.125% catechol solution and kept for 10~15min, then the seeds were covered with plastic membrane and kept for 4~6h. After drying, the seeds were graded in term of color display. There was a significant positive correlation between sprouting percentage and colored seed grade. Light colored seeds with low polyphenol oxidase activity had higher resistance to pre-harvest sprouting than those of dark colored seeds. The new wheat variety Baomai 8 with high resistance ability to pre-harvest sprouting was successfully selected and bred using this method. It was registered and released to commercial production in Hebei Province in 2004 (registered number: JS2004012).

Abstract: To evaluate the effects of a system of serum-free maturation culture on in vitro maturation of goat oocytes and development of parthenogenesis embryos，0.3 % BSA (m/v), 1% PVA (m/v) and 1% ITS (v/v) were added to basic medium of oocyte maturation as substitute of serum, 5% fetal bovine serum was added to control. Cumulus-oocyte complexes (COCS) derived abattoir ovaries were cultured in vitro with those culture medium respectively. Maturation rate of PVA supplementation group (50.00%) was significant lower than that of control (83.23%)(P<0.01), and that of BSA group (71.19%)and ITS group(76.79%) had no difference compared to control; The matured oocytes were actived by ionoycin and 6-dimethylaminopurine, then, parthenogenesis embryos were cultured in SOFaa medium to observe development of embryos each group. Cleavage rate from BSA, PVA, ITS and FBS group was 61.90%, 46.97%, 79.07% and 83.58% respectively, and blastocyst rate respectively was 25.00%, 19.70%, 55.81% and 60.45%. The results show that substitute PVA and BSA for FBS in elemental maturation medium decrease significantly the abilities of maturation and early development of embryos(P<0.05 or P<0.01),substitute ITS for FBS decrease the abilities of maturation ,but has no effect on early development of embryos(P>0.05).Conclusion: ITS may be applied to in vitro maturation of goat oocyte as substitute of FBS.

A fragment of 1018 bp of ACO gene cDNA sequence was cloned using RT-PCR with two PCR primers designed according to the sequence of tomato cDNA clone (E11). The result of BLAST showed the sequence presented a very high match with the ACO genes from other plants; the homologue is from 83% to 99%. Using the sequence, an RNA interference (RNAi) transformation vector (pD311) was constructed through the way of BP cloning. The transformation was performed to tomato. 27 regenerated plants with kanamycin resistance were obtained. And the transgene integrated into tomato genome was proved with PCR and Southern blotting. The ethylene respiration amount of the RNAi transgenic tomato plants was measured by gas chromatography. The results showed that ethylene evolution was specifically inhibited in the leaves and fruits of transgenic plants.

Using UV mutant breeding method for improving the herbicidal activity of HGE which was evaluated as a potential bioherbicide for biological Echinochloa species control in rice, we screened one mutant strain M1 which might be utilized as the industrial strain in liquid fermentation. It was clearly observed that the morphological characteristics of M1 were obviously different from the wild strain HGE that included M1 colony color was gray to white but HGE was dark green. Furthermore the mycelia growth rate of M1 was faster than wild strain HGE and inhibition rate of fermentation liquid to barnyardgrass radicle and plumule was also superior to HGE. After pass-generation tests for 7 times, M1 still showed fairly excellent genetic stability. Further RAPD-PCR analysis DNA of M1 and HGE suggested that their genetic substance had been actually varied. For a mass of mycelia yield and metabolized toxin products of mycoherbicide, the optimal liquid fermentation technics was confirmed with submerged culture experiments. The highest mycelial yield was up to 14.37g/L when cultured in No. 3 medium at pH7, 150r/min and 24℃ for 6 days. The optimal fermentation technics of toxin products was obtained as following instance: stationary culturing in No. 4 medium at pH7 and 24℃ for 2 weeks.

S-layer protein CTC surface display system of Bacillus thuringiensis was used to test the possibility of displaying H5N1 avian influenza virus haemagglutinin HA1 on cell surface. Two fusion genes were constructed by replacing the central part below the surface anchor sequence slh of S-layer protein gene ctc with part of ha1 gene (ha1p). Two recombinant B. thuringiensis strains were constructed by transferring recombinant plasmids to B. thuringiensis plasmid-free derivative strain BMB171, such as BCCH (harboring ctc-ha1p as well as a plasmid carrying surface display helper csaAB operon) and CH (harboring csa-ctc-ha1p). Hemagglutination assay showed recombinant HA1 proteins were displayed on the surface of two recombinant strains. Hemagglutination inhibition assay showed recombinant HA1 proteins were specific to standard HI (Hemagglutination Inhibition test) serum of type H5 AIV. After immunizing mice with recombinant strains, the recombinant HA1 proteins elicited a humoral respone to HA1 and exhibited immunogenicity as assayed by enzyme-linked immunosorbent assay. Meanwhile these assay showed recombinant strain CH exhibited the higher activity than BCCH. This demonstrated that the construction way of fusion gene csa-ctc-ha1p was more appropriate to S-layer protein displaying heterologous antigen on cell surface. The strategy developed in this study gives a possibility to generate heat stable, oral, veterinary vaccine against AIV with B. thuringiensis S-layer protein CTC surface display system.

In order to get the soluble expression products, Shiga-like toxin IIe A subunit was analyzed by TMpred software. Then a pair of primers were designed and the expression plasmid pGEX-A was constructed before being transformed into BL21 (DE3). SDS-PAGE and Western blotting results showed the gene fragment was expressed abundant soluble protein and the expression products had immunogenicity. The assay of it’s bio-characteristic toxicity showed that the soluble fusion protein lost the toxicity as SLT-IIe. Polyclonal antiserum produced by immunizing the New Zealand rabbits with the purified expression product restrained the toxicity of SLT-IIe to Vero E6 cells because it had significantly high neutralizing activity (90.4℅) against SLT-IIe. But the protective efficacy test showed that the immunization with the expression products could not reduce mortality of the mice. The ELISA method was established using the expressed protein to detect anti-SLT-IIe A IgG. The assay had excellent specificity, sensitivity and reduplication and could be applied to detect the edema disease in piglets.

The fused gene of Enterotoxigenic Esherichia coli mSTⅠ-linker-LTB was construction and was cloned into plasmid pET32a(+).The recombinatant plamsid was transformed into the expression host strain E.coli BL21（DE3）. After 5 hour induced by 1mM IPTG at 30℃, the fusion protein which molecular weight was about 37K was at the highest level. Its content was about 30% of that of total bacterial proteins. The fusion protein was mainly in inbody and was positive by Western blot. It was no the biological toxicity of the native STI demonstrated by the Suckling Mouse Assay. The purified recombinant protein emulsificated with mineral oil offered ICR mouse 83.3%、100%、66.7% and 66.7% protection against the challenge of Enterotoxigenic Esherichia coli K88、K99、987P and F41,respectively.

To test immune protection of SjIR3 against challenge infection of Schistosoma japonicum(Sj) in mice.The special primers were designed according the sequence of SjIR3 in GeneBank (Ay25148). SjIR3 was amplified using Sj adult worm cDNA library as temple. The obtained sequence of SjIR3 was analyzed in internet. The recombinant plasmid pET-32(a) / SjIR3 was constructed by routine methods. The recombinant protein rSjIR3 was expressed by induced with IPIG. rSjIR3 was analyzed by SDS-PAGE and western-blot. Forty-four Kun-Ming mice (female, 20g) were randomly divided into 4 group each with per 11. Group A.B were control immunized with PBS and FCA respectively. Groups C,D were immunized with 50µg rSjIR3,50µg rSjIR3 + FCA respectively in 0-2-4-6th week. Levels specific antibody were detected by ELISA. The mice were challenged with 40±1 S.j cercariae per mouse 2 after the fourth vaccination. Forty-five days later, mice were killed and perfused, and the adult worms and eggs were counted. Results showed DNA segment of SjIR3 is 890 bp long and 100% identify with SjIR3 in GeneBank (AY251481). Protein expressed by SjIR3 is trans-membrane, the worm reduction rate was 26.63% and 34.79%. The egg reduction rate was 36.38% and 44.09%.

A rabbit monoclonal antibody(RabMab) was generated toward theβ-Adrenergic agonist Ractopamine. Bovine serum albumin(BSA) and ovalbumin(OVA) were used as carrier proteins and Ractopamine-glutarate- BSA was used as the antigen. The WHB dark-eye rabbits were used as antibody generation animals. B lymphocytes from immuned rabbits and 240E myeloma cells were fused to generate hybridoma cells. Supernatant from clone A3-2-6 was used for the development of an immunoassay. The selected antibody was specific for ractopamine with an IC50 of 4.09ng/ml. The linear range of the inhibition rate vs log ractopamine competition curve generated in PBST was 0.1～100ng/ml. β-adrenergic agonists which had similar structures to ractopamine such as clenbuterol hydrochloride, isoxsuprine, epinephrine, norepinephrine, isoprenaline, salbutamol and dopamine showed no cross-reactivity. In conclusion, a highly specific RabMab to ractopamine hydrochloride was developed that could be used in indirect competitive ELISA screening assays.

By immunohistochemistry and in situ hybridization，the localizations of heat shock protein 70 (HSP70) and HSP70 mRNA in heart，liver，lung，kidney，spleen，thymus and bursa of Fabricius of 6 h heat stressed broilers were studied. The result showed that the positive signals of HSP70 mRNA at 6 h heat stress were localized in the liver and lung，especially in the wall of vessel. The weak positive signals were seen in the myocardial cells. No significant signals were observed in spleen，thymus and bursa of Fabricius；HSP70 signals were localized in the wall of vessel of all detected tissues at 6 h heat stressed broilers. The localizations of HSP70 and HSP70 mRNA suggest that HSP70 is correlation with the cardiovascular function.

Abstract: the RT-PCR method was established. In this test, total RNA was extracted from the fresh liver of rabbit haemorrhagic disease rabbits, a pair of primers were designed according to the sequences of RHDV published in GenBank, which can amplified a 269 bp segment. The product was sequenced and there was 95.8％～98.5％homology between the product and the relevant sequence published in GenBank. RHDV viscera distribution experiment shows that liver, kidney, spleen, blood, lung were HA positive, and all the viscera except feces were positive by RT-PCR. The sensitivity of RT-PCR for RHDV is 4×104times that of HA. All those experiments above showed the established RT-PCR method detecting RHDV has strong speciality, high sensitivity and good repetition. This RT-PCR can not only be used in RHDV clinical diagnosis and epidemiology study but also in quarantine of rabbit product ,example for meat.

Abstract: The avian reovirus (ARV) σ3 gene DNA fragment was labeled with Digoxigenin as cDNA probe for detection of ARV in chicken tissue and feather tips in Dot blot hybridization. As little as 1.6pg of ARV RNA could be detected with the DIG-labeled probe. Chickens were infected with ARV by inoculating at one-day old and in-contact infection. ARV could be detected in most tissues of inoculated chickens 24 hr post infection(PI) and spread to un-inoculated birds by contact rapidly. Positive rates in feather tips were correlated with other tissues. The result demonstrated that the dot blot with DIG probe to detect ARV in feather tips is a useful method for determination of ARV infection and spread in chickens, and it could be helpful when a large number of samples should be tested.

In order to better understand the proliferation and differentiation of intramuscular preadipocyte and to clarify factors influence its proliferation and differentiation, cells from adipose tissues within muscles between the sixth and the seventh rib were cultured by means of using collagenaseⅠdigestive method. The results were as follows: the separated cells were highly homogeneous, highly proliferative and with a high differentiation rate. Their dynamic morphological changes, growth curve, oil O staining, and reaction to insulin and dexamethasone all verified their preadipocyte identity. Under controlled conditions, the intramuscular preadipocytes replayed their proliferation and differentiation process in vitro, especially, their euploid status was proved by the analysis of chromosome. In conclusion, the study confirmed there are functionally active preadipocytes in adult cattle’s adipose tissues within muscles, which gives a theoretical basis for improving meat quality by further studying the deposition of intramuscular adipose tissues.

Using the gus gene as reporter gene, inducible expression activity in wheat calli of Prd29A promoter was measured by transient expression. The results showed that Prd29A promoter had expression activity under the salt-stress conditions in wheat. Based on the experiment above, we introduced Escherichia coli trehalose-6-phosphate synthase gene(otsA)into NC239 wheat variety through pollen tube pathway, and some otsA transgenic plants were obtained by PCR、Southern blot and trehalose amount analysis.

Abstract: Using six pairs of specific primers, DNA fragments containing the promoter of 16S rRNA gene and the promoter and terminator of psbA gene, were obtained by PCR and sequenced from Populus tomentosa chloroplast genomic DNA. Sequence analyzing results showed that the functional domains such as -35 domain, -10 domain and the translation control sequence motif in the two cloned promoters and the “TAA” sequence in the cloned terminator were also confirmed. New primers were synthetized, the promoter Prrn, PpsbA and the terminator TpsbA were cloned correctly and the prokaryotic expression vectors of GFP were constructed by using Prrn with rbcL RBS sequence, PpsbA and T7 promoter, and the terminator TpsbA, respectively. Prokaryotic expression result showed that the promoter Prrn, PpsbA and the terminator TpsbA we cloned were functional and were constitutive expressed in E.coli BL21. It slso showed that the RBS sequence was very important in gene expression.

The expression of many induced genes is controlled by specific transcription factors and cis-acting element. Dehydration-responsive element (DRE) is very important in the regulation of the expression of drought and low-temperature responsive genes. DRE-binding transcription factors from Arabidopsis have been described. While research on DRE-binding transcription factors tree plants is scarce. Yeast one-hybrid approach is a powerful method for the searching for sequence-specific DNA binding transcription factors. Construction of DRE-containing report yeast strain is the first step for finding DRE-binding transcription factors from tree plants through yeast one-hybrid approach. This paper reports the construction of the DRE-containing report yeast strain. Our preliminary data show that the report strain we constructed is suitable for the screening for DRE-binding transcription factors from tree plants. Our work should be helpful for the improving of the stress-resistance features of tree plants.

The regulative sequence (Posrbcs) of ribulose-1 ,5-bisphosphate carboxylase small subunit genes ( rbcS ) of rice was cloned by PCR amplification. The cloned Posrbcs was fused with gus gene, and then introduced into rice by Agrogacterium-mediated transformation. The result of GUS histochemical staining showed that gus gene driven by Posrbcs expressed in leaf , culm, sheath and glume, and not in endosperm. Also, 5’ end difference length deletions of Posrbcs were fused with gus gene and the fused genes transformed rice respectively. Quantitative analysis of GUS activity showed that the expression level of gus gene was reduced with the deletion of promoter. The result of this experiment also showed that light induction could enhance GUS activity. Furthermore, the deletion of light inducible elements such as I box, GT1binding factors, GATA box, T box could reduce GUS activity and postpone the light-induction time. Electrophoretic mobility shift assays showed I box, GT1 factor, T box and GATA box could bind with the nuclear protein.

In this paper, we detected the Papaya Ringspot Virus (PRSV) on the papaya plants with nucleic acid hybridization by digoxgenin labeling; alkphos direct labeling cDNA probes and RT-PCR. The specific primers were designed according to published sequences of PRSV CP genes of Chinese Sm strain in Genebank to perform a RT-PCR, total RNA extracted from PRSV-infected samples，Carica papaya L.cv. Meizhonghong, cloned cDNA was linked to pGEM-T easy plasmid vector and sequence determinate. Recombinant plasmids were used as the template for labeling with PCR DIG Probe Synthesis Kit. Recovery the cDNA fragments by electrophoresis and labels cDNAs with AlkPhos Direct Labeling method. The result indicated that A. the sequence of the Meizhonghong’ No.2 isolate showed a 94.7 % identity with the Chinese PRSV-Sm genome. B. Blot hybridization obtained identical hybridization result when 861bp，455bp and 215bp probes labeled by DIG, and that 861bp probe lead to the clearest hybridization signal, above detected result were accord with that of RT-PCT, nucleic acid hybridization technique resulted in a sensitivity and special detection of PRSV in papaya, which is sufficient for routine diagnosis routine. C.The AlkPhos Direct Labeling 861bp probes could make for ideal blot hybridization result, and that the 455bp was invalidation. D. DIG labeling probe was performed in the vein tissue print hybridization in the determinations of PRSV.

The open reading frame (ORF) 27 of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus is 768bp in length and potential to encode a polypeptide of 255 amino acids, with a molecular weight of about 29.5kD. The coding region of ORF27 was amplified from HaSNPV genome DNA by PCR, and cloned into pGEM-T easy vector, and then subcloned into the expression vector pGEX-4t-2 and the recombinant plasmid was named pGEXORF27. DNA sequencing was performed to confirm the correct amplification and orientation of the target sequence. pGEXORF27was transformed into competent cells of Escherichia coli BL21, and expression was induced for 3 hours in 1 mmol/L IPTG at 37℃. The molecular weight of fusion expression protein was 55.5kD, and it was produced to an amount about 9.2% of the total bacterial cellular protein. Western blot analysis showed the recombinant protein could react with GST antibody. Cutting gel and dialysis purified ORF27 protein. ORF27 gene was also inserted into pFastBacHTb and constructed pFastBacHTORF27. The recombinant donor plasmids were transformed to DH10 competent cells and constructed recombinant BacmidORF27. Mediating by Lipofectin, BacmidORF27 was transfected into Tn-5B1-4 cells of cabbage looper, Trichopolusia ni. SDS-PAGE analysis showed the expression product of ORF27 gene was 32kD that was identical to the predicted molecular weight. SDD-PAGE analysis demonstrated that the protein was produced to an amount about 2.9% of the total insect cellular protein. Western blotting analysis with anti-ORF27 further confirmed the preciseness of the expression product. Immunoelectron microscopy demonstrated ORF27-His fusion protein was localized in the cytoplasm and hardly present in the nucleus. The expression of ORF27 and its localization would lay the foundation for its function study.

In order to obtain the more thermostable β-glucanase, directed evolution was used to evolve the β-glucanase gene, and the enzyme properties were analyzed. The two mutants, i.e., EGs1 and EGs2, were obtained. The results of the enzyme properties analysis indicated that the melting temperatures of EGs1 and EGs2 were increased by 3℃ and 5℃, and approached 65.5 ℃ and 67.5℃; The optimum temperatures of the two mutants were increased by 5℃, and approached 60℃;The abilities to produce reducing sugar from lichenin were either much higher (28%) for the former or much lower (21.6%) for the latter ;The affinities of the two mutants for the lichenin were basically unchanged in comparison with their parental enzymes. The optimum pH were 6.5 for the EGs1 and wild-type, and 7.0 for EGs2; The pH stability for all enzymes was good, the activity of parental enzyme and the two mutant enzymes have no loss at pH 6.0-8.0 and pH 6.5-8.0 for 48 hours, respectively; The plasmids stability of the two mutants were good and their expression ability of β-glucanase kept stable after 50 generations. The results demonstrate that directed evolution is an effective approach to improve the properties of a mesophilic enzyme.

Having known the Xyn II cDNA sequence (Genbank accession number DQ114485),a pair of primers (with the EcoR I and Sal I sites) were designed.With the recombinant plasmid pUCm-T-xyn II as a template,Xyn II cDNA fragment (555 bp) encoding mature peptide was then amplified,inserted into the plasmid pET-28a.The resultant recombinant plasmid pET-28a- xyn II was transformed into E. coli BL21- CodonPlus(DE3)-RIL,and finally the recombinant strain B21/xyn II was obtained.Induced by IPTG,a maximum activity of 35.6 U/mg was obtained from cellular extract of E. coli B21/xyn II.The recombinant protein was purified by immobilizing metal affinity chromatography.The optimum temperature and pH for recombinant enzyme were 45℃ and 4.6,respectively.

It is known that the performance of meat sheep is controlled by some major genes or quantitative trait lacus(QTL). So a field in these genes and their mechanism on some important traits is a focus for genetic and animal breeding specialists all over the world. Some recent advances on major genes or quantitative trait lacus (QTL) for carcass traits were reviewed in the article ,such as Callipyge ,Carwell, double muscling and other QTL affecting muscle development. Furthermore, some opinions and resolution of some problems in the research concerned was put forward in the article.

Transgenic animals have become the development hotspot of life sciences ,and transgenic sheep has already been the important parts of transgenic animas.Since the first transgenic sheep was produced by microinjection in 1985,more and more transgenic sheep has been born successfully by different transgenic techniques .At the same time,the application of transgenic sheep has been moving towards mammary gland bioreactor from transgenic breeding,and the outcomes are great.Here, author makes a summary for this.

The technology of ultrasound-guided ovum pick-up (OPU) was reserched in bovine. The effects of different opu pressure、different opu interval and injecting of FSH were evaluated.These results demonstrate that: 110mmHg is the best pressure in opu. with this pressure, recovery rate was 66.03%,6.90±2.08 oocytes were obtained, of wich 68.84% were of good quality.With the frequency of twice weekly OPU, 5.29±1.19 oocytes were obtained and the recovery rate was 68.65±4.19%.there are more oocytes can be obtained under twice weekly OPU than once weekly and once two weekly opu.FSH treatment result in a higher oocytes yield and a better oocytes quality: 12.36±2.39 oocytes were yield in once opu and 75.19% with good quality,it was significantly different to non FSH group.

The AFLP markers was applied to analyze the genetic structures of Oreochromis niloticus, Oreochromis aureus and their hybrids F1, as well as the orange-colored Oreochromis mossambicus, Oreochromis hornorum and their hybrids F1. 8 pairs of primers had been chosen from ninety pairs of primer combinations for selection amplification. A total of 250 loci had been produced using these 8 pairs of primers (100-500bp in length for each segment). The number of the amplified bands detected by each AFLP primer combination for each individual ranged from 20 to 47. The analysis indicated that the intraspecies genetic similarity of the four tilapia, arranged from high to low, were Oreochromis aureus (0.913), O. niloticus (0.871), O. hornorum (0.829) and O. mossambicus(0.784). The genetic similarity and genetic distance between O. niloticus and Oreochromis aureus were 0.811 and 0.189 respectively, while the genetic distance of the parent fish, O. niloticus, O. aureus and their hybrids F1 (Ni-ao) were 0.165 and 0.142 respectively. The genetic similarity and genetic distance between O. mossambicus and O. hornorum were 0.756 and 0.244 respectively, while the genetic distance between the parents and their hybrids F1 (Mo-he) were 0.148 and 0.162 respectively. Specific segments were produced by primer combination E-AGG/M-CTT in O. niloticus and E-ACA/M-CAC in the Ni-ao, which would work as molecular markers to identify O. niloticus, O. aureus and their hybrids F1.

To study the changes of immunity activity after the hemopoietic organs damaged, silkworm larvae were radiosurgery with carbon ion beams, each us the change of body weight in 5th instar, phenolxidase activity in hemolymph and the quantity of induced bacterial challenge-related proteins were investigated. The results showed: after the hemopoietic organs being damaged, the activity of phenolxidase was down, the quantity of bacterial challenge-related proteins induced by E. coli significantly decreased and the survival rate of larvae fed with Bacillus thuringiensis dropped obviously, the body weight of larvae fed with mulberry leaves hardly decreased, while for larvae fed with commercial artificial diet, the body weight increased and almost had no significant different with that of control’s. It is considerated: after the hemopoietic organs of silkworm being destroyed, they dropped body immune activity; raise by no bacteria condition can meliorate the effect caused by the hemopoietic organs functional obstacle.

Fe(II) transporter gene MxIrt1 was cloned from Malus xiaojinensis Cheng et Jiang. MxIrt1 has the conserved domain of ZIP gene family. It was supposed to be involved in the iron absorption. Full length of MxIrt1 gene under the regulation of 35S promoter was introduced into Arabidopsis mutant irt1-1 by Agrobacterium tumefaciens. GUS staining and Southern analyses indicated that MxIrt1 gene has been integrated into Arabidopsis genome. The transgenic Arabidopsis plants showed increased tolerance to iron stress than the mutant.

This experiment applies technique to analyze the polymorphism of the Fusarium strains. Using SDS alkaline lysis method extract the genomic DNA of eleven Fusarium which from the different part of Shanxi province. Amplifying the rDNA ITS region of these eleven Fusarium, which used one pair of differential primer, and betaken four restriction enzymes (TaqI, AluI, HaeⅢ, EcoRI) to digest the PCR amplification .Totally it gains sixteen polymorphic fragment which include eight special band, it’s molecular weight is 70-900bp. Finally adopted NTSYS-PC(2.1) package analyze the DNA restriction fragment polymorphism among every strains, which is classified five big groups and is coincident with the traditional classification. The result shows that this method can be used to analyze polymorphism among the Fusarium strains.

A keratinase-producing gram-positive bacterium, Bacillus sp.YYW-1, isolateded from feather waste, was identified as Bacillus subtilis and named as Bacillus subtilis YYW-1, based on its morphology, physiological and biochemical characteristics, growth and 16S rDNA sequence analysis. The optimum temperature and the optimum pH of the keratinase were determined to be 60℃ and 7.2, respectively, and its activity could be completely inhibited by EDTA and PMSF.

Bacteria have developed a variety of secretion pathways to secrete toxins and enzymes into the extracellular medium. Each system has its own advantages and limitations. There are four different forms of secretion systems enumberated as types I-IV. This review focuses on the relationship between the type II secretion system and pathogenicity of plant phathogenic bacteria.