Global transgenic herbicide-resistant crops accounted for more than 80% of the total transgenic crops plant area 125 million hm2 in 2008. Herbicide-resistance has been becoming the most important application of transgenic traits. The cost of weed control, herbicidal safety issue and residual injury problems contributed to quick development of transgenic herbicide-resistant crops. It is analyzed that large-scale commercialization of transgenic herbicide-resistant crops may result in uniform of herbicidal market and harmfully impact on herbicidal industry. In addition, it may face to other severe challenges that herbicide-resistant weed may evolve and the environmental safety issue of gene-flow may occur. Although transgenic herbicide-resistant crops are not commercialized yet, China is facing a good opportunity for development because of potential huge market and policy support. This article also addresses how to use herbicide-resistant gene reasonably, and the research and development strategy for transgenic herbicide-resistant crop. The purpose of this paper is to provide beneficial information with study and development of transgenic herbicide-resistant crops in our country.

Abstract: The object of this study is to screen the genes differentially expressed in flower induction by ethylene on bromeliads using suppression subtractive hybridization (SSH). SSH was performed in two directions to isolate the differentially expressed genes between exposed to ethylene for 1,6,24 h and the contract on Guzmania‘Attila’. The cDNA obtained from the final nested PCR were directly inserted into T/A cloning vector to establish subtractive hybridization library of specifically or highly expressed genes in flower induction by ethylene on bromeliads. The three forward SSH libraries contained 1063 clones. Reverse Northern dot blot was performed to screen the truly differentially expressed genes. Sequence analysis of these clones identified 990 differentially expressed clones, including 205 clones which were homologious to the reported genes in GenBank, and 313 clones were unknown genes. Information of this study could be used to isolate the Guzmania‘Attila’flower genes and understand the molecular mechanism of flowering.

Abstract: A full-length cDNA library of chinese dwarf cherry leaves was constructed by using λTriplEx2 vector according to the SMART cDNA library kit protocol. The titer of primary cDNA library was 6.17×107 pfu/mL containing 3.7×107 clones and the rate of recombinant was 95%. The average size of insert fragments was about 1.2Kb. 400 clones from the cDNA library were sequenced and the first 364 valid Chinese dwarf cherry ESTs were generated. Most of the ESTs were between 500~1300 bp with an average of 877 bp. 181 known or hypothetical functional genes、5 unidentified genes and 14 novel genes were annotated by BLASTX and BLASTN searches against the NCBI non-redundant protein and nucleotide databases.

Abstract: A suppression subtractive hybridization (SSH) cDNA library was constructed by using the storage roots of sweetpotato (Ipomoea batatas (L.) Lam.) cv. Lushu 3 resistant to sweetpotato stem nematode. A total of 1379 positive clones were screened and sequenced, and then clustered by CAP3 software, 185 unique expressed sequence tags (ESTs) were obtained. After blasted in Genbank, innotated GO and matched metabolism pathway in KEGG by Blast2Go software, it was found that 151 unigenes, accounting for 81.6% of all the unigenes, showed high homology with the function known genes or ESTs, which were related to energy metabolism, signal transduction, defense response and protein synthesis metabolism. Results of analysis on the function known genes suggest that mitogen-activated protein kinase, serine/threonine protein kinase, catalase, chitinase and glucanase may involve in the process of resistance reaction to sweetpotato stem nematode. Four defense-related genes wereconfirmed to have different expression profile at different times after inoculated with the stem nematodes by real-time quantitative reverse-transcriptase PCR.

To identify the genes controlling water stress response in Malus sieversii Roem. seedlings, suppression subtractive hybridization (SSH) was applied to construct forward and reverse subtractive cDNA libraries from Malus sieversii Roem cv. Xinyuan 1 treated with 20% PEG 6000. After sequencings 1000 clones from two cDNA libraries at random, 268 UniESTs were isolated. Homology search and sequences annotation were completed by Blastx and Blastn NCBI network service. According to MIPS function catalogue, all UniESTs were classified into 16 classifications. They were ascribed to proteins which were taken part in metabolism, transcription, protein biosynthesis and proteolytic, cell fate, cellular communication transduction, energy metabolism and so on.

In this paper, to study the molecular evolution patterns of polygalacturonases and the feasibility of using polygalacturonases gene to study species phylogeny, we collected 88 polygalacturonases sequences of bacteria, fungi, and plants from GenBank、EMBL and DDBJ database and constructed the phylogenetic tree. All sequence alignments were performed using the program CLUSTAL W, and the methods used for building the evolutionary trees was the neighboring-joining (N-J) method. The amino acid sequence alignment of all polygalacturonases sequences confirmed that there were four strictly conserved sequence segments (175NTD, 197DD, 218GHG, 250RIK). And conserved cysteines were also found. However, the conservation of cysteines reflects taxonomy, and the corresponding sites could be conserved only in the frames of the respective bacterial, fungal and plant groups. There is no one cysteine residue conserved out all the polygalacturonases. The evolutionary tree reflects both the taxonomy and specificity so that bacterial, fungal and plant enzymes form their own clusters, the endo- and the exo-mode of action being respected, too.

Abstract：Through analysis of the databank of Inspection America on field trials of genetic modified crops during the last 20 years between 1986 and 2007, the basic rule and characteristics in research and development of genetic modified crops were studied．Data were processed with the software “data perspective table” method imbedded in Microsoft Excel．Results showed that the field trials undertaken in America by the end of September, 2008 totaled 13782, involving158 crops, more than 1114 novel traits and about 98 companies or institutions．During 1987 and 2008, the number of field trials approved were firstly rising in a straight line, then stabilized at around 1,000 a year．The main test species include corn, soybean and cotton, accounting for 45.8％、9.7%、6.4% respectively． The main characters are involved in the anti-herbicide, insect resistance and quality traits． About 62.7% field trials were undertaken by 5 private companies such as Monsanto and Pioneer Hi-Bred International, Inc．The genetic modified crops which have a high economic value, mature technology and easier accepted have broader prospects．China should continue strengthen non－food crops genetic modified research and pay more attention to the genetic modified food at the same time．We also should build a sound safety of genetic modified production systems and use of transgenic technology for the service of socialist construction．

Mitochondrial DNA(mtDNA) was extracted by tender root tissue and leaf tissue from seedlings of four materials such as PolA and its maintainer line PolB, Shaan 2A and its maintainer line Shaan 2B in Brassica napus L., and the extracted mtDNA was examined. The results showed that the mean content of mtDNA extracted from tender root tissue was about 3250ng/g.FW, significantly higher than that from leaf tissue in the same conditions. That is, the mean content of mtDNA extracted from tender root tissue was about 6.5 times of leaf tissue, and the mtDNA extracted from tender root tissue had a high purity. The agrose gel electrophoresis also showed that mtDNA extracted from tender root tissue had clearer bands than that from leaf tissue in Brassica napus L.. Moreover, the PCR result of mitochondrial genes showed that the repetition and stability of bands expanding from mtDNA extracted by tender roots were much better than that by leaves. Therefore, mtDNA extracted from tender root tissue have good quality and high concentration. Tender root tissue not only can easily be acquired, but also had no interference with chloroplast and its cpDNA. Tender root tissue is an ideal material for extracting mtDNA. It is a better method that mtDNA is extracted by tender root tissue in Brassica napus L..

The objective of this experiment was to establish an effective JIVET technology system on prepubertal dairy Cattle. In this experiment, we set up an effective gonadotropic hormone stimulating protocol on calf, and 31 oocytes per calf on average were obtained. After in vitro maturation and in vitro fertilization, the fertilization rate of calf oocytes reached 63.2%, which was no significant difference from matured bovine oocytes (59.7%). After embryo in vitro culture, the blastocyst rate of calf embryos was 31.6%, which was still significantly lower than matured bovine embryos (48.0%). After embryos transferred to 3 recipients, 2 of them became pregnant, and 1 recipient completed the whole course of pregnancy.

Abstract: The objective was to determine the effect of estrous cow serum on porcine oocyte maturation. In the experiment, various sera was added to maturation medium and observed matured results. The effect of serum to oocyte quality was tested by in vitro developmental abilities of parthenactivation and reconstructed embryo. Moreover, in vivo development ability of nuclear transfer embryos constructed by oocytes matured with medium supplemented estrous cow serum was tested. Results indicated that there was no significant difference in rate of maturation among three sera (estrous cow serum, cow serum, and fetal cow serum) supplementation medium (P>0.05). In vitro developmental capacities of oocyte matured in medium supplemented various sera were not significantly different (P>0.05). Embryos reconstructed with fetal fibroblast cell and oocyte derived estrous cow serum medium completed full-term development. Key words: porcine; oocyte; in vitro maturation; estrous cow serum; parthenactivation; nuclear transfer

为研究特定基因或蛋白质在牛卵母细胞体外成熟过程中的角色与作用，探索牛卵母细胞体外成熟的分子机理，需要建立一个能够检测基因或蛋白质在牛卵成熟过程中作用的技术平台。本研究在牛卵母细胞体外成熟培养的不同时间（0h和10h），剥除卵母细胞外周的颗粒细胞后继续培养，观察其与正常体外成熟培养卵母细胞之间有何差异。研究结果显示：成熟培养0h脱颗粒细胞的卵母细胞，与正常成熟培养的卵母细胞在成熟率（19.51%vs80.14%,P<0.05），卵裂率（27.08%vs82.61%,P<0.05），孤雌激活胚胎囊胚率（7.69%vs21.71%,P<0.05）方面差异显著；成熟培养10h后脱颗粒细胞的牛卵母细胞，与正常体外成熟培养的卵母细胞相比，在成熟率（82.71%vs80.14%, P>0.05），卵裂率（83.01%vs82.61%,P>0.05），孤雌激活胚胎囊胚率（19.30%vs21.71%, P>0.05）等方面无显著差异。在此基础上，向成熟培养10小时脱颗粒细胞的卵母细胞内注射pVenus－H1foo的mRNA、pVenus质粒、pDsRed1—N1质粒和pDsRed1-H1e质粒等都能够得到表达。非功能标记基因显微注射组与对照组在卵母细胞成熟率（81.42%vs82.03%, P>0.05），卵裂率（75.24%vs78.15%, P>0.05），孤雌激活胚胎囊胚率（17.42%vs18.82%, P>0.05）上差异不显著。本研究可以得到如下结论：在成熟培养10小时，剥离颗粒细胞不会影响牛卵母细胞的发育潜能，这一技术平台可以用于牛卵母细胞体外成熟分子机制的研究。

To investigate the effect of feeding Bacillus subtilis natto on the structure of Holstein calves rumen bacterial community, eight calves were randomly assigned into two groups: a group fed with starter (control), and another group supplemented with Bacillus subitilis natto N1 (1×1010 CFU), which was mixed with starters at a dose of 1% of the daily dry matter intake. DNA from rumen samples were extracted and two rumen 16S rDNA libraries were constructed. The results showed that the vast majority of the two clone libraries were represented by sequences related to the Bacteroidetes and Firmicutes. Of the two clone libraries, the diversity index was high, and the ACE and Chao1 collector’s curves didn’t reach the stable libraries were represented by sequences related to the Bacteroidetes and Firmicutes. Of the two clone libraries, the diversity index was high, and the ACE and Chao1 collector’s curves didn’t reach the stable trend. There was significant difference between the structure of 16S libraries from control and treatment group. Real-time PCR quantification of Ruminobacter albus and Ruminococcus flavefaciens showed that we can detect the Ruminococcus flavefaciens in TRT group, but not in CK group. Compared with CK group, Ruminobacter albus increased to 11.3% in TRT group. These results indicated that feeding Bacillus subtilis natto to Holstein calves could promote the establishment of rumen microbial community, especially the colonization and growth of fiber degradation-related bacteria.

In June 2007, an infectious disease outbreak was occured at a mink（Mustela vison）farm in Dalian, China. The sick minks developed clinical symptoms of ocular discharges , nasal-dry and others, and some minks died from the diseases. A virus was isolated from collected samples of nasal swabs, ocular swabs and other organs. The virus isolate grew well in Vero cell and other cell cultures. Identification of the virus isolate was conducted by a series of methods and procedures, such as morphological observation, physical and chemical analysis, identification of the type of nucleic acid and nucleic acid sequence analysis. Physicochemical identification results show that the virus has resistance to chloroform, ethyl ether and sodium deoxycholate, can tolerate the acidic environment till pH3.0, reserves infection capacity at 50℃ for 1 hour, and does not sensitive to trypsin. Virus morphology assay results show that the diameter of the virus is 30 ~ 35nm, no capsule. Nucleic acid identified it as RNA-type virus. Nucleic acid sequence analysis with the GenBank recorded on the United States in 1980 isolated from the mink calicivirus（MCV） AF338405, AF338406 and AF338407 showed that the nucleotide homology is 99.3% ~ 99.5%, and amino acid homology is 95.4% ~ 100%. The results suggested that the virus isolate from the died mink is a Mink calicivirus. It is the first time to report the isolation and identification of the calicivirus from mink in China.

The total 48 Haemophilus parasuis strains isolated from Guangdong province in 2007-2008 were serotyped by Indirect Haemagglutination Assay, and respectively belonged to 7 serotypes, 5 serotype with 23.68%. Furthermore, the genotypes and phylogenetic relationship of all isolates were analysed by PHYLIP-3.68 after Enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR), the results indicated that the 48 isolates distributed to 31 genotypes among 8 clusters, especially B cluster with 66.67%. The findings suggested that the genetic polymorphism and serotypes of H.parasuis parasuis isolates are diverse in Guangdong province, and ERIC-PCR is an effective method of researching molecular epidemiology of animal infective diseases.

Abstract: The total RNA was extracted from leaves of Zaosu pear（Pyrus bretschneideri cv. Zaosu). Using RT-PCR method, the full-length cDNA of NPR1 gene was cloned from the Zaosu pear leaves by a pair of specific primers designed according to the Japanese pear （Pyrus pyrifolia ）NPR1 gene sequence released on the GenBank（EF079077）. Bioinformatics analysis showed that the obtained 1771bp fragment contained a 1761bp full open reading frame encoding 586 amino acid residues（GenBank accession No. FJ769372）. The homology rates of amino acid sequences of NPR1genes between Zaosu pear and Pyrus pyrifolia (ABA62792), Pyrus ussuriensis, Malus x domestica (ACC77697), Nicotiana tabacum (AAT57641) and Arabidopsis thaliana (AAM98084) were 99%、98%、98%、67% and 59% respectively. The gene was sub-cloned into the expression vector pGEX-4T-1 and expressed a fusion protein of approximately 91kD in Escherichia coli BL21 cells induced by IPTG. The result indicated that the fusion protein was expressed in inclusion body. In order to further analysis the expression profile of the NPR1 protein and study subcellular localization of the NPR1 protein in transformed pears, our future work will be the purification of the NPR1 protein and preparation of its polyclone antibodies.

Abstract：Sweet Sorghum (Sorghum bicolor (L.)Moench) accumulates large amount of sugars in the culm parenchyma. Culm extracts can be used for manufacture of sucrose and syrup. Furthermore, sweet sorghum has been identified as a good biomass crop for fermentation to ethanol. Sucrose Synthase (SuSy) plays an important role in plant growth and development. It can regulate sucrose contents in storage tissue of many plants. In this paper, a novel full-length sweet sorghum sucrose synthase gene (Susy2, GenBank Accession, FJ513325) DNA sequence was cloned by blasting the sorghum bicolor EST database with the sugarcane Susy2 (GenBank Accession, AY118266) cDNA sequence information as a querying probe. It was also verified by genome-PCR. The full-length DNA of 4587bp contains fifteen exons that were separated by fourteen introns and a 2409 bp open reading frame (ORF). Protein encoded by Susy2, molecular weight of 91.7 kD, is comprised of 802 amino acids. Its isoelectric point (pI) is 6.15. Conserved domain searching results showed that this protein contains a GT1 family of glycosyl transferases which have four ADP-binding pockets and catalyzes the synthesis of sucrose 6-phosphate from fructose 6-phosphate and uridine 5'-diphosphate-glucosewas. SuSy is very conservative among sorghum, sugarcane, maize, rice, bamboo, wheat and ryegrass. The results will provide a help for improving of characters by biotechnology and researching of molecular mechanism of sucrose accumulation in sweet sorghum. Key words： sweet sorghum；sucrose synthase；clone；PCR；EST database

K+ channel is a kind of important ways for K+ uptake and transport and plays key roles in ion balance in plant cells. In this study a K+ channel gene was isolated from Hevea brasiliensis by RT-PCR and RACE-PCR technuques and charactirized, showing its cDNA has a total length of 1480bp with an open reading frame(ORF) of 1059bp encoding a precursor protein of 353 amino acid residues. Homology and cluster analysis indictaed that this gene belongs to known KCO family in plants, designated HbKCO1. Semi-quantitive RT-PCR demonstrated that HbKCO1 was present in all plant organs, but preferentially in leaves, strong in stem, latex and bark, and very lower in root. The expression of HbKCO1 in leaves could be significantly induced by high K+, salt(NaCl) and high Ca2+stresses but inhibited by low K+ stress and ABA treatment. In addition, the HbKCO1 transcripts in latex were weakly affected by ET and JA treatments.

Study on in vitro cultures of ten cassava cultivars cultivated mainly in China demonstrated the significant different capacities of somatic embryogenesis (SE) and shoot organogenesis (SO) among tested genotypes. Using apical and axillary buds as explants, SE was induced on induction medium containing 12 mg/L picloram under the dark condition. After 2 weeks culturing, their SE frequencies were recorded. Among tested cultivars, NZ188, SC124 and SC205 could be over 80%. For SE induction, the effect of 12 mg/L picloram was better than that of 6 mg/L 2,4-D. Secondary SE could be established during cyclic subculturing of primary somatic embryos. Mature somatic embryos with green cotyledons could germinate and develop plantlets. The maturation frequency of NZ188 and SC8 could reach more than 80%. Using the pieces of somatic cotyledons as explants, secondary SE and SO could be induced after culturing on SE and SO induction media, respectively. The SE frequencies from somatic cotyledons of all tested cultivars achieved more than 77%. On SO medium, shoot meristems and adventitious shoots were developed after 2 weeks. The induction frequencies are between 34.7% and 72.2% among tested cultivars, and the performance of NZ188, NZ199 and SC124 are better than other cultivars. Adding ethylene inhibitor AgNO3 to the regeneration medium significantly reduced callus formation and improved the development of shoots, including regeneration rate and average shoot number per explant. AgNO3 at 4 mg/L has been considered the most suitable concentration to be used in this study. The shoots regenerated from AgNO3-containing medium grew much better and were easy for planting into soil.

Preadipocytes were obtained from newborn Chinese Holstein calves and induced differentiation for 9 days. The cells became larger and circular accompanied by the small lipid droplets confluenting into large ones and increased mRNA expression of PPAR-γ. At same time ,the expression of Pref-1 was decreased and cells differentiated ultimately into typical adipocytes. Using semi-quantitative RT-PCR method, 1 nmol/L insulin did not affect the expression adiponectin mRNA, but 10, 100 and 1000 nmol/L insulin decreased expression adiponectin mRNA 48.7%,61.4% and 61.9%, respectively (P<0.05). Meanwhile, 100 nmol/L insulin in time-dependent fashion inhibits expression adiponectin mRNA within 24 h. Furthermore, inhibition of PI3K/Akt with LY294002 reverses the inhibitory effect of insulin on adiponectin mRNA expression. These results suggest that insulin at least partially suppressed the expression of Adiponectin mRNA via the PI3K/Akt pathway.

The production of xylanase (XylA) by a newly isolated Aspergillus niger JL15 strain in solid-state fermentation (SSF) on orange peel was optimized by response surface methodology (RSM). Results revealed that four factors of concentrations of added glycerin and ammonium sulfate, moisture content, fermentation time had significant effect on the XylA production (P<0.05). The maximum xylanase activity (917.7 U/g dry fermentation product) obtained at 4.2% glycerin, 3.1% (NH4)2SO4 by employing orange peel as the solid substrate, 61% moisture content and 73.4-h fermentation, which was close to the predicted one (922.9 U/g), and was 3.2 times higher than that of the basic medium (218.5 U/g). The optimum temperature and pH for XylA activity were 55°C and pH5.0, respectively. XylA exhibited Km and Vmax values of 9.24 mg/ml and 54.05 μmol/min/ml, respectively. HPLC analysis showed that XylA liberated mainly xylotriose from birchwood xylan and wheat bran, which suggested the XylA might be an endo-acting xylanase.

In order to study the origin of germ stem cells, the biological characteristics and the mechanism of cell differentiation . The CR4 and CR1 Fragments of Oct4 promoter were inserted into EGFP eukaryotic report vector and to reconstruct a mouse germ cell-specific enhance green fluorescent protein (EGFP) report vector. The recombinant plasmid was subsequently transfected into a variety of cells using Liposome 2000. Expression of the EGFP were detected under Fluorescent microscopy . the germ cell-specific gene expression was semi-quantitative detected in transfected cells by RT-PCR. The results showed that the green fluorescent protein expressed in testicular and P19 cells. In mouse embryonic stem cells, mouse embryonic fibroblasts (MEF) and C2C12 cells GFP did not express. RT-PCR showed that P19 and testicular cells express Oct4 and other germ cell-specific markers. The results suggested that the recombinant vector specificity expressed in germ cells and it will be an available tracer for the study of germ stem cell by fluorescent imaging,also it could be used to show the pluripotential state of germ stem cells.

Molecular Cloning and Bioinformatic Analysis of Three C-Type Lysozyme Genes from Oreochromis aureus

Abstract: A pair of primers P-in1 were designed according to the sequences of Adipocyte Fatty Acid-Binding Protein (A-FABP) gene mRNA in duck and A-FABP gene in chicken to amplified duck A-FABP intron-1, then 4 pairs of primers were designed according to cloned gene. The SNPs were detected by technique of PCR-SSCP, confirmed by sequencing and to study the. genetic discrepancy in different groups. The results indicated that the product of P-in1 included partial exon-1, exon-2 and completely intron-1 after cloned and sequenced and homologous comparison was done between duck and chicken A-FABP gene, the results showed the homology was 75.1%. Three and six nucleotides mutation were found in the products of primer P1 and P3, which resulted in 3 and 6 genotypes, separately. The results of Chi-Square test showed that Cherry Valley duck and Sumu duck deviated from the Hardy-Weinberg equilibrium (P<0.01) except Jingding duck in P1 locus , but all of these three groups fit with it in P3 locus (P>0.05) and the distribution of them was different or very different in various groups (P<0.05 or P<0.01).Genetic analysis showed there had a high level of polymorphism in different duck groups, and could be used as a candidate gene to analysis the correlation between its polymorphism and fat traits in duck.

Sequencing, PCR-RFLP and dCAPS methods were used to study the polymorphism in exon 3 and 3′UTR of B-cell translation gene 2 (Btg2), which was associated with the multi-vertebra traits of Hebei Small Tail Han Sheep (Ovis aries). 3 SNPs (A150G, C243T and A607G) were found in the amplified fragments. The restriction endonuclease digestion analysis was used to determine genotypes and then haplotype analysis was conducted. The results of the independence Chi-square test showed that genotype frequencies at Site 150 had no significant difference among different vertebral patterning sheep (P>0.05). And at Site 243, the genotype frequencies had extremely significant difference among different vertebral patterning sheep (P<0.05), while at Site 607, the result was of significant difference. The result of haplotype frequencies among different phenotypes showed a significant difference as well by independence Chi-square test (P<0.05). These results implied that Site 243, Site 607 and GTA haplotype may have associations with the multi-vertebrae traits of Hebei Small Tail Han Sheep.

The purpose of this study was to explore whether the bovine enucleated oocytes can be activated and its development capacity after activation.We activated the enucleated oocytes using chemical methods,and cultured the embryos with granulosa cell feeder layer in vitro. To observe dynamic changes of microtubules in the enucleated and normal oocytes, we dyed the α-tubulin using immunofluorescence technology at different stages. The results showed that after activation the enucleated oocytes can developed to morula in vitro, and the distribution of α-Tubulin was very different with that of the normal oocytes.

Abstract: To study the effect of Caspase and Bcl-2 families on apoptosis HL-60 cells induced by insecticidal crystal proteins from Bacillus thuringiensis Bt9875. The activation of Caspases and the effect of Caspase inhibitors on apoptosis of HL-60 cells were detected. The cleavage of poly ADP-ribose polymerase (PARP), Bcl-2/Bax regulation and the release of cytochrome C were observed by Western blot analysis. Insecticidal crystal proteins from Bacillus thuringiensis Bt9875 can activate Caspase-3, 8 and 9. Moreover, the cell death can be significantly suppressed by family Caspase inhibitor (Z-VAD-FMK), Caspase-3 inhibitor (Z-DEVD-FMK) and Caspase-9 inhibitor (Z-LEHD-FMK). Futhermore, up-regulation of Bax and down-regulation of Bcl-2 have also been detected by Western blot in HL-60 cells which treated with Bt9875. The results suggest that the progression of cell death induced by Bt9875 was mediated by both Caspase family and Bcl-2 family. The mitochondria pathway plays an important role on apoptosis of HL-60 cells induced by insecticidal crystal proteins from Bacillus thuringiensis Bt9875.

Lycopene, one kind of pigment carotenoids, performing the important function on antioxidant, disease prevention and so on. Lycopene cyclase-β(Lyc-β) is one of the key enzyme in the carotenoid biosynthesis pathway. Two fragment of 300 bp from Lyc-β gene were reverse transcribed and amplified from tomoto mRNA according to Genbank sequence X86452. One intron fragment of 170 bp was amplified from gusA gene. Two plant expression vectors to interference Lyc-β gene were constructed by ligation the intron fragment of gusA gene with two Lyc-β fragments which were designed in different direction separately. After transformation into Lycopersicun　esculentum through Agrobacterium tumefaciems, the interference vectors were used to analyses the interference effect. The twenty-two transgenic tomato plant were obtained to perform real time PCR assay. The results demonstrated the different interference efficacy to the mRNA abundance of Lyc-β gene in average of 11.78% and 13.86% separately Compared with the control of wild plant. The lycopene content of transgenic tomoto were analyzed by HPLC, and its highest content was 13.84 μg/g FW, 4.26 fold compared with control.

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most important wheat diseases worldwide. Application of resistance varieties is the most economical and effective method for controlling this disease. The common wheat line 3D256, a derivative of wild emmer (Triticum dicoccoides) with pedigree Yanda1817/WE27//Nongda015/3/941, is highly resistant to prevailing local wheat powdery mildew isolate E09 in Beijing area. Genetic analysis of the F2 population and their F3 families, developed from 3D256 and a susceptible common wheat cultivar Xuezao, indicated that the powdery mildew resistance in 3D256 was controlled by a single dominant gene, designated temporarily MlWE27. Molecular markers and the bulked segregant analysis were used to characterize the powdery mildew resistance gene MlWE27. Three SSR markers (Xwmc243、Xwmc154 and Xbarc318), one EST-SSR marker (Xdp358), one AFLP-derived SCAR markers (XCAUG1) and one RFLP-derived STS marker (XWG516-1) were found to be linked to the powdery mildew resistance gene MlWE27, with an order of Xdp357-MlWE27-Xwmc243-XCAUG1-XWG516-1-Xwmc154-Xbarc318 in the genetic linkage map. Using Chinese Spring nullisomic-tetrasomics, ditelosomics and deletion lines, MlWE27 was physically mapped on chromosome 2BS terminal bin 0.84-1.00. The new developed common wheat powdery mildew resistance line 3D256 and its resistance gene linked molecular markers provide useful germplasm and tool in disease resistance genes pyramiding and marker-assisted selection (MAS) in wheat breeding program.