不同氮素水平对7S亚基缺失大豆品质及相关性状的影响

为明确7S亚基缺失大豆品系—东富2的理想施氮水平,充分体现特用大豆品种的高附加营养品质,设置4种施氮水平(纯N),N0(0 mg·kg~(-1))、N1(25 mg·kg~(-1))、N2(50 mg·kg~(-1))、N3(75 mg·kg~(-1)),采用桶栽法研究氮素对7S亚基缺失大豆品质及其相关性状的影响。结果表明,N1水平下,7S亚基缺失大豆的光合速率、叶绿素含量、氮素吸收效率和籽粒理论产量较高,蛋白质含量和脂肪含量分别为40.36%和21.78%,其中脂肪酸组分中软脂酸为12.9%,油酸为16.6%。N2水平下,7S亚基缺失大豆的干物质重、氮素积累量和籽粒理论产量最高,蛋白质含量和脂肪含量分别为39.57%和21.23%,其中脂肪酸组分中软脂酸和油酸分别为14.4%和16.1%。综合籽粒理论产量、品质及其相关性状,大豆品系东富2在N1水平下,光合生理特性表现良好,氮素吸收率较高,籽粒产量未降低的同时,蛋白质含量、脂肪含量和有益脂肪酸含量均有所提高。本研究结果对实现节本增效优质的7S亚基缺失大豆生产具有重要意义。     

Characterization of a Novel 1Ay Gene and Its Expression Protein in Triticum urartu

High molecular weight glutenin subunit （HMW-GS） plays an important role in determining dough property and breadmaking quality, and the exploration of novel genes for HMW-GS will be crucial for quality improvement program. A gene coding the y type HMW-GS at Glu-A1 locus in Triticum urartu （AA, 2n=2×= 14） with an electrophoretic mobility similar to that of 1Dyl2, was cloned, sequenced, and heterologously expressed. This novel active lay gene FJ404595 was confirmed by structure analyses of nucleotide and deduced amino acid sequences combining with phylogenetic analysis. The open reading frame （ORF） of this gene was 1 830 bp, encoded a protein of 608 amino acid residues containing 46 hexapeptides and 14 nonapeptides, which was mostly similar to the lAy gene AM183223 at a high identity of 99.62% with the two substitutions of both leucine/proline and valine/glutamate, obviously different from the lAy gene EU984504 with 587 residues containing 44 hexapeptides and 13 nonapeptides in T. urartu. The amino acid （leucine） at 446 differed from that （proline） of all the eight compared active lAy subunits. The predicted secondary protein structure implied that this lAy subunit might also have positive impact on flour processing quality.     

转基因小麦1Dx5基因沉默的遗传表达和品质效应分析

为了明确通过RNA干扰技术（RNAi）获得的转基因小麦品系“Glu1Dx5RNAi”中1Dx5基因的遗传规律,以“Glu1Dx5RNAi”为供体亲本,以弱筋品种扬麦18和扬麦13为轮回亲本进行回交, 采用半籽粒SDSPAGE法检测亲本、F₁、F₂和BC₁F₁籽粒的高分子量谷蛋白亚基（HMWGS）组成。结果表明,1Dx5基因的沉默效应在杂交后代中表现为显性遗传,在F₂和BC₁F₁世代1Dx5亚基不表达的种子数与1Dx5亚基表达的种子数的比例分别符合13∶3和3∶1的分离比例,说明转小RNA基因导致1Dx5基因沉默是单基因显性遗传。对转基因小麦“Glu1Dx5RNAi”及其受体品种Bobwhite进行品质测试表明,Glu1Dx5RNAi蛋白质含量（13.28%）比转化受体Bobwhite（12.83%）高0.45个百分点,硬度指数、微量SDS沉降值比受体品种分别降低3.13和3.7 mL。1Dx5基因沉默对弱筋小麦育种可能会有一定的利用价值。     

Low expression of nicotinic acetylcholine receptor subunit Mdα2 in neonicotinoid‐resistant strains of Musca domestica L.

BACKGROUND: Neonicotinoid action as well as resistance involves interaction with nicotinic acetylcholine receptors (nAChRs). In the housefly, neonicotinoid resistance also involves cytochrome P450, as indicated by bioassay with synergist as well as altered expression. In bioassay, synergism was only partial and indicated possible target‐site resistance. The nAChR α2 subunit is important in neonicotinoid toxicity to insects, and gene expression of the Mdα2 subunit was investigated in field populations and laboratory strains of neonicotinoid‐resistant and insecticide‐susceptible houseflies, Musca domestica L. The genomic sequence covering exon III–VII of Mdα2 was analysed for mutations. RESULTS: Gene expression profiling of Mdα2 revealed notable differences between neonicotinoid‐resistant and insecticide‐susceptible houseflies. On average, the neonicotinoid‐resistant field population 766b and the imidacloprid selected strain 791imi had 60% lower copy numbers of Mdα2 compared with the susceptible reference strain. Sequencing of exon III–VII of the Mdα2, encoding acetylcholine binding‐site regions and three out of four transmembrane domains, did not reveal any mutations explaining the increased neonicotinoid tolerance in the strains examined. CONCLUSION: Previous discoveries and the results of this study suggest that the neonicotinoid resistance mechanism in Danish houseflies involves both cytochrome P450 monooxygenase‐mediated detoxification and reduced expression of the nAChR subunit α2. Copyright © 2010 Society of Chemical Industry     

拟南芥SWR1复合体亚单位编码基因ARP6和SWC6的水稻同源基因功能研究

用RT-PCR的方法分别克隆出拟南芥SWR1复合体亚单位编码基因ARP6和SWC6在水稻中的同源基因OsARP6和OsSWC6,对其组织表达特异性进行了分析。构建这两个基因的表达载体,分别转化拟南芥突变体arp6和swc6,它们在阳性植株表达后均能在一定程度上使突变性状回复,表明在拟南芥和水稻之间,这些基因在功能上具有一定的保守性。本研究结果为水稻中存在SWR1功能复合体提供了重要的证据,也有助于探明该复合体调控基因表达的分子机制。     

牛新孢子虫NcSAG1基因工程亚单位疫苗的免疫试验

为了解新孢子虫NcSAG1表面蛋白基因工程亚单位疫苗对实验动物的免疫效果,本试验提取延边黄牛新孢子虫野毒株基因组DNA,用PCR技术扩增新孢子虫NcSAG1表面蛋白基因,并在大肠杆菌中进行原核表达,将Western-blot鉴定具有免疫活性的重组蛋白与弗氏佐剂混合制备NcSAG1基因工程亚单位疫苗,免疫BALB/c小鼠,应用间接ELISA方法测定体液免疫水平,应用流式细胞技术测定细胞免疫水平,以此评价疫苗对实验动物的免疫应答反应。结果显示,制备的NcSAG1基因工程亚单位疫苗免疫BALB/c小鼠后,在三免后第3天时,检测抗体的OD450nm值达0.688;CD4+/CD8+值达3.650,均显著高于重组蛋白免疫组和PBS对照组,说明制备的基因工程亚单位疫苗能够提高实验动物的体液免疫和细胞免疫水平。本试验为牛新孢子虫NcSAG1表面蛋白基因工程亚单位疫苗的深入研究奠定了基础。     

豫南黑猪、杜洛克、长白和大白猪FSHβ基因多态性与产仔性能的关联分析

采用PCR-RFLP技术测定了豫南黑猪、杜洛克、长白、大白猪FSHβ亚基基因多态性,测序发现豫南黑猪BB型的128与129 bp位点上插入277 bp的片段而产生多态性,限制性内切酶BamHⅠ酶切后共产生6种基因型,运用最小二乘模型将不同基因型与产仔数进行关联分析,结果显示:豫南黑猪就初产而言,基因型CC型比BC型的TNB和NBA分别多2.67和2.79头(P<0.05);经产基因型CC型比AC型的TNB和NBA分别多2.03和2.14头(P<0.05);而引进猪种初产和经产都没有显著差异。     

Norovirus分子生物学特征及其亚基疫苗研究进展

诺如病毒(norovirus, NV)是导致人类非细菌性急性胃、肠炎的主要病原,该病毒可以感染各年龄段人群,全年均可爆发,特别是婴幼儿、老年人和免疫系统缺陷病人属于易感人群。目前临床防治或控制norovirus疫情爆发的重要手段之一是使用疫苗。基于此,对norovirus发现、流行病学特征、分子生物学特征和norovirus疫苗的研究进展加以综述。同时,在比较norovirus的病毒类似颗粒(virus-like particles, VLPs)和P粒子(P particles)与人组织血型抗原亲合能力的基础上,对利用植物系统特别是番茄果实生产适于口服norovirus的P粒子作为亚基疫苗优势和存在难点以及本实验室的研究进展进行了讨论,以期为利用植物系统表达口服疫苗研究提供一些借鉴。     

Effects of HIF-1α/SDF-1 signaling axis on hypoxia-induced migration and adhesion of progenitor cells

AIM: To observe the expression of hypoxia-inducible factor-1alpha (HIF-1α) and stromal cell-derived factor 1 (SDF-1) in pulmonary artery endothelial cells (PAECs) of SD rats and to investigate the role of HIF-1α/SDF-1 signaling axis on hypoxia-induced migration and adhesion of progenitor cells to PAECs. METHODS: Immunomagnetic beads were used to separate and purify the CD34⁺/CXCR4⁺ progenitor cells derived from the peripheral circulation of SD rats. The expression of HIF-1α and SDF-1 in PAECs exposed to hypoxia (1% O₂, 5% CO₂ and 94% N₂) was detected by immunofluorescence, Western blotting and ELISA. The migration index and adhesion rate were measured in the progenitor cells, which were subjected to the following different treatments: (1) normoxia (21% O₂); (2) hypoxia 12 h; (3) hypoxia 12 h +HIF-1α inhibitor (2ME2); (4) hypoxia 12 h+SDF-1 neutralizing antibody; (5) hypoxia 12 h+2ME2+SDF-1 neutralizing antibody.RESULTS: The expression of HIF-1α and SDF-1 in PAECs was effectively induced by the hypoxic exposure, and both of them reached the peak levels after 12 h of hypoxic treatment (P<0.01), while administration of 2ME2 decreased the hypoxia-induced SDF-1 expression (P<0.05). Treatment of the PAECs with 2ME2 or SDF-1 neutralizing antibody attenuated the migration index and adhesion rate of progenitor cells to the PAECs (P<0.05). CONCLUSION: There is a HIF-1α/SDF-1 signaling axis in hypoxia-exposed PAECs, which may play a crucial role in the migration and adhesion of progenitor cells to PAECs.     

基于线粒体序列特征的茶树菇及杨柳田头菇分离物快速鉴定

此试验拟探寻特异的DNA序列,快速鉴定茶树菇（Agrocybe aegerita）及其形态上易混淆种,给种质资源收集及利用提供有效工具。本研究以云南采集到的18个菌株为材料,其中5株为A. aegerita,13个为A. salicacola,并对线粒体小亚基V9区进行序列分析。结果发现A. aegerita比A. salicacola多1个约50碱基的插入片段,而其他序列组成一致。根据此特点,在插入区筛选到1个特异DraⅠ酶切位点。通过对18个菌株V9区PCR产物进行酶切,发现A. aegeritaV9片段能完全被切开,而A. salicacola V9片段则为完整。根据此酶切方法,可以高效鉴定A. aegerita及A. salicacola 2个近似种,为食用菌遗传及育种研究提供了重要的基础。