Synechocystis sp. strain PCC 6803定量PCR模板制备方法的改进

以Synechocystis sp.strain PCC 6803为实验菌株,设计改良制备定量PCR模板的方法;同时,对Synechocystis sp.strain PCC 6803总RNA的Trizol抽提法进行优化.结果显示,利用改良方法抽提得到的RNA结构更完整;同时,利用改良方法制备的模板可以用于后续定量PCR实验;其中,只有高表达量基因适用于半定量PCR,低表达量基因则需要实时荧光定量PCR检测.     

枯草芽胞杆菌TR21防控粉杂1号香蕉枯萎病的效果和对根系抗性相关信号物质累积的影响

本文研究了枯草芽胞杆菌Bacillus subtilis TR21在大田对抗病品种粉杂1号枯萎病的防控效果及其与病原菌不同组合处理种苗根系抗性相关信号物质累积情况,评估了菌株TR21与粉杂1号结合减轻大田发病率的应用价值。田间试验设置清水对照区、菌株TR21可湿性粉剂喷叶区、菌株TR21可湿性粉剂喷叶+叶腋接种1次胶囊剂型区、菌株TR21可湿性粉剂喷叶+叶腋接种1次栓剂剂型区,并统计田间发病率、抽蕾率、断蕾率和产量。室内采用清水处理、菌株TR21叶腋接种、香蕉枯萎病菌FOC004孢子根系接种、菌株TR21叶腋接种后再在根系接种FOC004孢子的挑战接种4种处理方式,测定接种后不同时间点根系NO、H₂O₂和水杨酸（SA）变化。结果表明,TR21可湿性粉剂的不同处理均能降低粉杂1号发病率,其中喷叶区防效达72%,此外增加1次栓剂处理可以显著增加单株产量,并显著缩短粉杂1号生育期。对抗性信号物质测定发现,只有挑战处理能够在接种早期（4 h）激发NO的显著升高,并维持较长时间（72 h）;3种处理均能够在早期（4 h）显著减少根系产生H₂O₂;病原菌和挑战处理在早期（4 h）能够激发根系SA含量显著升高。TR21叶腋喷施通过提高粉杂1号对枯萎病的抗性来降低田间发病率。     

1株生防枯草芽孢杆菌作为活体菌剂在小麦种子贮藏中的应用研究

生物防治是植物病害防治的重要手段之一,为了研究生物防治措施在小麦种子贮藏中的应用,试验以生防枯草芽孢杆菌JGA2(-5)27-2为研究对象,对其作为活体菌剂在小麦种子贮藏中的应用进行了初探。生防枯草芽孢杆菌JGA2(-5)27-2能产生植物激素细胞分裂素,并具有较广的真菌病害抗菌谱。小麦发芽试验表明,其作为活体菌剂处理小麦后发芽率提高了10.0%。模拟小麦种子吸湿回潮受真菌侵染的试验显示,活体菌剂处理过的小麦种子受侵染的程度低于对照。试验证明：利用生防菌株JGA2(-5)27-2作为活体菌剂在小麦种子贮藏中的应用具有一定的应用价值。     

Shear Deformation Properties of Sliding Zone Soil and Its Effect on Sliding Mechanism of a Landslide in the Three Gorges Reservoir Area

The mechanical properties of slip soil play a key role in landslide stability. In order to study this control mechanism, multigroup experiments on the slip soil of a landslide were conducted in the Three Gorges reservoir in Zigui county involving consolidated quick direct shear test and consolidated drained direct shear test, through which the strain hardening property of this slip soil has been studiedas well as mechanism of landslide deformation resulting from the strain hardening slip soil. The results show that from plastic deformation to shear failure, the shear stress applied to this slip soil increases all the way. After repeated shearing, its shear strength increases, and the greater the normal stress, the more the shear strength increases, in which process the angle of internal friction contributes the most to its shear strength variation. Through the field investigation and numerical modeling, how the strain hardening slip soil affects the landslide stability was studied. In the process of sliding, the frictional strength of the strain hardening slip soil increases after being shearing, leading to the consumption of the landslide’s kinetic energy, following the phenomenon that a temporary steady state occurs after a short distance sliding.     

口蹄疫病毒Hankou/99株基因组全序列测定与基因特征分析

本研究主要对口蹄疫病毒(FMDV) Hankou/99株全基因序列进行测定,通过基因序列分析,确定其基因型,丰富了FMDV基因库,为研究猪源FMDV的分子变异、感染性分子克隆及致病机理奠定基础.从感染FMDV Hankou/99株的细胞液中提取RNA,通过RT-PCR技术,获得猪FMDV Hankou/99分离株覆盖全基因组的5个cDNA片段(S、L、C、D、E),分别对这些片段进行克隆和序列测定.结果显示,FMDV Hankou/99株全基因组长8099 bp;5'NCR长1040 bp,开放阅读框长6966 bp;3'NCR长93 bp,其后是30个碱基的连续poly(A)结构.通过与参考株基因组结构比较分析,显示其在分类地位上属于O型FMDV,并与猪源FMDV毒株OLZ、TW/97同源性较高,特别是3A区域上都有30 bp的缺失.另外,通过与9个参考株的VP1系统发生树分析,显示其与OLZ、TW/97、O/Akesu/58、O/OMⅢ 4个毒株为同一基因型.     

天人菊菌核病病原鉴定及其拮抗木霉菌株的筛选

通过致病性测定、病害症状、病原菌培养性状及形态特征等对天人菊菌核病病原菌进行鉴定;通过室内拮抗作用测定和盆栽防治效果测定筛选生防木霉菌株。结果表明:将天人菊菌核病的病原菌鉴定为核盘菌(Sclerotinia sclerotiorum(Lib)de Bary);筛选到对天人菊菌核病菌拮抗作用最强,盆栽防治效果最好的菌株是哈茨木霉H5,其拮抗指数为Ⅰ级,防治效果为93.2%。在测试的5个木霉菌株中,哈茨木霉H5对天人菊菌核病的防治效果最好。     

利用简单重复序列分子标记分析我国侧耳栽培菌株的遗传多样性(英文)

采用FIASCO方法,从糙皮侧耳(Pleurotus ostreatus)菌株ACCC 50838分离简单重复序列分子标记(Simple sequence repeat,SSR),所得标记用于分析我国52个侧耳(平菇)栽培菌株多样性,对于SSR标记相同的菌株再进行ISSR PCR分析。结果表明:18个SSR标记在供试菌株中的等位基因的数量为3～12个,多态性指数(PIC)为0.34～0.84;除了2组(ACCC50618,ACCC 50866和ACCC 50915;ACCC 50249和ACCC50476)共5个菌株之外,其他47个菌株都能利用这18个SSR标记进行有效的鉴别。SSR标记相同的的菌株,其ISSR、体细胞不亲合性和出菇性状(温型、菌盖颜色)也相同,这些菌株可能是同物异名。利用这18个SSR标记分析52个菌株的遗传多样性,相似性指数为0.755～0.9995。根据相似性指数进行聚类分析,52个菌株分成两个大的类群,分别为糙皮侧耳(P.ostreatus)和肺形侧耳(P.pulmonarius)。结合这些栽培菌株的农艺性状和SSR分子标记的聚类分析结果,我国平菇栽培菌株的遗传多样性非常丰富。研究表明SSR分子标记技术在平菇遗传多样性和菌株鉴别中具有广泛的应用前景。     

密度和施肥量对辣椒新品系ZJ1-12产量和经济效益的影响

为探索辣椒新品系ZJ1-12的高产优化农艺措施,运用4因素(完全实施)5水平的二次正交旋转回归设计研究了密度、氮、磷、钾肥对辣椒新品系ZJ1-12产量和经济效益的影响。结果表明:各因子表现正向增加作用,作用大小排序为密度>K20>N>P2O5肥用量。单产达156.93kg/667m2以上的优化农艺措施为密度10235～10813株/667m2,纯N量13.525～14.855kg/667m2,P2O58.232～9.748kg/667m2,K2010.875～12.6kg/667m2。当最高产量为278.84kg/667m2时,纯收益高达4149.04元/667m2。     

Application of a real-time PCR-based system for monitoring of O26, O103, O111, O145 and O157 Shiga toxin-producing Escherichia coli in cattle at slaughter

Faecal samples were collected from 573 slaughtered cattle aged between three and 24 months in seven abattoirs. After enrichment (mTSB with novobiocin), samples were screened by real‐time PCR first for stx and if positive, tested for the top‐five Shiga toxin‐producing Escherichia coli (STEC) serogroups using PCR assays targeting genes specific for serogroups O26, O103, O111, O145 and O157. Of 563 samples with available results, 74.1% tested positive for stx genes. Amongst them, the serogroups O145, O103, O26, O157 and O111 were detected in 41.9%, 25.9%, 23.9%, 7.8% and 0.8%, respectively. From 95 O26, 166 O145 and 30 O157 PCR‐positive samples, 17 O26, 28 O145 and 12 O157 strains were isolated by colony hybridization after immunomagnetic separation. The 17 O26 strains were eae‐positive, but only nine strains harboured stx (eight possessing stx1 and one stx2). Of the 28 O145 strains, ten were eae‐positive including four harbouring stx1 or stx2, whereas 18 were negative for stx and eae. Five of the 12 O157 strains harboured stx2 and eae, did not ferment sorbitol, and were identified as STEC O157:H7/H^?. The other seven O157 strains were negative for stx and eae or positive only for eae. Shiga toxin genes and the top‐five STEC serogroups were frequently found in young Swiss cattle at slaughter, but success rates for strain isolation were low and only few strains showed a virulence pattern of human pathogenic STEC.     

夏秋用雄蚕品种限7×平48的育成

饲养雄蚕品种是提高蚕丝品质和蚕茧生产效益的重要途径之一。以引进的性连锁平衡致死系S-14为母本,用普通家蚕品种夏6为优良性状基因供体回交改良性连锁平衡致死系,经2轮杂交、自交、回交组成的循环改良和纯化,育成经济性状优良的家蚕性连锁平衡致死系平48,将该品种与家蚕限性皮斑品种限7组配成夏秋用雄蚕杂交组合限7×平48。经浙江省家蚕新品种实验室联合鉴定,该雄蚕品种的雄蚕率达99%以上,在农村生产鉴定试验中表现出强健好养、产量高、丝质优等特点:张种产茧量和50 kg桑产值分别较对照品种秋丰×白玉提高8.4%和9.4%;茧丝长、解舒丝长、解舒率和干茧出丝率分别较对照种提高7.9%、12%、2.57个百分点、1.54个百分点。新品种2009年通过浙江省家蚕新品种审定,适合在长江中下游蚕区的夏秋季饲养。