Foc4 2个谷胱甘肽S-转移酶基因的克隆、鉴定及其在外源氧化胁迫下的表达

为鉴定香蕉枯萎病菌(尖孢镰刀菌古巴专化型4号生理小种,Fusarium oxysporum f.sp.cubense race 4,Foc4)中的2个假想谷胱甘肽S转移酶(GSTs),采用RT-PCR方法克隆了这2个GSTs基因cDNA编码序列,随后分别将2个基因定名为Fogst1和Fogst2。其中,Fogst1的开放阅读框长609 bp,编码202个氨基酸残基,Fogst2的开放阅读框长693 bp,编码230个氨基酸残基。进化树分析表明:Fogst1属于GSTs超家族的sigma(σ)亚型成员,Fogst2属于GSTs超家族中目前未知的亚家族成员。为了验证Fogst1和Fogst2的表达,分别构建了Fogst1和Fogst2的原核表达重组载体pET28a-Fogst1和pET28a-Fogst2,并将pET28a-Fogst1和pET28a-Fogst2转化到大肠杆菌表达菌株BL21,经IPTG诱导后获得以可溶形式表达的重组蛋白Fogst1和Fogst2。GSTs活性分析表明,以CDNB为底物检测,2个重组蛋白均具有GSTs酶活性。分别取外源氧化胁迫处理后1、5、12、24 h菌丝样品进行相对荧光定量PCR分析,结果表明:Fogst1和Fogst2在前5 h表达量均大幅上调,表达量随后下调并恢复正常水平。这些结果均暗示Fogst1和Fogst2可能参与了Foc4抗外源氧化胁迫过程。     

锦鲤4个人工雌核发育家系的微卫星标记研究

利用Crooijmans et al.(1997)分离的包含CA重复单元的普通鲤鱼（Cyprinus carpio L.)的8个微卫星DNA标记,对从锦鲤（Cyprinus carpio L.)的红白,大正和昭和3个不同品系中所获得的4个不同人工雌核发育家系的20尾个体进行PCR扩增。电泳结果表明,8对引物在20尾个体中均能重复稳定地扩增出相应的同源序列。随引物不同,各等位基因数为1－11个,大小在68－264bp。在MFW4,MFW7,MFW19,MFW20,MFW23和MFW24 6个微卫星的扩增结果中,20尾个体的扩增图谱呈现了高度的遗传多态性,不同雌核发育家系内个体的遗传异质性也较大。其中大正（TaS)和红白1（RW1）的个体不仅花色分化显著,而且个体间的平均遗传距离分别高达0．28。通过对微卫星等位基因和基因型分析发现,由于锦鲤品系中的每一个体是通过不断地杂交选育而获得,基因组来源复杂,基因高度杂合。因此,只进行1代的人工雌核发育,其家系内仅部分个体的部分座位出现纯合。所获得的人工雌核发育锦鲤为后续的色素遗传调控机制研究提供了必要的实验材料;同时,所鉴定的微卫星分子标记为进行锦鲤的分子标记育种的基因组作图提供了理想的工具。     

水稻矮缩病毒昆明分离物抗血清制备及免疫捕捉PCR检测

由水稻矮缩病毒(Rice dwarf viru S,RDV)引起的水稻矮缩病害,最早由日本报道,随后在东南亚等国以及我国的福建、云南等南方稻区普遍发生,云南主要发生于中部及南部地区[1].水稻在苗期至分蘖期感病后,植株矮缩,分蘖增多,叶片浓绿,僵直,出现白斑,生长后期病稻不能抽穗结实,在暴发流行年份可以引起水稻的严重减产.     

非人灵长类基因修饰模型研究进展

非人灵长类动物在生命科学基础研究和生物医药研究领域具有非常重要的地位。近年来随着慢病毒载体转染及靶向核酸酶(ZFN,TALEN,CRISPR/Cas9)等基因操作技术的出现,科学家们成功地获得了外源基因过表达的转基因猴和目的基因定点切割的基因编辑猴。文中对目前利用慢病毒载体获得转基因猴和利用靶向核酸酶获得基因编辑猴的研究进展进行了综述,并讨论了基因修饰猴的嵌合体现象、脱靶现象及非人灵长类动物较长性成熟时间这几个影响非人灵长类基因修饰模型推广应用的因素,最后展望了非人灵长类基因修饰模型构建技术的研究热点及发展趋势。     

一株广谱中和抗原性出血热病毒株的发现

一株分离自杭州市褐家鼠的出血热病毒Gou_3株的免疫血清对10株I型病毒的中和滴度除二株为160外均为320,而对4株Ⅱ型病毒的滴度为320—640,说明Gou_3株免疫血清对两型毒株中和效价大多数无差异或只差2倍,是一株中和抗原广谱的毒株。用I型和Ⅱ型毒株免疫血清对Gou_3株进行型别检定结果表明Gou_3株是Ⅱ型病毒。     

缺失宿主Vta1蛋白的MIT结构域对杆状病毒AcMNPV复制的影响

【目的】克隆草地贪夜蛾(Spodoptera frugiperda)Vta1基因,检测Vta1在苜蓿银纹夜蛾核多角体病毒(Autographa californica multiple nucleopolyhedrovirus,AcMNPV)复制中的作用。【方法】利用反转录-PCR与PCR方法筛选草地贪夜蛾Vta1基因及缺失Vta1N-端MIT结构域的突变体并构建其瞬时表达质粒,通过转染Sf9细胞检测表达;构建Vta1及其突变体的双分子荧光互补表达质粒,并通过瞬时转染检测其与Vps4及ESCRT-III亚基Vps46与Vps60的相互作用;共转染gp64与Vta1及其突变体瞬时表达质粒,检测瞬时表达Vta1突变体对AcMNPV出芽型病毒产量及病毒基因启动子指导报告基因表达的影响。【结果】获得了草地贪夜蛾Vta1基因。氨基酸序列相似性分析表明,昆虫、酵母与人类Vta1同源蛋白的相似性分别约为20%与50%。Western blotting分析表明GFP标签的Vta1及其突变体均能在瞬时转染的Sf9细胞中表达。双分子荧光互补分析发现,缺失第1个或第2个MIT结构域显著降低Vta1突变体与Vps4、Vps46或Vps60的相互作用。此外,瞬时表达Vta1突变体显著降低了AcMNPV感染性出芽型病毒的产量,但并未影响AcMNPVie1基因早期启动子和p6.9基因晚期启动子指导的LacZ和GUS报告基因的表达。【结论】Vta1可能参与杆状病毒AcMNPV子代病毒粒子的组装和/或出芽释放过程。     

A dynamic, architectural plant model simulating resource-dependent growth

BACKGROUND AND AIMS: Physiological and architectural plant models have originally been developed for different purposes and therefore have little in common, thus making combined applications difficult. There is, however, an increasing demand for crop models that simulate the genetic and resource-dependent variability of plant geometry and architecture, because man is increasingly able to transform plant production systems through combined genetic and environmental engineering. MODEL: GREENLAB is presented, a mathematical plant model that simulates interactions between plant structure and function. Dual-scale automaton is used to simulate plant organogenesis from germination to maturity on the basis of organogenetic growth cycles that have constant thermal time. Plant fresh biomass production is computed from transpiration, assuming transpiration efficiency to be constant and atmospheric demand to be the driving force, under non-limiting water supply. The fresh biomass is then distributed among expanding organs according to their relative demand. Demand for organ growth is estimated from allometric relationships (e.g. leaf surface to weight ratios) and kinetics of potential growth rate for each organ type. These are obtained through parameter optimization against empirical, morphological data sets by running the model in inverted mode. Potential growth rates are then used as estimates of relative sink strength in the model. These and other 'hidden' plant parameters are calibrated using the non-linear, least-square method. KEY RESULTS AND CONCLUSIONS: The model reproduced accurately the dynamics of plant growth, architecture and geometry of various annual and woody plants, enabling 3D visualization. It was also able to simulate the variability of leaf size on the plant and compensatory growth following pruning, as a result of internal competition for resources. The potential of the model's underlying concepts to predict the plant's phenotypic plasticity is discussed.     

Allometric growth and carbon storage in the mangrove Sonneratia apetala

Allometric growth reflects different allocation patterns and relationships of different components or traits of a plant and is closely related to ecosystem carbon storage. As an introduced species, the growth and carbon storage of Sonneratia apetala are still unclear. To derive allometric relationships of the mangrove S. apetala and to estimate carbon storage in mangrove ecosystems, we harvested 12 individual Sonneratia apetala trees from four different diameter classes in the Futian National Nature Reserve, Guangdong, China. Allometric growth models were fitted. The results showed that diameter at breast height (DBH) and wood density were better variables for predicting plant biomass (including above- and below-ground biomass) than plant height. There were significant power function relationships between biomass and DBH, with a mean allometric exponent of 2.22, and stem biomass accounted for 97% of the variation in S. apetala total biomass. Nearly isometric scaling relationships were developed between stem biomass and other biomass components. To better understand the carbon stocks of the S. apetala ecosystem, we categorized all trees into five age classes and quantified vegetation carbon storage. The S. apetala vegetation carbon storage ranged from 96.48 to 215.35 Mg C ha^?1, and the carbon storage significantly increased with stand age. The allometric equations developed in this study are useful to estimate biomass and carbon storage of S. apetala ecosystems.

     

玉米及马齿苋叶片SOD活性诱导研究

以玉米幼苗及马齿苋作材料,通过甘露醇、H2O2、臭氧和强光等胁迫后,用NBT光化还原抑制法测定叶中SOD活性的变化。臭氧和强光能诱导玉米叶片SOD活性增加。0.5mol／L甘露醇处理玉米叶片12h,SOD活性上升,至48h后下降;在该甘露醇溶液中另外加入10^-2mol/L H2O2;处理12h后SOD活性基本不变。对玉米叶片单独用10^-2mol/L H2O2诱导,30min内SOD活性上升到最高值,随着处理时间的延长又逐渐下降。用耐强光、耐干旱的野生马齿苋作为材料,与玉米相比,其叶片SOD基础活性低于后者;若予以正午强光结合渗透胁迫2h,其叶中SOD活性增幅超过玉米,从种间比较的角度旁证了SOD在抗逆性中的作用。推测植物中存在比活性氧更为直接的物理诱导机制。