一个水稻根时空差异表达启动子的克隆及功能分析

本研究旨在分离克隆水稻根特异表达的启动子,为育种提供资源.本实验结合本室芯片数据库和RT-PCR验证得到一个水稻根特异表达的基因(TIGR Locus:Loc_Os05g19570),采用PCR技术从MH63的基因组DNA中扩增其上游启动子P12,长度为1950 bp,将该启动子片段与报告基因gus相连,构建植物表达载体,农杆菌(Agrobaterium tumefaciens)介导转化水稻(Oryzative ssp.japonica)中花11.组织化学分析及荧光定量测定显示该启动子具有根特异性及时间差异性.根据P12启动子序列特征,利用PCR对其进行有目的的缺失,将5个长度不同的缺失片段分别与gus相连,构建植物表达载体,转化水稻中花11.荧光定量测定结果表明:-1059～-819bp区域缺失后,报告基因在根里的表达量显著降低,说明这个区域可能存在顺式元件.本研究成功的分离克隆到一个水稻根特异的启动子P12,进一步分析发现其还具有时间差异性.     

水稻OsHKT基因表达模式分析

【目的】OsHKT(High-affinity K~+transporter)是与水稻耐盐胁迫有关的一类Na~+或K~+转运体或Na~+-K~+共转运体,对水稻体内Na~+再循环,维持植株地上部特别是叶片中的低Na~+浓度和低Na~+/K~+比具有重要作用。本研究的目的是分析OsHKT基因家族表达的组织特异性、昼夜节律性以及盐、ABA对其表达的影响。【方法】利用荧光定量PCR技术分析OsHKT家族基因的表达模式。【结果】OsHKT基因家族中不同成员的表达具有明显的组织特异性,OsHKT1;1、OsHKT1;3、OsHKT2;3以及OsHKT2;4主要在水稻的叶片中表达,其余基因在根中表达水平较高。分析盐处理后OsHKT家族基因在水稻根和叶中的表达模式,结果表明OsHKT家族基因的表达均受盐胁迫的影响,但是不同OsHKT家族成员对盐胁迫反应不同。盐处理后OsHKT1;5、Os HKT2;1和OsHKT2;2基因在根和叶中的表达模式是一致的,而其他家族成员在根和叶中存在差异。所有OsHKT基因的表达在根或叶片中均受ABA调控,其中只有OsHKT1;3和OsHKT1;5在根和叶片中表达模式相似。光周期表达模式结果表明,OsHKT基因家族成员的表达具有相似的昼夜节律性。【结论】本研究结果表明水稻OsHKT家族不同成员的表达具有组织特异性和相似的昼夜节律性,并且不同成员对盐胁迫和ABA应答反应不同。这种表达模式可能与水稻在不同环境下的盐胁迫反应相适应。本研究结果为进一步揭示OsHKT基因的作用机制奠定了基础。     

病毒诱导的基因沉默在番茄组织中的特异性分析

 病毒诱导的基因沉默（VIGS）是植物基因功能研究的新技术。以组成型表达gusA基因的转基因番茄为材料,通过VIGS技术沉默gusA基因,对VIGS在番茄中的组织特异性进行了初步分析。结果表明,VIGS技术能有效地诱导gusA基因在番茄植株的叶片、花、果实、侧枝、主茎及茎分枝点等部位和组织中沉默,证明VIGS在番茄中具有较强的系统性沉默特征。     

鸡传染性支气管炎病毒S1蛋白抗原表位多肽的预测、鉴定及免疫效果的初步研究

旨在对鸡传染性支气管炎病毒（IBV）H120株S1蛋白的B细胞及T细胞可能的表位进行预测并合成相应的多肽,然后免疫小鼠,并分析其免疫效果,以验证候选表位多肽的免疫原性。利用表位预测软件筛选并化学合成针对IBV H120株S1蛋白的5条表位多肽,然后免疫小鼠,通过间接ELISA、中和试验和流式细胞技术检测各表位多肽诱导的特异性抗体、中和抗体和外周血T细胞亚群。ELISA检测结果显示,5条表位多肽均具有良好的反应原性,免疫小鼠的血清效价高低顺序依次为Pep76-106 > Pep240-257 > Pep511-537 > Pep403-421 > Pep135-172;中和试验结果表明,5条多肽免疫小鼠的血清中和滴度均高于空白对照组,其高低顺序依次为Pep240-257=Pep403-421=Pep511-537>Pep76-106=Pep135-172;流式检测结果表明,5条多肽免疫小鼠在CD3⁺、CD4⁺CD8^-、CD8⁺CD4^- T淋巴细胞水平上均极显著高于空白对照组（P<0.01）,CD3⁺T及CD4⁺CD8^-T淋巴细胞数大小顺序依次为Pep403-421 > Pep240-257 > Pep76-106 > Pep511-537 > Pep135-172,CD8⁺CD4^-T淋巴细胞数大小顺序依次为Pep403-421 > Pep76-106 > Pep511-537 > Pep240-257 > Pep135-172。合成的5条表位多肽中,Pep240-257、Pep76-106和Pep403-421可以诱导体液免疫,Pep403-421可以诱导细胞免疫,其中,Pep403-421可以同时诱导细胞免疫及体液免疫。本研究结果为深入了解S1蛋白的免疫学特性以及研发诊断试剂和有效表位疫苗奠定了基础。     

Larval abundance across the European eel spawning area: An analysis of recent and historic data

The abundance and distribution of leptocephalus larvae of the European eel (Anguilla anguilla, Anguillidae) were examined using ten historic and recent Sargasso Sea expeditions that were selected on the basis of having the largest number of sampling stations and highest catches. The surveys cover the period 1920–2014. Station data were recalculated to the same unit of larval density per unit area, and the irregular station positions were transformed to a regular spatial grid to allow calculation of comparable measures of abundance of the youngest (0+) leptocephalus cohort. The result is that the mean and maximum densities of 0+ leptocephali after 2007 on average have decreased by 70%–80% from the densities during the period before the drastic decrease in glass eel recruitment, which started in the 1980s. This is of the same magnitude as the change in spawning stock, if the total continental commercial landings are used as a proxy. In the same period, the glass eel recruitment in Europe has decreased by more than 95%. The conclusion is that a major cause for the recruitment decrease may be an increased leptocephalus mortality during the oceanic phase or a large geographic shift in glass eel arrival. Combining the survey data, the spatial distribution of 0+ leptocephali was concentrated south of the northernmost front in the Subtropical Convergence Zone, but high densities were also found far south of the front in the western part of the distribution area and leptocephali were present also north of the average frontal position.     

风信子鹿角珊瑚的胚胎和幼虫发育研究

采集异株风信子鹿角珊瑚Acropora hyacinthus自然排放的精卵,进行人工授精,并在显微镜下连续观察其胚胎和幼虫的发育过程。结果表明：风信子鹿角珊瑚是雌雄同体,体外受精,在月圆后2~3d同期排卵;其胚胎发育分为6个阶段,即受精卵期、卵裂期、囊胚期、原肠期、浮浪幼虫期、珊瑚幼体;受精卵经过48 h发育为浮浪幼虫,再经7 d着床变形发育为珊瑚幼体;其浮浪幼虫附着在附着基上后逐渐膨大,由圆形变成蝶形完全附着在基底上,口盘朝上,随后幼虫发生附着变形,长出触手,形成珊瑚幼体。本研究为国内对文昌云龙湾风信子鹿角珊瑚的胚胎和幼虫发育过程的首次报道,研究结果可为珊瑚礁的生态修复提供参考依据。     

稻瘟菌激发子CSBI专化性及相关性质研究

以一套水稻抗稻瘟病近等基因系为材料 ,接种稻瘟菌 ( Magnaporthe grisea)细胞壁来源的糖蛋白激发子 CSBI,其诱导植保素的积累在高度非亲和性互作水稻远高于亲和性互作水稻 ;研究同时表明 ,CSBI可专化性诱导完全非亲和性互作和高度非亲和性互作水稻的过敏性坏死反应 ;表明该激发子具有小种 -品种专化性。经热、胰蛋白酶和过碘酸钠处理后的生物活性检测结果表明 ,CSBI的活性位点为糖基部分。经 p H稳定性检测 ,CSBI在酸性及相对弱碱性条件下较稳定 ;而在强碱性条件下 ,激发子活性下降较多 ,甚至完全丧失。对 CSBI诱导活性的有效浓度测定表明 ,激发子诱导水稻叶片酶活性升高的最低有效浓度为 0 .0 7～ 0 .70 nmol/L。     

幼虫接种密度对桃小食心虫生长发育的影响

为了对桃小食心虫的人工饲养提供参考,室内条件下[温度（23±1）℃,RH 80％±7％,光周期15L∶9D],观察苹果不同接虫密度（10头／果,20头／果,30头／果,40头／果,50头／果）桃小食心虫的生长发育情况。结果表明：各密度的蛀果率无显著差异,在95％以上;随着虫口密度的增加,脱果率呈先下降后上升再下降趋势,羽化率逐渐下降,脱果历期呈逐渐升高再下降,脱果幼虫体重呈先升高再下降。综合比较,10～40头／果的饲养密度能得到较高的饲养回报。     

New insight on vertebral anomalies in cultured Senegalese sole (Solea senegalensis,Kaup) at early stages of development

Senegalese sole (Solea senegalensis, Kaup) is a promising flatfish species in aquaculture. However, skeletal anomalies are still a great concern in sole farming. Investigation of this issue is crucial to improving larval quality and optimizing production. The aim of this study was to thoroughly assess anomalies in the rachis of reared sole at early developmental stages. Sole (n = 507) were sampled at 31 or 32 days after hatching (dah). The specimens were stained with alcian blue and alizarin red and evaluated for the detection of vertebral deformities. Most fish presented 9:34:3 vertebrae in abdominal, caudal and caudal complex regions, respectively. Remarkably, all specimens showed at least one spinal anomaly. Alterations of neural/haemal elements, as well as deformities of hypurals, parhypural and epural, were recurrent. Vertebral body anomalies and/or vertebral column deviations were identified in 52% of the individuals. Vertebral deformations and fusions were common, especially in caudal complex. ‘Minor’ anomalies were predominant, and some of the detected disorders might be a result of non‐/low‐pathological processes. These results contribute a new insight into the main skeletal anomalies affecting cultured sole larvae. Further research is required to determine their impact on fish welfare and external appearances at commercial stages.     

Culture possibilities of the razor clam Ensis arcuatus (Pharidae: Bivalvia)

This study investigated razor clam Ensis arcuatus culture from larval rearing to commercial size to assess it's aquaculture potential. Broodstock was spawned several times in hatchery facilities to demonstrate that larval availability is not a constraint for hatchery production. Larval culture lasted 20 days, showing an average survival of 14.35%. The razor clam seed reached a size of 28.5 mm at 4 months. Seed culture is feasible; however, it is constrained by the fact that razor clams need to be buried. Higher survival was observed when culturing razor clam seed with a substrate (82–83%), irrespective of the substrate grain size, while containers held without a substrate showed a significantly lower survival (56.5%). We showed that it is possible to rear E. arcuatus juveniles in bottles, with survival as high as 94–95% being achieved. The length increase of the hatchery‐produced E. arcuatus over 600 days was described by the equation Ln L=4.5+(?141.17/t), where L is length in mm and t is time in days. On‐growing showed adequate survival rates and it took 2–3 years to reach commercial size.