黄瓜抗黑星病相关基因的差异表达分析

以抗黑星病黄瓜材料HX1为试材,接种黑星病菌（Cladosporium cucumerinum）2h、8h、20h、32h和72h的叶片作为试验方（Tester）,相应的未接种叶片作为对照方（Driver）,利用SSH技术,构建了黑星病菌侵染初期的正向和反向cDNA-SSH文库。用巢式引物PCR检测插入片段,获得了200个阳性克隆,通过测序,除去重复序列,共得到105个Unique ESTs。与非冗余蛋白数据库进行BLASTx比对,结果显示,17条ESTs未找到同源序列,88条非重复序列和已知基因的同源性较高,占全部ESTs序列的83.8%,其中86条ESTs与非冗余蛋白数据库已知功能的蛋白具有高度的相似性。结合高密度点阵膜杂交差异筛选,阳性率为75.0%。经初步分析这些序列的功能,差异表达的ESTs功能涉及能量和基础代谢、信号转导、蛋白和核酸代谢、光合作用及逆境中特异表达的基因等方面。为研究黄瓜抗黑星病基因提供了依据。     

Differences in Incubation Length and Hatchling Morphology among Five Species of Oviparous Phrynocephalus Lizards （Agamidae） from China

We incubated eggs of five Phrynocephalus species （P. albolineatus, P. axillaries, P. grumgrzimailoi, P. helioscopus and P. przewalskii） at three constant temperatures （24℃, 28℃ and 32℃） to examine differences in incubation length and hatchling morphology among species and among temperature treatments. We combined data from this study with those reported previously for P. frontalis and P. versicolor to examine whether embryonic stage at laying is a causal factor for interspecific variation in incubation length, and whether the phylogenetic relationship inferred from hatchling morphology is consistent with the relationship based on mitochondrial DNA data. Mean values for incubation length differed among the five species studied herein and, in all these five species, incubation length decreased at a decreasing rate as temperature increased. In none of the five species did hatchling size （snout-vent length and body mass） and other morphological variables differ among the three temperature treatments. The seven oviparous Phrynocephalus lizards found in China differ from each other in hatchling morphology, and embryonic stage at laying is a causal factor of inter- and intra-specific variation in incubation length. The phylogenetic relationship inferred from hatchling morphology is not always consistent with the currently known relationship based on mitochondrial DNA data. Data from this study and those reported previously allow the conclusion that any Phrynocephalus species may have its unique position along the axis defined by hatchling morphology.     

抑制差减杂交筛选禽致病性大肠杆菌基因组差异片段及其分析

采用抑制差减杂交技术(Suppression subtractive hybridization,SSH)对禽致病性大肠杆菌E037株(血清型O78)与非致病菌株K-12MG1655以及同一O2血清型高致病菌株E058与低致病菌株E526进行基因组差异片段克隆与分析。从E037株中共检出17个特异性差异片段,E058株中共检出32个特异性差异片段。经同源分析,这些序列可分为4类:质粒相关序列、噬菌体相关序列、已知功能序列、未知功能序列。这些差异片段包含许多重要的大肠杆菌毒力相关基因,如大肠杆菌素、气杆菌素受体、铁基因簇等。49个片段中,14个片段与其它微生物基因组同源性较高。结果表明,大肠杆菌高致病株与低致病菌株或非致病菌株基因组间存在较多差异基因,其中包括毒力、毒力相关基因、代谢以及噬菌体等基因成分。     

新疆3个民族3对性状的遗传学研究

通过对新疆地区汉族、维吾尔族和哈萨克族的利手、扣手及优势足这3对遗传性状的调查,比较这3对性状在3个民族间基因频率的分布。结果显示：除了汉族和维吾尔族在优势足上存在明显差异,这3对遗传性状在3个民族间差异较小。     

褐飞虱flightin的原核表达及其差异表达检测

flightin最早发现于果蝇Drosophila melanogaster的间接飞行肌中,并且定位于粗肌丝。这种蛋白对维持肌节的结构和功能起到了重要的作用,但其在具有长短翅型分化的褐飞虱Nilaparvata lugens Stl的不同翅型间差异并不清楚。本研究以长翅型雌虫褐飞虱cDNA为模板,通过PCR扩增得到褐飞虱flightin基因ORF全长,将其连接到表达载体pGEX-6P-1中以与谷胱甘肽S-转移酶(GST)融合表达。将表达载体转入大肠杆菌表达株Rosseta,在不同温度、不同浓度IPTG的条件下诱导表达flightin,得到了最优表达条件,获得了高水平可溶性表达。在用GST抗体进行Western blotting验证GST-flightin融合重组蛋白表达的正确性后,我们通过GST柱纯化了的GST-flightin,进而用纯化后的蛋白免疫新西兰兔制备了高特异性的多克隆抗体。最后,我们用制备的多克隆抗体检测了长、短翅型雌成虫和不同发育阶段的褐飞虱体内flightin的表达差异。结果显示,flightin仅在长翅型成虫中表达,在短翅型雌成虫中未检测到其明显表达,而且flightin只在成虫期表达。本研究为进一步研究褐飞虱的flightin与其它蛋白互作、翅肌发育和翅型分化打下了基础。     

濒危鱼类稀有白甲鱼沅江种群与西江种群形态度量学性状的差异性

分别以采自沅江水系142例和西江水系92例稀有白甲鱼标本为材料,采用差异系数法、方差分析法、判别分析法和主成分分析法,研究了稀有白甲鱼沅江种群、西江种群包括10个可数性状和31个比例性状等形态度量学性状的差异性.方差分析表明,2个种群之间共有6个可数性状和27个比例性状存在着显著性差异(P＜0.05).判别分析筛选出了9个对区分2个种群贡献较大的比例性状,并建立了2个种群的判别函数式.主成分分析显示,稀有白甲鱼种群鱼体沿背腹轴、头尾轴分别有16和6个比例性状表现出了明显的变化,在尾部有2个比例性状差异明显.可见2个种群之间形态度量学性状已显示了明显的分化.这可能是两个种群长期隔离,在不同水系生态环境中产生了适应性进化的结果.然而,差异系数的计算结果表明,两个种群之间形态度量学性状的差异尚未达到亚种分化的水平.     

黄海鳀鱼时空分布季节差异分析

利用1986-2010年的底拖网调查数据,基于GIS的空间分析和地统计分析技术,从不同角度阐明在捕捞压力和气侯变化的双重胁迫下,黄海鳀鱼资源密度时空分布及其变动的季节差异.结果表明:不同季节鳀鱼空间分布趋势变化明显,冬季,资源密度从调查区域东北部向岸边和南部水域递减,春季,与冬季相反,资源密度由岸边向东部水域递减,夏季,整体上资源密度呈北高南低的趋势,但高值点散乱分布,秋季,与冬季相似,资源密度由东部水域向岸边环形递减,但变化梯度较冬季缓和;鳀鱼资源密度重心季节变化明显,但不同年代变化规律并不相同;1986-2010年,春、夏和秋3个季节,鳀鱼资源密度重心均向北移动,冬季,资源密度重心年间变化明显,但无整体迁移规律;不同季节,鳀鱼资源密度高值聚集区空间分布位置不同.     

不同生态型摩西球囊霉菌株对蜈蚣草砷吸收的影响

砷超富集植物——蜈蚣草无论是在野外或是在室内均能被丛枝菌根真菌(AM真菌)侵染,但其对蜈蚣草砷吸收及转运的机理尚不清晰.本研究将分离于湖南省郴州市金川塘某铅锌尾矿蜈蚣草根际土壤(Glomus mosseae BGC GD01,简称污染菌株)和云南省未污染土壤(G.mosseae BGC YN05,简称非污染菌株)的2种摩西球囊霉菌株分别接种于非污染生态型和污染生态型蜈蚣草根际,8周后利用菌根化蜈蚣草幼苗在浓度为100 μmol·L-1砷(Na2HAsO4·7H2O)营养液中进行为期24 h的水培试验.结果表明,2种生态型摩西球囊霉菌株分别与蜈蚣草形成中等程度侵染,侵染率为25.2％ ～31.3％.无论是接种污染菌株或是非污染菌株,均明显促进了蜈蚣草根部对磷的吸收.在24 h水培试验期间,接种非污染菌株显著促进了蜈蚣草根部砷的吸收,但接种污染菌株对蜈蚣草根部砷吸收的促进作用有限,说明AM真菌对蜈蚣草砷吸收存在种内差异.     

Genome-wide analysis of Nilaparvata lugens nymphal responses to high-density and low-quality rice hosts

The brown planthopper （BPH） Nilaparvata lugens is an economically impor- tant pest on rice plants. In this study, the higher population density and yellow-ripe stage of rice plants were used to construct adverse survival conditions （ASC） against BPH nymphs. Simultaneously, the low population density and tillering stage of rice plants were used to establish a suitable survival condition （SSC） as a control. Solexa/Illumina sequencing was used to identify genes of BPH nymphs responding to ASC. Significantly longer duration development of BPH nymphs and significantly lower brachypterous ratio of BPH adults were observed by ASC compared with SSC. A total of 2 544 differentially expressed genes （DEGs） were obtained and analyzed by BLASTx, Gene Ontology and KEGG Orthology. Gene ontology analysis revealed that the DEGs were mainly involved in categories of cell, cell part, cellular process, binding, catalytic, organelle and metabolic processes. 1138 DEGs having enzyme commission numbers were assigned to different metabolic pathways. The largest clusters were neurodegenerative diseases （137, 12.0%）, followed by carbohy- drate metabolism （113, 9.9%）, amino acid metabolism （94, 8.3%）, nucleotide metabolism （76, 6.7%）, energy metabolism （64, 5.6%）, translation （60, 5.3%）, lipid metabolism （58, 5.1%）, and folding, sorting and degradation （52, 4.6%）. Expressing profile of 11 DEGs during eight nymphal developmental stages of BPH were analyzed by quantitative real- time polymerase chain reaction. The 11 genes exhibited differential expression between ASC and SSC during at least one developmental stage. The DEGs identified in this study provide molecular proof of how BPH reconfigures its gene expression profile to adapt to overcrowding and low-quality hosts.