柱状黄杆菌强毒株和弱毒株胞外蛋白中差异蛋白的初步鉴定

柱状黄杆菌(Flavobacterium columnare)是一种世界范围的水产动物致病菌,是中国重要养殖鱼类草鱼(Ctenopharyngodon idellus)、鳜(Siniperca chuatsi)等烂鳃病的病原。本研究以1972年从患"烂鳃病"草鱼上分离的两株冻干柱状黄杆菌G4和G18菌株为研究对象,并将G4株再次分离纯化得纯化菌株,命名为G4R3。对草鱼鱼苗浸泡攻毒结果显示,G4R3的LD50至少比G18的高3个数量级,因此G4R3为"强毒株",G18为"弱毒株"。利用蛋白质组学方法分析柱状黄杆菌强毒株G4R3和弱毒株G18的胞外蛋白,经过双向电泳并结合图像分析,共发现了34个点是差异表达的蛋白。胶内酶解、肽质量指纹图谱和串联质谱分析后,鉴定出其中的7个蛋白点,代表滑动蛋白K、腺酐甲硫氨酸合成酶和一种可能的膜蛋白等3种蛋白,它们可能是柱状黄杆菌的毒力因子。     

鲫源嗜水气单胞菌毒力基因多重PCR检测及ERIC-PCR分子分型

为快速了解鲫源嗜水气单胞菌株毒力基因携带情况及与菌株基因型的相关性,建立多重PCR法和ERIC-PCR分子分型,为临床快速检测、菌株分型和菌株致病性分析提供依据。通过单重PCR法检测出标准菌株ATCC7966内5个毒力基因气溶素(aerolysin,aer)、溶血素(hemolysin,hly)、细胞毒性肠毒素(cytotoxic enterotoxin,alt)、胞外蛋白酶(extracellular protease,ahp)和细胞肠兴奋性肠毒素(intestinal cells of excitatory enterotoxin,act),其扩增产物长度依次为300 bp、592 bp、442 bp、856 bp和500 bp。在此基础上,优化并建立特异性高,敏感度达7.2×102cfu·m L-1多重PCR法,用于检测从江苏射阳地区患病水产动物体内分离的17株嗜水气单胞菌5个毒力基因携带率。结果显示,毒力基因act的携带率为100%,而80%的菌株5个毒力基因均有检出。采用ERIC-PCR分子分型技术,以标准菌株ATCC7966为对照,对17株鲫源致病性嗜水气单胞菌进行基因分型,获得两种基因型,分别描述为A型和B型,其中B型菌株14株,带型与ATCC7966一致,认为是当地的主要流行株。探究菌株基因型与毒力基因分布相关性,携带5个毒力基因的均为B型菌株,而所有A型菌株存在一或多个毒力基因缺失,有可能是此类菌株更易发生毒力基因漂变,但还需进一步研究。     

两种鸡艾美耳球虫冰箱保存后的毒力观察

鸡艾美耳球虫（柔嫩艾美耳球虫和毒害艾美耳球虫）卵囊分成两部分处理,一部分常规冰箱保存6个月,到期后增殖1次;另一部分每月繁殖1次,为新鲜对照虫株。之后,进行致病性试验。结果显示：冰箱保存6个月后,虫株毒力明显下降,仅增殖1次不能达到新鲜虫株毒力水平。如果两种球虫卵囊冰箱保存6个月后,用于试验研究前需至少继代繁殖2次。     

毛里塔尼亚外海乌贼的繁殖力特性研究

利用2015年7-8月毛里塔尼亚外海渔业生产所得的乌贼(Sepia officinalis)样本,研究该属种的繁殖力特性,并利用数学模型分析个体繁殖力的生物学指标表达关系。结果显示,乌贼个体胴长、体质量等生物学指标均呈单峰型区间分布,个体潜在繁殖力PF为1 306~3 375粒,平均值为2 466±545粒,潜在繁殖力随着性腺发育增加显著(P0.05);胴长相对繁殖力PF_(ML)为8.70~18.56粒/mm,平均为14.17±2.12粒/mm;体质量相对繁殖力PFBW为2.72~7.81粒/g,平均为5.18±1.23粒/g;PFBW比PF_(ML)更为稳定(δ2PFBWδ2PFM L)。PF、PFBW均与胴长、胴宽、头宽、鳍长、鳍宽、体质量、净质量和头质量等生物学指标关系密切(P0.05),PF_(ML)与这些生物学指标的数学模型拟合相关系数较低,函数关系不显著(P0.05)。多元线性逐步回归分析结果进一步显示,PF与鳍长呈显著的相关关系,而PFBW则与体质量关系显著。研究表明,鳍长和体质量分别是衡量个体潜在繁殖力、体质量相对繁殖力的可靠的生物学指标。     

草地螟的生物学特性及室内毒力测定研究

通过田间调查和室内饲养观察,对草地螟Loxostege sticticalis的生物学特性及发生危害特点进行了研究.选择4种农药用浸叶片法对草地螟进行室内毒力测定,结果表明,4种农药对草地螟的毒力为7.2%苦参碱.烟碱乳油》3%高渗苯氧威》25%灭幼脲Ⅲ号悬浮剂》5%吡虫啉乳油,其中以苦参碱.烟碱的毒力最强,半致死浓度(LC50)为10.499 mg/L,致死浓度(LC95)为16.093 mg/L.     

Long‐distance dispersal and its influence on adaptation to host resistance in a heterogeneous landscape

Small propagules like pollen or fungal spores may be dispersed by the wind over distances of hundreds or thousands of kilometres, even though the median dispersal may be only a few metres. Such long‐distance dispersal is a stochastic event which may be exceptionally important in shaping a population. It has been found repeatedly in field studies that subpopulations of wind‐dispersed fungal pathogens virulent on cultivars with newly introduced, effective resistance genes are dominated by one or very few genotypes. The role of propagule dispersal distributions with distinct behaviour at long distances in generating this characteristic population structure was studied by computer simulation of dispersal of clonal organisms in a heterogeneous environment with fields of unselective and selective hosts. Power‐law distributions generated founder events in which new, virulent genotypes rapidly colonized fields of resistant crop varieties and subsequently dominated the pathogen population on both selective and unselective varieties, in agreement with data on rust and powdery mildew fungi. An exponential dispersal function, with extremely rare dispersal over long distances, resulted in slower colonization of resistant varieties by virulent pathogens or even no colonization if the distance between susceptible source and resistant target fields was sufficiently large. The founder events resulting from long‐distance dispersal were highly stochastic and exact quantitative prediction of genotype frequencies will therefore always be difficult.     

Response of five dry tropical tree seedlings to elevated CO2: Impact of seed size and successional status

The impact of seed size and successional status on seedling growth under elevated CO₂ was studied for five dry tropical tree species viz. Albizia procera, Acacia nilotica, Phyllanthus emblica, Terminalia arjuna and Terminalia chebula. Seedlings from large (LS) and small seeds (SS) were grown at two CO₂ levels (ambient and elevated, 700–750 ppm). CO₂ assimilation rate, stomatal conductance, water use efficiency and foliar N were determined after 30 d exposure to elevated CO₂. Seedlings were harvested after 30 d and 60 d exposure periods. Height, diameter, leaf area, biomass and other growth traits (RGR, NAR, SLA, R:S) were determined. Seedling biomass across species was positively related with seed mass. Within species, LS seedlings exhibited greater biomass than SS seedlings. Elevated CO₂ enhanced plant biomass for all the species. The relative growth rate (RGR), net assimilation rate (NAR), CO₂ assimilation rate, R:S ratio and water use efficiency increased under elevated CO₂. However, the positive impact of elevated CO₂ was down regulated beyond 30 d exposure. Specific leaf area (SLA), transpiration rate, stomatal conductance declined due to exposure to elevated CO₂. Fast growing, early successional species exhibited greater RGR, NAR and CO₂ assimilation rate. Per cent enhancement in such traits was greater for slow growing species. The responses of individual species did not follow functional types (viz. legumes, non-legumes). The enhancement in biomass and RGR was greater for large-seeded species and LS seedlings within species. This study revealed that elevated CO₂ could cause large seeded, slow growing and late successional species to grow more vigorously.     

辽宁省猪链球菌流行株主要血清型毒力因子检测及其致病性

为了解辽宁省猪链球菌流行菌株主要血清型毒力因子分布特征及致病性,依据猪链球菌属保守基因及血清型特异性基因,从辽宁地区采集猪链球菌疑似病例肺脏、肝脾、脑、关节液等样品进行猪链球菌及其血清类型鉴定;应用猪链球菌6种主要毒力因子特异性基因扩增检测方法,检测所分离到的不同血清类型猪链球菌的毒力因子分布情况,并应用小鼠攻毒试验和病理学技术对其致病性进行观察研究。结果显示:从辽宁地区采集的72个样品中共分离到23株猪链球菌,主要血清型有1,2,7型,其中SS1型4株,SS2型2株,SS7型5株。其余12株未确定血清型。SS1、SS2、SS7型阳性率分别为17.39%,8.69%,30.43%。毒力因子检测结果显示,上述6种毒力因子检出率分别为100%,66.6%,25%,16.7%,75.0%,58.3%,其中SS2均具备6种毒力因子,SS1、SS7和血清型阴性株猪链球菌均具有部分毒力因子,与SS2的差异毒力因子为epf。SS2可引起小鼠的急性败血症以及脑膜炎,SS1、SS7均可引起小鼠发病,但各器官的损伤则较轻。SS1、SS2、SS7脑内接种均可引起小鼠脑膜炎,炎症反应SS2最重,SS1次之,SS7最轻。     

Is virulence phenotype evolution driven exclusively by Lr gene deployment in French Puccinia triticina populations?

Puccinia triticina is a highly damaging wheat pathogen. The efficacy of leaf rust control by genetic resistance is mitigated by the adaptive capacity of the pathogen, expressed as changes in its virulence combinations (pathotypes). An extensive P. triticina population survey has been carried out in France over the last 30 years, describing the evolutionary dynamics of this pathogen in response to cultivar deployment. We analysed the data set for the 2006–2016 period to determine the relationship between the Lr genes in the cultivars and virulence in the pathotypes. Rust populations were dominated by a small number of pathotypes, with variations in most of the virulence frequencies related to the corresponding Lr gene frequencies in the cultivated landscape. Furthermore, the emergence and spread of a new virulence matched the introduction and use of the corresponding Lr gene (Lr28), confirming that the deployment of qualitative resistance genes is an essential driver of evolution in P. triticina populations. However, principal component analysis (PCA) revealed that certain pathotype–cultivar associations cannot be explained solely by the distribution of Lr genes in the landscape. This conclusion is supported by the predominance of a few pathotypes on some cultivars, with the persistence of several other compatible pathotypes at low frequencies. Specific interactions are not, therefore, sufficient to explain the distribution of virulence in rust populations. The hypothesis that quantitative interactions between P. triticina populations and bread wheat cultivars—based on differences in aggressiveness—is also a driver of changes in pathotype frequencies deserves further investigation.