奶牛难产激发应激性急性肾上腺皮质功能不全——心衰、水电解质严重失调辩析

应激性急性肾上腺皮质功能不全症的病理机制十分复杂,目前人医的各类文献中大都以“垂体—肾上腺轴”功能不全、功能衰竭、或发生广泛性出血坏死性病演作解释。本病在奶牛分娩难产过程中并不少见,病牛的症状表现以心衰和失钠性低血症为主征。但在兽医临床上由于认识不足,曾有不少中外学者将奶牛产后卧地不起的各类疾病,含糊地统称为“母牛睡倒爬不起来综合征”;更由于救治不当,病牛常以淘汰或死亡告终。本文通过对一具体典型病例辨析,着重讨论了本病的基本病理反应和确立诊断依据,强调对本病救护施治时应遵循的基本原则,供同道们在分析和解决这一实际问题时参考。     

从海南省杂草胜红蓟和假马鞭上检测到粉虱传双生病毒

 利用三抗体夹心ELISA(TAS-ELISA)及PCR检测的方法对采自海南的胜红蓟(Ageratum conyzoides)和假马鞭(Stachytarphetajamaicensis)的5个病样进行了检测,表明均为粉虱传双生病毒。PCR扩增产物克隆后进行序列测定,结果表明存在2类双生病毒,其中样品Hn2存在2类病毒的复合侵染。这是在海南首次报道存在有粉虱传双生病毒。     

Spread of potato virus Y in potato cultivars (Solanum tuberosum L.) with different levels of sensitivity

The aim of this work was to correlate the appearance of the symptoms, multiplication and spread of virus after mechanical inoculation of potato (Solanum tuberosum L.) cultivars showing different levels of susceptibility and sensitivity to Potato virus Y^NTN (PVY^NTN). The potato cultivars used were the resistant cultivar Sante and susceptible cultivars Igor, Pentland squire and Désirée. The spread of the virus PVY^NTN in infected plants was monitored using different methods: DAS-ELISA, tissue printing, immuno-serological electron microscopy and real-time PCR. In all three susceptible cultivars, the virus was detected in the inoculated leaves 4–5 days after inoculation. From there virus spread rapidly, first into the stem, then more or less simultaneously to the upper leaves and roots. Real-time PCR was shown to be very sensitive and enabled viral RNA to be detected in non-inoculated leaves of susceptible cultivar Igor earlier than other methods. Therefore, for exact studies of plant–virus interaction, a combination of methods which detect viruses on the basis of their different properties (coat protein, morphology or RNA) should be used to monitor the spread of viruses.     

sikFBA4基因启动子的克隆及低温表达模式分析

天山雪莲生长于常年积雪覆盖高海拔处的高山区域,具有很强的极端低温生境适应能力,属于研究植物低温适应的良好模式植物。前期的研究表明,sikFBA4基因可以显著提高番茄的抗寒能力。为进一步研究sikFBA4基因的耐寒响应模式,以天山雪莲为材料,采用实时荧光定量PCR分析sikFBA4在低温胁迫下的表达模式;利用高效热不对称PCR法(Hi-Tail PCR)克隆sikFBA4启动子序列并进行生物信息学分析。为分析PsikFBA4序列克隆的完整性及转录表达特性,将PsikFBA4与GUS基因融合在烟草中进行瞬时表达。结果表明,sikFBA4在低温胁迫条件下的表达发生瞬时显著上调,并在1 h达到峰值,而后表达下调。通过启动子克隆分析,在PsikFBA4序列-1648 bp位置处具有冷响应元件LTRE;进一步的GUS染色和酶活力测定结果表明,低温能够显著提高GUS基因的表达活性,说明sikFBA4属于低温诱导型基因。SikFBA4基因启动子的克隆、序列分析以及表达分析,为进一步探究雪莲果糖-1,6-二磷酸醛缩酶(sikFBAs)基因的表达与调控机制奠定了基础。     

牡蛎中GⅡ型诺如病毒巢式RT-PCR检测方法的优化与评价

为了解决目前巢式PCR法检测牡蛎样品中GII型诺如病毒时存在的非特异性扩增、假阴性等问题,本研究对NCBI数据库中在线获得的所有GII型诺如病毒序列进行系统分析,重新设计出针对GII型诺如病毒检测的通用型巢式PCR引物(NG2OF/NG2OR),并从基因型覆盖度、特异性、灵敏度和实际样品检测的稳定性几个方面对新设计的引物进行评估。结果显示：(1)新引物的特异性好,只对GII型诺如病毒进行有效扩增,且不受牡蛎样品本底的影响。(2)新引物灵敏度高,最低可检测至26.4个拷贝。(3)在对92个牡蛎样品进行检测时,新引物的阳性检出率为28.3%,比经典引物高出7.5%。因此,新建立的GII型诺如病毒巢式PCR检测方法在检测牡蛎样品时特异性更强,灵敏度更高,为牡蛎中诺如病毒的检测提供了一种新的、可靠的技术手段。     

白背飞虱酚氧化酶原PPO基因特性及其免疫应答

【目的】酚氧化酶(phenoloxidase,PO)是昆虫体内的免疫蛋白,在昆虫体内起重要的调节作用,主要以无活性的酚氧化酶原(prophenoloxidase,PPO)形式存在。本研究在转录组测序获得白背飞虱(Sogatella furcifera)3条PPO序列的基础上,通过分析白背飞虱体内不同SfPPO的发育和组织表达模式,并进一步抑制SfPPO的表达以及病原菌诱导,探讨SfPPO在白背飞虱体内的生物学功能。【方法】以白背飞虱3条SfPPO序列为研究对象,利用生物信息学方法分析其蛋白结构以及与其他昆虫之间的同源性。并以注射绿色荧光蛋白(green fluorescent protein,GFP)的dsRNA作为对照组,通过注射合成的ds SfPPO后采用实时荧光定量PCR(qRT-PCR)检测基因表达抑制效果;此外,为了探究SfPPO在白背飞虱生长发育和免疫应答方面的作用,利用qRT-PCR检测SfPPO在不同发育阶段及组织中的表达情况,以及注射大肠杆菌(Escherichia coli)和枯草芽孢杆菌(Bacillus subtilis)后3个SfPPO的诱导表达情况。【结果】...     

实时荧光PCR检测马铃薯白线虫技术研究

马铃薯胞囊线虫是马铃薯上最重要的有害生物之一,也是我国特别关注的重要植物检疫性线虫.针对马铃薯白线虫ITS序列,我们设计了引物和TaqMan探针,使用15种马铃薯胞囊线虫群体和4种其它胞囊线虫样品进行验证,可高度灵敏地检测单个马铃薯白线虫的胞囊或幼虫,最高检测灵敏度达到10fs;同时开展了混合样品和未知样品的检测,证明了引物的专化性和TaqMan探针特异性.该检测方法可自动化检测马铃薯白线虫并进行定量,适合进行标准化的常规检测.     

Use of real-time PCR to examine the relationship between disease severity in pea and <Emphasis Type="Italic">Aphanomyces euteiches</Emphasis> DNA content in roots

Aphanomyces euteiches causes severe root rot of peas. Resistance is limited in commercial pea cultivars. Real-time fluorescent PCR assay specific for A. euteiches was used to study the relationship between disease severity and pathogen DNA content in infected peas. Five pea genotypes ranging in levels of resistance were inoculated with five isolates of A. euteiches. Plants were visually rated for disease development and the amount of pathogen DNA in roots was determined using the PCR assay. The susceptible genotypes Genie, DSP and Bolero tended to have significantly more disease and more pathogen DNA than the resistant genotypes 90-2079 and PI 180693. PI 180693 consistently had less disease, while 90-2079 had the lowest amount of pathogen DNA. The Spearman correlation between pathogen DNA quantity and disease development was positive and significant (P < 0.05) for three isolates, but was not significant for two other isolates. This suggests that the real-time PCR assay may have limited application as a selection tool for resistance in pea to A. euteiches. Its utility as a selection tool would be dependent on the correlation between disease development and pathogen DNA content for a given pathogen isolate. The accuracy and specificity of the real-time PCR assay suggests considerable application for the assay in the study of mechanisms of disease resistance and the study of microbial population dynamics in plants.