薄壳山核桃疮痂病菌TaqMan荧光定量PCR检测方法的建立及应用

疮痂病是薄壳山核桃上最具毁灭性的病害,带菌植物材料是传播疮痂病的重要来源。准确、灵敏、快速的检测方法可为该病害流行规律调查和防控提供有力的依据。本文通过比较薄壳山核桃疮痂病菌Venturia effusa及其近似种之间的ITS序列差异,设计了特异性引物和TaqMan探针,建立了薄壳山核桃疮痂病菌的荧光定量PCR检测方法。特异性检测结果表明,该方法可以检测不同地区的薄壳山核桃疮痂病菌菌株,而对其近似种以及薄壳山核桃上的其他真菌均没有信号。本研究建立的检测方法对薄壳山核桃疮痂病菌DNA的最低检测限可达0.5 pg/μL。该方法用于田间样品检测时,检测时间仅需1 h,远快于常规的分离培养法。本研究建立的基于TaqMan探针的荧光定量PCR检测方法为薄壳山核桃疮痂病菌的快速检测和监测提供了有力工具。     

荧光定量PCR法检测点带石斑鱼神经坏死病毒研究

根据己公布的神经坏死病毒全基因序列,应用引物设计软件依据Gene Bank所收录的各种石斑鱼神经坏死病毒RNA2基因组全序列选择保守区域,根据Taqman探针引物设计原则,设计Taq Man探针及其引物。采用Taq Man探针技术,并且将假病毒技术构建的含有神经坏死病毒RNA2基因组的MS2噬菌体假病毒作为阳性质控品,建立Taq Man探针检测点带石斑鱼神经坏死病毒的荧光定量PCR方法,拟合标准曲线,能够在病毒检测中进行绝对定量,能检测到10个病毒拷贝模板数,在临床检测中得到很好运用。本研究建立实时荧光定量RT-PCR,有助于日常检验工作,为鱼类神经坏死病毒的筛选和监测提供基础,有助于控制神经坏死病的疫情传播。     

猪瘟病毒和猪繁殖与呼吸综合征病毒高致病性/经典毒株多重RT-PCR方法的建立及应用

为了建立鉴别猪瘟病毒(Classical swine virus,CSFV)、高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)及经典猪繁殖与呼吸综合征病毒(C-PRRSV)的快速检测方法,根据GenBank中公布的已知序列,设计合成了针对CSFV、HP-PRRSV和C-PRRSV特异性引物。经过引物筛选及对退火温度、引物比例等扩增条件的优化,建立了用2对引物同时检测CSFV、HP-PRRSV和C-PRRSV的多重RT-PCR方法。该方法可同时扩增出287bp的CSFV、374bp的HP-PRRSV和464bp的C-PRRSV目的核苷酸片段,与猪圆环病毒2型(PCV2)、猪细小病毒(PPV)、伪狂犬病毒(PRV)、猪流行性腹泻病毒(PEDV)和传染性胃肠炎病毒(TGEV)等无交叉反应,最低可以检测到11.3pg的CSFV、4.7pg的HP-PRRSV和2.8pg的C-PRRSV总RNA。分别用多重和单项RT-PCR检测了从河北省采集的31份血清或组织样品,其中CSFV、HP-PRRSV、C-PRRSV以及CSFV与HP-PRRSV混合感染的检出率分别为19.4%、22.6%、0%与9.7%,多重与单项RT-PCR的符合率为100%。本试验建立的多重RT-PCR方法可同时鉴别CSFV、HP-PRRSV与C-PRRSV,适于猪瘟病毒和猪繁殖与呼吸综合征病毒的快速检测与流行病学调查。     

Quantitative CT assessment of bone mineral density in dogs with hyperadrenocorticism

Canine hyperadrenocorticism (HAC) is one of the most common causes of general osteopenia. In this study, quantitative computed tomography (QCT) was used to compare the bone mineral densities (BMD) between 39 normal dogs and 8 dogs with HAC (6 pituitary-dependent hyperadrenocorticism [PDH]; pituitary dependent hyperadrenocorticism, 2 adrenal hyperadrenocorticism [ADH]; adrenal dependent hyperadrenocorticism) diagnosed through hormonal assay. A computed tomogaraphy scan of the 12th thoracic to 7th lumbar vertebra was performed and the region of interest was drawn in each trabecular and cortical bone. Mean Hounsfield unit values were converted to equivalent BMD with bone-density phantom by linear regression analysis. The converted mean trabecular BMDs were significantly lower than those of normal dogs. ADH dogs showed significantly lower BMDs at cortical bone than normal dogs. Mean trabecular BMDs of dogs with PDH using QCT were significantly lower than those of normal dogs, and both mean trabecular and cortical BMDs in dogs with ADH were significantly lower than those of normal dogs. Taken together, these findings indicate that QCT is useful to assess BMD in dogs with HAC.     

关岭牛MyoD Ⅰ基因在不同组织和时期中的表达分析

为研究MyoDⅠ基因在关岭牛不同组织表达量的差异及在肌肉发育过程中的表达情况,试验采用实时荧光定量PCR技术对关岭牛背最长肌、大腿肌、心脏、肝脏、脂肪、小肠6个组织的MyoDⅠ基因相对表达量进行检测。同时分析关岭牛胎儿、18月龄、30月龄时期背最长肌组织中MyoDⅠ基因的表达规律。结果显示,MyoDⅠ基因在背最长肌中表达量最高,在出生后的关岭牛肌肉组织中也具有较高的表达量,且随着出生时间的推移呈上升趋势。推测可能由于背最长肌是肌肉富集地方,因此基因表达量较高,随着年龄增大,肌肉丰满度增加,基因表达量也随着增加。     

云南地方栽培木豆群体数量性状变异分析

对云南省8个地方栽培木豆群体进行调查,对木豆株高、地径、冠幅、单株荚数、虫荚率、单株粒质量及百粒质量等数量性状变异进行分析,结果表明7个性状不论是群体间还是群体内个体间均存在丰富变异。性状相关性分析表明,单株粒质量与单株荚数呈高度正相关,与株高、地径呈弱正相关。单株荚数、虫荚百分率、百粒质量对产量构成贡献最大,通径系数分别为0．8551、-0．2374、0．1799。若将三个性状相结合,选择单株荚数多、粒大、虫荚少的单株,则后代产量可望有明显增加,有可能育成高产高抗虫性品种。     

Multi‐temporal Digital Photogrammetric Analysis for Quantitative Assessment of Soil Erosion Rates in the Landola Catchment of the Upper Orcia Valley (Tuscany,Italy)

This paper focuses on the analysis of the slope and catchment erosion dynamics in a typical Mediterranean context and its sensitivity to recent climatic and socio‐economic changes. The main objective of the present study is to test the reliability of digital photogrammetric analysis to evaluate the time and space evolution of erosion processes mainly triggered by surface running waters and landslides for about the last 60 years. The selected test area is the Landola catchment, a minor tributary of the Upper Orcia River Valley (Southern Tuscany). The Upper Orcia valley is a key site for the comprehension of denudation processes typically acting in Mediterranean badland areas thanks to the following: (i) the availability of long‐lasting erosion monitoring datasets (20 years' long direct measurements at erosion ‘hot spots’); (ii) its representativeness as a sub‐humid Mediterranean badland area; and (iii) the rapidity of development of erosion processes, which makes it suitable as an open‐air laboratory for the study of badland dynamics. The outputs of this work highlight that the application of high‐resolution digital photogrammetric methodologies can represent a powerful and low‐cost tool to evaluate rates and spatial–temporal distribution of denudation processes, as confirmed by the validation through field point monitoring in areas close to the study area. The results obtained for the study area confirm that high erosion rates are a persistent environmental problem for the Upper Orcia Valley, which is not solved up to now, despite various land conservation interventions. The performed multi‐temporal analysis shows a slight decrease in the average water erosion rate during the last 60 years and a parallel increase in the frequency of mass wasting events. These variations are most likely related to a complex interplay between land use changes that have affected the study area during the studied period, revealing a very delicate equilibrium between farming activities and erosion processes. Copyright © 2014 John Wiley & Sons, Ltd.     

北京市森林生态效益功能定位及其经营对策

系统分析北京市森林的作用和经营目的,包括对社会和谐和文明发展的作用,对建设社会主义新农村和解决三农问题的作用,及为市民的生产和生活提供生态安全屏障等.分析认为,实现森林多资源经营,并以发挥其生态功能为主是北京市森林经营的关键,对森林生态效益进行计量评价是实现此目标最好的手段.提出北京市森林经营仍需要从政策上、管理上和技术上等方面去考虑.     

雾灵山森林公园旅游资源评价

先采用定性方法，从直观上描述了雾灵山森林公园的自然资源、人为景观、旅游环境和开发条件；然后根据游客的调查问卷，又对其进行了定量评价。结果表明：雾灵山的生态旅游资源丰富，区位条件良好，适宜扩大开发生态旅游。并根据评价结果，对公园开发生态旅游提出了几点建议。     

运用数量化方法估测普洱县森林蓄积量

根据普洱县森林遥感调查资料 ,利用卫星影像特征判读数据与地面实测数据建立数学类型 ,估测森林蓄积量。首先将 2 0 0块有、疏林地数据输入数据本中 ,然后分析其与优势树种、龄组、郁闭度、海拔、坡向、坡度的关系 ,结果把样地分为针叶林、阔叶林、针阔混交林三个类型分别建立数学模型 ,以估测每公顷蓄积量。数学模型的建立是根据数量化模型的原理来编程 ,并通过计算机的不断调试 ,直到可行为止。蓄积量比较精度达到 90 3%。