实时荧光定量PCR在植物病害流行学中的应用

实时荧光定量PCR广泛应用于植物病理学研究的各个领域。近年来已开始应用于植物病害流行学的定量研究中,本文对近年来实时荧光定量PCR技术在病菌初始菌源的定量分析、病害流行动态的监测、寄主抗病性的快速鉴定和植物病原菌的抗药性监测等方面的应用进行了综述。     

花生HyPGIP基因克隆及抗叶腐病表达分析

本文采用基因克隆和生物信息学方法研究了花生Hy PGIP基因及编码蛋白的基本性质和生物学功能,并通过测定接种叶腐病菌后花生植株内PG酶的活性变化和PGIP蛋白的生成量变化,以及Hy PGIP基因瞬时表达分析,探讨了PGIP蛋白与寄主抗病性关系。结果表明,受叶腐病菌侵染的花生植株中,抗病品种的PG酶活性都明显低于感病品种,而生成的PGIP蛋白量都比感病品种显著多,暗示PGIP蛋白与花生叶腐病抗性有关。经基因克隆和测序获得HyPGIP基因全长序列为1 029 bp,编码342个氨基酸,无内含子,终止密码子为TGA,与已报道的五个花生PGIP序列的同源性都很高;结构预测发现该序列存在8个LRR结构域,存在信号肽,疏水性较强,定位于细胞壁上;RT-PCR显示,经接种叶腐病菌处理后,花生植株中的PGIP基因表达量明显增加。     

黄土丘陵植被恢复过程中土壤种子库萌发数量特征及动态变化

采用野外定点采样与室内土壤种子萌发试验相结合的研究方法,选取黄土丘陵区典型草原带宁夏固原地区不同植被恢复年限(2,5,12,24,35,55,75 a)及当年农地(对照)的8个样地,研究该区土壤种子库萌发数量特征及动态变化.结果表明,8个不同植被恢复年限样地土壤中可萌发的种子的平均数量为3542.5粒/m2,可萌发种子以双子叶植物为主,约占萌发总数的90%.0～5 cm土层可萌发种子数量变化于166.7～9100.0粒/m2;5～10 cm土层变化于133.38～2120.02粒/m2.土壤中可萌发种子数量随着恢复年限的增加表现先增加后下降的趋势.土壤中可萌发的种子数量随时间的变化近似单峰型,双子叶植物峰值主要出现在第4～6周,而单子叶植物萌发高峰在5～6周.表明在环境,气候等条件适宜时,黄土丘陵区典型草原带可萌发的土壤中的种子可形成萌发高峰.     

基于GIS与RS的土壤侵蚀变化定量监测——以黄土高原水保二期世行贷款庆城项目区为例

在GIS软件ArcMap和ArcView的支持下,建立研究区的空间数据库,利用GIS软件的栅格数据空间分析功能,将研究区空间离散化为10 m×10 m的栅格,生成通用土壤流失方程RUSLE所需的各因子栅格图;借助GIS软件的地图代数运算,将各因子连乘,得到土壤侵蚀量栅格图,在此基础上对侵蚀栅格图进行分类,获得土壤侵蚀等级图.将得到的土壤侵蚀图与土地利用图和坡度图叠加,获得不同土地利用方式下和不同坡度下的土壤侵蚀量,据此对土壤侵蚀与土地利用以及土壤侵蚀与坡度之间的关系进行分析,以期为水土流失的防治和保护规划方案的制定提供科学依据.     

我国草坪学研究文献的统计分析

以《中文科技期刊全文数据库》为统计分析源,采用文献计量方法对1997-2006年我国草坪学研究论文进行分析。通过对文献量年代变化、期刊分布情况以及论文主题等方面的统计分析,结果表明,我国的草坪学研究在1997-2006年间文献量呈线性增长趋势,正处于快速发展阶段;《草业科学》、《草原与草坪》、《四川草原》、《中国草地》、《草业学报》、《草地学报》6种期刊在1997-2006年发文量较高,是我国草坪学研究的核心期刊群;草坪草的生态学、生物学、生理生化、解剖学、细胞学、分子学、基因水平研究已经成为我国草坪学研究的重点和热点。     

Shedding and Seroprevalence of Pathogenic Leptospira spp. in Sheep and Cattle at a New Zealand Abattoir

A cross‐sectional study was carried out on sheep and cattle slaughtered at a New Zealand abattoir from September to November 2010 to investigate the supplier‐specific shedding rate, renal carriage rate and seroprevalence of leptospires. In the 2008/2009 season, this abattoir experienced three human leptospirosis cases from 20 staff, of which two were hospitalized. Urine, kidney and blood samples were collected from carcasses of 399 sheep (six suppliers, 17 slaughter lines) and 146 cattle (three suppliers, 22 slaughter lines). The urine and kidney samples were tested by quantitative real‐time PCR (qPCR), while serum samples (from coagulated blood samples) were tested by microscopic agglutination test (MAT). In total, 27% (73/274; 95% CI: 18–37) of urine samples tested positive by qPCR. Species‐specific shedding rates (prevalence of positive urine qPCR) were 31% (95% CI: 17–48) for sheep and 21% (95% CI: 14–30) for cattle. For 545 kidney samples tested, 145 were qPCR positive (27%; 95% CI: 17–39). The average prevalence of kidney qPCR positivity was 29% (95% CI: 17–45) for sheep and 21% (95% CI: 15–28) for cattle. Three hundred and thirty of 542 sampled sheep and cattle had antibodies against Leptospira borgpetersenii serovar Hardjobovis (Hardjobovis) and/or Leptospira interrogans serovar Pomona (Pomona), based on reciprocal MAT titre ≥1 : 48 (overall seroprevalence of 61%; 95% CI: 48–73). Seroprevalence was 57% (95% CI: 40–72) for sheep and 73% (95% CI: 59–83) for cattle. Among the seropositive animals, 41% (70/170; 95% CI: 30–54) were shedding (tested positive by urine qPCR) and 42% (137/330; 95% CI: 30–54) had renal carriage (tested positive by kidney qPCR). Some risk management options for abattoirs or farms to prevent human leptospirosis infections include vaccination of maintenance hosts, the use of personal protective equipment, and the application of urine qPCR to detect shedding status of stock as surveillance and as an alert.     

用于鸡生长和肉质性状定位资源群体的构建

采用远交F-2设计,以杏花鸡(A系)为亲本,分别与隐性白洛克鸡(B系)、泰和丝羽乌骨鸡(C系)进行全同胞和半同胞的正反交,产生了包含A♂×B♀、B♂×A♀、A♂×C♀、C♂×A♀4个杂交组合的各两组F2群体,并进行详细的性状表型记录,建立了可用于定位生长和肉质的数量性状座位(quantitative trait loci,QTL)的4个资源群体。对F2群体的各种性状表型值进行分析,结果显示,F2群体各生长性状和肉质性状都分离得很好。资源群体有足够的群体规模,并有利于校正母体效应和环境影响,保证有效单核苷酸多态性(single nucleotide poly-morphism,SNP)检测数量和QTL定位的准确性,使资源群体的构建达到了预期的效果。     

杧果种质资源果实数量性状评价指标探讨

为了更好地对我国杧果种质资源进行评价研究,尽快建立我国数量化、规范化的果种质资源描述系统,2002-2004年,在广东省湛江市中国热带农业科学院南亚热带作物研究所杧果种质资源圃内进行了48-57份杧果品种(种质)资源果实的平均单果重、果核质量、可溶性固形物、可滴定酸含量及维生素C含量等5项数量性状指标的测定。结果表明,以上5个经济性状均存在14%以上的变异系数。可溶性固形物呈正态分布,其他4项指标均呈偏态分布。根据分析结果,提出单果重、果核质量、可溶性固形物、可滴定酸含量及维生素C含量5个数量性状的评价标准,建议将5个数量性状分为1-5个等级,每个级别提出2个参照品种,1个国外品种,1个国内品种。该项研究为我国杧果种质资源描述系统数量化、规范化的建立奠定了基础。     

Development and initial validation of a sensory threshold examination protocol (STEP) for phenotyping canine pain syndromes

Objective

To study the feasibility and test–retest repeatability of a sensory threshold examination protocol (STEP) and report the quantitative sensory threshold distributions in healthy dogs.

Development and validation of a quantitative PCR assay method of assessing relative resistance of oat (Avena sativa) to crown rust (Puccinia coronata f. sp. avenae)

Evaluation of oat crown rust resistance is usually based on visual assessment of disease severity or infection types. Visual assessment is subjective, prone to rater bias and requires expert knowledge. PCR-based quantitative assays can overcome challenges associated with visual assessment. New TaqMan primers and probes were designed from Puccinia coronata f. sp. avenae (Pca) sequences. The primer–probe sets were specific to Pca, amplified using as little as 0.5 pg fungal DNA (fDNA) and allowed for scaling to variation in sample total DNA quantity. The quantitative PCR (qPCR) assay was validated using oat recombinant inbred lines (RILs) from the Provena × 94197A1-9-2-2-2-5 cross evaluated under a controlled environment. For comparison with fDNA load, inoculation with the Pca race LCBB provided segregation data on the hypersensitive response, while Pca race LSLG provided data on segregation for reduced pustule number. fDNA content was positively correlated with both pustule number and infection type (IT). Composite interval mapping identified two quantitative trait loci (QTLs) on oat linkage groups Mrg12 and Mrg20 using visual and qPCR assessments (pustule number, IT and fDNA). In this study a qPCR assay method that can be used to assess the relative resistance of oat to crown rust was refined and validated, and single nucleotide polymorphisms (SNPs) closely linked with two QTLs derived from the crown rust resistant line 94197A1-9-2-2-2-5 were identified.