Detection of the Defoliating Pathotype of Verticillium dahliae in Infected Olive Plants by Nested PCR

Spread of Verticillium wilt into newly established olive orchards in Andalucía, southern Spain, has caused concern in the olive industry in the region. This spread may result from use of Verticillium dahliae-infected planting material, which can extend distribution of the highly virulent, defoliating (D) pathotype of V. dahliae to new areas. In this study, a molecular diagnostic method for the early in planta detection of D V. dahliae was developed, aimed especially at nursery-produced olive plants. For this purpose, new primers for nested PCR were designed by sequencing a 992-bp RAPD marker of the D pathotype. The use of the specific primers and different nested-PCR protocols allowed the detection of V. dahliae pathotype D DNA in infected root and stem tissues of young olive plants. Detection of the pathogen was effective from the very earliest moments following inoculation of olive plants with a V. dahliae pathotype D conidia suspension as well as in inoculated, though symptomless, plants.     

Seasonal distribution of phytoplasmas in Australian grapevines

The distribution and persistence of phytoplasmas were determined in Australian grapevines. Phytoplasmas could be detected using the polymerase chain reaction (PCR) from shoots, cordons, trunks and roots throughout the year, and phytoplasmas appear to persistently infect Australian grapevines from year to year. Phytoplasmas were not always detected in samples from the same sampling area from one sampling period to the next. Phytoplasma detection by PCR was improved by sampling from shoots, cordons and trunks, especially during October (early spring). The diseases expressed by the 20 grapevines used in the distribution and persistence studies were monitored. Australian grapevine yellows disease (AGY) was expressed by 17/20 grapevines at some time during the study, whilst only 4/20 and 15/20 grapevines expressed restricted growth disease (RG) and late season leaf curl disease (LSLC), respectively. All grapevines with RG and LSLC also had AGY. The three diseases were persistently expressed in some grapevines and remission of disease was observed in others. The results of PCR detection in the same grapevines indicated that phytoplasmas were more frequently detected in AGY-affected grapevines that also expressed RG and LSLC compared with grapevines expressing AGY alone. Phytoplasmas were detected in symptomless plant material but less frequently compared with AGY-affected material.     

Production and characterization of a monoclonal antibody specific to the M serotype of plum pox potyvirus

A monoclonal antibody to an Albanian isolate of plum pox potyvirus (PPV) was obtained (MAbAL), that specifically recognized strain M of this virus. The specificity of MAbAL, assessed by comparative ELISA on 130 PPV isolates of different geographical origin, 22 of which were also tested by comparative IC-PCR, gave consistent and highly reproducible results. MAbAL seems to be elicited by a stable surface determinant that makes it particularly suitable for successful use under a wide range of conditions. MAbAL is an useful addition to the panel of PPV-specific MAbs available to date.     

Simple diagnosis using ethanol immersion of strawberry plants with latent infection by Colletotrichum acutatum, Dendrophoma obscurans, and Fusarium oxysporum f. sp. fragariae

Simple diagnosis by ethanol immersion (SDEI) to detect Glomerella cingulata was used to detect three other fungi that also cause latent infection of strawberry plants. Signs on strawberry leaves with asymptomatic latent infection by Colletotrichum acutatum became visible using SDEI. Salmon-pink conidial masses were produced in the acervuli on the treated leaves 5 days after incubation at 28°C. In the case of Dendrophoma obscurans, pycnidia with amber conidial masses formed 5 days after incubation at 28°C. The pycnidia were observed mainly on the ribs, and conidial masses exuded from the ostiole. These macroscopic conidial masses were similar to those of G. cingulata and C. acutatum. When water was dripped onto a lesion caused by D. obscurans, the pycnidia exuded white filamentous conidial masses, making the distinction of D. obscurans from G. cingulata or C. acutatum. On petioles with latent infection by Fusarium oxysporum f. sp. fragariae, white aerial hyphae grew out from the vascular tissues on the cut surface 3 days after incubation at 28°C and were easily observed by eye or with a loupe. Thus, SDEI was also useful for diagnosing latent infection of strawberry plants by C. acutatum, D. obscurans, and F. oxysporum f. sp. fragariae.     

通过自制PCR试剂盒确诊雏鹅细小病毒感染

通过 PCR条件选择 ,组装了可用于确诊鹅细小病毒 (goose parvovirus,GPV)感染的试剂盒。首先根据 Gen Bank中已发表的鹅细小病毒不同株的核苷酸序列 ,设计并合成了 GPV特异性简并引物 ,利用该引物确定 PCR扩增条件 ,并鉴定了扩增产物的特异性。根据上述扩增条件 ,组装 GPV诊断试剂盒。试剂盒共有 3个反应管 ,贮于 - 2 0℃ ,应用时取 1号管 ,加入煮沸的肝脏悬液上清 ,2、3号管分别为阴性和阳性对照。按固定条件扩增 ,结果显示 ,待检病料的检出率在 96 %以上。该试剂盒操作简单 ,效果稳定 ,可用于 GPV感染的确诊     

水貂大肠埃希氏菌病的诊治试验

通过对患病水貂进行病理剖检,细菌纯培养,病毒检测,生化鉴定,确定是水貂大肠埃希氏菌病。再应用多种抗生素作药敏试验,筛选出了高度敏感的抗菌药。并对山东省威海地区水貂大肠埃希氏菌病的发生规律进行了探讨。     

基于Harris和卡尔曼滤波的农业机器人田间稳像算法

针对田间颠簸环境影响农业机器人采集实时稳定图像问题，提出了基于Harris和卡尔曼滤波的农业机器人田间稳像算法。首先，利用摄像头获取田间抖动视频图像序列，进行图像子区域划分并计算各区域灰度均方差，进而确定各区域Harris角点阈值；通过自适应角点阈值设置，增加角点距离约束，完成图像角点检测。然后，对检测出的角点进行光流跟踪，计算出帧间运动估计参数。最后，利用自适应卡尔曼滤波算法对运动估计参数进行平滑操作并动态调整滤波平滑性能，获得精确运动估计矢量。测试结果表明，改进后的Harris角点检测算法区域平均分布标准差减小；自适应卡尔曼滤波算法在保证平滑随机运动前提下，跟踪主动运动性能平均提升30.75个百分点；稳像后的图像峰间信噪比提升15.93%,单帧处理时间为25.66 ms,满足农业机器人30 f/s高速图像采集时同步稳像对实时性要求。     

2016/2017冬中国气温偏暖的基本特征及可能成因

利用中国740余站观测的月平均2 m气温资料以及NCEP/NCAR再分析资料,通过动力学诊断的方法,对2016—2017年中国冬季异常偏暖现象进行了研究。结果发现：中国2016—2017年冬季气温偏暖主要是由于西伯利亚高压和阿留申低压偏弱导致东亚冬季风偏弱,在中国存在异常的偏南风。阿留申低压偏弱由两个因素引起,一是由于阿留申群岛地区的海温偏低,冷源对应异常高压导致阿留申低压偏弱;二是阿留申低压在东亚副热带高空西风急流出口区以北,然而该冬季高空西风急流出口区位置偏南,导致阿留申地区偏离急流出口减压区,从而导致阿留申低压偏弱。此外,西风带偏北,中高纬亚洲大陆上空以纬向环流为主,不利于冷空气南下。