亚洲不同生态区水稻群体遗传多样性与特异性分析

研究不同生态区水稻群体的遗传多样性及其遗传结构分化、群体特异性及其相互关系,为水稻起源及亲本遴选提供遗传基础。本研究利用119对SSR标记对太湖、黑龙江省和越南的440种水稻品种进行分析。结果在三个不同生态区中的太湖流域水稻中检测到783个等位基因,平均多态信息含量（PIC）为0.49;在黑龙江水稻中检测到296个等位基因,平均多态信息含量（PIC）为0.21;在越南水稻中共检测到374个等位基因,平均多态信息含量（PIC）为0.28,这表明越南水稻和黑龙江水稻的多样性低于太湖水稻。特缺等位变异数最多的是黑龙江亚群（132）,最少的是太湖亚群（4）;特有等位变异数最多的是太湖亚群（284）,最少的是黑龙江亚群（25）。从亚群间补充等位变异数来看：其补充等位变异数最多的是太湖亚群对黑龙江亚群的补充（447）,最少的是黑龙江亚群对太湖亚群的补充（29）。利用Structure2.2软件将三个地区的440个品种分为7类血缘,发现每个血缘都不是独立的而是相互渗透的。     

Design standards for experimental and field studies to evaluate diagnostic accuracy of tests for infectious diseases in aquatic animals

Design and reporting quality of diagnostic accuracy studies (DAS) are important metrics for assessing utility of tests used in animal and human health. Following standards for designing DAS will assist in appropriate test selection for specific testing purposes and minimize the risk of reporting biased sensitivity and specificity estimates. To examine the benefits of recommending standards, design information from published DAS literature was assessed for 10 finfish, seven mollusc, nine crustacean and two amphibian diseases listed in the 2017 OIE Manual of Diagnostic Tests for Aquatic Animals. Of the 56 DAS identified, 41 were based on field testing, eight on experimental challenge studies and seven on both. Also, we adapted human and terrestrial‐animal standards and guidelines for DAS structure for use in aquatic animal diagnostic research. Through this process, we identified and addressed important metrics for consideration at the design phase: study purpose, targeted disease state, selection of appropriate samples and specimens, laboratory analytical methods, statistical methods and data interpretation. These recommended design standards for DAS are presented as a checklist including risk‐of‐failure points and actions to mitigate bias at each critical step. Adherence to standards when designing DAS will also facilitate future systematic review and meta‐analyses of DAS research literature.     

不同PCR引物对柑桔黄龙病菌的特异性检测

本文对国内外研究者设计的11对PCR引物检测柑桔黄龙病菌的特异性进行了比较。其中包括5对一步法PCR引物(A2/J5、GB1/GB3、MP1/MP2、CQULA03F/CQULA03R和CLA-F2/CLA-R2)和6对巢式PCR引物(P1/P2、P3/P4、LP1/LP2、LP3/LP4、DP1/DP2和DP3/DP4)。结果发现,MP1/MP2和CLA-F2/CLA-R2 2对引物具有非特异性,不能区分亚洲柑桔黄龙病菌和非洲柑桔黄龙病菌,其余的9对引物特异性较好。本文旨在为研究者选择柑桔黄龙病菌扩增引物提供参考。     

Baseline toxicity and stage specificity of recently developed reduced‐risk insecticides chlorantraniliprole and spinetoram to obliquebanded leafroller,Choristoneura rosaceana (Harris) (Lepidoptera: Tortricidae)

BACKGROUND: Studies were designed to assess baseline toxicity of the two recently developed reduced‐risk insecticides chlorantraniliprole and spinetoram to obliquebanded leafroller (OBLR), Choristoneura rosaceana (Harris), and to determine stage‐specific effects of these products on OBLR, so that these new chemicals could be judiciously incorporated into IPM programs by targeting only the most susceptible stages of OBLR. RESULTS: Both chlorantraniliprole and spinetoram were highly effective against neonate OBLR larvae at much lower doses than conventional standard azinphosmethyl. Most of the mortality caused by spinetoram occurred during the 4 day exposure period, while significant delayed mortality was observed as a result of chlorantraniliprole treatment during the 7 day recovery period, indicating that chlorantraniliprole is a slower‐acting compound compared with spinetoram. Unlike broad‐spectrum predecessors, the toxicity of chlorantraniliprole significantly increased with larval age. Similarly, the toxicity of spinetoram increased as larvae grew from neonates to third instars. CONCLUSION: Chlorantraniliprole and spinetoram are highly effective against OBLR larvae, regardless of larval age. Incorporation of these reduced‐risk chemistries into IPM programs for OBLR would lead to effective management of this pest in tree fruit systems. Copyright © 2011 Society of Chemical Industry     

梨轮纹病和炭疽病病原菌PCR检测

为建立梨轮纹病原菌和炭疽病病原菌的分子检测方法,根据GenBank上炭疽病原菌和轮纹病原菌不同种的ITS序列差异设计2对引物TJ1/TJ2和LW1/LW2,对2种病原菌菌株进行扩增,比较了常规PCR和巢式PCR的检测灵敏度,并对果园中接种梨轮纹病原菌的梨果实进行检测。结果表明,建立的PCR体系可分别从炭疽病原菌和轮纹病原菌菌株扩增出317 bp和325 bp的特异性条带,对其他相似或相近的病原真菌则无扩增条带。巢式PCR技术的灵敏度比常规PCR提高了约105倍,且可以在接种较低浓度(1 ml 10个)轮纹病原菌孢子液7 d后的梨果实组织中检测到病原菌。说明使用巢式PCR技术,可以准确、灵敏地监测梨轮纹病原菌在果园的种群消长动态。     

蔗糖密度梯度法定量口蹄疫完整病毒粒子（146S）的特异性研究

为验证蔗糖密度梯度法检测口蹄疫完整病毒粒子（146S）的特异性,选择pH值6．0、5．0、4．0PBS酸解处理3批提纯后的146S灭活抗原液,并将酸解后样品进行146S检测,发现8至11级份OD259峰值消失,146S含量由酸解前的1．77、6．60、3．67μg／mL至酸解后完全不能检测到。同时对3批不含病毒的细胞液按抗原纯化灭活工艺处理后进行146S检测,结果显示3批细胞液在8至11级份均没有明显吸收峰,而且其OD259值与PBS空白对照OD259值基本平行一致,说明不含病毒的细胞液对检测结果没有明显干扰。试验证明,蔗糖密度梯度法定量FMD病毒液中146S含量具有很好的特异性。     

马鼻肺炎双重荧光定量 PCR 检测方法的建立与应用

【目的】建立快速、准确检测马疱疹病毒 1 型（EHV1）和 4 型（EHV4）的双重荧光定量 PCR 检 测方法。【方法】以 EHV1、EHV4 的全基因组序列为基础,针对糖蛋白 B（gB）基因的保守序列,分别设计 EHV1 与 EHV4 特异性引物和探针,筛选引物,优化反应条件,分型检测马疱疹病毒（EHV1、EHV4）。【结果】 检测到 EHV1、EHV4、EIV、EAV、EVJ 和 EIAV 标准毒株,EHV1、EHV4 为阳性结果,其余病毒为阴性结果; EHV1、EHV4 DNA 模板的最低检出限为 1.47×102 copies/μL;检测 64 份临床样本,阳性检出率为 14.06%,病 毒分离证实 100% 符合。【结论】双重荧光定量 PCR 检测方法可直接应用于检测并区分 EHV1、EHV4, 并可在 短时间内获得对 EHV1/EHV4 特异性及敏感性的检测结果。     

菌根真菌对3种兰花幼苗生长作用研究

菌根真菌在兰科植物生活史中具有重要作用。为获得促进兰科植物生长的共生真菌,阐明菌根真菌对兰科植物的作用,本研究对从野生建兰中分离获得的3株共生真菌进行形态学和系统进化分析,并将3株真菌与建兰、硬叶兰和铁皮石斛幼苗在不同浓度燕麦琼脂培养基(OMA)条件下共培养,进行比较研究。结果表明,3株真菌分属于胶膜菌属(Tsc)、角担菌属(Cej3)和蜡壳菌属(Sec),且菌丝和菌落特征存在明显差异;系统进化分析发现,Tsc菌株与Cej3菌株亲缘关系较近,与Sec菌株亲缘关系较远;与兰科植物共培养结果表明,Tsc菌株可在全部OMA浓度条件下与3种兰科植物幼苗建立共生关系,且定殖率均为100%;Cej3菌株和Sec菌株与3种兰科植物在不同OMA浓度条件下定殖时间及定殖率存在差异,且具有一定的专一性;与对照组相比,与真菌共生的兰科植物幼苗的平均鲜重、干重、苗高、根长增长率均明显提高。本研究结果为深入研究兰科植物与真菌共生的分子机制奠定了一定的理论基础,并为兰花种质资源的保护提供了参考。