菊花绿变病植原体在中国的发生及分子鉴定

对内蒙古农业大学校园内表现花器绿变症状的菊花样品进行采集和DNA提取,应用植原体16S rRNA基因和rp基因的引物进行巢式PCR扩增,从感病样品中分别扩增得到了长度均约为1.2 kb的片段。序列一致性分析表明,菊花绿变植原体16S rRNA基因与翠菊黄化植原体匈牙利风信子株系(GenBank登录号MN080271)、印度玉米株系(KY565571)、印度繁缕株系(KC623537)和印度马铃薯株系(KC312703)的核酸一致性最高,为99.9%,rp基因序列与翠菊黄化植原体立陶宛洋葱株系(GU228514)的核酸一致性最高,为99.8%。基于16S rRNA基因和rp基因构建系统进化树时发现,菊花绿变植原体均与16SrI-B亚组成员聚为一起。16S rRNA基因相似性系数分析表明,菊花绿变植原体与洋葱黄化植原体(AP006628)的相似性系数最高为1.00,洋葱黄化植原体(AP006628)在分类上属于16SrI-B亚组。因此,我们可以确定该菊花绿变植原体属于16SrI-B亚组。这是我国首次报道菊花绿变病的发生。     

青海省不同生态区蚕豆根瘤菌16S rDNA分析

为了阐明青海省蚕豆根瘤菌的遗传多样性和亲缘关系,同时为发现和获得新的根瘤菌物种提供资源,采集和分离了青海省不同生态区蚕豆主栽品种青蚕14中的根瘤菌,并对其中分离到的6个菌株(CD1~6)进行16S rDNA鉴定分析。将6株蚕豆根瘤菌的全序列结果与NCBI中已报道的序列进行相似性比对,发现其相似度较高,其中供试菌株CD4与KF008225.1相似度最高,达到98%,其余均在95%到96%之间。6株供试菌分属4个菌属,CD1和CD2共属节细菌属,CD3属于分枝杆菌属,CD4和CD5属于快生根瘤菌属,CD6属于中慢生根瘤菌属,表明来自不同地区的蚕豆根瘤菌存在较大差异。聚类分析结果显示,蚕豆根瘤菌的6个菌株分属5个不同系统发育分支,证明青海省不同生态区蚕豆根瘤菌种类多样性较为丰富。     

黄土丘Ⅴ区典型小流域坝控区重力侵蚀分析与预测

黄土丘Ⅴ区因其独特的土壤及地形和水文地质条件,使淤地坝坝控区的重力侵蚀严重,以至于毁坏耕地、道路和桥梁等,严重影响淤地坝效益的发挥。针对黄土丘Ⅴ区典型小流域坝控区在运行期间因沟岸崩塌和岸坡滑塌等产生的重力侵蚀导致淤地坝泥沙淤积量计算失真的问题,采用实测库容与3S技术生成库容资料和原设计库容曲线复核率定的三重复核分析方法,对运行期淤地坝坝控区域的重力侵蚀来源、诱发因素及预测进行分析。结果表明:(1)反映淤地坝淤积形态的形态判别系数α'为0.711~208.517,均值48.238,说明除阳山上小型坝小于临界数2.2为椎体淤积外,其他淤地坝为三角洲淤积;(2)坝控区形态参数如淤积面面积、周长、淤积体积及淤积形态判别系数等对重力侵蚀影响较大,给出了重力侵蚀量及侵蚀模数的计算关系;(3)重力侵蚀模数的范围184.79~812.80t/(km~2·a),均值360.68t/(km~2·a),坝控区淤积产沙模数的范围225~2 719t/(km~2·a),均值1 382t/(km~2·a)。重力侵蚀模数约占淤积产沙模数的26.1%,说明在黄土丘陵沟壑区第Ⅴ副区淤地坝的库区拦沙既有上游因水力侵蚀产沙又有库区自身重力侵蚀所致,淤地坝的运行管理既要防止重力侵蚀破坏现有耕地,又要防止淤地坝淤积出的土地被沟道基流苦咸水破坏,从而导致淤地坝淤地无法高效利用。     

小麦-玉米轮作体系中土壤细菌菌群结构及多样性分析

合理的轮作可以改善土壤微生物群落结构，提高土壤肥力，减少病虫害的发生。为了探讨小麦和玉米轮作对土壤细菌的影响，采用传统PCR克隆文库构建、克隆测序并结合EzBioCloud细菌分类鉴定，对河南小麦-玉米轮作体系下农田土壤细菌的菌群结构变化及多样性特征进行了分析。结果表明，变形菌门（33.73%）和拟杆菌门（31.50%）细菌是小麦-玉米轮作体系土壤中最主要的优势菌群，其次为酸杆菌门、芽单胞菌门、放线菌门和疣微菌门等。在小麦和玉米生长期内细菌的丰度和多样性明显高于二者的间隔期；玉米生长期间细菌菌群丰度和物种多样性更高。不同的微生物群落在土壤中担负不同的生态功能。     

坛紫菜叶状体的抗生素联合除菌效果

为研究坛紫菜及其外生菌群对氯霉素、竹桃霉素和头孢噻肟3种抗生素的敏感性,通过将其与氨苄青霉素、卡那霉素和庆大霉素的组合,优化坛紫菜叶状体的除菌方法,并利用涂布平板、qRT-PCR及16S rRNA测序分析其除菌效果。结果显示,当坛紫菜先以氨苄青霉素(300 mg/L)、卡那霉素(100 mg/L)和庆大霉素(100 mg/L)混合处理18 h,再以氯霉素(50 mg/L)、头孢噻肟(200 mg/L)和竹桃霉素(50 mg/L)混合处理4 d,叶状体的健康率保持在96.3%以上,而对可培养细菌的抑制率达到99.9%。根据qRT-PCR和16S rRNA的结果可知,该种组合方法较前3种抗生素的组合,其菌总数下降41.5%,微生物的丰度和多样性指数明显下降。其中,假单胞菌、交替赤杆菌、交替单胞菌以及海杆菌等得到针对性的抑制。研究表明,多种抗生素的优化组合能够对坛紫菜叶片的菌群结构产生显著影响,使其受到抑制,但仍无法达到绝对除菌的效果。     

Lactococcus lactis,causative agent of an endocarditis valvularis and parietalis thromboticans in the allis shad,Alosa alosa (L.)

Since the 1940s, the anadromous allis shad, Alosa alosa (L.), has suffered population declines throughout its distribution range in Europe. In context of EU‐LIFE projects for the reintroduction of the allis shad in the Rhine system, a comprehensive study was started in 2012 to investigate infectious diseases occurring in allis shad. In course of the study, 217 mature and young‐of‐the‐year allis shad originating from the wild population from the Gironde–Garonne–Dordogne system (GGD‐system) and the Rhine system as well as 38 allis shad from the breeding population were examined by use of bacteriological and histological methods. In 2012 and 2014, an endocarditis valvularis thromboticans caused by a coccoid bacterium was detected in 16% and 25% of mature allis shad originating from the GGD‐system. Results of microbiologic examinations, including biochemical characteristics, matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) and 16S rRNA gene sequence analysis, revealed Lactococcus lactis as causative agent of this infection. This is the first report of an endocarditis valvularis and parietalis thromboticans caused by Lactococcus lactis in fish. Possible sources of infection as well as the impact for the reintroduction programme are discussed.     

应用iTRAQ定量蛋白质组学技术筛选无乳链球菌鱼源株与人源株差异表达蛋白

以斑马鱼和巨噬细胞作为体内、外感染模型,比较无乳链球菌鱼源株GD201008-001与人源株A909的致病性差异,利用iTRAQ技术和质谱分析技术进行差异表达蛋白鉴定,以期为揭示无乳链球菌不同宿主来源株致病机制提供新思路。实验通过测定菌株GD201008-001、A909的斑马鱼半数致死量和巨噬细胞吞噬率,比较二者致病性差异;提取全菌蛋白,经iTRAQ试剂标记后进行质谱鉴定,质谱数据用软件Mascot 2.2和Proteome Discoverer 1.4进行查库(Uni Prot数据库)鉴定及定量分析,并对差异蛋白进行GO功能注释和KEGG通路分析。结果显示,GD201008-001毒力显著高于A909;通过iTRAQ分析两株菌差异表达蛋白,发现差异蛋白涉及的生物学功能较为广泛,在鉴定出的368个差异表达蛋白中,GD201008-001中上调表达蛋白193个(比值1.5),下调表达蛋白175个(比值0.667)。生物信息学分析预测这些蛋白主要涉及26个生物学功能,14个通路,推测Clp X、Glm S和Cps IVK可能在两株菌致病性差异中发挥重要作用。本研究为阐明无乳链球菌不同宿主来源株致病性差异奠定了基础。     

基于深度学习服务的遥感影像农作物分类系统设计

【目的】深度学习在图像分类方面效果显著,但对机器的硬件配置要求高,将深度学习的技术应用于作物分类的同时,降低客户端的IT成本,开发基于深度学习服务的遥感影像农作物分类系统。【方法】系统采用C/S架构,服务器端部署Caffe的深度学习框架,通过PaaS提供计算服务,统一处理客户端模型训练、影像分类等任务;客户端提供用户界面,负责数据输入、结果解析和可视化操作;客户端和服务器之间通过异步RPC实现网络通信,利用FTP进行数据的上传和下载;考虑到农作物分类在实际应用中的时效性,系统还提供了一个简化的Alexnet模型,在保证分类精度的前提下加速了模型的收敛速度。【结果】通过对采集的428144个农作物样本数据集的应用表明,服务器训练的时间比单机训练缩短了近3倍,该系统不仅能够快速地完成深度学习模型训练的任务,还能实时准确得到遥感影像的农作物分类结果。【结论】该研究进一步推动了深度学习在农业分类方面的应用,同时为遥感技术在农业应用中的发展、农作物面积统计工作和对农业资源进行优化配置提供了重要的科学指导。     

Construction and Partial Biological Characteristics Analysis of Listeriolysin S llsB Gene Deletion Strain

In order to analyze the effect of listeriolysin S (LLS) llsB gene deletion on the biological characteristics of Listeria monocytogenes (LM),this study used homologous recombination to construct the llsB gene deletion strain LM90-ΔllsB,and the biological characteristics of growth characteristics,median lethal dose (LD₅₀) and organ-borne bacteria were studied in healthy Kunming male mice at 8 weeks old and weighing 40 g±5 g.The llsB gene deletion strain was successfully constructed,and the deletion strain had good genetic stability through continuous passage to 20 generations in vitro.Based on its growth curve examination,we found that the growth rate of the mutant strain was slightly higher than that of the parent strain.The results of mice infection test showed that the LD₅₀ of the parent strain and the deletion strain were 10^6.17 and 10^6.50 CFU,respectively.Compared with the parent strain,the amount of bacteria load of the deletion strain in the liver and spleen of the mice was extremely significantly decreased (P<0.01),the results showed that the infection ability of the mutant strain on mice was obviously weakened.No Listeria monocytogenes was detected in the brain.The results suggested that llsB gene might have direct or indirect regulatory effect on some biological characteristics of LM90,and it would provide a theoretical basis for further understanding of the pathogenic mechanism of LLS,prevention and control of listeriosis.