16S rRNA在兽医病原菌分类鉴定中的应用

介绍16S rRNA基因的特点，阐述其用于病原菌分类鉴定的基本原理，综述其在兽医病原菌分类鉴定中的应用。     

Phytotoxicity on cotton ex-plants of an 18.5 kDa protein from culture filtrates of Verticillium dahliae

A phytotoxic protein that evokes the typical symptoms of Verticillium wilt disease in seedlings of Gossypium hirsutum L. (Upland cotton) was isolated from culture filtrates of Verticillium dahliae. The protein was purified by ammonium sulfate precipitation, Sephadex-G100 fractionation, and native PAGE. The 18.5 kDa protein, designated VD18.5, appears to be a single subunit protein with an isoelectric point between 3 and 5. VD18.5 induces symptoms of leaf dehydration, chlorosis, necrosis and stem discoloration in seedlings of the disease susceptible cotton cultivar Siokra 1–4. The LD₅₀ of VD18.5 on protoplasts of Siokra 1–4 was 18 μg mL⁻¹. VD18.5 had no noticeable effect on Pima S-7, which is a disease resistant cultivar. Phytotoxic activity was partially destroyed at high temperature and was abolished by digestion with proteinase K. Mass spectrometry fingerprinting and protein sequence data from VD18.5 yielded no significant matches when submitted to the Mascot search engine and NCBI non-redundant protein databases, respectively. These results suggest that VD18.5 is a novel protein that may be involved in the development of some of the symptoms associated with Verticillium wilt disease in the cotton plant.     

Biological and Physical Constraints on Maize Production in the Humid Forest and Western Highlands of Cameroon

The aim was to identify biological and physical factors responsible for reducing maize yield in Cameroon. Two surveys were conducted in 137 fields in two agroecological zones in 1995–1997. In the Humid Forest (HF), Bipolaris maydis, Stenocarpella macrospora, Puccinia polysora, Rhizoctonia solani and soil fertility were factors that reduced maize production in 1995 and 1996. In the Western Highlands (WHL), Cercospora zeae-maydis, and the interaction between soil fertility and maize variety were the most important constraints to maize production in 1996. In 1997, C. zeae-maydis, S. macrospora, physiological spot and stem borer damage (Busseola fusca) were negatively related to ear weight. The combination of these biological factors (diseases and insects), and the physical parameter of soil fertility were responsible for reducing maize yield in these selected benchmarks of Cameroon. Maximum potential yield reductions were estimated at 68% due to B. maydis and 46% due to S. macrospora, respectively, in the HF in 1995. In 1996, maximum potential yield reductions in the HF were estimated at 34%, 41% and 30% due to S. macrospora, P. polysora and R. solani, respectively. In the WHL, C. zeae-maydis had the potential to cause a yield reduction of 79% in 1996. In the WHL in 1997, the interaction between C. zeae-maydis and B. fusca, stem diseases and the physiological spot caused potential reductions of 52%, 34% and 39%, respectively.     

接种茄病镰刀菌(Fusarium solani f.sp.glycines)对不同基因型大豆植株和须根的影响

摘要：茄病镰刀菌（Fusarium solani f．sp．glydne）是一种土生细菌,通过侵染大豆根系引发猝死综合症。利用温室盆栽试验和须根培养试验研究了接种茄孢镰刀菌对13个不同基因型大豆的影响。结果表明：接种后所有盆栽大豆主根都有明显深褐色的侵染病斑;移植后21d测定了盆栽植株叶部发病程度,Peking表现最为严重,然后依次为Spencer,Ripley,P3981,Williams82,Essex,Forrest,Iroquois,PI520733,Hartwig,PI567650B,Jack,和PI567374。叶部发病程度与冠高（r=-0．422,P=0．0018）、冠重（r=-0．857,P〈0．0001）和根重（r=-0．732,P〈0．0001）呈显著负相关。主根病斑长度与叶部发病程度没有相关性,表明大豆对病原菌的抗性不能仅通过根系得到充分控制。对培养的大豆须根接种茄病镰刀菌菌丝体10d后,不同基因型大豆的菌落直径的变化范围为17—40mm,差异显著（P=0．05）,其中Spencer和Peking须根上的菌落直径显著（P=0．05）大于PI567374和PI520733。对Spencer和PI567374的须根接种10灿茄病镰刀菌常量成分悬浮液,10d后菌落直径分别为50和38mm,差异显著（P=0．05）。通常,不同基因型大豆间茄病镰刀菌在培养须根上的生长与整株的症状间有一定的相关性,但不总是这样,这是因为即使根系对毒素产生抗性来减少叶部病害症状,但并不是所有的大豆都表现出明显的根系抗性。     

玉米小斑病抗性种质鉴定及生理生化分析

以200份玉米优良自交系为材料,2022-2023年夏分别进行小斑病菌的田间接种,综合两年数据共鉴定出高抗小斑病的玉米自交系3份,分别为K22、CML50和齐319;筛选到包括Mo17在内的抗性玉米自交系24份。选取K22和B73两个具有不同抗性水平的玉米自交系进行小斑病菌的室内接种。结果表明,48 h和72 h时,自交系K22叶片的病斑数、病斑面积和病斑面积比等各项指标均显著优于自交系B73,进一步验证田间鉴定的准确性。48 h或72 h时,高抗小斑病自交系K22与感病对照B73相比,5种防御酶活性增加14.32%～70.45%。     

用于食品安全检测的生物传感器的研究进展

生物传感器特异性好、分析速度快、成本低,在食品安全检测领域有着重要的应用价值。该文介绍了电化学、光学、压电和量热生物传感器在食品安全检测中的应用,包括致病菌、抗生素残留、生物毒素和农药残留检测,指出了目前研究中需要解决的问题并展望了未来发展方向,认为高灵敏度、集成化、微型化、多功能化等是未来用于食品安全检测的生物传感器的发展趋势和重点方向。它在食品污染物的快速实时及特异性检测方面有着广阔的应用前景。     

单端孢菌素对几种作物病原菌的室内毒力及田间防效

为明确单端孢菌素对主要作物病原菌的抑菌活性和田间防效,采用孢子萌发抑制测定法、菌丝生长速率法和田间药效常规试验方法,测定了单端孢菌素在不同质量浓度下对作物病原菌的室内毒力及田间药效。结果表明:单端孢菌素对供试16个病原菌的MIC为1.9~15.5mg/L,对辣椒炭疽病菌、番茄早疫病菌、水稻恶苗病菌、水稻稻瘟病菌、番茄灰霉病菌、玉米小斑病菌和马铃薯晚疫病菌等7个病原菌的EC50分别为5.93、7.90、9.27、10.78、13.34、14.95和16.50mg/L,500mg/L质量浓度对番茄灰霉病、番茄早疫病、辣椒炭疽病、马铃薯晚疫病、玉米小斑病和水稻稻瘟病的防治效果分别为78.98%、81.29%、85.28%、72.42%、78.99%和72.74%。单端孢菌素具有较强的抑菌活性和较广的抑菌谱,对几种主要作物真菌和卵菌病害具有较好的田间防治效果,应用前景广。     

Goat horns: Platforms for viroid transmission to fruit trees?

Mechanical inoculations with contaminating tools and propagation of infected budwood were considered the main causes for the omnipresence of multiple viroid species among citrus and other Middle Eastern and Mediterranean fruit trees and grapevines. However, neither means could explain viroid infections of wild trees — scattered on terrains inaccessible to humans — nor the finding of similar viroids among graft-incompatible plants. Northern hybridization of RNA extracts made of scrapings from the surfaces of goat (Capra hircus) horns that were rubbed against etrog (Citrus medica) stems infected with a citrus viroids complex, revealed accumulation of considerable amounts ofCitrus exocortis viroids (CEVd) andHop stunt viroids (HSVd). Experimental transmission of both CEVd and HSVd was obtained by rubbing healthy citrus plants with goat horns that had been rubbed 24 h earlier on infected etrog stems. These results implicate goats as possible vectors of viroids. Transmissionvia goats could have facilitated the long-range spread of viroids among cultivated and wild plants andvice versa and also among graft-incompatible plants.     

拮抗细菌BMY-1菌株对几种水果储藏病原真菌的抑制作用

采用平板对峙培养法测定从土壤中分离获得的拮抗细菌BMY-1菌株对柑桔绿霉病菌（Penicillium digatatum）、柑桔青霉病菌（Penicillium italicum）、苹果炭疽病菌（Glomerella cingulata）、苹果轮纹病菌（Physalospora piricola）、梨扩展青霉病菌（Penicillium expansum）、葡萄白腐病菌（Coniothyrium dlodiella）等6种水果产后病原真菌的抑制作用。结果表明，BMY-1菌株对苹果炭疽病菌（G.cingulata）和梨扩展青霉菌（P．expansum）的生长有较好的作用，抑菌率分别达48．43％和50．37％。采用液体共培养法观察到BMY-1菌株对病原真菌的菌丝有破坏作用，使苹果炭疽病菌菌丝扭曲和断裂，葡萄白腐病菌菌丝膨大畸形。对几种水果同时接种拮抗细菌BMY-1和病原真菌，BMY-1菌株对苹果炭疽病病斑和梨扩青病病斑扩展有较强的抑制作用，并能有效延迟葡萄白腐病发病时间。     

鉴别显色培养基在奶牛乳房炎病原菌快速鉴定中的应用

为研发用于奶牛乳房炎常见病原菌分离鉴定的鉴别显色培养基,并检验其与普通分离鉴定培养基辅以生化鉴定和16S rRNA核酸序列测定两种传统方法鉴定结果的一致性,笔者以生物信息学方法筛出各种属细菌的特异性酶、可作为唯一碳源发酵产酸的碳源,然后合成酶显色底物,连同碳源、酸碱指示剂制备显色培养基,将各种属标准菌株、野生型菌株涂布培养,评估菌落形态、菌落颜色及培养基基质颜色变化。采集隐性乳房炎(CMT法检测)及临床型乳房炎病例的牛奶样品(絮状物、凝块、清亮状或血乳)共计482份,用鉴别显色培养基和两种传统方法进行病原菌的分离、鉴定。鉴别显色培养基上菌落纯化后提取DNA用于16S rRNA序列扩增,产物送去测序(n=194)。以两种传统方法为参考,判定鉴别显色培养基鉴定病原菌的可靠性,用SAS 9.4的FREQ程序计算鉴别显色培养基的简单科恩κ系数。鉴别显色培养基上常见乳房炎病原菌不同种属的菌落形态和培养基基质颜色明显不同,肉眼即可辨别。鉴别显色培养基与两种传统方法鉴定结果的一致性参数分别为κ=0.70、κ=0.96。鉴别显色培养基能够作为常见奶牛乳房炎病原菌的标准鉴定方法。