大白菜腐霉根腐病的病原

在山东青岛发现一种新的大白菜根腐病,主根下部腐烂,根部维管束变褐变黑,地上部生长缓慢,矮小,甚至死亡。通过组织分离法和单菌丝分离法获得纯化菌株HMQAU170164。经柯赫氏法则验证,综合该病原菌形态学特征、生物学特性、ITS和coxⅠ序列分析,界定其为芸薹腐霉Pythium brassicum。该菌引起的大白菜腐霉根腐病为首次报道。     

软枣猕猴桃镰孢菌根腐病的病原

在辽宁丹东新发现一种软枣猕猴桃根腐病害,罹病植株叶片枯萎,老叶变干凋谢,最后干枯死亡。通过组织分离法和单孢分离法获得纯化菌株HMQAU150043和HMQAU150044。经形态学观察、ef‐1a序列分析、mt SSU序列分析及柯赫氏法则验证,确定两株病原菌均为多见镰孢Fusarium commune。这是该菌所致软枣猕猴桃根腐病的首次发现。     

蓝莓毛色二胞枝枯病的病原菌

近期在山东胶南的蓝莓种植区发现一种枝枯新病害,被侵染的枝条变色坏死,甚至引起全株干枯死亡。通过组织分离法和玻璃毛细管法获得纯化菌株HMQAU140073。通过形态学观察、rDNA-ITS和EF1-α序列分析及柯赫氏法则验证,确定该病原菌为假可可毛色二胞Lasiodiplodia pseudotheobromae。这是假可可毛色二胞所致蓝莓枝枯病的首次发现。     

内生真菌对羽茅抗病性的影响

为探讨不同种类内生真菌对宿主植物羽茅(Achnatherum sibiricum)抗病性的影响, 以感染不同内生真菌的天然禾草羽茅为实验材料, 进行了体外纯培养的内生真菌、感染内生真菌的离体叶片和在体叶片对3种植物病原真菌的抑菌实验。结果表明: 体外纯培养条件下, 分离自羽茅的内生真菌Neotyphodium sibiricum、Neotyphodium gansuensis和Epichloë gansuensis对新月弯孢霉(Curvularia lunata)、根腐离蠕孢(Bipolaris sorokiniana)和枝孢霉(Cladosporium sp.)等3种病原真菌都具有抑制作用, 其中N. sibiricum的抑制作用最强, 对新月弯孢霉、根腐离蠕孢和枝孢霉的抑菌率分别为47.8%、40.1%、39.4%; 内生真菌培养滤液也可以有效抑制这3种病原真菌的孢子萌发, 其中N. gansuensis的抑制作用最强, 新月弯孢、根腐离蠕孢和枝孢霉的孢子萌发率分别为9.8%、8.7%、8.5%。对于离体叶片, N. sibiricum和N. gansuensis感染可以有效降低叶片受3种病原真菌侵染后的病斑数和孢子浓度, 其中N. sibiricum对根腐离蠕孢的抑制作用显著高于N. gansuensis, 而E. gansuensis只降低新月弯孢和枝孢霉侵染的病斑数以及枝孢霉侵染的孢子浓度。在体条件下, 内生真菌均可以显著降低病原真菌侵染羽茅后的病斑数、病斑长度和孢子浓度, 其中E. gansuensis的抑菌作用趋于最弱, 而N. sibiricum的抑菌作用趋于最强。     

内蒙古天竺葵腐霉枯萎病病原菌

为了明确天竺葵腐霉枯萎病的病原菌种类,从内蒙古通辽采集天竺葵腐霉枯萎病样品,对病原物进行了分离、纯化和致病性测定,并对病原物进行形态学和rDNA-ITS序列分析鉴定。结果表明,从10个植物样品中分离得到16个腐霉菌株,分别属于终极腐霉Pythium ultimum var. ultimum、瓜果腐霉P. aphanidermatum和两个与P. ultimum var. ultimum相似的待定种。其中,P. ultimum var. ultimum的分离频率为75%、P. aphanidermatum的分离频率为12.5%,两个腐霉待定种的分离频率均为6.25%;P. ultimum var. ultimum是优势类群。致病性测定结果表明,4种腐霉菌都能引起天竺葵腐霉枯萎病,与自然发病症状相同,其中P. ultimum var. ultimum和P. aphanidermatum的致病性较强,发病率分别为71.4%和85.7%。     

生姜共生真菌的分离及其体外抑菌效应初探

为筛选得到优良植物病害生防菌,对广西生姜Zingiber officinale种植区健康生姜根系和叶片中的共生真菌进行了组织分离,以生姜茎腐病菌群结腐霉Pythium myriotylum和香蕉枯萎病菌尖孢镰刀菌古巴专化型4号生理小种Fusarium oxysporum f. sp. cubense race 4为指示菌,通过平板对峙培养法和发酵液菌落直径法试验进行筛选评价,并结合形态学观察及ITS序列分析对筛选出的生防效果最好的共生真菌进行了鉴定。结果表明,从生姜植株共分离得到34株共生真菌,其中根系分离22株,叶片分离12株;对峙培养发现有6株共生真菌对生姜茎腐病菌和香蕉枯萎病菌均有抑制作用;其中菌株SBM-11拮抗作用最强,对生姜茎腐病菌抑制率达到93%,对香蕉枯萎病菌抑制率达到82%;SBM-11的发酵液对生姜茎腐病菌和香蕉枯萎病菌抑制率分别为82%、73%,与其他菌株发酵液抑制效果相比差异明显;结合形态和分子鉴定结果表明SBM-11菌株为绿色木霉Trichoderma viride,极具生防潜力。     

燕麦紫斑病的病原

2013年8月在甘肃省通渭县的一块燕麦田中,叶片和小穗上出现大量紫色病斑,病株率达100%。采用组织分离法得到核腔菌属Pyrenophora（synonym:Drechslera）真菌。通过Koch’s法则明确了菌株ASA-13的致病性:在8-22℃下,接种植株产生紫色叶斑,在20-25℃下,接种植株产生中心黑色、周围黄褐色至淡黄色叶斑,一些病斑几乎成为条斑。在20℃下,菌株ASA-13人工接种可侵染皮燕麦、裸燕麦、高粱、小麦、玉米和青稞离体叶片。采用形态学和分子生物学方法对病原菌进行了鉴定,燕麦紫斑病病原为毛壳核腔菌Pyrenophora chaetomioides（syn.:D. avenae）。采用温度梯度法测定了试验菌株的适宜生长温度,在PDA培养基上菌株ASA-13的菌丝适宜生长温度为25-30℃。     

稻镰状瓶霉对水稻白叶枯病的生物防治

本文研究了野生稻暗色有隔内生真菌稻镰状瓶霉与水稻的共生关系,通过根部接种DsRED荧光标记菌株,观察稻镰状瓶霉在水稻根系内的定殖,并对其抗白叶枯病特性进行调查,结果发现：经叶部接种白叶枯病菌后,对照组发病严重,病级集中在7级和9级,分别占比43.33%和34.67%,病情指数为79.26;相比之下,接种了稻镰状瓶霉的水稻植株发病较轻,叶片病斑面积小,并伴随零星黑色过敏性坏死斑出现,未发病植株占8%,72%的植株病级为1级,病情指数仅为15.26。稻镰状瓶霉对水稻白叶枯病的防治效果达到80.75%。稻镰状瓶霉的定殖能够引起叶片中超氧化物歧化酶、过氧化氢酶和过氧化物酶活性极显著提高,分别是对照组的5.26、12.08和10.53倍;诱导PR1a和PR1b显著上调表达8.71和3.37倍,AOS、OsSAUR2和EL5基因显著下调表达0.28、0.57和0.65倍。利用野生稻内生真菌防控水稻白叶枯病,在国内外尚属首次,可为水稻白叶枯病的生物防治提供新途径。     

白及根部内生真菌多样性研究

白及是地生兰科多年生草本植物,是常见的中草药之一。白及根是白及植株吸收养分和水分的主要场所,开展其根部内生真菌的群落组成及多样性分析,对了解云南原生境白及与根部内生真菌间的共生关系具有重要意义。本研究采用形态学和ITS rDNA分子系统发育分析相结合的方法进行菌株鉴定。结果表明,3个采样点的白及根部分离到可培养内生真菌32株,包括9个目、13个科、16个属。其中木霉属Trichoderma、镰刀菌属Fusarium、拟盘多毛孢属Pestalotiopsis为优势类群,分别占总物种数的21.88%、18.75%、9.38%;中碳垫菌属Nemania、丛赤壳属Nectria、炭角菌属Xylaria、紫霉属Purpureocillium、刺盘孢属Colletotrichum、黑孢子菌属Nigrospora、Biscogniauxia、腐质霉属Humicola、脉孢菌属Neurospora、拟茎点霉属Phomopsis、植物腐霉Phytopythium、毛霉属Mucor、伞状霉属Umbelopsis为常见内生真菌类群。α多样性指数分析表明：各采样点的白及根部内生真菌物种多样性最高的是昆明市水源保护区采样点（D=0.799,H’=1.698）,保山市龙陵县采样点物种多样性最低（D=0.787,H’=1.580）。β多样性分析采样点之间的Jaccard（0.912-0.993）、Bray-Curtis（0.838-0.986）和UniFrac（加权0.618-0.631;不加权0.770-0.799）距离,结果表明3个采样点之间的内生真菌群落组成存在差异。3个采样点的不同地理环境和土壤类型给白及提供了不同的生境条件,是影响白及根部内生真菌物种丰度和群落组成差异的主要因素之一。     

丛枝菌根对喜树幼苗生长和氮、磷吸收的影响

喜树(Camptotheca acuminata)是我国特有的多年生亚热带落叶阔叶树种,因其次生代谢产物喜树碱具有良好的抗肿瘤活性而受到人们的广泛关注。该文通过温室盆栽接种试验,观察了2属6种丛枝菌根真菌即蜜色无梗囊霉(Acaulospora mellea)、光壁无梗囊霉(A. laevis)、木薯球囊霉(Glomus manihot)、地表球囊霉(G. versiforme)、幼套球囊霉(G. etunicatum)和透光球囊霉(G. diaphanum)对喜树幼苗生长和氮、磷养分吸收的影响。结果表明,丛枝菌根的形成对喜树幼苗的生长以及氮、磷营养的吸收均有影响。从生物量看,除幼套球囊霉和光壁无梗囊霉侵染形成的丛枝菌根喜树幼苗与无菌根幼苗(CK)差异不显著外,其余菌根幼苗的生物量均明显大于无菌根幼苗,透光球囊霉和蜜色无梗囊霉菌根幼苗尤为突出,分别达到无菌根幼苗的1.9和1.4倍。丛枝菌根的形成似乎不利于喜树幼苗的氮素营养吸收,并且主要体现在叶片的氮含量上。相反,丛枝菌根形成总体上促进喜树幼苗对磷素营养的吸收,并且主要体现在根的磷含量上。与无菌根幼苗比,所有菌根幼苗根的氮、磷分配比例增加,而叶片的氮、磷分配比例减少。     

进境美国大豆中值得关注的真菌Diaporthe novem

采用常规平板分离法,从一批进境的美国大豆样品中获得1株可疑的间座壳属菌株MDD57.经形态学观察发现,该菌株在PDA培养基上产生分生孢子器,且同时产生大量α型和β型分生孢子,未见有性阶段.经ITS和tef1α基因扩增、核酸序列比对分析,发现该菌株同GenBank中2株Diaporthe novem菌株的基因序列同源性达...     

锁阳及其寄主白刺内生真菌的生态分布及遗传关系

首次研究了寄生植物锁阳及其寄主白刺内生真菌的分布特征及遗传关系.采用组织块法分离天然白刺、寄生体中锁阳和白刺的内生真菌,利用ITS-rDNA分子序列并结合形态学方法鉴定菌种,研究内生真菌的分离率、定殖率、分离频率、多样性指数、均匀度指数及相似性系数等的差异,以及寄生关系中内生真菌的多样性、遗传关系及分布特征等.结果表明: 本次获得的49株内生真菌隶属于18个分类单元,95.9%为子囊菌,4.1%为担子菌;内生真菌总分离率为15.3%,总定殖率为25.0%;天然白刺中内生真菌Shannon多样性指数最大,为2.13;锁阳花序与锁阳茎的内生真菌相似性系数最大,为0.50;镰孢菌属为白刺的优势菌属,青霉属为锁阳的优势菌属.锁阳与白刺寄生体中真菌类群的差异性分布表明寄生关系对内生真菌群落存在一定影响.     

Plectosporium tabacinum Root Rot Disease of White Lupine (Lupinus termis Forsk.) and its Biological Control by Streptomyces Species

Plectosporium tabacinum root rot disease of white lupine is reported for the first time in Cairo Governorate provinces. Symptoms of the disease appeared on young seedlings as stunting, yellowing and wilting of the foliage. Rotted roots were first became light brown and the lesions progressively extended over the whole root system and became dark brown. Plants died when the whole root system had become infected. For successful biological control of the disease 70 rhizosphere actinomycetes were isolated by the use of normal standard isolation techniques. Three of these isolates were found to be strongly antagonistic against P. tabacinum in vitro . They also significantly reduced the incidence of white lupine root rot disease in soil infested with P. tabacinum in greenhouse trials. The three actinomycete isolates were identified as Streptomyces cyanoviridis , Streptomyces murinus and Streptomyces griseoplanus .     

草甘膦和百草枯对毛桃幼苗根系形态及地上部生长的影响

以砧木毛桃幼苗为研究对象,通过土施草甘膦和百草枯研究2种桃园常用除草剂对毛桃营养生长、根系结构、根尖细胞分裂、叶片光合特性等的影响,为除草剂在桃生产中的安全使用提供科学依据。结果表明: 草甘膦处理显著抑制毛桃地上部和根生长,与对照相比,株高降低31.5%,总根系长度、总根表面积、总根体积和总根尖数分别降低了39.5%、39.5%、49.8%和44.6%,而百草枯处理以上指标与对照差异均不显著;草甘膦和百草枯处理后毛桃根尖细胞有丝分裂指数分别降低38.0%和35.9%,且草甘膦处理分裂中期细胞数占分裂细胞总数的比例显著低于对照和百草枯处理;毛桃根尖细胞对2种除草剂响应迅速,从处理第2天开始根尖细胞电解质渗漏率始终显著高于对照。叶片细胞电解质渗漏率则从处理5 d后开始显著升高,且草甘膦处理出现幼叶基部变黄并向叶尖蔓延,同时部分叶尖逐渐焦枯的现象;2种除草剂处理导致毛桃叶片净光合速率、气孔导度、蒸腾速率有不同程度的降低,其中草甘膦处理下降更明显。综上,使用草甘膦和百草枯均会降低毛桃幼苗根尖细胞分裂指数,提高根尖细胞电解质渗透率,总体降低叶片净光合速率。草甘膦对毛桃营养生长、叶片光合作用影响更大,而且会造成幼叶变黄、叶尖焦枯等现象。     

竹叶对萝卜幼苗生长的影响

采用琼脂混粉法研究了22属83种竹叶对萝卜幼苗生长的影响,结果表明:在2g·L^-1的添加浓度下,供试竹叶对萝卜幼苗表现出不同程度的抑制作用,且对根生长的抑制作用大于对胚轴生长和鲜重的抑制作用。刚竹属、簕竹属、大明竹属、牡竹属中,对根长的抑制率在50%以上的竹种分别占70.83%、71.43%、62.50%、85.71%;其它竹属的30种竹叶,有73.33%的竹种对萝卜幼苗根长的抑制率超过50%。淡竹、桂竹、孝顺竹、宜兴苦竹、长叶苦竹、甜龙竹、铺地竹和铁竹等8种竹叶对萝卜幼苗的抑制作用显著,对萝卜根的抑制率均在80%以上。结果对于竹类植物化感作用研究具有参考作用,对拓展竹叶资源的化学利用途径具有一定的指导意义。     

Effects of Pseudomonas chlororaphis on Pythium aphanidermatum and Root Rot in Peppers Grown in Small-scale Hydroponic Troughs

The ability of Pseudomonas (Ps.) chlororaphis isolate Tx-1 to suppress Pythium aphanidermatum and control root rot was investigated in sweet peppers grown in small-scale hydroponic trough units with recirculating nutrient solution. The agent was introduced to the nutrient solution 3 days after the peppers were inoculated with P. aphanidermatum, or 4 days before and 3 days after inoculation, or 4 days before and 3 and 10 days after. Controls either received no agent or pathogen, or the agent only (applied once, 4 days before the time of pathogen inoculations), or the pathogen only. The experiment was conducted in a greenhouse with repetitions beginning in January, March, and May. Severity of root browning associated with P. aphanidermatum, in the absence of Ps. chlororaphis, increased in the first, second and third repetitions, respectively, to 74% at 21 days, 28% at 21 days, and 68% at 11 days. In treatments with one, two, or three applications of Ps. chlororaphis, respectively, areas under root browning progress curves were reduced by 18-48%, 50-73%, and 62-79% compared to the pathogen controls, and recovery incidence of the pathogen from roots at 21 days after inoculation was reduced by 14-47%, 62-82%, and 60-89%. Roots not inoculated with P. aphanidermatum were whitish and not colonized by the pathogen. Density of Ps. chlororaphis in treated plants was usually 2×10³-1.2×10⁵ cfu g^-1 fresh roots. The Ps. chlororaphis treatments almost invariably prevented reductions in plant height, leaf area, fresh mass and dry mass of the shoots and of the roots, and root volume associated with P. aphanidermatum and root browning. It is concluded that Ps. chlororaphis Tx-1 strongly antagonized P. aphanidermatum in the pepper roots and that the agent has considerable potential for controlling root rot and maintaining productivity in commercial hydroponic peppers.     

茄病镰刀菌单克隆抗体的制备及胶体金免疫层析试纸条的研发*

Objective: The cell lines secreting specific monoclonal antibodies (McAbs) were prepared by using Fusarium solani, one of the pathogenic fungi causing root rot of Fritillaria thunbergii, and the colloidal gold immunochromatographic test strip based on McAbs was developed to provide scientific basis for detecting root rot of F. thunbergii. Methods: Hybridoma technology was used to obtain cell lines that could secrete specific McAbs against F. solani using the whole protein extract of F. solani as the antigen. The specificity, titer, sensitivity and binding protein of McAbs were detected by indirect ELISA and Western blot. Colloidal gold particles were prepared by trisodium citrate reduction method and McAbs were labeled to prepare colloidal gold immunochromatographic strip. Results: Three cell lines secreting specific McAbs against F. solani were obtained, which were named as FsA3, FsG6 and FsD4. The detection sensitivity of FsA3 was 24.41 ng / mL, and that of both FsG6 and FsD4 was 12.21 ng / mL. FsA3, FsG6 and FsD4 had strong reactions to F. solani, and had no cross-reaction to Alternaria tenuissima, A. alternata, Botrytis cinerea, F. equiseti, F. incarnatum, F. oxysporum, Phoma sp., and Phomopsis oblonga. The colloidal gold immunochromatographic strip based on FsG6 showed only a quality control line when detecting the tissue culture seedlings of F. thunbergii. When 100 ng F. solani antigen or the samples of F. thunbergii infested with root rot disease were detected, there were visible quality control lines and test lines. Conclusion: The specificity and sensitivity of the McAbs and test strip are sufficient to detect F. solani isolated from diseased strains of F. thunbergii, which provides the technical support for the rapid detection of root rot of F. thunbergii in the field.     

绿色荧光蛋白标记下镰刀菌侵染地黄的组织学观察

为分析对地黄有较强致病性的尖孢镰刀菌的侵染机制,本研究采用携带有潮霉素抗性标记的强组成型表达载体pCT74,经PEG-CaCl₂介导的原生质体转化法导入镰刀菌,获得荧光信号强烈的sGFP标记菌株,并采用喷施接种和根部灌根接种。研究发现镰刀菌首先在地黄外部聚集繁殖,并通过伤口或气孔等通道侵入地黄维管组织,后逐渐导致周围海绵组织破裂,进而致使地下根茎逐渐变黑腐烂;由于地黄叶部气孔发达,使得镰刀菌由叶部侵入速度要快于根部灌根接种;不同孢子浓度接种实验表明镰刀菌对寄主的致害程度与其在叶部与根部的接种浓度并不呈相关性。进一步在接种处理后采集地黄叶部、茎部和根部组织提取真菌DNA后进行PCR扩增发现：在根部接种60h后即能检测到镰刀菌的侵入,经84h后侵入到地黄茎部组织,经96h后侵入到地黄叶部组织。该标记菌株可为今后探索地黄连作栽培中真菌病害大规模爆发的根际生物学过程提供研究材料。