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1.
秦楠  郝林  李鑫 《植物保护学报》2015,42(5):813-819
为探究解淀粉芽胞杆菌Bacillus amyloliquefaciens HRH317菌株所产抗菌蛋白物质对禾谷镰孢菌Fusarium graminearum的抑制机理,采用硫酸铵分级沉淀菌株发酵液,DEAE Sepharose Fast Flow柱和Sephadex G-200柱层析分离纯化抗菌蛋白,聚丙烯酰胺凝胶电泳检测抗菌蛋白分子量;并采用平板计数法和扫描电子显微镜观察病原菌孢子萌发和菌丝生长情况。结果表明,硫酸铵饱和度为50%时HRH317菌株发酵液的抑菌活性最大,抑菌圈直径可达2.02 cm,显著大于其它饱和度处理;2次柱层析可纯化出抗菌蛋白,分子量约为36.0 k D。禾谷镰孢菌经该抗菌蛋白处理后12h,病原菌孢子萌发率为55.8%,显著低于对照,且2~12 h间其孢子萌发率平均下降了57.3%;该抗菌蛋白还可造成病原菌菌丝变细、畸形与扭曲,且末端膨大等现象。表明解淀粉芽胞杆菌HRH317菌株产生的抗菌蛋白对禾谷镰孢菌孢子萌发和菌丝发育具有较强的抑制作用。  相似文献   

2.
野生药用植物内生真菌的分离及拮抗菌株筛选   总被引:1,自引:0,他引:1  
为挖掘生防新资源,采用组织分离法从21种野生药用植物不同组织部位分离纯化内生真菌,以6种植物病原菌为靶标菌筛选拮抗菌株,并根据形态学及分子生物学对菌株进行鉴定,在此基础上研究了高效拮抗菌株对靶标菌菌丝生长及孢子萌发的影响。结果表明:从分离纯化的478株内生真菌中筛选出11株高活性拮抗菌株,分属于青霉属Penicillium、平脐蠕孢属Bipolaris、棘壳孢属Pyrenochaeta、镰孢属Fusarium、粒毛盘菌属Lachnum、垫壳孢属Coniella和Neonectria 7个属,其中青霉属占比最高,达36.36%。平皿对峙试验表明, 棘壳孢菌12-R-5对灰葡萄孢表现出高效性和专一性,其对菌丝抑制率达66.67%。含药平皿试验表明,菌株12-R-5发酵滤液10倍稀释液对灰葡萄孢菌丝生长抑制率达100%,与其他处理差异显著。菌株12-R-5 10倍发酵滤液处理灰葡萄孢10 h,孢子不萌发;100倍稀释液处理对孢子萌发抑制率为91.01%。显微镜观察发现,棘壳孢菌使灰葡萄孢菌丝扭曲变形,膨大肿胀及断裂,部分菌丝发生抑缩、消融现象。说明棘壳孢菌12-R-5菌株具有很好的生防潜力及应用前景。  相似文献   

3.
氰烯菌酯(2-氰基-3-氨基-3-苯基丙烯酸乙酯,JS399-19) 是一种对禾谷镰孢菌Fusarium graminearum具有专化活性的新型杀菌剂。离体条件下,采用菌丝生长速率法测定了该药剂对禾谷镰孢菌抗性菌株和敏感菌株的生长抑制活性;同时采用孢子萌发法测定了其对禾谷镰孢菌分生孢子萌发的影响。结果表明,氰烯菌酯能够强烈地抑制禾谷镰孢菌敏感菌株菌丝的生长,EC50值分布在0.092~0.141 μg/mL之间;并可降低敏感菌株分生孢子的萌发速率,以及影响其萌发的方式,使芽管从分生孢子基部和中间细胞萌发的比率增加;同时氰烯菌酯使敏感菌株分生孢子膨大、畸形,并使其芽管肿胀、扭曲,明显抑制其芽管的伸长生长;但对抗性菌株的抑制作用和致畸作用不明显。  相似文献   

4.
辽细辛提取物对灰葡萄孢菌的抑制效果   总被引:1,自引:0,他引:1  
王桂清 《植物保护》2008,34(2):53-57
采用生长速率法和孢子萌发法测定了辽细辛根的不同溶剂(石油醚、氯仿、乙酸乙酯、乙醇)粗提取物和精油对引起黄瓜灰霉病的灰葡萄孢菌菌丝生长和孢子萌发的作用。结果表明,细辛不同溶剂提取物和精油对灰葡萄孢菌的菌丝生长和孢子萌发均有一定的抑制作用,且对孢子萌发的抑制效果好于对其菌丝生长的抑制效果;石油醚提取物和精油的抑菌效果较好,对菌丝生长的EC50分别为177.44 mg/L和159.98 mg/L,对孢子萌发的EC50分别为173.23 mg/L和125.29 mg/L;试验还显示细辛所含的主要抑菌活性成分易被极性小的有机溶剂所提取。  相似文献   

5.
为明确洛阳市牡丹灰霉病的病原菌种类,于2015年从该地区的4个牡丹Paeonia suffruticosa Andr.种植区采集灰霉病样品,采用组织分离法对病原菌进行分离和纯化,结合形态学和基因序列分析对分离物进行鉴定,采用针刺接种法测定不同分离物对牡丹离体叶片的致病性。结果表明,从灰霉病样品中分离获得40株分离物,这些分离物分别属于Ⅰ、Ⅱ、Ⅲ型菌,I型菌不易产孢但产菌核;Ⅱ型菌易产孢,后期产生少量菌核;Ⅲ型菌易产孢,且产生大量菌核。产孢分离物均形成直立状分生孢子梗,孢子梗分枝顶端聚生葡萄穗状分生孢子,分生孢子卵圆形或长卵圆形。供试菌株的ITS序列和G3PDH基因序列与灰葡萄孢Botrytis cinerea Pers.的同源性达到99%;致病性测定结果表明,各菌群代表菌株对牡丹均有致病性,但不同菌群间致病力有差异。研究表明,引起洛阳市牡丹灰霉病的病原菌为灰葡萄孢,且菌群类型多样。  相似文献   

6.
为明确我国藜麦灰霉病病原菌的归属及环境因子对其菌丝生长和分生孢子萌发的影响,在藜麦灌浆期从山西省忻州市原平市和五台县采集具典型症状的藜麦标本,应用直接挑取法分离病原菌,选取代表性菌株对其形态学特征、系统发育、致病性及生物学特性进行观察和测定。结果表明,综合形态学特征及基于G3PDH、HSP60和RPB2多基因系统发育分析结果,最终确定引起我国藜麦灰霉病的病原菌为灰葡萄孢Botrytis cinerea。致病性测定发现接种6 d时,接种处密生灰色霉层,籽粒霉变,与田间症状基本一致,说明分离菌株确为藜麦灰霉病的病原菌。灰葡萄孢在燕麦琼脂培养基、马铃薯葡萄糖琼脂培养基和察氏酵母培养基上培养后可产生菌核,而在麦芽浸粉琼脂培养基上培养可产生分生孢子;在察氏酵母培养基上培养的菌丝生长速率最快,平均为16.8 mm/d。适宜灰葡萄孢菌丝生长和分生孢子萌发的温度为15~25℃、水活度≥0.96、pH 5~6。  相似文献   

7.
为明确土壤来源的黄柄曲霉ASD次生代谢产物中的抑菌物质及其生物活性,利用硅胶柱层析、凝胶柱层析和半制备高效液相色谱对ASD发酵液的二氯甲烷粗提物进行纯化,运用核磁共振及高分辨质谱技术,结合文献报道数据对化合物的结构进行鉴定,利用X-射线单晶衍射验证鉴定结果;采用含药稀释平板法及菌丝生长速率法测定化合物对辣椒疫霉菌、大斑凸脐蠕孢菌和禾生刺盘孢菌等9种病原真菌的抑菌活性。结果表明,从ASD发酵液中分离获得已知活性抑菌物质为sulochrin和4,5-二甲基间苯二酚。其中,sulochrin对辣椒疫霉菌、大斑凸脐蠕孢菌和禾生刺盘孢菌有较好的抑制作用,4,5-二甲基间苯二酚只对辣椒疫霉菌具有较好的抑制作用,两者对灰葡萄孢菌、玉蜀黍尾孢菌和新月弯孢菌等6株致菌抑菌效果不明显或无抑制作用。  相似文献   

8.
本文通过孢子萌发检测、平板生长抑制法和果实病害防治试验,测定了淡紫紫孢菌Purpureocillium lilacinus PL36-1防治储藏期番茄果实灰霉病的效果。结果表明,菌株PL36-1发酵滤液不仅能够抑制灰葡萄孢分生孢子萌发和芽管伸长,而且可以抑制灰葡萄孢菌丝生长。对峙培养的菌株PL36-1显著抑制灰葡萄孢的菌丝生长,菌丝生长抑制率可达到74.45%,菌株PL36-1产生的挥发性物质也对灰葡萄孢菌丝生长有一定的抑制作用。菌株PL36-1发酵液处理樱桃番茄果实后,能有效控制番茄果实灰霉病的发生,含有2×10~8孢子/m L的发酵液10 d的防治效果达70.64%。  相似文献   

9.
为进一步扩宽拟康宁木霉Trichoderma koningiopsis T-51菌株的生防应用范围,该研究测定T-51菌株菌丝生长、产孢和分生孢子萌发的最适温度、光照和碳氮源条件及其对灰葡萄孢Botrytis cinerea菌核的重寄生能力和对12种植物病原真菌的生防潜力。结果表明,拟康宁木霉T-51菌株在26℃下菌丝生长最快,生长速度达到2.5 cm/d,在20℃光照条件下产孢量最大,达到1.64×109个/皿,26~28℃光照条件下适宜T-51菌株分生孢子萌发;供试氮源中硫酸铵和硝酸铵适宜T-51菌株菌丝生长、产孢和分生孢子萌发,供试碳源中葡萄糖和可溶性淀粉适宜T-51菌株菌丝生长和分生孢子萌发,果糖和乳糖适宜T-51菌株产孢;T-51菌株能够重寄生灰葡萄孢菌核,当分生孢子悬浮液浓度为1×108个/mL时,重寄生效果最好;T-51菌株对核盘菌属Sclerotinia和丝核菌属Rhizoctonia植物病原真菌重寄生能力较强,对镰刀菌Fusarium spp.、暹罗刺盘孢菌Colletotrichum siamense、桃褐腐病菌Monilinia fructicola和稻瘟菌Mag...  相似文献   

10.
以多菌灵为对照药剂,采用生长速率法、悬滴法和浸渍法分别测定了不同温度下纳他霉素对灰葡萄孢Botrytis cinerea菌丝生长、分生孢子萌发和菌核萌发的毒力,研究了常温下两种药剂对菌株产孢和菌核形成的影响。结果表明:纳他霉素及多菌灵对灰葡萄孢不同生育阶段菌体的抑制作用随温度降低而有不同程度增强;分生孢子对药剂最为敏感;温度对药剂对菌丝毒力的影响最显著;多菌灵对菌株不同发育阶段的抑制活性均高于纳他霉素。供试药剂对菌株产孢时间和菌核产生时间无显著影响,但多菌灵可显著刺激菌株产孢和菌核形成。  相似文献   

11.
枯草芽孢杆菌G3菌株的抗菌物质及其特性   总被引:29,自引:3,他引:29  
 产几丁质酶枯草芽孢杆菌G3菌株的固体培养物在黄瓜灰霉病菌和番茄叶霉病菌抑菌试验中证实,抑菌活性物质存在于过滤上清液中,它们是从酸沉淀物中提取出的伊枯草菌素、生物表面活性素和存在于盐析粗蛋白中的几丁质酶。在叶霉孢子萌发试验中,伊枯草菌素微弱地抑制孢子萌发但强烈破坏芽管和新生菌丝;生物表面活性素和几丁质酶则强烈抑制孢子萌发并长久性地抑制芽管伸长。在PDA平板上的灰霉菌丝抑菌试验中,伊枯草菌素抑制菌丝生长,引发菌丝顶端膨大,形成泡囊,泡囊破裂后原生质外泄;几丁质酶抑制菌丝生长,引发产生不规则的菌丝团;生物表面活性素在平皿上对菌丝则不显示出抑菌活性。  相似文献   

12.
ABSTRACT Botrytis blight (gray mold), caused by Botrytis cinerea, is one of the most widely distributed diseases of ornamental plants. In geranium plants, gray mold is responsible for important losses in production. The mold Aspergillus giganteus is known to produce and secrete a basic low-molecular-weight protein, the antifungal protein (AFP). Here, the antifungal properties of the Aspergillus AFP against various B. cinerea isolates obtained from naturally infected geranium plants were investigated. AFP strongly inhibited mycelial growth as well as conidial germination of B. cinerea. Microscopic observations of fungal cultures treated with AFP revealed reduced hyphal elongation and swollen hyphal tips. Washout experiments in which B. cinerea was incubated with AFP for different periods of time and then washed away revealed a fungicidal activity of AFP. Application of AFP on geranium plants protected leaves against Botrytis infection. Cecropin A also was active against this pathogen. An additive effect against the fungus was observed when AFP was combined with cecropin A. These results are discussed in relation to the potential of the afp gene to enhance crop protection against B. cinerea diseases.  相似文献   

13.
枯草芽孢杆菌CAB-1抑菌蛋白对黄瓜白粉病的防治作用   总被引:1,自引:0,他引:1  
采用子叶喷雾法分别测定了枯草芽孢杆菌Bacillus subtilis CAB-1所产抑菌蛋白粗提物对黄瓜白粉病的保护和治疗生物活性。结果表明该抑菌蛋白粗提物能显著降低黄瓜白粉病的病情指数,其保护和治疗作用的效果分别为73.33%和76.85%;EC50分别为175μg.mL-1和125μg.mL-1。利用曲利苯兰染色法检测抑菌蛋白粗提物对黄瓜白粉菌Sphacrotheca fuliginea生长的影响,结果表明该抑菌蛋白粗提物能有效降低白粉菌分生孢子的萌发率及萌发的芽管个数,造成菌丝顶端畸形膨大呈球状并减少新生分生孢子的个数。接菌3 d后,对照的孢子萌发率为36.98%,而经抑菌蛋白粗提物处理后的孢子萌发率仅为4.8%;接菌7 d后对照每个分生孢子梗上串生孢子个数最多为7个,多数5~6个;而抑菌蛋白粗提物处理后每个分生孢子梗上串生孢子个数最多为2个,大部分为1个。  相似文献   

14.
啶菌噁唑对番茄灰霉病菌的抑菌作用研究   总被引:3,自引:2,他引:1  
采用生物测定方法研究了啶菌噁唑对番茄灰霉病菌Botrytis cinerea不同发育阶段的影响。结果表明:其对菌落扩展、芽管伸长、分生孢子产生和菌核形成均具有显著的抑制活性,其中啶菌噁唑对P-9菌株菌丝生长和芽管伸长的EC50值分别为0.193和0.154 μ g/mL,0.5 μ g/mL的啶菌噁唑对菌丝生长和芽管伸长的抑制率大于77%,对分生孢子和菌核萌发的抑制率分别低于10%和40%,表明药剂对分生孢子萌发无明显抑制作用。紫外吸收分析显示,啶菌 NFDA1 唑可破坏细胞的膜结构,促使菌体核酸和蛋白外渗。离体叶片毒力测定结果表明,120 μ g/mL的啶菌 NFDA1 唑可有效抑制番茄灰霉病病斑扩展,抑制率达83.74%。  相似文献   

15.
采用大孔吸附树脂提取和硅胶柱层析分离技术,从黄瓜炭疽病菌Colletotrichum orbiculare代谢物中获得一个抗真菌化合物,采用高分辨质谱、核磁共振波谱和红外光谱等技术鉴定了其化学结构,并评价了其对番茄灰霉病菌、西瓜枯萎病菌、烟草赤星病菌、苹果炭疽病菌和玉米弯孢病菌的孢子萌发和菌丝生长的抑制作用。结果表明:该化合物为间二羟基苯甲酸大环内酯类抗生素——根赤壳菌素;其对所有供试真菌的孢子萌发均未表现出明显的抑制活性,但对番茄灰霉病菌Botrytis cinerea的菌丝生长表现出很强的抑制作用。  相似文献   

16.
短肽LD-1是从牛心朴子草根中新分离的具有抗菌活性的多肽。本文以番茄灰霉病为供试菌株,研究了短肽LD-1对番茄灰霉病的抑制作用及其抗病性。结果表明:短肽LD-1对番茄灰霉病菌菌丝生长及孢子萌发具有较好的抑制作用。离体试验中,短肽LD-1对番茄灰霉病菌菌丝生长抑制作用的EC50在24、48、72h分别为28.3051、30.2134、32.7427mg/L,对孢子萌发的EC50为16.6919mg/L;活体试验中,短肽LD-1对番茄灰霉病具有较好的防治效果,90mg/L短肽LD-1对番茄灰霉病防治效果达到74.24%。短肽LD-1能导致病菌芽管发育畸形,不能正常发育。短肽LD-1对番茄种子的萌发和根系生长具有明显的促进效果,并且经短肽LD-1处理后的番茄须根发达,须根数目明显增多,促进了番茄的生长发育,提高了抗病性。  相似文献   

17.
石榴皮萃取物对番茄灰霉病菌抑制作用及防病效果   总被引:3,自引:0,他引:3  
试验测定了石榴皮萃取物对番茄灰霉病菌室内抑菌作用和对番茄灰霉病的温室盆栽防治作用。室内抑菌试验分别采用生长速率法和孢子萌发法测定对番茄灰霉病菌菌丝生长和孢子萌发的抑制作用,结果表明:正丁醇萃取物对病原菌菌丝的抑制活性最高,EC50为50.28 g/L;浓度为200 g/L对番茄灰霉病菌分生孢子的产生具有显著的抑制作用,抑制作用与浓度呈正相关关系。在温室盆栽试验中, 正丁醇萃取物稀释100倍的处理对番茄灰霉病的保护防治效果达到46.41%,治疗防治效果达到48.00%。  相似文献   

18.
Huang R  Li GQ  Zhang J  Yang L  Che HJ  Jiang DH  Huang HC 《Phytopathology》2011,101(7):859-869
A study was conducted to identify volatile organic compounds or volatiles produced by Candida intermedia strain C410 using gas chromatography-mass spectrometry, and to determine efficacy of the volatiles of C. intermedia in suppression of conidial germination and mycelial growth of Botrytis cinerea and control of Botrytis fruit rot of strawberry. Results showed that, among 49 volatiles (esters, alcohols, alkenes, alkanes, alkynes, organic acids, ketones, and aldehydes) identified from C. intermedia cultures on yeast extract peptone dextrose agar, two compounds, 1,3,5,7-cyclooctatetraene and 3-methyl-1-butanol, were the most abundant. Synthetic chemicals of 1,3,5,7-cyclooctatetraene; 3-methyl-1-butanol; 2-nonanone; pentanoic acid, 4-methyl-, ethyl ester; 3-methyl-1-butanol, acetate; acetic acid, pentyl ester; and hexanoic acid, ethyl ester were highly inhibitory to conidial germination and mycelial growth of B. cinerea. Inhibition of conidial germination and mycelial growth of B. cinerea by volatiles of C. intermedia was also observed. Meanwhile, results showed that incidence and severity of Botrytis fruit rot of strawberry was significantly (P < 0.01) reduced by exposure of the strawberry fruit to the volatiles from C. intermedia cultures or C. intermedia-infested strawberry fruit. These results suggest that the volatiles of C. intermedia C410 are promising biofumigants for control of Botrytis fruit rot of strawberry.  相似文献   

19.
新杀菌剂对番茄灰霉病菌的室内毒力及田间防效   总被引:4,自引:0,他引:4  
室内采用菌丝生长速率法、孢子萌发法和黄瓜子叶法测定了咯菌腈、抑霉唑、福美双、吡唑醚菌酯和嘧霉胺对番茄灰霉病菌( Botrytis cinerea)的抑制作用。结果表明,菌丝生长速率法和孢子萌发法的测定均以咯菌腈毒力最高,EC 50分别为0.005 2 μg/mL和0.087 6 μg/mL;黄瓜子叶法测定以抑霉唑毒力最高,其EC 50为0.675 3 μg/mL。田间药效试验结果表明:50%咯菌腈可湿性粉剂90 g/ hm2对番茄灰霉病的田间防效达到90%以上,显著高于对照药剂50%啶酰菌胺水分散粒剂300 g/hm2和40%嘧霉胺悬浮剂 480 g/hm2的防效。  相似文献   

20.
Zhang Z  Yuen GY 《Phytopathology》2000,90(4):384-389
ABSTRACT The role of chitinase production by Stenotrophomonas maltophilia strain C3 in biological control of leaf spot on tall fescue (Festuca arundinacea), caused by Bipolaris sorokiniana, was investigated in vitro and in vivo. The filtrate of a broth culture of C3, with chitin as the carbon source, was separated into fractions. A high molecular-weight fraction (>8 kDa) was chitinolytic and more inhibitory than a low-molecular-weight, nonchitinolytic fraction to conidial germination and hyphal growth by B. sorokiniana and to leaf spot development. A protein fraction derived by ammonium sulfate precipitation and a chitinase fraction purified by chitin affinity chromatography also were chitinolytic and highly antifungal. The chitinolytic fractions caused swelling and vacuolation of conidia and discoloration, malformation, and degradation of germ tubes. When boiled, the chitinolytic fractions lost chitinase activity along with most of the antifungal properties. Two chitinase-deficient and two chitinase-reduced mutants of C3 were compared with the wild-type strain for inhibition of germination of B. sorokiniana conidia on tall fescue leaves and for suppression of leaf spot development in vivo. The mutants exhibited reduced antifungal activity and biocontrol efficacy, but did not lose all biocontrol activity. An aqueous extract of leaves colonized by wild-type C3 had higher chitinase activity than that of noncolonized leaves and was inhibitory to conidial germination. The addition of chitin to leaves along with the wild-type strain increased both chitinase and antifungal activity. The chitinase activity level of extracts from leaves colonized by a chitinase-deficient mutant of C3, with and without added chitin, was no higher than the background, and the extracts lacked antifungal activity. Chitinolysis appears to be one mechanism of biological control by strain C3, and it functions in concert with other mechanisms.  相似文献   

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