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1.
苹果树腐烂病内生拮抗放线菌A-2的鉴定及其活性评价   总被引:4,自引:1,他引:3  
以苹果树腐烂病菌Valsa mali为靶标菌,从健康苹果树枝条中获得一株高效内生放线菌A-2。采用生长速率法、对峙培养法和玻片法测定了菌株A-2的抑菌谱及对苹果树腐烂病菌菌丝生长和孢子萌发的影响;利用离体枝条烫伤接种法测定了菌株A-2对腐烂病的防治效果。结果表明:A-2发酵滤液对腐烂病菌的抑制率达90%以上,对其他9种常见的植物病原真菌也具有良好的拮抗作用;菌株A-2可导致腐烂病菌菌丝畸形、分支增多和局部膨大;不同稀释倍数的A-2发酵滤液对苹果离体枝条腐烂病均有明显的防效,接种病原菌前后24 h和同时施用500倍发酵滤液的防效均达70%以上。根据培养特征、生理生化特性和16S rDNA 序列分析,将菌株A-2鉴定为卡伍尔链霉菌Streptomyces cavourensis。  相似文献   

2.
采用菌丝生长速率法和显微镜观察,研究了咪鲜胺对水稻恶苗病菌Fusarium fujikuroi生长发育及菌丝形态的影响。结果表明:咪鲜胺对水稻恶苗病菌敏感菌株及其抗药性突变体的菌丝生长、孢子萌发、芽管伸长均有明显的抑制作用,而对产孢量无影响。当咪鲜胺浓度为0.5 μg/mL时,能够完全抑制敏感菌株菌丝的生长,而不能抑制不同抗性水平抗性菌株的菌丝生长。随抗性倍数从24倍增加到112倍,药剂对抗药突变体菌落扩展的影响逐渐减小。咪鲜胺对敏感菌株菌丝生长的EC50 值为0.047 μg/mL,最小抑制浓度(MIC) 为 0.5 μg/mL。 20.00 μg/mL咪鲜胺可完全抑制亲本敏感菌株的孢子萌发,而对不同抗药突变体孢子萌发和芽管伸长的抑制率分别为84.1% ~89.0%和58.5% ~65.8%,表明抗药突变体在菌丝生长、孢子萌发、芽管伸长等阶段对药剂的敏感性与亲本菌株相比都有了不同程度的降低。10 μg/mL咪鲜胺处理菌丝12 h后,亲本菌株菌丝顶端膨大畸形,抗药突变体的菌丝顶端膨大,并在菌丝上长有许多细小分枝,表明药剂对抗药突变体次生菌丝生长的抑制作用不明显。  相似文献   

3.
不同杀菌剂对黄瓜靶斑病菌的毒力作用特性比较   总被引:4,自引:2,他引:2  
为筛选防治黄瓜靶斑病的有效药剂,采用菌丝生长速率法、孢子萌发法和芽管伸长法测定了19种杀菌剂对黄瓜靶斑病菌的毒力,探究了4种毒力较高杀菌剂对病原菌菌丝形态的影响。结果表明:氟啶胺、吡唑萘菌胺、咯菌腈和四霉素对黄瓜靶斑病菌菌丝生长、孢子萌发和芽管伸长各生长发育阶段均有较强的抑制活性,其中,4种药剂抑制菌丝生长的平均EC50值分别为0.516 3、1.538、1.605和0.648 2 mg/L,抑制孢子萌发的平均EC50值分别为0.164 4、1.359、1.127和0.002 5 mg/L,抑制芽管伸长的平均EC50值分别为0.189 6、0.144 7、0.060 4和0.001 5 mg/L;啶酰菌胺、百菌清和肟菌酯3种不同作用机制杀菌剂对黄瓜靶斑病菌孢子萌发和芽管伸长的抑制活性较强,3种药剂抑制孢子萌发的平均EC50值分别为1.564、0.373 0和0.021 3 mg/L,抑制芽管伸长的平均EC50值分别为0.629 5、0.233 4和0.405 0 mg/L,但对菌丝生长的抑制作用较弱,平均EC50大于16 mg/L;氟啶胺、吡唑萘菌胺、咯菌腈和四霉素可引起黄瓜靶斑病菌菌丝不同程度的异常生长,主要表现为菌丝隔膜间距变小、菌丝节间肿大、主菌丝上二次分枝发生频繁、菌丝变形黏连、菌丝新分枝处生长受抑制或分生孢子生长变形。研究表明,氟啶胺、吡唑萘菌胺、咯菌腈和四霉素对黄瓜靶斑病菌具有较高的毒力,在黄瓜靶斑病的田间防治中具有较好的开发应用潜力。  相似文献   

4.
林木腐烂病是苹果树、梨树和杨树等林木枝干的重要真菌性病害。为了筛选出对苹果树腐烂病菌Valsa mali var. mali、梨树腐烂病菌V. mali var. pyri和杨树腐烂病菌V. sordida等3种不同寄主腐烂病菌都能有效防控的杀菌剂,本研究开展室内毒力试验比较了7种杀菌剂对3种腐烂病病原菌菌丝生长和分生孢子萌发的抑制效果,并进一步通过田间活性测定试验比较7种杀菌剂对梨树腐烂病病斑扩展和分生孢子发生的防治效果,同时测定了增效剂8.6%聚乙二醇(PEG)对7种杀菌剂的增效作用。毒力测定结果表明,苯醚甲环唑、戊唑醇、吡唑醚菌酯和丙唑·多菌灵对3种腐烂病病原菌菌丝生长和分生孢子萌发的抑制作用较强,其中EC50平均值最低的是苯醚甲环唑,而戊唑醇的MIC平均值最低,在0.33 mg/L浓度下对3种腐烂病病原菌的菌丝生长和分生孢子萌发抑制率均达到100%。田间试验结果表明,45%苯醚甲环唑SC、43%戊唑醇SC和35%丙唑·多菌灵SE对梨树腐烂病病斑扩展和分生孢子萌发的防治效果突出,其中45%苯醚甲环唑SC 30.00 mg/L对病斑扩展防治效果达到82.23%,孢子萌发抑制效果达到85.96%,田间防治效果最好。10%丙硫唑SC+8.6% PEG处理组对病斑扩展防治效果提高了15.39百分点,达到73.46%,分生孢子萌发抑制率提高了23.75百分点,达到83.06%,增效作用显著。本研究为苹果树、梨树和杨树等3种寄主腐烂病的化学防控提供了科学依据。  相似文献   

5.
 采用电子显微镜技术对青杨叶锈病菌(Melampsora larici-populina Kleb.)的侵染过程进行了研究。发现该菌夏孢子萌发产生1~3个芽管,且具较多的树杈状分枝。芽管由气孔侵入,侵入前不形成明显的附着胞或仅个别芽管形成附着胞。芽管侵入气孔后在气孔腔内形成气孔下囊,再分化出圆形的膨大体而产生1~2支初生菌丝。初生菌丝在寄主细胞间扩展,与叶肉细胞壁接触后分化出吸器母细胞,吸器母细胞中的细胞器与胞间菌丝相同,双核。吸器母细胞产生侵入钉侵入叶肉细胞内部形成吸器,成熟吸器由细长具颈环的管状颈部和膨大的吸器体组成,此时胞间菌丝在吸器母细胞处分化出次生菌丝,在叶肉细胞间扩展形成次生菌落,产生孢子堆。病菌在寄主细胞间隙或沿寄主细胞壁延伸时,寄主细胞仍保持正常状态。  相似文献   

6.
腔孢叶斑病是牡丹生产上出现的一种新病害,近几年在洛阳及菏泽牡丹种植园危害越来越重。为了筛选防治该病的化学药剂,本文采用菌丝生长速率法测定了3类8种杀菌剂对病菌菌丝生长的抑制活性,采用涂布平板法测定了多菌灵、戊唑醇及嘧菌酯对病菌孢子萌发和芽管伸长的影响,并在含药平板上测定了3种药剂对病菌产孢量的作用。结果表明:三唑类药剂对病菌菌丝生长的毒力最强,EC50为0.020~0.096μg/mL,且戊唑醇对孢子萌发和产孢也有很强的抑制活性,在0.1μg/mL时孢子即不能萌发;甲氧基丙烯酸酯类药剂对菌丝生长的毒力较强,EC50为0.408~0.939μg/mL,嘧菌酯对芽管伸长和产孢量均表现出很强的抑制活性,10μg/mL时,芽管几乎不再伸长,对产孢的抑制率可达到99%以上;而苯并咪唑类药剂对菌丝生长的毒力较弱,EC50为12.167~30.104μg/mL,多菌灵对孢子萌发的抑制作用弱,100μg/mL时孢子萌发率仍达到90%以上,对产孢的抑制作用也较弱,为63.02%。根据药剂对病菌生长和发育各阶段的影响,生产中应采用适宜的施用方法:戊唑醇和嘧菌酯可作为保护剂在病害发生前期喷施,也可作为治疗剂在病害发生流行期应用,但为防止抗药性菌株的出现,两者应当轮换或复配使用。  相似文献   

7.
多堆柄锈菌侵染玉米的细胞学及超微结构特征   总被引:2,自引:1,他引:1  
为明确玉米对多堆柄锈菌Puccinia polysora侵染后病理反应的细胞学特征,利用扫描和透射电镜技术分析了玉米自交系与多堆柄锈菌互作中二者的细胞变化过程。多堆柄锈菌对玉米的侵染主要以直接穿透叶片表皮侵入为主,少量可从气孔和细胞间隙侵入。接种后,病菌夏孢子在感病自交系叶片上快速并大量萌发,在叶表生长蔓延并侵入表皮组织细胞,7 d后形成夏孢子堆;在抗病自交系上,病菌萌发、菌丝生长均受到明显抑制,少量入侵的病菌也由于寄主细胞死亡而导致菌丝和夏孢子干瘪死亡。侵染早期在感病寄主细胞间隙出现菌丝并穿透细胞壁,在胞内产生分枝菌丝,此时寄主细胞结构正常;随着菌丝进一步扩展,叶绿体等结构发生紊乱,被侵染细胞逐渐死亡。在抗病自交系上,接菌24 h后寄主即出现过敏性坏死反应,侵入位点与周围细胞快速坏死,抑制菌丝生长蔓延;叶绿体中清晰可见深色颗粒状物质;72 h后细胞壁外侧产生大量致密的深色结晶体,应为与抗病反应相关的酚类物质。表明抗多堆柄锈菌的玉米材料可能存在2种抗病途径,即寄主与病菌互作中由分子识别引起的免疫反应和病菌侵入后的系统防卫反应。  相似文献   

8.
多堆柄锈菌侵染不同抗性玉米的组织病理学研究   总被引:2,自引:0,他引:2  
[目的] 了解不同抗感玉米自交系对南方锈病的组织病理学反应,为今后筛选不同类型抗病自交系提供参考。[方法] 采用曲利苯蓝透明染色法,以4个玉米自交系为材料,研究了玉米南方锈病致病菌—多堆柄锈菌在不同抗性材料上侵染过程的组织学特征。[结果] 多堆柄锈菌侵入和定殖可以分为5个阶段:孢子萌发与芽管形成、附着胞形成、侵入细胞、胞内吸器产生、菌丝在细胞间扩展。在不同抗性的玉米材料上,病菌孢子萌发和芽管形成差别不明显,但侵入后病菌在不同抗性材料内的发育进程和发育程度具有显著差异。在抗病玉米材料上,病菌初生菌丝、吸器母细胞、次生菌丝的形成时间推迟,胞内吸器少,菌丝分枝少,菌丝生长缓慢。[结论] 这些抗性特征与田间表现出的细胞过敏性坏死、叶片上夏孢子堆少的特征具有一致性。  相似文献   

9.
石斛内生细菌DEB-2对5种辣椒病原真菌的抑制作用   总被引:3,自引:2,他引:1  
为了探明石斛内生细菌DEB-2菌株对贵州辣椒病原真菌的抑制作用,采用生长速率法测定了菌株DEB-2发酵液对辣椒黑斑病病原菌Alternaria alternate、辣椒疫病病原菌Phytophthora capsici、辣椒黑点炭疽病病原菌Colletotrichum capsici、辣椒红色炭疽病病原菌Colletotrichum gloesporioide和辣椒早疫病病原菌Alternaria solani的毒力。结果显示,内生菌DEB-2菌株能显著抑制5种辣椒病原真菌菌丝生长和孢子萌发,菌丝尖端均有不同程度变粗、膨大、崩解或者菌丝分枝增多的现象;分生孢子及芽管产生畸形。DEB-2菌株发酵液对5种病原菌菌丝生长的抑制中浓度EC50值分别是28.5、76.2、80.7、79.9、72.0 μL/mL;对辣椒疫病病原菌和早疫病病原菌分生孢子萌发的EC50分别为73.5 μL/mL和68.7 μL/mL。研究表明,DEB-2菌株能有效地同时抑制5种辣椒病原真菌菌丝生长和孢子萌发,对该5种辣椒真菌病原菌具有一定的生防潜力,可作为生防菌进行开发和应用。  相似文献   

10.
七株植物内生放线菌对苹果树腐烂病的防治作用   总被引:7,自引:5,他引:7  
通过分生孢子萌发试验、菌丝抑制试验、接种离体枝条试验及田间病斑涂抹药剂防治试验,研究了7株植物内生放线菌对苹果树腐烂病的防治效果。结果表明,7株植物内生放线菌无菌发酵滤液对病原菌分生孢子萌发及菌丝生长均有明显的抑制作用,2倍稀释液对孢子萌发抑制率均达85%以上,20倍稀释液对菌丝生长抑制率均达80%以上。显微观察发现,Hhs.015 (BAR1-5)菌株无菌发酵滤液可导致病菌芽管、菌丝畸形。在人工接种条件下,无菌发酵滤液原液对离体枝条病斑扩展具有明显的抑制作用,其中AR1-14、Hhs.015 (BAR1-5)和TGYXCSA-7处理5天后防治效果分别达83.2%、69.7%和85.6%。田间刮除病斑后涂抹放线菌发酵原液也可明显抑制病斑复发,防效均达80%以上。  相似文献   

11.
The effects of tebuconazole, a systemic fungicide, on the morphology, structure, cell wall components and toxin production of Fusarium culmorum were investigated in vitro. Treatment was by application of four filter paper strips (0.75 cm × 5.0 cm) soaked in 20 µg ml ?1 fungicide placed around a point inoculum in Petri dishes. Mycelial growth was strongly inhibited by fungicide treatment. Scanning electron microscopic observations showed that the fungicide caused irregular swelling and excessive branching of hyphae. The morphological changes induced by the fungicide at the ultrastructural level included considerable thickening of the hyphal cell walls, excessive septation, the formation of the incomplete septa, extensive vacuolisation, accumulation of lipid bodies and progressing necrosis or degeneration of the hyphal cytoplasm. Non‐membrane inclusion bodies were often detected in the hyphal cytoplasm. Furthermore, the formation of new hyphae (daughter hyphae) inside collapsed hyphal cells was common following treatment. The daughter hyphae also displayed severe alterations such as irregular thickening of the cell walls and necrosis of the cytoplasm. Using cytochemical techniques, the labelling densities of chitin and β‐1,3‐glucan in the cell walls of the fungicide‐treated hyphae were more pronounced than in those of the control hyphae. Moreover, immunogold labelling with antiserum against deoxynivalenol (DON) revealed that Fusarium toxin DON was localized in the cell walls, cytoplasm, mitochondria and vacuoles of the hyphae from the control and the fungicide treatment, but the labelling density in the fungicide‐treated hyphae decreased dramatically compared with the control hyphae, indicating that tebuconazole reduced Fusarium toxin production of the fungus. © 2001 Society of Chemical Industry  相似文献   

12.
内生菌螺旋毛壳抗生素和水解酶的协同抗真菌作用   总被引:2,自引:0,他引:2  
 生防因子螺旋毛壳(Chaetomium spirale) ND35生长于含立枯丝核菌(Rhizoctonia solani)菌丝细胞壁制备物的SM培养基中,从培养滤液提取的粗酶液强烈抑制Valsa sordidaValsa maliGlomerella cingulataCurvulavia lunata病原真菌的菌丝生长和孢子萌发。经DNS法检测,粗酶液同时具有β-1,3-葡聚糖酶(包括内切和外切酶)和几丁质酶活性,分别为0.19 U.mg-1和0.09 U.mg-1。生长于3%玉米粉浸渍液的螺旋毛壳ND35的培养滤液也能抑制上述病原菌的菌丝生长和孢子萌发。离体条件下检测了ND35产生的一纯化的具有分子量为73 kD的内切β-1,3-葡聚糖酶(GLUC73)和一纯化的抗生素对苹果炭疽病菌的分生孢子萌发和芽管延长的抑制效果。当内切β-1,3-葡聚糖酶使用浓度为180 μg·mL-1时,抑制了测试真菌的孢子萌发,造成细胞壁的改变,导致了菌丝顶端的破裂;抗生素的有效抑菌中浓度ED50为1.75 μg·mL-1。单独应用时,1.0 μg·mL-1的抗生素或40 μg·mL-1的内切β-1,3-葡聚糖酶没有或仅有很低的抑菌效果;两者组合导致孢子萌发约78%受抑制。甚至10 μg·mL-1的内切β-1,3-葡聚糖酶也使1.5 μg·mL-1抗生素的抑菌作用从低于25%增加到超过50%的抑菌效果。此外,80 μg·mL-1的内切β-1,3-葡聚糖酶使1.0 μg·mL-1抗生素的抑菌活性从0%提高到90%。抗生素和β-1,3-葡聚糖酶的协同抗菌活性可能在内生菌螺旋毛壳的植病生防中起重要作用。  相似文献   

13.
平板培养结果表明:枯草杆菌B-921菌株对苹果树腐烂病菌、泡桐腐烂病菌和杨树烂皮病菌,有较强抗生作用。共同培养到第5天,能形成1.5~4.5cm的抑菌圈。在显微镜下观察到,菌丝体壁被溶解,原生质外溢;有的菌丝体扭曲、生长点膨大,胞壁断裂、菌体崩溃。发酵液有同样作用。  相似文献   

14.
香蕉枯萎病拮抗菌的筛选及其作用机制研究   总被引:15,自引:0,他引:15  
通过分离和筛选,从香蕉园或者其他果园的土壤中分离获得13株对香蕉枯萎病菌(Fusarium oxysporum f.sp.cubense)具有抑制作用的拮抗菌,并对部分拮抗菌抑制病菌菌丝生长和孢子萌发进行了试验。结果表明,拮抗菌株d4、d5、B3和p发酵液对香蕉枯萎病菌生长具有显著的抑制作用,在平板上产生的抑菌圈直径为21.75~34.75 mm,抑菌效果具有持续稳定性,对孢子萌发的抑制率为90.49%~97.18%;拮抗菌对病菌的作用表现为对菌丝的消融、菌丝细胞的泡囊化、抑制病菌分生孢子的萌发、孢子芽管的扭曲。  相似文献   

15.
黄腐酸作为一种广谱性植物生长调节剂,具有稳定可靠的抗逆增产效果。本试验以黄腐酸为生长调节剂,研究其对苹果树腐烂病菌(Valsa mali)的抑制作用及对苹果树皮抗腐烂病的影响。试验结果表明,黄腐酸能抑制苹果树腐烂病菌菌丝的生长,浓度越大抑制效果越明显。经黄腐酸处理后,苹果树腐烂病组织中SOD、POD、PAL和CAT均呈现先升高后降低的趋势。其中黄腐酸处理后SOD活性于14d达到峰值,而其他酶活性均在21d达到最大值。说明黄腐酸能显著提高苹果树体内相关防御酶系的活性,很大程度上提高了植物的免疫力,即增强了苹果树的抗病性。  相似文献   

16.
ABSTRACT The mechanism of the effects of flumorph (a novel fungicide) was investigated by analyzing alterations of hyphal morphology, cell wall deposition patterns, F-actin organization, and other organelles in Phytophthora melonis. Calcofluor white staining suggested that flumorph did not inhibit the synthesis of cell wall materials, but disturbed the polar deposition of newly synthesized cell wall materials during cystospore germination and hyphal growth. After exposure to flumorph, zoospores were able to switch into cystospores accompanied with the formation of a cell wall, whereas cystospores failed to induce the isotropic-polar switch and did not produce germ tubes but continued the isotropic growth phase. In flumorph-treated hyphae, the most characteristic change was the development of periodic swelling ("beaded" morphology) and the disruption of tip growth. Newly synthesized cell wall materials were deposited uniformly throughout the diffuse expanded region of hyphae, in contrast to their normal polarized patterns of deposition. These alterations were the result of F-actin disruption, identified with the fluorescein isothiocynate (FITC)-phalloidin staining. The disruption of F-actin also was accompanied by disorganized organelles: each swelling of subapical hyphae was associated with a nucleus. Vesicles did not undergo polarized secretion to the apical hyphae, but diffused around nuclei for the subapical growth; thus, the cell wall was thickened with periodic expansion along the hyphae. Upon removing flumorph, normal tip growth and organized F-actin were observed again. These data, as well as data published earlier, suggest that flumorph may be involved in the impairment of cell polar growth through directly or indirectly disrupting the organization of F-actin. The primary site of action by flumorph in the disruption of the F-actin organization is under investigation.  相似文献   

17.
An important mechanism for genetic diversity in filamentous fungi is hyphal anastomosis and the formation of heterokaryons. In this study, we observed fusion of germ tubes in germinating urediniospores of Phakopsora pachyrhizi resulting in a complex hyphal network. Staining of the germ tubes derived from P. pachyrhizi urediniospores with 4’, 6-diamidino-2-phenylindole (DAPI) showed migration of nuclei through the network resulting in multinucleate hyphae. Short bridges connecting the hyphal network tubes were also observed. Our study provides the first evidence of germ tube and hyphal anastomosis, and nuclear migration in P. pachyrhizi. Considering the lack of a known sexual stage of P. pachyrhizi, this hyphal anastomosis followed by the parasexual cycle may explain the genetic diversity in virulence among populations of P. pachyrhizi.  相似文献   

18.
Germinating teliospores of Uromycladium tepperianum formed penetration pegs, which directly penetrated host epidermal cells, on the tip cell of three-celled germ tubes. Diploid nuclei within teliospores appeared to divide soon after germination commenced. Only one of the daughter nuclei moved out of the teliospores with the developing germ tubes. Reactions of seedlings inoculated with teliospores from different host species and differences in teliospore dimensions indicated that distinct genotypes of U. tepperianum adapted to particular host species occur in Australia. Limited hyphal growth occurred in two of 20 African Acacia spp. inoculated with teliospores from Acacia saligna. Fungal development was, however, restricted by necrosis of surrounding host cells. U. tepperianum appears to be suitably host-specific for use as a biological control agent against Ac. saligna , a weed in South Africa.  相似文献   

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