内生细菌B504的鉴定及对黄瓜枯萎病的生防作用

B504是从黄瓜植株中分离到的内生细菌。平板对峙试验表明,该菌株对黄瓜枯萎病菌等10种植物病原真菌具有拮抗作用,对黄瓜枯萎病菌的生长抑制率达44.1%。该菌株造成黄瓜枯萎病菌的分生孢子畸形,形成泡状物,尔后细胞壁消解,引起细胞内含物外溢,明显影响孢子萌发。用B504对黄瓜种子进行催芽处理,对黄瓜苗期、成株期枯萎病的防治效果分别为54.0%和41.7%。B504的16S rDNA序列与枯草芽胞杆菌具有高度相似性。根据菌体形态、培养性状、生理生化特性及16S rDNA序列分析等指标,确定B504为枯草芽胞杆菌（Bacillus subtilis）。     

拮抗放线菌B1菌株鉴定及其防治番茄灰霉病的初步研究

为明确放线菌B1菌株在植物病害生物防治中的应用潜力,采用平板对峙培养法、离体叶片和果实及温室盆栽苗测定其抑菌作用,通过多相分类法研究其分类地位,并且采用单因子试验与正交试验相结合的方法,筛选出该菌株的液体发酵培养基配方.结果表明:菌株B1对10种植物病原真菌和3种植物病原细菌有抑制作用,其中对番茄灰霉病菌、番茄早疫病菌、小麦纹枯病菌、甘蓝黑腐病菌和西瓜果腐病菌的抑制作用较强,对番茄灰霉病菌的生长抑制率达86.25%;菌株B1的代谢产物对番茄灰霉病菌菌丝有致畸作用;菌株B1发酵上清液在离体叶片和活体植株上对番茄灰霉病具有良好的生防作用,其防治效果分别为68.80%和64.03%.根据形态特征、培养性状、生理生化特性和16S rDNA序列分析结果,将菌株B1初步鉴定为淡紫灰链霉菌Streptomyces lavendulae.通过正交试验筛选出菌株B1的液体发酵培养基组成为:葡萄糖15 g、玉米浆20g、NH4H2PO40.5 g、MgSO41.0 g、自来水1 000mL,pH 7.2～7.4.     

香蕉枯萎病拮抗细菌的分离筛选与鉴定

从采集的香蕉根际土壤中经分离、纯化获得细菌菌株74个,采用对峙培养法,获得对香蕉枯萎病菌4号小种具有强抑制作用的菌株S-1,相对抑制率达76%。抑菌谱测定表明,菌株S-1对尖孢镰刀菌古巴专化型1号小种、番茄枯萎病菌、玉米弯孢叶斑病菌、胶孢炭疽病菌、棉花枯萎病菌、棉花黄萎病菌、小麦赤霉病菌、西瓜枯萎病菌等病原菌具有抑制作用。经Biolog系统鉴定及16S rDNA序列同源性分析,菌株S-1为枯草芽孢杆菌。该菌株理想的培养条件为：PYTG或PDA培养基,26-30℃,pH值7.0-8.0,振荡培养40-48h。     

西瓜枯萎菌拮抗细菌的筛选、鉴定及防效测定

从内蒙古、新疆、安徽、河南、江苏、江西6省的西瓜、黄瓜和茄子3种作物根际土壤和病健组织中,共获得样本64份,分离得到细菌1050株。室内抑菌活性测定结果表明：9株细菌分离物对西瓜枯萎病菌Fusariumoxysporumf.sp.niveum、甘蓝黑斑病菌Alternaria brassicicola、辣椒疫霉病菌Phytophthora capsici和番茄灰霉病菌Botrytis cinerea均具有很强的拮抗作用,其中菌株NBT-15和PL-82对西瓜枯萎病菌菌丝生长的抑制作用比较明显,抑制率分别为60.5%和55.3%。盆栽防病试验结果表明：NBT-15和PL-82对西瓜枯萎病的防治效果分别为80.4%和75.6%。形态特征、生理生化特性及16S rDNA序列分析结果确定菌株NBT-15为枯草芽孢杆菌Bacillus subtilis,PL-82为铜绿假单胞菌Pseudomonas aeruginosa。     

生防细菌C-4的特性鉴定及其对香蕉枯萎病的防治效果

采用平板对峙法从香蕉根际土壤中共分离到74株细菌,其中菌株C-4对香蕉枯萎病菌1号小种和4号小种均有强抑制作用,相对抑制率分别为81.25%和83.75%。抑菌谱测定结果表明,菌株C-4具有较广的抑菌范围,对番茄枯萎病菌、玉米弯孢叶斑病菌、胶孢炭疽病菌、棉花枯萎病菌、棉花黄萎病菌、小麦赤霉病菌和西瓜枯萎病菌等均有抑制作用。根据Biolog系统鉴定及16S rDNA序列分析结果,将菌株C-4鉴定为地衣芽孢杆菌Bacillus licheniformis,其在YPGA或PDA培养基、32~35℃、pH 7.8~8.0、40~52h培养条件下生长和拮抗作用最为理想。室内盆栽试验结果显示,菌株C-4对香蕉枯萎病具有良好的防效,接种30天后病情指数明显低于对照。     

植物土传病原菌拮抗细菌的筛选与鉴定

从作物根际土壤中分离到1056株细菌，筛选出7个具有较强拮抗活性的菌株。室内测定对水稻纹枯病菌、辣椒疫病菌、瓜果腐霉、油菜菌核病菌、棉花枯萎病菌、茄根腐病菌、棉花黄萎病菌、番茄青枯病菌、甘蓝黑腐病菌等重要土传病原菌有较强拮抗作用；温室盆栽试验对番茄青枯病表现出较好防效，其中以BOH2和OH11效果较为明显，防效分别为90．9％和86．4％。通过形态观察、生理生化试验和16SrDNA序列分析，确定OH11为产酶溶杆菌。BOH3为荧光假单胞菌，其余5个菌株为不同芽孢杆菌。     

解淀粉芽胞杆菌YU-1对水稻纹枯病的防治作用

本文旨在筛选对水稻纹枯病有生防作用的菌株,并初步探索其生防作用机理。收集水稻、甘蓝、黄瓜等不同植物根际土壤,采用稀释分离和对峙培养法筛选对水稻纹枯病菌Rhizoctonia solani Kühn有抑菌作用的菌株;通过离体接种防效、盆栽防效、抑菌谱、对水稻纹枯病菌菌核萌发及形成的抑制作用等方面评价其生防潜力,并对生防菌株进行鉴定。结果表明,从采集的37份根际土壤中共分离获得细菌297株,其中4株对纹枯病菌具有较好的拮抗作用,菌株YU-1对水稻纹枯病菌菌丝生长的抑制率达89.8%;对西瓜枯萎病菌、草莓灰霉病菌、番茄早疫病菌、油菜菌核病菌、小麦赤霉病菌菌丝生长的抑制率均在85%以上;对水稻纹枯病的离体和盆栽防效分别为73.1%和66.3%;对水稻纹枯病菌菌核萌发的抑菌率在92%以上;经生理生化和分子鉴定为解淀粉芽胞杆菌Bacillus amyloliquefaciens。由此可看出,菌株YU-1对水稻纹枯病的防治具有较强的应用价值,具有进一步开发成生物农药的潜力。     

芹菜软腐病拮抗芽胞杆菌筛选及防治效果

本研究从芹菜根际土壤分离筛选得到一株对芹菜软腐病具有良好防治效果的菌株ZF75。通过菌落形态观察、生理生化特性及16S rDNA序列分析，初步将该菌株鉴定为解淀粉芽胞杆菌Bacillus amyloliquefaciens。平板对峙结果表明，菌株ZF75对番茄疮痂病菌等10种病原菌具有较强的拮抗效果。用温室栽培试验对菌株ZF75进行防病效果初探，试验结果表明，菌株ZF75对芹菜软腐病防效可达89.62%。同时，该菌株对芹菜具有显著的促生效果，经菌株ZF75菌悬液处理后的芹菜平均株高和茎粗分别达24.18 cm和0.51 cm。综合分析表明，菌株ZF75具有良好的田间应用开发潜力。     

对甲氧基肉桂醛的抗真菌活性及田间药效分析

通过对植物香料产物对甲氧基肉桂醛的抗真菌活性研究,发现该化合物对鞭毛菌亚门卵菌纲、子囊菌亚门和半知菌亚门的8种植物病原真菌,番茄晚疫病菌(Phytophthora infestans)、水稻纹枯病菌(Rhizoctonia solani AG-1IA)、油菜菌核病菌(Sclerotinia sclerotiorum)、西瓜枯萎病菌(Fusarium oxysporum)、番茄灰霉病菌(Botrytis cinerea)、草莓褐色轮斑病菌(Phomopsis obscurans)、草莓炭疽病菌(Colletotrichum gloeosporioides)和烟草赤星病菌(Alternaria alternata)具有高效、广谱的特性;田间药效试验表明其对黄瓜霜霉病和水稻纹枯病具有较高的防治效果。这为其在植物病害防治中的应用以及进一步开发提供了依据。     

鸭粪中1株水稻纹枯病菌拮抗细菌的鉴定

为探索稻鸭复合生态系统抑制水稻纹枯病发生的作用机理,以从鸭粪中分离到的1株拮抗细菌A168对水稻纹枯病菌的拮抗效果进行了研究。测定了其对水稻纹枯病菌的拮抗作用及其抑菌谱;根据培养性状、形态特征、生理生化测定、扫描电镜观察及16S rDNA序列分析对其进行了鉴定;构建了包括其在内的13株相关种属菌株的系统发育树。A168菌株对水稻纹枯病有明显的拮抗作用,且对小麦赤霉病菌(Fusarium graminearum)、稻曲病菌(Ustilaginoidea virens)和油菜菌核病菌(Sclerotinia sclerotiorum)有抑制作用,抑菌谱广;A168菌体杆状,革兰氏染色阳性、芽孢中生、周生鞭毛、16S rDNA分析与其亲缘关系较近菌株Bacillus cereus(AJ577288.1)的同源性达99%,结合形态和生理生化特征,鉴定A168菌株为蜡状芽孢杆菌(Bacillus cereus),其16S rDNA序列已在GenBank中注册,登录号为GQ118339。A168为开发防治水稻纹枯病生防制剂提供了依据。     

Isolation of Nonpathogenic Mutants of Fusarium oxysporum f. sp. melonis for Biological Control of Fusarium Wilt in Cucurbits

ABSTRACT Two nonpathogenic mutant strains 4/4 and 15/15 of Fusarium oxysporum f. sp. melonis (race 1,2) were isolated by a continuous dipinoculation technique following UV mutagenesis of the virulent wild-type isolate FOM1.2. No disease symptoms or detrimental effects were observed following inoculation of muskmelon seedlings by strain 4/4. In contrast, strain 15/15 caused mortality of susceptible cultivars although to a lesser extent than the wild-type isolate. Strain 4/4 colonized a variety of muskmelon and watermelon cultivars. In muskmelon cv. Ein Dor, seedlings were dipped in a conidial suspension of strain 4/4 and planted in medium amended with the mutant to achieve 100% colonization of roots and between 30 to 70% of the lower stem tissues 7 days after planting. Similar percent colonization of watermelon seedlings by strain 4/4 was recorded. In cross-protection experiments with muskmelon cultivars, significant reduction in seedling mortality was observed between 4/4-colonized FOM1.2. challenged plants compared with that of wild-type challenged plants alone. Similarly, strain 4/4 was able to significantly reduce mortality of watermelon seedlings caused by F. oxysporum f. sp. niveum race 2. This novel approach of generating nonpathogenic mutants for biological control in Fusarium spp. and other fungal pathogens from virulent wild-type isolates may be beneficial for control, because the mutant strains, lacking only in pathogenicity, may compete more efficiently than other biocontrol organisms against the pathogen of origin.     

AM真菌和镰刀菌对西瓜根系膜脂过氧化作用和膜透性的影响

 在温室盆栽条件下研究了丛枝菌根(arbuscular mycorrhiza,AM)真菌Glomus versiforme (Karsten) Berch和西瓜枯萎镰刀菌(Fusarium oxysporum f. sp. niveum)对西瓜根系中膜脂过氧化产物丙二醛(MDA)含量和细胞膜透性的影响。结果表明,接种AM真菌的西瓜根系中MDA含量、组织自动氧化速率和膜透性均低于对照,先接种G. versiforme,后接种F. oxysporum f. sp. niveum处理的MDA含量、组织自动氧化速率和细胞膜透性均低于只接种F. oxysporum f. sp. niveum的处理。接种G. versiforme感枯萎病西瓜品种"郑杂5号"MDA含量、组织自动氧化速率和膜透性降低幅度大于抗病品种"京欣1号"的接种处理,说明G. versiforme可降低感病西瓜品种的膜透性和MDA的产生,从而有效地保护细胞膜系统,减轻F. oxysporum f. sp. niveum对西瓜的为害程度。     

农杆菌介导的西瓜枯萎病菌遗传转化

为获得带GFP标记的西瓜枯萎病菌转化株,用于后期观察病原菌侵染过程,采用农杆菌介导的方法,对西瓜枯萎病菌1号生理小种进行了遗传转化。结果表明:共培养时间为36h,枯萎病菌孢子和农杆菌AGL1比例为1∶1时该菌株的遗传转化效率最高,可以达到117.33个转化子/107个孢子。转化株的孢子、菌丝体及萌发的孢子均能发出稳定而强的绿色荧光。转化株的致病力检测显示其致病力与转化前的野生菌株致病力无明显差异。结果表明本研究获得的带GFP标记的西瓜枯萎病菌转化株可用于观察病菌在西瓜根系的侵染过程。     

Quantification of Root and Stem Colonization of Watermelon by Fusarium oxysporum f. sp. niveum and Its Use in Evaluating Resistance

ABSTRACT Colonization of watermelon root and stem tissues by Fusarium oxysporum f. sp. niveum race 1 and its relationship to the apparent resistance to Fusarium wilt was investigated. In each of 2 years, 17 differentially susceptible watermelon cultivars and one accession were tested in the greenhouse, and 7 cultivars also were tested in the field. Colonization by a chlorate-resistant marked isolate of the fungus was assayed by plating homogenized tissue samples on a selective medium. Six days after inoculation, seedlings of highly resistant, moderately resistant, and susceptible cultivars had F. oxysporum f. sp. niveum race 1 CFU counts in the lower stems of 10(2), 10(3), and 10(4) CFU/g of fresh tissue, respectively. Percent wilt (Y) of the seedlings was positively correlated with colonization (X) by F. oxysporum f. sp. niveum race 1 in roots (Y = 21.2 ln [X + 1] - 140.7, R(2) = 0.85) or lower stems (Y = 17.3 ln [X + 1] - 78.6, R(2) = 0.86). Percent wilt (Y) also was correlated with the ratio (X(r), 0 to 1 values) of lower stem to root colonization (Y = 34 ln X(r) + 112, R(2) = 0.36). Field evaluations confirmed these relationships, and a link between cultivar resistance and a reduced rate of spread of the fungus in primary stems during a season was observed. Fruit yield decreased with increased tissue colonization at linear rates of 9.9 to 12.7 t/ha per ln (CFU/g + 1) (R(2) >/= 0.58). The greenhouse seedling stem colonization assay described may be utilized as a collaborative method to quantify Fusarium wilt resistance in watermelon.     

万寿菊根提取物对西瓜枯萎病菌的抑菌活性成分及作用机理研究

本文研究了万寿菊根提取物中西瓜枯萎病菌的抑菌活性成分及其抑菌作用机理。结果表明,在4类主要化学物质中,精油类含量最高(59.8%),且抑菌效果最佳;10mg/mL精油提取物的抑制率在24、36和48h分别为62.83%,58.31%和56.30%。在西瓜枯萎病的3个主要发病期(苗期、伸蔓开花期和座果期)施用精油提取液,能有效抑制西瓜枯萎病菌,促进植株生长,同时提高西瓜植株的POD和SOD酶活性,并维持CAT活性,有效减轻了西瓜枯萎病菌对植株的毒害作用。     

生防菌株B-3对辣椒枯萎病的防治及其鉴定

为了明确生防菌株B-3对辣椒枯萎病的防效及其分类地位,采用管碟法测定了生防菌株B-3对辣椒枯萎病菌的抑制作用,考察了其对辣椒枯萎病的防效,并采用表型特征培养观察法、生理生化测定法和16SrDNA序列分析法对菌株B-3进行了鉴定。结果表明,生防菌株B-3的发酵液及其滤液对辣椒枯萎病菌均表现出较好的抑制作用,对辣椒枯萎病的盆栽与大田试验的防效都比较好,分别为73.6%和64.8%,显著优于100mg/L多菌灵。菌株B-3的形态与生理生化特性与枯草芽孢杆菌很接近,由16SrDNA序列分析的系统发育树发现,菌株B-3的序列与Bacillussubtilis菌株MO5单独构成一个分支,进化上的距离最近,由此可将其鉴定为枯草芽孢杆菌Bacillussubtilis。     

农抗120防治西瓜枯萎病的机制

研究了农抗120对西瓜枯萎病菌和西瓜幼苗的作用。结果表明,农抗120对西瓜枯萎病菌孢子的萌发和菌丝的形态无明显影响,对孢子的形成和菌丝生长有一定抑制作用,能够提高西瓜幼苗抗枯萎病菌毒素致萎的能力和幼苗体内过氧化物酶活性。说明农抗120的防病机制是通过抑制病原菌和提高植株抗病性而起作用的。     

产铁载体PGPR菌筛选及其对病原菌的拮抗作用

采用改进的CAS定性、定量方法,从16株PGPR菌株中初步筛选出具有抗病原真菌作用的菌株,再利用平板对峙法将筛选出的5株PGPR菌与3种病原真菌进行拮抗试验。结果表明,LHS11对黄瓜枯萎病菌（Fusarium oxysporum f. sp. cucumerinum）,西瓜枯萎病菌（Fusarium oxysporum f. sp. niveum）的抑制效果最好,抑菌率达到80%以上;其次是191对黄瓜枯萎病菌,抑菌率为74%,LHS11、191对立枯丝核菌（Rhizoctonia solani）的抑制效果相近,抑菌率分别为64%和65%。191和LHS11是抑菌效果较好的生防PGPR菌株,具有较好的应用潜力。     

Characterization of a Regional Population of Fusarium oxysporum f. sp. niveum by Race, Cross Pathogenicity, and Vegetative Compatibility

ABSTRACT Eighty-eight isolates of Fusarium oxysporum f. sp. niveum, collected from wilted watermelon plants and infested soil in Maryland and Dela-ware, were characterized by cross pathogenicity to muskmelon, race, and vegetative compatibility. Four isolates (4.5%) were moderately pathogenic to >/=2 of 18 muskmelon cultivars in a greenhouse test, and one representative isolate also was slightly pathogenic in field microplots. The four isolates all were designated as race 2, and were in vegetative compatibility group (VCG) 0082. Of the 74 isolates to which a VCG could be assigned, 41 were in VCG 0080, the VCG distributed most widely; 27 were in VCG 0082, and were distributed in half of the 20 watermelon fields surveyed; and 6 were in the newly described VCG 0083, and were restricted to three fields. Among the isolates in VCG 0080, 8 were designated as race 0, 21 as race 1, and 12 as race 2. Of the isolates in VCG 0082, 6 were designated as race 0, 11 as race 1, and 10 as race 2. All isolates in VCG 0083 were designated as race 2. Isolates from more than one race within the same VCG or isolates from more than one VCG were recovered from single plants and fields. No differences in aggressiveness on differential watermelon cultivars were observed among isolates from different VCGs of the same race. A diverse association between virulence and VCG throughout the Mid-Atlantic region suggests that the pathotypes of F. oxysporum f. sp. niveum may be of local origin or at least long existent in the region.     

土壤施加氧化钙对西瓜枯萎病的影响及其机理初探

为探明土壤撒施石灰对西瓜枯萎病发生的影响,本试验设计5个浓度剂量的氧化钙对连作西瓜5年的土壤进行消毒处理,通过栽培感病品种并调查植株生长情况、枯萎病发病率等指标,结合土壤中尖孢镰刀菌西瓜专化型1号小种病原菌孢子量和土壤pH的变化动态,以此评估土壤施加石灰防治西瓜枯萎病的效果,并利用不同浓度的钙对尖孢镰刀菌西瓜专化型菌丝生长和产孢量的影响进行防治机理的初步研究。试验结果表明,当土壤施加氧化钙后,西瓜发病率显著降低,各处理的相对防效分别为14%、57.8%、96.3%、94.7%、94.7%。在定植10d后,各处理均有促进植株生长发育的作用,其中1.34g/kg最显著。而随着氧化钙施加量的增加,土壤pH值也不断提高,30d趋于稳定。对土壤病原菌孢子数量检测表明:各处理土壤中病原菌数量均呈现显著降低的趋势。PDA平板生测结果表明,当钙离子浓度高于80mmol/L时,病原菌的生长及产孢量开始受到抑制,随着浓度的增加,抑制作用加强。土壤施加氧化钙不仅能改善土壤的酸化,还能有效防控西瓜枯萎病,其机理可能是土壤pH值的提高改变了土壤微环境,不利于病原菌侵染;施加的钙离子抑制了病原菌菌丝的生长及孢子的萌发。研究结果对揭示氧化钙防控西瓜枯萎病机制,具有一定的参考价值。