小麦孢囊线虫病生防真菌08F04菌株的鉴定及防效测定

小麦禾谷孢囊线虫病已成为我国黄淮麦区的重要病害,目前生产上尚缺乏有效的防控方法。菌株08F04是作者从禾谷孢囊线虫孢囊上分离的1株寄生真菌。本研究通过形态学和分子生物学方法,将其鉴定为球孢白僵菌Beauveriabassiana（Bals．）Vuill。2011和2012年对该菌株进行了盆栽和田间防效试验,菌株08F04麦粒砂培养物6％（w／W）土壤处理的盆栽试验孢囊减退率分别达到65．03％和50．95％;田间试验中,菌剂08F0420g·m-2沟施处理土壤,2011年在许昌和荥阳的孢囊减退率分别为58．28％和32．13％,2012年分别达到58．49％和44．50％。结果表明,菌株08F04是防治小麦禾谷孢囊线虫病具有较大潜在开发价值的生防菌株。     

豫北及冀南地区4个小麦禾谷孢囊线虫群体的种类鉴定

禾谷孢囊线虫病(Cereal Cyst Nematode,CCN)是世界上为害小麦等禾谷类作物的重要根部线虫病害,许多国家受害严重[1].该病的病原禾谷孢囊线虫(Heterodera avenae group)是一个复合种群组,包括12个有效种和几个未命名的种,其中发生普遍、危害严重的有燕麦孢囊线虫(H.avenae)、菲利普孢囊线虫(H.filipjevi)、大麦孢囊线虫(H.hordecalis)和麦类孢囊线虫(H.latipons)[1].我国1987年在湖北省天门县首次在小麦上发现禾谷孢囊线虫病,Wang等[2]鉴定病原为燕麦孢囊线虫(H.avenae);后陆续在河南、安徽、山东等多个省份发现该病,病原均为该种线虫.2010年Li等[3]在河南许昌报道发现了菲利普孢囊线虫(H.filipjevi).     

鲁豫皖交界地区四个小麦禾谷孢囊线虫群体致病型鉴定

为了明确鲁豫皖交界地区小麦禾谷孢囊线虫的致病类型,利用23个国际鉴别寄主品种和1个当地小麦品种温麦19对采自山东菏泽、安徽颍上、河南商丘和淮阳的4个小麦禾谷孢囊线虫群体致病型进行了鉴定。4个群体的致病型不同于国际上已命名的16个小麦禾谷孢囊线虫致病型。13个A组鉴别寄主对山东菏泽与安徽颍上群体具有相同的抗性反应,这2个线虫群体属于同一致病型;河南商丘群体与上述2个群体的毒性相似,但大麦寄主KVL191对山东菏泽群体和安徽颍上群体表现为抗病,而对河南商丘群体表现感病;河南淮阳群体的致病谱较宽,明显不同于其它3个群体。对照小麦品种温麦19对4个小麦禾谷孢囊线虫群体均表现感病。     

北京地区小麦禾谷孢囊线虫病发生动态调查

小麦禾谷孢囊线虫病是小麦生产上的重要病害.2010-2011年对北京地区小麦禾谷孢囊线虫的发生规律进行了定期定点调查.结果表明,禾谷孢囊线虫在北京地区全年只发生1代,夏季滞育,卵孵化高峰为4月初;2龄幼虫侵染高峰为4月上旬,3龄幼虫发育高峰为4月下旬至5月初,4龄幼虫发育高峰为5月上旬,白雌虫发育高峰为5月下旬至6月上旬,10月份播种后部分2龄幼虫就可以发生侵染并且冬前发育至3龄幼虫.本研究结果可为北京地区禾谷孢囊线虫的防治提供理论依据.     

青海省小麦孢囊线虫病发生和分布特点

小麦孢囊线虫病是青海省小麦青稞产区的主要病害之一。2007-2010年,采用随机取样方法对青海省小麦孢囊线虫病的发生、分布情况进行了调查。调查结果表明,小麦孢囊线虫病主要分布在青海省西宁、海东、海北、黄南、玉树、海西州6个地区,在不同海拔、不同生态区均有分布,但不同生态区以及同一生态区不同地块间发生量差异较大。垂直分布调查结果表明,同一地块中禾谷孢囊线虫主要分布在20 cm以上的土层中。     

江苏省小麦孢囊线虫病发生情况初步调查

2009年5月对江苏省5个地区的22个县市小麦孢囊线虫病发生情况进行了初步调查,共采集76份含根系和根际土壤的样本,经分离与鉴定表明,小麦禾谷孢囊线虫(Heterodera avenae,CCN)在徐州、连云港、宿迁和盐城等地区都有分布,检出率为35%。徐州的沛县、丰县、邳州和睢宁等4个县市的小麦孢囊线虫病检出率超过90%。其中,丰县华山镇、沛县河口镇和丰县城东的田块孢囊密度较大,每100 mL土壤的平均孢囊数分别为161、159个和112个,每个孢囊的平均含卵量分别为187、190粒和174粒。这是小麦孢囊线虫病在江苏省发生的首次报道。     

M-14复合生物肥料抗小麦孢囊线虫及增产效果

本试验将复合生物肥料M-14与基质、常规肥料一次性施入各小区,采用对越冬期小麦根部胞囊线虫侵染情况统计、成熟期小麦基本生物学性状测量、收获期土壤小麦禾谷孢囊线虫有效孢囊数的筛查统计以及测产结果分析等方法对复合生物肥M-14在田间条件下抗小麦禾谷孢囊线虫及增产效果进行了研究。结果表明,施750 kg/hm~2复合微生物肥料M-14,不仅能有效增加小麦株高、茎粗、穗长、公顷穗数及千粒重,并且与空白对照相比增产14.3%,增产效果达极显著水平(P0.01),而且还能显著地减少胞囊线虫侵染率和收获时土壤中有效孢囊数,与空白对照、常规复合肥、基质处理组相比其侵染率分别减少43.4%、70.8%、65.9%,与对照相比有效孢囊数减少了97.59%,与其他各处理相比差异极显著(P0.01),从而降低小麦胞囊线虫发病级别和发病指数,有效地防止小麦禾谷孢囊线虫病发生。因此,施复合生物肥料M-14既能抗小麦孢囊线虫病又能增产,为复合生物肥料M-14的推广应用奠定了理论基础。     

甘肃省马铃薯镰刀菌干腐病优势病原的分离鉴定

 为明确甘肃马铃薯镰刀菌干腐病的优势病原,2006 年12 月~ 2007 年3 月由西至东从甘肃张掖、天祝、永登、临洮、渭源和西和等6 县市的马铃薯贮藏窖中采集表现有镰刀菌干腐病症状的马铃薯薯块,以组织分离法分离病原,单孢纯化镰刀菌(Fusarium spp. )菌株后,以形态学为基础,参照Nelson 镰刀菌分类系统进行鉴定。结果表明:6 个采样区共分离到293株镰刀菌菌株,其中以接骨木镰刀菌(F. sambucinum)和茄病镰刀菌(F. solani)出现频率高,是优势种。分析发现第一优势种随采样区而异,张掖、天祝和渭源采集的样品中茄病镰刀菌分离频率分别为42. 6% 、42. 1% 和32. 4% ,接骨木镰刀菌分离频率分别为14. 8% 、5. 3% 和26. 5% ,茄病镰刀菌为第一优势种;永登、临洮和西和采集的样品中接骨木镰刀菌分离频率分别为52郾1% 、50. 9% 和55. 2% ,茄病镰刀菌的分离频率分别为23. 3% 、32. 7% 和20. 7% ,接骨木镰刀菌为第一优势种。本文进一步对其在PDA、CLA 上的培养特征进行了观察和描述。按照柯赫氏法则用混合菌株接种法对大西洋(Atlantic)、夏波蒂(Shepody)以及一地方品种进行致病性测定,证实了优势菌种对块茎的致病性。利用EF鄄1琢基因引物(EF鄄1H 和EF鄄2T)对接骨木镰刀菌菌株GAUF鄄F12 进行基因组DNA 的PCR 扩增,将PCR 产物回收测序后在GenBank 上比对,菌株GAUF鄄F12 与GenBank 登记的接骨木镰刀菌5 个菌株的同源性均达99% ;用DNASTAR 分析软件将同源性较高的登记菌株的序列与GAUF鄄F12 菌株构建同源性树,结果表明:该菌株与以上5 个接骨木镰刀菌菌株均位于同源性树的同一分支,聚为一类,与形态学的鉴定结果一致。     

河北廊坊大豆枯萎病病原镰刀菌的分子鉴定

 为明确河北廊坊中国农科院植保所试验基地大豆孢囊线虫病田内大豆枯萎病病原镰刀菌的种类,对362份罹病枯萎大豆植株进行病原真菌分离,得到335株真菌;使用镰刀菌通用引物鉴定出镰刀菌(Fusarium spp.) 279株,占分离菌株83.3%;镰刀菌特异性引物、测序等分子生物学技术结合形态学特征进一步鉴定镰刀菌种类,鉴定出尖孢镰刀菌(F. oxysporum)189株,占分离菌株56.4%;茄病镰刀菌(F. solani)67株占20.0%、禾谷镰刀菌(F. graminearum)16株占4.8%、木贼镰刀菌(F. equiseti)3株、层出镰刀菌(F. proliferaum)2株、燕麦镰刀菌(F. avenaceum)和厚孢镰刀菌(F. chlamydosporum)各1株;致病性测试结果表明数量最多的尖孢镰刀菌(F. oxysporum)中约92.8%菌株具有不同程度的致病力;这些结果表明该试验基地大豆枯萎病的优势病原菌为尖孢镰刀菌(F. oxysporum);研究结果可为大豆枯萎病的防治提供科学依据,并为大豆孢囊线虫与尖孢镰刀菌复合侵染大豆的研究奠定基础。     

小麦赤霉病拮抗菌株筛选及其抑制作用研究

从高寒草甸野牦牛牛粪中分离得到细菌菌株27株。采用平板对峙法筛选小麦赤霉病高效拮抗菌株,研究其对病原菌禾谷镰刀菌(Fusarium gramminearum)的抑制作用。结果表明,筛选得到的8株小麦赤霉病拮抗中,以菌株LM3403拮抗能力最佳,抑菌带宽(13.2±0.8)mm。通过形态观察、生理生化特征分析、16S r DNA序列分析和gyr B基因序列分析,将其鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。研究发现,菌株LM3403可明显抑制禾谷镰刀菌菌丝体生长和分生孢子萌发,使病菌的菌丝体畸形、细胞膜损伤。该菌株还具有合成surfactin、iturin、fengycin 3种抗菌脂肽的能力。     

防治斜纹夜蛾蛹和2龄幼虫的棒束孢菌株筛选

为获取对斜纹夜蛾Spodoptera litura(Fabricius)具致病力的生防真菌资源,通过对8株寄主为鳞翅目蛹的棒束孢属Isaria spp.真菌进行形态学和分子系统学鉴定;以斜纹夜蛾的蛹和2龄幼虫为试虫,分别采用拌土法和浸渍法测试菌株的致病力。结果显示:菌株GZUIFR08XS1、GZUIFR04XS8、GZUIFR08XS13为环链棒束孢I.cateinannulata,菌株GZUIFR08LYS4、GZUIFR04XS5、GZUIFR04XS7、GZUIFR08XS12、GZUIFR04XS16为粉棒束孢I.farinose。8株菌对斜纹夜蛾的蛹和2龄幼虫均有致病力。土壤接种5 mL浓度为2×10~8个/mL的分生孢子,环链棒束孢GZUIFR08XS1、GZUIFR04XS8对蛹的致死率均为100%。接种浓度为1×10~8个/mL的分生孢子,环链棒束孢GZUIFR04XS8对2龄幼虫致死率为(78.21±2.22)%,较其他菌株有差异显著(P0.05)。将环链棒束孢GZUIFR04XS8分别接种在2～5龄幼虫时,对同一龄期幼虫的致死率随接种浓度的增大而上升,其中,接种孢子浓度为1×10~9个/mL可致2龄幼虫100%死亡;同一接种浓度下,对幼虫致死率随幼虫龄期的增大而下降,其中,2龄幼虫的致死中浓度LC_(50)为1.28×10~5个/mL。环链棒束孢GZUIFR04XS8有较好防治斜纹夜蛾蛹和低龄幼虫的生防潜力。     

Non‐pathogenic rhizosphere bacteria belonging to the genera Rhizorhapis and Sphingobium provide specific control of lettuce corky root disease caused by species of the same bacterial genera

Lettuce corky root (CR) is caused by bacteria in the genera Rhizorhapis, Sphingobium, Sphingopyxis and Rhizorhabdus of the family Sphingomonadaceae. Members of this family are common rhizosphere bacteria, some pathogenic to lettuce. Sixty‐eight non‐pathogenic isolates of bacteria obtained from lettuce roots were tested for control of CR caused by Rhizorhapis suberifaciens CA1^T and FL1, and Sphingobium mellinum WI4^T. In two initial screenings, 10 isolates significantly reduced CR induced by one or more pathogenic strains on lettuce seedlings in vermiculite, while seven non‐pathogenic isolates provided significant CR control in natural or sterilized field soil. Rhizorhapis suberifaciens FL11 was effective at controlling all pathogenic strains, but most effective against R. suberifaciens CA1^T. The other selected isolates controlled only pathogenic strains belonging to their own genus. In a greenhouse experiment, a soil drench with selected biocontrol agents (R. suberifaciens FL11, Sphingomonas sp. NY3 and S. mellinum CA16) controlled CR better than seed treatments or application of alginate pellets. In microplots infested with R. suberifaciens CA1^T, seed treatment with R. suberifaciens FL11 provided complete control and a soil drench with FL11 significantly reduced the disease. Pathogenicity tests with FL11 on 23 plant species in 10 families resulted in slight yellowing on roots of lettuce and close relatives; similar yellowing appeared on some roots of non‐inoculated lettuce plants. This research showed that biocontrol agents can be genus‐specific. Only one isolate, FL11, provided more general control of various pathogenic strains causing CR even in field soil in pots and microplots.     

Molecular Approaches to Assist the Screening and Monitoring of Postharvest Biocontrol Yeasts

Nineteen yeast isolates obtained from the surface of several fruits and vegetables grown in Southern Italy and Israel were compared by molecular analysis using arbitrarily primed polymerase chain reaction (AP-PCR) and random amplified polymorphic DNA technique (RAPD-PCR). Genetic analysis made it possible to distinguish between closely-related genitically different strains which had the same morphological characteristic, and to discard isolates which were genetically identical. Following PCR characterisation, 6 isolates were selected and tested for their biocontrol activity against major postharvest pathogens (Penicillium digitatum on grapefruit, Botrytis cinerea, Rhizopus stolonifer and Aspergillus niger on table grape and B. cinerea and R. stolonifer on cherry tomato). All the isolates showed a good biocontrol efficacy on both wounded and non-wounded fruits. Furthermore, the preharvest application of the most effective antagonist (LS15) on table grape resulted in a significant reduction in grey mold ranging from 28.3% to 38.2% compared to the untreated control. The RAPD-PCR technique was also useful for identifying and monitoring the survival of the antagonist after field application.     

Isolation and selection of Hemileia Vastatrix antagonists

Organic coffee growing is rapidly increasing in Brazil, and many diseases, especially coffee leaf rust (Hemileia vastatrix), are threatening its production. This study is a first step towards a biocontrol program for coffee rust on organically grown plants. In three organic coffee farms in the state of Minas Gerais, 393 microbial strains including 154 bacterial and 239 fungal strains were isolated from leaves, leaf debris, and soil samples, and in 6 month-old coffee plants, 17 of these isolates reduced both the infection frequency (IF) and the number of H. vastatrix urediniospores produced per leaf (UPL) by more than 70 %. The isolates were identified as eight bacteria isolates, seven Bacillus spp. and one Pseudomonas sp., and nine fungal isolates, four Fusarium spp., two Penicillium spp., one Aspergillus sp., one Acremonium sp. and one Cladosporium sp. Each fungal and bacterial isolate was applied 0, 4, 8, 12 or 16 days before and 0, 4, 8, 12 or 16 days after H. vastatrix inoculation, and the efficiency in reducing both IF and UPL was evaluated. The efficiency was higher and lasted longer when the bacterial isolates were applied before H. vastatrix inoculation. Six Bacillus (B10, B25, B143, B157, B171, B175), two Fusarium (F205, F281), and one Pseudomonas (B286) isolates are potentially efficient as biocontrol agents of H. vastatrix and will be tested using field experiments.     

Suppression of Fusarium wilt of spinach with hypovirulent binucleate <Emphasis Type="Italic">Rhizoctonia</Emphasis>

 Four isolates of hypovirulent binucleate Rhizoctonia (HBNR) were evaluated for their ability to control Fusarium wilt of spinach (FWS) caused by Fusarium oxysporum f. sp. spinaciae (FOS). Fourteen-day-old spinach seedlings grown in paper pots with HBNR-amended soil (1% w/w ground barley grain inoculum) were transferred to artificially pathogen-infested soil. Treatments with HBNR isolates significantly (P = 0.05) reduced disease and discoloration severity by 56%–100% and 52%–100%, respectively. The numbers of colony-forming units of FOS per gram fresh weight in petioles or roots were reduced significantly (P = 0.01) in the plants treated with HBNR. HBNR isolates were well reisolated from the roots inside paper pots where they were inoculated, whereas inconsistent colonization of HBNR was recorded from the roots outside paper pots where only pathogen was inoculated. Root extracts from HBNR-treated and pathogen-challenged plants significantly inhibited germination and germling length of FOS. The fresh weight of spinach leaves in the HBNR-treated plants increased significantly (P = 0.01), as much as 53%–63%, over the untreated and pathogen-challenged plants. This is the first report of biocontrol of FWS by HBNR. Received: July 18, 2002 / Accepted: October 22, 2002 Acknowledgments We are grateful to Dr. Komada for providing nonpathogenic Fusarium F13. The senior author (A.M.) thanks the Ministry of Education, Culture, Sports, Science, and Technology (Monbukagakusho) Japan, for financial assistance.     

生物源杀菌剂与化学药剂协调防控番茄病害

为了阐明化学药剂与生防菌剂及植物源杀菌剂交替使用对番茄病害发展的影响,田间试验检测了交替施药的防病效果及病害发展曲线下面积.结果表明,枯草芽孢杆菌BAB-1水剂及化学药剂交替喷施7次对番茄灰霉病、叶霉病、晚疫病和早疫病的防效分别为86.9%,61.0%,72.3%和77.2%.植物源杀菌剂1-氧-乙酰基大花旋覆花内酯(ABL)乳油与化学药剂及生防菌剂枯草芽孢杆菌BAB-1水剂交替喷施9次对上述病害的防效分别为88.6%,87.4%,85.4%和83.2%,相当于不同作用机制的化学药剂交替喷施9次的防效.此外,化学药剂与BAB-1水剂及ABL乳油交替喷施明显延缓了这4种病害的发展,减少了化学药剂施用次数.     

Efficacy of some rhizospheric and endophytic bacteria <Emphasis Type="Italic">in vitro</Emphasis> and as seed coating for the control of <Emphasis Type="Italic">Fusarium culmorum</Emphasis> infecting durum wheat in Tunisia

Sixty two rhizospheric and endophytic bacterial strains were evaluated for their biocontrol effect on two aggressive Fusarium culmorum isolates (Fc2 and Fc3). We observed that 35 % and 23 % of the tested strains inhibited the in vitro growth of Fc2 and Fc3 respectively. The observed antagonism was due to inhibition by contact (13–19 % of the strains) or at distance (10–16 % of the strains) for both fungal isolates. Some of the antagonistic bacteria showed the ability to produce diffuse and/or volatile compounds that inhibit the growth, the sporulation and macroconidia germination of F. culmorum. None of the tested antagonistic bacteria showed chitinase activity on synthetic medium. The sequencing of the 16S rDNA genes of some antagonistic bacteria showed that they belong to the genera Bacillus, Pseudomonas and Microbacterium. The double inoculation of durum wheat seeds by the antagonistic bacterial strains (B13, B18, BSE1, BSE3 and B16E) and the two F. culmorum isolates showed that germination and seedling vigor were generally improved in vitro. The percentage of infected seeds was also reduced. In greenhouse trials, the biocontrol effectiveness of F. culmorum was dependant from the virulence of the fungal strain and the specificity of the antagonistic interaction between bacterial and fungal strains. The bacterial strains B18 and B16E reduced F. culmorum infection on durum wheat plants probably due to their antagonistic and plant growth promoting activities and they may be used in a mixture as seed biopriming inoculum for plant growth bio-promoting and Fusarium wheat diseases biocontrol.     

A novel rhizobacterium Bk7 for biological control of brown sheath rot of rice caused by Pseudomonas fuscovaginae and its mode of action

The study presents the first report on biocontrol of brown sheath rot disease of rice caused by Pseudomonas fuscovaginae using rhizo-bacterial isolate Bacillus amyloliquefaciens Bk7. Four potential bioactive antagonists were selected from 120 Bacillus isolates. Results obtained from in vitro laboratory assay showed that rhizosphere bacterial strain Bk7 and its metabolites significantly suppressed the growth of Pseudomonas fuscovaginae with 93 % efficacy. In glasshouse experiments, strain Bk7 exhibited biocontrol efficacy of 76.6 % by reducing the disease incidence to 16.9 %, compared to 72.8 % observed in control treatment. In addition, the isolate Bk7 showed the growth promotion efficacy of plant height (GPE, 46.4 %) and fresh weight (GPE, 84.3 %). Characterization of isolate Bk7 revealed its strong capability for biofilm formation, inorganic phosphate solubilization and production of high amounts of Indole-3 acetic acid, siderophores and ammonia in vitro. Results obtained from multiplex PCR assay confirmed the presence of five lipopeptide biosynthetic gene markers (srfAA, fenD, bmyB, bacA and ituC) in the genome of strain Bk7. Moreover, Real-time qPCR of these genes demonstrated that surfactin, iturin and bacylisin coding genes were highly expressed in response to P. fuscovaginae exposure in vitro. Rhizosphere bacterial strain Bk7 was identified as B. amyloliquefaciens strain Bk7 based on the analysis of 16S rDNA internal transcribed spacer sequences and a fatty acid methyl ester analysis. The results obtained from this study showed the potential usefulness of Bk7 as a biocontrol agent in disease control of rice brown sheath rot.     

Sensitivity of Rhizoctonia solani causing rice sheath blight to fluxapyroxad in China

Fluxapyroxad,3-(difluoromethyl)-1-methyl-N-(3’,4’,5’-trifluorobiphenyl-2-yl) pyrazole-4-carboxamide (C₁₈H₁₂F₅N₃O, Fig. 1), is a SDHI fungicide, which is a new active ingredient that interferes with succinate ubiquinone reductase in the electron transport chain of fungi. Between 2008 and 2010, a total of 128 isolates of Rhizoctonia solani from Anhui Province of China were characterized for the baseline sensitivity to fluxapyroxad. The isolates collected between 2008 and 2010 all showed similar sensitivity to fluxapyroxad. Baseline sensitivity was distributed as unimodal curves with an average EC₅₀ value of 0.054?±?0.014 μg ml^?1. However, EC₅₀ values of boscalid for inhibition of mycelial growth of R. solani isolates ranged from 1.89 to 2.68 μg/ml and the average (±SE) EC₅₀ value was 2.212?±?0.228 μg/ml, indicating that the R. solani isolates were less sensitive to boscalid than that of fluxapyroxad. Fluxapyroxad exhibited excellent protective and curative activity against rice sheath blight and provided 82.6–94.2 % protective or curative control efficacy. In field trials, control efficacy of fluxapyroxad at 100 g a.i/ha 15 days and 30 day after second application was 83.4–88.0 %, suggesting excellent activity against sheath blight. Control efficacy of boscalid at a dosage of 600 g a.i/ha 15 days and 30 day after second application was about 51.7–57.0 %. There was a significant difference in the efficacy between fluxapyroxad and boscalid or jinggangmycin. These results suggested that fluxapyroxad is a good alternative fungicide to jinggangmycin for the control of rice sheath blight.