2种昆虫病原线虫共生菌的代谢产物对苹果病原菌真菌的抑菌作用

嗜线虫致病杆菌(Xenorhabdus nematophila HB310)和发光杆菌(Photorhabdus luminescens HB140)是分别从小卷蛾斯氏线虫和异小杆线虫中分离到的2株昆虫病原线虫共生菌。为了明确这2种共生菌的抑菌活性,分别测定了其对苹果上6种主要病原真菌的抑菌活性。结果表明,这2种共生菌对6种苹果病原真菌均有不同程度的抑制作用。嗜线虫致病杆菌HB310菌株发酵液对苹果轮纹病病原菌抑制效果最好,抑制率达76.9%,对其他病原菌的抑制作用依次为苹果树腐烂病病原菌(74.1%)>苹果斑点落叶病病原菌(60.9%)>苹果灰霉病病原菌(46.7%)>苹果青霉病病原菌(44.5%)>苹果炭疽病病原菌(23.0%);发光杆菌HB140菌株发酵液对苹果灰霉病病原菌的抑制作用最好,抑制率达到86.7%,对其他病原菌的抑制作用依次为苹果树腐烂病病原菌(76.6%)>苹果轮纹病病原菌(58.8%)>苹果斑点落叶病病原菌(46.0%)>苹果青霉病病原菌(44.2%)>苹果炭疽病病原菌(39.9%)。2种共生菌发酵液对苹果轮纹病菌均有较强的抑制作用,其EC50分别为42.28和42.92 mL/L。     

50％多·锰锌可湿性粉剂防治苹果主要病害的应用技术研究

50%多·锰锌可湿性粉剂500、600、700倍液室内对苹果轮纹病、炭疽病菌菌丝生长及孢子萌发的抑制效果均为100% ,对斑点落叶病菌菌丝生长及孢子萌发的抑制效果在70.21%以上优于单剂代森锰锌、多菌灵的效果 ;其对上述3种病菌的抑制中量 (EC50)小于单剂 ,共毒系数均>200,增效显著。田间对斑点落叶病和褐斑病的防效在81.12%以上 ,显著优于单剂 ,对轮纹病、炭疽病生长期及贮藏期的防治效果分别为97%～99.2%、70.9%～87.6%和87.1%～95.5%、50.5%～86.2% ,优于单剂代森锰锌的效果 ,优于或相当于多菌灵的效果。     

苹果树的几种常见病害及其防治对策

在泰安地区，苹果栽培品种主要有富土、新红星、乔纳金、嘎啦和金冠。据1997～1999年调查发现苹果斑点落叶病、轮纹病和炭疽病等几种病害发生严重且呈逐年上升趋势。一般果园平均发病率为30％～50％，严重者可达70％，造成了巨大经济损失。1病原及为害　　1)斑点落叶病：该病病原菌主要是轮斑病菌(Alternariamali)的强毒株系。此病病原性很强，无伤接种可直接侵染叶片。菌丝体发育最适温度28℃，最适pH4.5～6。以菌丝体在病落叶上越冬，还可在枝上病斑、皮孔及芽鳞等处越冬。该病菌可以为害叶片、枝条和果实。在叶片上形成具紫色晕圈的病斑…     

一株内生拮抗细菌的分离鉴定及其抗菌机理研究

本研究通过平板稀释法、平板对峙法从核桃根部皮层筛选出一株对核桃根腐病有拮抗作用的内生菌株1A,通过形态特征测定、理化特性分析和16SrDNA序列分析鉴定该菌为芽胞杆菌(Bacillus sp.),GenBank序列登录号为KJ865856;其抑菌特性研究结果表明:该拮抗菌对核桃根腐病菌有显著的抑制效果,抑菌率达63.33%;此外,该菌对板栗疫病菌等几种不同的林木病原菌均有较强的抑制作用。抗生机理研究结果表明,该菌株通过分泌蛋白酶和纤维素酶降解真菌细胞壁中蛋白质和纤维素,破坏病菌菌丝,病菌生长受到抑制。该拮抗菌能够显著抑制核桃根腐病菌菌丝生长和孢子萌发,孢子萌发抑制率达74.89%,病菌菌丝表现为菌丝扭曲、断裂,分支增多并缠绕,菌丝颜色加深等。盆栽生防效果研究显示,该拮抗菌对各组核桃根腐病具有显著的防治效果,采用该拮抗菌预处理具有相对较好的生防效果。     

拮抗细菌BRF-1对几种植物病原真菌的抗生效果

从大豆根际土壤分离到一株具有拮抗作用的细菌BRF—l,经扫描电镜观察和耐高温试验,初步鉴定为芽孢杆菌。对峙培养试验表明,该菌对大豆根腐病菌、南瓜疫霉病菌、大豆立枯病菌、烟草赤星病菌、小麦赤霉病菌、水稻立枯病菌和小麦根腐病菌等7种病原其菌具有较强的抗菌活性。BRF-1的培养滤液可抑制病原菌菌丝生长和孢子萌发,具有一定的耐酸、碱及高温特性。盆栽接种结果表明,该菌可促进大豆、小麦幼苗生长;在豆长连、重一年和正茬土壤上,对大豆根腐病的防治效果分别达53．9％、34．4％和15．8％。     

稻曲病菌拮抗菌的筛选及拮抗活性测定

从对稻曲病菌具有拮抗能力的18株芽孢杆菌中选出拮抗作用较强的2个菌株A1、H-51,分别对稻曲病菌进行抑菌率、致畸性、孢子萌发影响以及对水稻促生性试验。结果表明,拮抗菌2个菌株的发酵液对稻曲病菌最大抑制率分别为67.66％和62.87％,对病菌分生孢子的萌发抑制率分别为77.37％和59.74％,但对水稻促生作用不明显。     

枯草芽孢杆菌BS80-6对苹果采后炭疽病的控病效果及作用机制

为提高枯草芽孢杆菌的生防效果,通过离子束诱变筛选出BS80-6菌株.利用生物测定方法研究BS80-6菌株及不同处理方法在不同贮藏温度下对采后苹果炭疽病的控制效果和作用机制.结果表明,BS80-6能显著提高对苹果炭疽菌的抑制作用.枯草芽孢杆菌BS80-6对苹果果实炭疽病的防治效果以活菌液效果最好,菌丝生长抑制率为80.14%,孢子萌发抑制率为98.65%,控病效果为60.34%;灭菌液和过滤液对苹果炭疽菌也有一定的抑制效果.贮藏温度明显影响BS80-6对苹果炭疽病的防治效果;接种方式对控病效果影响显著,先接拮抗菌再接病菌的防效好于先接病菌再接拮抗菌.     

短小芽胞杆菌AR03对烟草炭疽病的抑制作用

为测定短小芽胞杆菌AR03菌株对烟草炭疽病菌的室内拮抗活性和盆栽防治效果,采用平板拮抗法初步筛选菌株拮抗活性,采用抑制菌丝生长法及抑制孢子萌发法筛选该菌株菌悬液的抑菌浓度,采用喷雾法进行防治效果测定。AR03菌株对烟草炭疽病菌的拮抗活性很强,能显著抑制菌丝生长、孢子萌发和病害发生,浓度为1×107 cfu/mL的AR03发酵液能有效抑制菌丝的生长和孢子萌发,AR03菌悬液(1×108 cfu/mL)对炭疽病的盆栽防治效果为83.03%,与对照药剂25%三唑酮可湿性粉剂的防治效果相当。AR03菌株是一株优良的广谱高效生防菌株。     

海南辣椒炭疽病病原菌鉴定及其对嘧菌酯的敏感性分析

2015年3月调查了海南省海口、琼海、万宁、儋州等市(县)的辣椒炭疽病的发生与病情,对采集的样本分离出的7株纯菌株进行了致病性测定及形态学观察,通过生长速率法及孢子萌发抑制法分别测定嘧菌酯对辣椒炭疽病病原菌菌丝和孢子的影响。结果表明,菌株DM4-2和DZ1-3为黑点炭疽菌(Colletotrichum capsici),菌株NP3-9、NP5-3、DZ2-7、HN5-11和HN2-6均为胶胞炭疽菌(C.gloeosporioides)。各菌株对嘧菌酯的敏感性显示,相对于菌丝,其病菌孢子对嘧菌酯的敏感性更高;相对于胶胞炭疽菌,黑点炭疽菌菌株DM4-2和DZ1-3对嘧菌酯更为敏感。     

海南省不同寄主植物炭疽病菌侵染油茶研究

为了明确海南省不同寄主植物炭疽病菌侵染油茶的潜力,本研究分离、鉴定海南省4个油茶林中不同寄主植物的炭疽病菌种类及其致病性。结果从油茶、葛根、山麻杆、飞机草、蒲葵、鲫鱼胆、桃花心木、黄兰、羊蹄甲、山矾、银柴、降香黄檀和檀香等13种植物上,共获得135株炭疽病菌,其中果生刺盘孢菌(Colletotrichum fructicola)85株、暹罗刺盘孢菌(C.siamense)45株,盘长孢状刺盘孢菌(C.gloeosporioides)3株,君子兰刺盘孢(C.cliviae)和山茶刺盘孢(C.camelliae)各1株;4个采样地的油茶树与葛根等当地其他12种植物具有相同的炭疽病菌C.fructicola和C.siamense,这两种菌占总分离炭疽菌的96%;通过致病性测定,不同寄主来源的这5种炭疽菌均能引起油茶炭疽病。海南省当地多种植物的炭疽病菌是油茶的潜在侵染源。     

Differential Inhibition of Sphaeropsis sapinea Morphotypes by a Phenolic Compound and Several Monoterpenes of Red Pine

ABSTRACT The in vitro effects of a red pine phenolic compound (pinosylvin), a phenolic compound common to other species (tannic acid), and the major red pine monoterpenes (alpha-pinene, beta-pinene, and delta-3-carene) on spore germination and mycelial growth of Sphaeropsis sapinea were examined. Two A and two B morphotype isolates were used. At 88 mug/mm(2), pinosylvin inhibited spore germination of all four isolates (98 to 100%). At 8.8mug/mm(2), spore germination of B isolates was inhibited more than that of A isolates (73 versus 30%). Pinosylvin also inhibited mycelial growth of B isolates more than that of A isolates (84 versus 13% at 88 mug/mm(2)). Tannic acid stimulated or had little affect on spore germination and had little affect on mycelial growth of either morphotype. Spore germination of B isolates was inhibited more than that of A isolates by beta-pinene at saturation (79 versus 37%). Spore germination of B isolates was inhibited and germination of A isolates was stimulated by delta-3-carene below saturation (49 versus -7%). Mycelial growth of B isolates was inhibited more than that of A isolates by all monoterpenes at saturation. Differences observed between morphotypes below saturation were significant only for beta-pinene. These results demonstrate the biological activity of a phenolic compound and monoterpenes that occur in red pine. The differential responses might provide means of distinguishing morphotypes and offer a potential explanation for ecological specialization.     

模拟酸雨对小麦条锈病流行学组分的影响

为明确酸雨对小麦条锈病菌孢子萌发及病害流行学组分的影响,采用不同酸度的模拟酸雨(p H值分别为2.5、3.5、4.5和5.6)处理小麦条锈病菌Puccinia striiformis f.sp.tritici生理小种CYR32和CYR33的夏孢子,并将不同酸度模拟酸雨处理后的夏孢子接种小麦感病品种铭贤169,采用组分分析法研究酸雨对病害发展过程的影响。结果表明,随着模拟酸雨酸度的增强,生理小种CYR32和CYR33的夏孢子萌发率均显著降低,萌发24 h后对照组生理小种CYR32和CYR33的夏孢子萌发率分别为93.7%和79.8%,在p H 3.5模拟酸雨处理后萌发率仅为13.1%和8.6%。温室内接种试验结果显示,不同酸度模拟酸雨对生理小种CYR32和CYR33的侵染概率、潜育期和病情指数均有显著影响,对生理小种CYR32和CYR33病斑扩展率影响不显著;与对照相比,p H 3.5模拟酸雨处理后生理小种CYR32和CYR33的侵染概率分别下降79.9%和79.8%。结果表明强酸雨(p H 3.5)显著延长小麦条锈菌潜育期,减少病斑产孢量,降低病情指数和病害进展曲线下面积。     

种子细菌处理防治番茄苗期病害

11984～1986年从杭州市郊黄瓜、番茄、青椒,大白菜等健苗根表及根围土壤分离得1000多株菌株,从中筛选出8个拮抗细菌防治番茄苗期病害。1986～1987年在温州市扩大试验,种子细菌处理后,能抑制发瘸率,提高出苗率拈抗效果在60％以上,最高达100％;还能促进植株生长,提高单株产量。用化学农药甲霜灵与代森锌混配处理,得到同样防治效果。     

Effect of Microsphaeropsis sp. Strain P130A on Germination and Production of Sclerotia of Rhizoctonia solani and Interaction Between the Antagonist and the Pathogen

Microsphaeropsis sp. strain P130A was evaluated for the control of tuber-borne inoculum of Rhizoctonia solani based on the viability of sclerotia produced in vitro and on both the viability and production of tuber-borne sclerotia. The interactions between the antagonist and the pathogen, as well as the effect of the toxins produced by the antagonist on mycelial growth of R. solani were studied using transmission electron microscopy. On sclerotia produced in vitro, for all incubation periods (1 to 42 days), Microsphaeropsis sp. significantly reduced germination. Percent germination of sclerotia treated with Microsphaeropsis sp. decreased with increasing incubation period from an average of 82.0% after 1 day to stabilize at an average of 5.8% after 35 days. Similarly, percent germination of tuber-borne sclerotia was significantly lower when tubers were treated with Microsphaeropsis sp. Both 2% formaldehyde and Microsphaeropsis sp. treatments significantly reduced sclerotia germination to approximately 10% after 42 days of incubation at 4 degrees C. Furthermore, on tubers treated with the antagonist, the number of sclerotia per square centimeter decreased from 1.6 to 0.5 during the 8 months of storage at 4 degrees C, whereas an increase from 1.2 to 7.8 sclerotia per square centimeter was observed on untreated tubers. Microsphaeropsis sp. (strain P130A) colonized hyphae of R. solani within 4 days after contact on culture media. Transmission electron microscopic observations showed that the antagonist induced a rupture of the pathogen plasma membrane and that a chitin-enriched matrix was deposited at sites of potential antagonist penetration. Host penetration was not associated with pathogen cell wall alterations, which occurred at the time of progress of the antagonist in the pathogen cytoplasm. In the presence of a crude extract of Microsphaeropsis sp., cells of R. solani showed cytoplasm disorganization and breakdown of plasma membranes. Antibiosis and mycoparasitism were involved in the antagonism of R. solani by Microsphaeropsis sp., but the sequence by which these events occur, as well as the significance of wall appositions produced by R. solani, is yet to be established.     

东亚飞蝗肠道细菌鉴定及其对金龟子绿僵菌拮抗作用分析

通过对室内饲养的东亚飞蝗5龄若虫肠道细菌的分离纯化培养,共获得8个细菌菌株,16s rDNA鉴定分别与绿脓杆菌(Pseudomonas aeruginosa)、克雷伯氏菌属(Klebsiellasp.)、地衣芽孢杆菌(Bacillus licheniformis)、阴沟肠杆菌(Enterobacter cloacae)、节杆菌属(Arthrobactersp.)、木糖葡萄球菌(Staphylococcus xylosus)、蜡状芽孢杆菌(Bacillus cereus)和芽孢杆菌属(Bacillussp.)有较高的同源性,其中克雷伯氏菌和阴沟肠杆菌检出率为100%,检出量最大,均达到109cfu/g肠道,为常驻的优势菌群。肠道细菌在东亚飞蝗体内的数量分布为后肠>中肠>前肠,种类分布为中肠(8种)>后肠(6种)>前肠(4种)。常驻细菌克雷伯氏菌和阴沟肠杆菌对金龟子绿僵菌则无拮抗作用,表明肠道常驻菌不是影响金龟子绿僵菌在蝗虫肠道内萌发侵染的主要因素,但同时也说明了在不同环境条件下使用绿僵菌防治蝗虫在防效上可能会有很大的差异。     

Influence of native endophytic bacteria on the growth and bacterial crown rot tolerance of papaya (Carica papaya)

The native plant microbiome is composed of diverse microbial communities that influence overall plant health, with some species known to promote plant growth and pathogen resistance. Here, we show the antibacterial and growth promoting activities of autoclaved culture metabolites (ACM) from native endophytic bacteria (NEB). These NEB were isolated from a papaya cultivar (var. Cariflora) that is tolerant to bacterial crown rot (BCR) caused by Erwinia mallotivora. In this cultivar, bacterial colonization in tissues recovering from the disease was observed before onset of tissue regeneration or ‘regrowth’. We further isolated and characterized these bacteria and were able to identify two culturable stem NEB related to plant endophytic genera Kosakonia sp. (ex. Enterobacter sp., isolate EBW), and to Sphingomonas sp. (isolate EBY). We also identified root NEB under genus Bacillus (isolates BN, BS, and BT). Inhibition assays indicated that ACM from these NEB promptly (within 18-30 h) and efficiently inhibited (60–65% reduction) E. mallotivora proliferation in vitro. When surface-sterilized papaya seeds were soaked in ACM from isolates EBY and EBW, germination was variably retarded (20–60% reduction) depending on plant genotype, but plant biomass accumulation was significantly stimulated, at around two-fold increase. Moreover, greenhouse experiments show that ACM from all isolates, especially isolate EBW, significantly reduced BCR incidence and severity in a susceptible genotype (var. Solo), at around two-fold. In general, our observations of pathogen antagonism and plant growth promotion leading to disease reduction, suggested the influence of native endophytic bacteria to increased fitness in plants, and tolerance against the re-emerging crown rot disease of papaya.

     

Effect of surface wettability on germination and gene expression in Cronartium quercuum f. sp. fusiforme basidiospores

Cronartium quercuum f. sp. fusiforme is an obligate pathogen of pine and oak. Its basidiospores are specifically adapted to recognize and establish infections on the pine host. Depending on environmental cues, the basidiospores can germinate directly, which typically leads to infection of pine, or indirectly, which usually results in formation of secondary basidiospores. We investigated how changes in surface wettability, or hydrophilicity, affect basidiospore germination. When we decreased surface wettability, the direct germination and total germination (direct+indirect) frequencies increased. Additionally, there was a critical threshold between 42% and 54% wettability, at which the basidiospores switched from direct to indirect germination. We conclude that the germination type of C. q. fusiforme basidiospores is influenced by the hydrophilicity of the surface upon which they land. To gain insight into gene expression during basidiospore germination, we made two suppression subtraction hybridization (SSH) libraries enriched for genes expressed during each type of germination. A total of 172 unique gene sequences were recovered from the two expression libraries. Annotation of these libraries indicates that they include several clones that may encode rust-specific or basidiomycete-specific functions.     

香蕉枯萎病拮抗菌的筛选及其作用机制研究

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。     

Association of the pitch canker fungus, Fusarium subglutinans f.sp. pini, with Monterey pine seeds and seedlings in California

The causal agent of pitch canker disease of pines, Fusarium subglutinans f.sp. pini , is a recent introduction to California. The work presented here demonstrates that this pathogen is seed-borne in Monterey pine ( Pinus radiata ). The pathogen was isolated from up to 83% of seeds collected from cones on recently infected branches. Seedling emergence from infested Monterey pine seeds was 9%, compared with 67% for uninfested seeds. The fungus was isolated more frequently from seedlings originating from diseased branches than from symptomless branches. However, more than 50% of seeds from symptomless Monterey pine branches produced symptomless seedlings from which the pathogen was isolated. It is hypothesized that F. s . pini is carried within the seeds, where it may remain dormant until germination. The implications of these findings for the regeneration of native and landscape-planted Monterey pines, and for the transport of this non-native pathogen in pine seeds and seedlings to previously uninfested areas, are discussed.     

胡杨根际可培养细菌的分离及抑制瓜列当种子萌发研究

从胡杨生存的干旱强光照逆境中获得新的细菌资源,从中筛选新的可抑制瓜列当种子萌发的生防菌。采用3种分离培养基,以稀释涂布法对塔里木河流域3处胡杨根系样品进行可培养细菌分离,通过分析分离菌株16S rRNA基因序列确定其种属分类。采用皿内培养法测定其对瓜列当种子萌发的抑制活性,利用盆栽试验检测分离菌株M2-3对瓜列当寄生的防治效果。结果分离获得98株细菌,分属于28个属,马赛菌属Massilia、Pontibacter、玫瑰单胞菌属Roseomonas为优势菌属。其中,菌株M2-3与Pontibacter salisaro HMC5104T的相似度最高,为97.92%,其菌体发酵液10倍稀释液对瓜列当种子萌发抑制率为87.8%,盆栽试验表明该菌株对瓜列当防效可达36.9%,具备进一步开发可能性。