Digital image analysis to measure lesion area of cucumber anthracnose by Colletotrichum orbiculare

A method was developed using scanned computer images and software programs to measure the lesion area on leaves with cucumber anthracnose caused by Colletotrichum orbiculare. After cucumber plants were inoculated with various concentrations of conidia, lesions on diseased leaves were scanned, manipulated with Adobe Photoshop 6.0, and measured using the blob analysis feature in Matrox Inspector 2.2. The method requires relatively low-cost equipment including a scanner, a personal computer, and two programs: Adobe Photoshop 6.0 and Matrox Inspector 2.2. Because stored images are used, the lesion area can be measured as time permits. Processing the images requires about 3 min per sample. The image assessment accurately detected anthracnose lesions on cucumber leaves and could be applied to other foliar necrotic or spot diseases.     

A promising strain of endophytic <Emphasis Type="Italic">Streptomyces</Emphasis> sp. for biological control of cucumber anthracnose

In a survey for effective biocontrol agents of Colletotrichum orbiculare, causal agent of cucumber anthracnose, 43 (MBCu series) and 135 (MBPu series) endophytic actinomycete strains were recovered from surface-sterilized organs of cucumber (Cucumis sativus) and pumpkin (Cucurbita moschata) plants, respectively. The strains were cultured with C. orbiculare on IMA-2 agar medium to determine their in vitro antagonistic ability. Eleven strains that strongly inhibited hyphal growth of the pathogen were selected as potent antagonists. Detached cotyledons of cucumber were soaked in a spore suspension of an antagonistic strain 1 day before challenge-inoculation with the pathogen, and six of these antagonists (MBCu-32, 36, 42, 45, and 56, and MBPu-75) significantly reduced the number and size of the lesions on the cotyledons compared to the untreated control. In the same way, these six strains inhibited lesion development on attached leaves of 3-week-old cucumber seedlings. Strain MBCu-56, the most suppressive of the strains, was selected for further tests. Its suppressiveness increased as concentrations increased; pretreatment of leaves with the strain at 10⁷, 10⁸ and 10⁹ cfu/ml suppressed disease by 72, 79 and 93%, respectively. These results strongly indicated that MBCu-56 has strong potential for controlling cucumber anthracnose. Based on the taxonomic characteristics and 16S rDNA sequence, MBCu-56 was identified as Streptomyces sp. With scanning electron microscopy, the substrate mycelia of the strain were seen to colonize the surface of leaves above the cuticle. Some hyphae also penetrated and grew underneath the cuticle.     

黄瓜幼苗内生真菌诱导植株对南方根结线虫的抗性

在2007-2008年的一项温室测验中,10株黄瓜幼苗内生真菌表现出了对南方根结线虫较好的防效,为了阐明其作用方式,用裂根试验设计研究了10个菌株诱导黄瓜产生根结线虫抗性的能力.同一植株的根系被分为彼此隔离的挑战根系和应答根系,镰刀菌Fusarium spp.菌株Fu234和Fu654、毛壳菌Chaetomium sp.菌株Ch1001、叶点霉Phyllosticta sp.菌株Ph511在挑战根系中接种后,应答根系中的线虫入侵总数显著降低,较对照分别降低了42.4％、35.7％、38.4％、23.6％(第1次测定)和63.6％、45.2％、51.0％、37.0％(重复测定),这4个菌株诱导了黄瓜对根结线虫入侵的抗性.同时,Fu234、Fu654、Ch1001和拟青霉Paecilomyces sp.菌株Pa972接种后应答根系中的卵与雌虫的比值显著减少,相比对照其比值分别降低了24.3％、37.7％、48.3％、21.2％(第1次测定)和21.6％、39.8％、30.0％、46.2％(重复测定),这4个菌株诱导了黄瓜对根结线虫繁殖的抗性.     

Note:In vitro antagonism of actinomycetes isolated from fungus-growing ants against plant pathogenic fungi

Fungus-growing ants have been found recently to be symbiotic with actinomycetes living on the ant’s cuticle; these bacteria are inhibitory to soil fungi that are detrimental to the ants’ fungus gardens. In order to investigate whether actinomycetes found on the cuticle of attine ants also had inhibitory properties against plant pathogenic fungi, we isolated 32 strains of actinomycetes from fungus-growing ants (Atta, Trachymyrmex, andCyphomyrmex), from the Mexican states of Coahuila, Nuevo León and Tamaulipas. Of the actinomycetes tested against selected plant pathogenic fungi (Alternaria solani, Aspergillus flavus, Colletotrichum lindemuthianum, Rhizoctonia solani, Sclerotium sp.) on Czapek-Dox agar medium, 13 isolates inhibited at least one of the fungi.C. lindemuthianum was inhibited by 11 actinomycetes, andRhizoctonia by three. An actinomycete strain isolated fromCyphomyrmex rimosus inhibited all five fungi tested. http://www.phytoparasitica.org posting July 30, 2008.     

Systemic resistance to bacterial leaf speck pathogen in Arabidopsis thaliana induced by the culture filtrate of a plant growth-promoting fungus (PGPF) Phoma sp. GS8-1

The culture filtrate (CF) from the plant growth-promoting fungus Phoma sp. GS8-1 was found to induce systemic resistance in Arabidopsis thaliana against the bacterial leaf speck pathogen Pseudomonas syringae pv. tomato DC3000 (Pst), and the underlying mechanism was studied. Roots of A. thaliana were treated with CF from GS8-1, and plants expressed a clear resistance to subsequent Pst infection; disease severity was reduced, and proliferation of pathogen was suppressed. Various mutants of A. thaliana were used to test whether the CF induced resistance through one of the known signaling pathways: salicylic acid (SA), jasmonic acid (JA), and ethylene (ET). The CF was fully protective against Pst in Arabidopsis mutants jar1 and ein2 similar to wild-type plants. However, its efficacy was reduced in plants containing transgene NahG. Examination of systemic gene expression revealed that CF modulates the expression of SA-inducible PR-1, PR-2 and PR-5 genes, the JA/ET-inducible ChitB gene, and the ET-inducible Hel gene. Moreover, the expression of these genes was further enhanced upon subsequent stimulation after attack by Pst. Our data suggest that in addition to a partial requirement for SA, the signals JA and ET may also play a role in defense signaling in Arabidopsis.     

黄瓜枯萎病拮抗菌的拮抗促生因子检测及生物效应研究

以黄瓜枯萎病病原菌,尖孢镰刀菌黄瓜专化型(FOC,Fusarium oxysporum f.sp.cucumerinum)为靶标筛选拮抗菌。通过初筛和复筛,获得2株对尖孢镰刀菌黄瓜专化型拮抗效果稳定、拮抗能力较强的细菌QHZ-Yb1和QHZ-Yb2(以下简称,Yb1和Yb2),采用平板对峙法测定其拮抗作用,抑菌带宽度达3.37 mm和3.42 mm,抑菌圈直径2.7 cm,抑菌率分别达到61.9%和62.3%;通过形态学结合生理生化及菌株的16S rDNA序列分析,初步确定这两株菌均为萎缩芽孢杆菌(Bacillus atrophaeus);潜在拮抗促生因子的测定结果表明:菌株Yb1和Yb2具有多种拮抗促生因子,其脂肽类粗提取物的抑菌率分别达到57.4%和58.0%,生长素IAA的分泌量可达1.56 mg·L~(-1)和3.69 mg·L~(-1),溶P量为0.77 mg·L~(-1)和1.19 mg·L~(-1),两个菌株均可产嗜铁素、分泌酪蛋白酶;种子萌发试验结果表明:Yb1和Yb2菌液均可促进黄瓜种子的萌发;菌株生物效应的盆栽试验结果表明:Yb1、Yb2以及Yb1+Yb2菌悬液的处理不仅对黄瓜枯萎病有明显的生防效果,而且可显著增加黄瓜植株的生物量,增加黄瓜植株根际土中细菌和放线菌的数量,减少真菌和尖孢镰刀菌的数量,尤其以Yb1+Yb2菌悬液的处理效果最显著,对黄瓜枯萎病的发病率降低11.11个百分点,植株生物量增加56.7%。结论:菌株Yb1和Yb2具有生防潜力,两菌株配合运用效果更好。     

苯并烯氟菌唑对尖孢炭疽菌的毒力及对其所致辣椒炭疽病的防效

为评价苯并烯氟菌唑对辣椒炭疽病的防治潜力,采用菌丝生长速率法和孢子萌发法分别测定苯并烯氟菌唑对108株辣椒炭疽病病原菌尖孢炭疽菌Colletotrichum acutatum菌株菌丝生长和孢子萌发的抑制活性,通过辣椒活体接种试验比较苯并烯氟菌唑对辣椒炭疽病的相对保护防效和相对治疗防效,测定其对辣椒炭疽病的室内相对防效和田间相对防效。结果表明,苯并烯氟菌唑对尖孢炭疽菌孢子萌发的抑制活性高于对其菌丝生长的抑制活性,对孢子萌发及对菌丝生长的有效抑制中浓度EC₅₀分别为0.005～0.196μg/mL和0.073～3.398μg/mL;苯并烯氟菌唑对辣椒炭疽病的相对保护防效优于相对治疗防效,240 g (a.i.)/hm²苯并烯氟菌唑对辣椒炭疽病的相对保护防效可达85.64%。240 g (a.i.)/hm²苯并烯氟菌唑喷雾处理对辣椒炭疽病的田间相对防效均在75.31%以上,显著高于相同剂量的氟吡菌酰胺、氟啶胺和多菌灵的相对防效。总之,苯并烯氟菌唑对尖孢炭疽菌有较高毒力,对辣椒炭疽病有稳定的田间防效,有开发为防治辣椒炭疽病新药剂...     

AMF+PGPR组合提高黄瓜抗枯萎病的作用机制

对黄瓜(Cucumis sativus)品种香翠16接种尖孢镰刀菌黄瓜专化型(Fusarium oxysporum f.sp.cucumerinum,Foc)、丛枝菌根真菌(AMF)Funneliformis mosseae(Fm)+Foc、丛枝菌根真菌Glomus versiforme(Gv)+Foc、根围促生细菌(PGPR)PS2-6+Foc、PS3-2+Foc、Fm+PS3-2+Foc以及Gv+PS2-6+Foc,以不接种处理作为对照。结果表明,接种Foc条件下,Fm+PS3-2和Gv+PS2-6组合处理的抗病效果显著优于单接种AMF或PGPR处理;镜检观测到靠近AMF菌丝的Foc菌丝纤细,生长不良。AMF菌丝能缠绕Foc菌丝,并阻挡其侵入根系;AMF侵染率与其防效均呈正相关(P0.05);Fm+PS3-2+Foc和Gv+PS2-6+Foc处理的根系活力是单接种Foc的4.6倍;与不接种对照相比,Fm+PS3-2+Foc和Gv+PS2-6+Foc处理的黄瓜叶片中吲哚乙酸(IAA)含量分别提高了205%和166%,赤霉素(GA3)提高了33%和22%,玉米素核甘(ZR)提高了100%和80%,JA平均提高了65%,ABA降低23%和17%;而与单接种Foc处理相比,IAA提高了667%和568%,GA3提高了223%和196%,ZR提高了254%和219%,茉莉酸(JA)含量平均提高了215%,而脱落酸(ABA)含量只有单接种Foc处理的53%;Fm+PS3-2+Foc和Gv+PS2-6+Foc处理显著提高了黄瓜叶内苯丙氨酸解氨酶(PAL)防御酶活性和PAL基因表达量。综上推断AMF+PGPR组合是通过拮抗病原物、促进合成抗病信号物质、上调防御基因表达、提高防御酶活性和降低植株体内有毒物质等机制降低病害和改善植物抗病性。     

贝莱斯芽胞杆菌菌株HY19对多种作物病原真菌的拮抗和对烤烟赤星病的防治作用

为研制有效防治烤烟赤星病的高效安全生物农药,以云南省玉溪市烟草上筛选的贝莱斯芽胞杆菌Bacillus velezensis HY19为研究对象,通过对峙试验、发酵液和挥发性物质抑菌试验以及温室烤烟赤星病防治试验,研究该菌株对多种作物病原真菌的拮抗和对烤烟赤星病的防治作用。结果显示,在对峙培养中,菌株HY19能不同程度抑制烤烟黑胫病菌Phytophthora parasitica、松苗立枯病菌Rhizoctonia solani、辣椒疫霉病菌P. capsici、灰霉病菌Botrytis cinerea、小麦根腐病菌Bipolaris sorokiniana、黄瓜蔓枯病菌Mycosphaerella melonis、烤烟赤星病菌Alternaria alternata、柑橘炭疽病菌Colletotrichum gloeosporoides和油菜核盘菌Sclerotinia sclerotiorum的生长,使病菌菌丝分枝增多、膨大、畸形、扭曲,抑菌率为56.75%～100.00%。无菌发酵液显著抑制供试病菌生长和孢子萌发,增加菌丝丙二醛含量,降低蛋白质、超氧化物歧化酶和过氧化氢酶活性。菌株...     

Siderophore-mediated competition for iron and induced resistance in the suppression of fusarium wilt of carnation by fluorescent Pseudomonas spp

The mechanisms of suppression of fusarium wilt of carnation by two fluorescentPseudomonas strains were studied.Treatments of carnation roots withPseudomonas sp. WCS417r significantly reduced fusarium wilt caused byFusarium oxysporum f. sp.dianthi (Fod). Mutants of WCS417r defective in siderophore biosynthesis (sid^–) were less effective in disease suppression compared with their wild-type. Treatments of carnation roots withPseudomonas putida WCS358r tended to reduce fusarium wilt, whereas a sid^– mutant of WCS358 did not.Inhibition of conidial germination of Fod in vitro by purified siderophores (pseudobactins) of bothPseudomonas strains was based on competition for iron. The ferrated pseudobactins inhibited germination significantly less than the unferrated pseudobactins. Inhibition of mycelial growth of Fod by bothPseudomonas strains on agar plates was also based on competition for iron: with increasing iron content of the medium, inhibition of Fod by thePseudomonas strains decreased. The sid^– mutant of WCS358 did not inhibit Fod on agar plates, whereas the sid^– mutants of WCS417r still did. This suggests that inhibition of Fod by WCS358r in vitro was only based on siderophore-mediated competition for iron, whereas also a non-siderophore antifungal factor was involved in the inhibition of Fod by strain WCS417r.The ability of thePseudomonas strains to induce resistance against Fod in carnation grown in soil was studied by spatially separating the bacteria (on the roots) and the pathogen (in the stem). Both WCS417r and its sid^– mutant reduced disease incidence significantly in the moderately resistant carnation cultivar Pallas, WCS358r did not.It is concluded that the effective and consistent suppression of fusarium wilt of carnation by strain WCS417r involves multiple mechanisms: induced resistance, siderophore-mediated competition for iron and possibly antibiosis. The less effective suppression of fusarium wilt by WCS358r only depends on siderophore-mediated competition for iron.     

Tobacco cultivars vary in induction of systemic resistance against <Emphasis Type="Italic">Cucumber mosaic virus</Emphasis> and growth promotion by <Emphasis Type="Italic">Pseudomonas chlororaphis</Emphasis> O6 and its <Emphasis Type="Italic">gacS</Emphasis> mutant

The colonization of plant roots with certain rhizosphere bacteria promotes plant growth and induces long lasting systemic protection against a broad spectrum of plant pathogens. The role of the global regulator, GacS, in the rhizosphere colonist Pseudomonas chlororaphis O6 in stimulating growth promotion and induced resistance against Cucumber mosaic virus was examined in tobacco. Responses were compared in tobacco cvs Samsun and GX3. Root colonization of Samsun with wild-type O6 and the gacS complemented mutant-elicited reduced viral symptoms and viral titre. On GX3, there was little affect on symptoms when roots were colonized by the wild-type, gacS mutant or complemented mutant but colonization by both the wild-type and the gacS mutant lowered viral titre. Wild-type O6 and the gacS mutant caused plant growth to be maintained in both tobacco cultivars after viral infection, although the affect was stronger with GX3 than Samsun. In contrast, although a chemical inducer, benzothiadiazole, reduced symptoms and viral titre in both cultivars, plant growth was suppressed. Our results indicate rhizobacteria-elicited induced viral resistance without a negative impact on growth but there was a differential response between cultivars. Detailed knowledge regarding the mechanisms inherent to these differences between cultivars requires further investigation.