PGPR and entomopathogenic fungus bioformulation for the synchronous management of leaffolder pest and sheath blight disease of rice

BACKGROUND: The biological control of plant pests and diseases using a single organism has been reported to give inconsistent and poor performance. To improve the efficacy, bioformulations were developed possessing mixtures of bioagents. RESULTS: Bioformulations combining Pseudomonas fluorescens Migula strains Pf1 and AH1 and Beauveria bassiana (Balsamo) Vuill. isolate B2 were developed and tested for their efficacy against leaffolder pest and sheath blight disease on rice under glasshouse and field conditions. The combination of Pf1, AH1 and B2 effectively reduced the incidence of leaffolder insect and sheath blight disease on rice compared with other treatments. An in vitro assay of leaffolder preference to rice leaf tissues treated with Pf1 + AH1 + B2 biformulation showed variation from normal growth and development of leaffolder larvae. Plants treated with the Pf1 + AH1 + B2 combination showed a greater accumulation of enzymes, lipoxygenase and chitinase activity against leaffolder insect compared with other treatments. Similarly, the plants showed a higher accumulation of defence enzymes, peroxidase and polyphenol oxidase activity against sheath blight pathogen in Pf1 + AH1 + B2 treatment compared with the untreated control. The bioformulation mixture attracted the natural enemy population of leaffolder under field conditions. In addition, a significant increase in rice grain yield was observed in Pf1 + AH1 + B2 treatment compared with the untreated control. CONCLUSION: The combination of P. fluorescens strains and B. bassiana isolate effectively reduced the incidence of leaffolder insect and sheath blight disease on rice plants and showed the possibility of controlling both pest and disease using a single bioformulation. Copyright © 2010 Society of Chemical Industry     

Plant growth promoting rhizobacteria (PGPR) and entomopathogenic fungus bioformulation enhance the expression of defense enzymes and pathogenesis-related proteins in groundnut plants against leafminer insect and collar rot pathogen

The bioformulation of Pseudomonas fluorescens (Pf1 and TDK1) and Beauveria bassiana (B2) strains was evaluated individually and in combinations with and without chitin for their efficacy against leafminer insect and collar rot disease and the effect of the interaction between Pseudomonas, Beauveria and groundnut leafminer insect and collar rot pathogen in the expression of defense enzymes and pathogenesis-related proteins (PR-proteins) in groundnut. Among the various bioformulations, B2 + TDK1 + Pf1 (amended with or without chitin) formulation significantly reduced the incidence of leafminer and collar rot disease when compared to untreated control. A significant increase in the enzymatic activity of phenylalanine ammonia-lyase, peroxidase, polyphenol oxidase, chitinase, β-1,3-glucanase, superoxide dismutase, catalase, lipoxygenase, and phenolics in groundnut plants treated with B2 + TDK1 + Pf1 bioformulation (amended with or without chitin) and challenge inoculated with Aproaerema modicella and Sclerotium rolfsii. Native gel electrophoresis also revealed the expression of more isoforms of pathogenesis-related proteins and other defense enzymes viz., polyphenol oxidase and superoxide dismutase in plants treated with B2 + TDK1 + Pf1 mixture challenged with A. modicella and S. rolfsii. The present study reveals that sustained and timely induction and accumulation of these defense enzymes and PR-proteins enhance the resistance in groundnut against leafminer insect and collar rot disease.     

Induction of systemic resistance in banana (<Emphasis Type="Italic">Musa</Emphasis> spp.) against <Emphasis Type="Italic">Banana bunchy top virus</Emphasis> (BBTV) by combining chitin with root-colonizing <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis> strain CHA0

Pseudomonas fluorescens strains CHA0 and Pf1 were investigated for their biocontrol efficacy against Banana bunchy top virus (BBTV) in banana (Musa spp.) alone and in combination with chitin under glasshouse and field conditions. Bioformulation of P. fluorescens strain CHA0 with chitin was effective in reducing the banana bunchy top disease (BBTD) incidence in banana under glasshouse and field conditions. In addition to disease control, the bioformulation increased the economic yield significantly compared to the untreated control. Increased accumulation of oxidative enzymes, peroxidase (PO), polyphenol oxidase (PPO), phenylalanine ammonia lyase (PAL), pathogenesis-related (PR) proteins, chitinase, β-1,3-glucanase and phenolics were observed in CHA0 bioformulation amended with chitin-treated plants challenged with BBTV under glasshouse conditions. Indirect ELISA indicated the reduction in viral antigen concentration in P. fluorescens strain CHA0 with chitin-treated banana plants corresponding to reduced disease ratings. The present study revealed that induction of defence enzymes by P. fluorescens with chitin amendment reduced the BBTD incidence and increased bunch yield in banana.     

河南省水稻穗期稻纵卷叶螟的危害损失与防治指标的研究

1979～1983年在郑州采用田间笼罩接虫和大田调查的方法,进行了穗期稻纵卷叶螟危害损失的研究。试验资料经协方差分析导出:虫量(X)与卷叶率(R)之关系式为:R=0.95+0.082X±0.77;卷叶率(R)与损失率(Y)之关系式为:Y=-13.09+21.69lgR±3.99;虫量(X)与损失率(Y)之关系式为:Y=-30.92+20.63lgX±4.65在经济允许损失水平为2%时,其防治指标为百丛三龄幼虫40～50头,卷叶率为5%,并根据各地经济允许损失水平和所用农药种类的防效不同,建立了计算防治指标的数学模型。     

水稻病虫危害损失和防治效益评估研究初报

2005-2008年笔者在早稻、晚稻生长期间进行防治与不防治对比试验。分析试验数据得出:全程不防治平均减产51.26%,常规防治平均增产113.14%,常规防治平均收益4498.58元/hm2,比全程不防治增86.12%,平均投入产出比1:5.18。拟合出单虫或单病在定局时的发生量(或发生率)与其危害损失之间的线性回归模型:稻飞虱每丛虫量(X)与损失率(Y)之间关系式为Y=8.7654+0.0881X;稻纵卷叶螟卷叶率(X)与损失率(Y)之间关系式为Y=1.9341+0.2851X;二化螟发生率(X)与损失率(Y)之间关系式为Y=1.4182+1.0407X;稻瘟病白穗率(X)与损失率(Y)之间关系式为Y=0.3765+0.9004X;纹枯病病株率(X)与损失率(Y)之间关系式为Y=-2.4469+0.2829X,相关分析均达极显著水平。     

Fusarium oxysporum f. sp. lycopersici retardation through induction of defensive response in tomato plants using a liquid formulation of Pseudomonas fluorescens (Pf1)

A liquid based Pseudomonas fluorescens (Pf1) bioformulation was found to contribute the restriction of Fusarium oxysporum f. sp. lycopersici in tomato roots by inducing defence enzymes. Induction of defence enzymes such as phenylalanine ammonia lyase (PAL), peroxidase (PO), polyphenoloxidase (PPO), catalase, β-1,3 glucanase and super oxide dismutase (SOD), was studied in tomato plants pretreated with liquid as well as a talc based formulation of Pf1 challenged with F. oxysporum f. sp. lycopersici in glasshouse vegetable production systems. There were increased activities of PAL, PO, PPO, catalase and β-1 3-glucanases in tomato plants treated with a combined application of seedling dip?+?soil application?+?foliar spray of liquid and talc formulation of Pf1 when compared to pathogen inoculated and untreated healthy controls. The activities of the above enzymes started to increase at 3rd day, reached maximum levels on 8-9th day and thereafter declined gradually. Similarly, native polyacralamide gel electrophoresis (PAGE) analysis revealed that one to six isoforms of the defence enzymes each with a higher intensity were expressed in these treatments, whereas fewer isoforms with less intensity were noticed in inoculated controls. These results suggest that the retardation of the invasion of F. oxysporum f. sp. lycopersici in tomato roots resulting from treatment with the liquid formulation of Pf1 was due to enhancement of activities of enzymes involved in the phenylpropanoid pathway. These results suggest that induced systemic resistance occurred in the treated tomato plants.     

Dominant Colonisation of Wheat Roots by Pseudomonas fluorescens Pf29A and Selection of the Indigenous Microflora in the Presence of the Take-all Fungus

Increases in populations of fluorescent pseudomonads on wheat roots are usually associated with take-all decline, natural control of take-all, a disease caused by the fungus Gaeumannomyces graminis var. tritici (Ggt). Colonisation by Pseudomonas fluorescens strain Pf29A was assessed on the roots of healthy plants and of plants with take-all, and the effect of this bacterium on indigenous populations of fluorescent pseudomonads was studied. The efficacy of Pf29A as an agent for the biocontrol of take-all on five-week-old wheat seedlings was tested in non-sterile conducive soil in a growth chamber. RAPD (random amplification of polymorphic DNA) fingerprinting with a decamer primer was used to monitor strain Pf29A and culturable indigenous rhizoplane populations of fluorescent pseudomonad. Pf29A decreased disease severity and accounted for 44.6% of the culturable fluorescent pseudomonads on healthy plant rhizoplane and 75.8% on diseased plant rhizoplane. Fewer RAPD patterns were obtained when Pf29A was introduced into the soil with Ggt. In the presence of Ggt and necrotic roots, Pf29A became the dominant root coloniser and dramatically changed the diversity and the structure of indigenous fluorescent pseudomonad populations. The results show that Ggt and reduced lesion size on roots can trigger a specific increase in antagonist populations and that the introduction of a biocontrol agent in soil influences the structure of indigenous bacterial populations.     

甲胺磷对稻纵卷叶螟及其天敌的影响

实验室、田间试验表明 ,甲胺磷对稻纵卷叶螟及其寄生蜂、捕食性天敌等均有不同程度的杀伤作用。各代推迟喷药、放宽防治指标可提高纵卷叶螟幼虫被寄生率 ,对产量并无影响。结合前期施药对褐飞虱再增猖獗的促进作用 ,建议在水稻生长前期谨慎用药 ,尤其应限制甚至禁止在稻田使用甲胺磷。     

Interaction ofPseudomonas fluorescens withRhizobium for their effect on the management of peanut root rot

A biocontrol agent (Pseudomonas fluorescens) and a phytostimulator (Rhizobium) have been shown to have beneficial effects on plant growth and health. The study of plants inoculated withPseudomonas andRhizobium requires special attention because of the possibility that these agents may influence each other. Our study was conducted to test the effect of these inoculants on co-inoculation in peanut to control root rot, a severe soilborne disease caused byMacrophomina phaseolina. One fluorescent pseudomonad strain, Pf 1, which effectively inhibited the mycelial growth ofM. phaseolina underin vitro conditions, was studied for its compatibility with the biofertilizer bacterial strainRhizobium TNAU 14. Dual culture and colorimetric studies indicated the existence of a positive interaction between the microbial inoculants. However, glasshouse and field studies showed seed treatment and soil application ofPseudomonas fluorescens Pf 1 to be the most effective treatment in reducing root rot incidence and improving the crop vigor index, in comparison with treatments in which both inoculants were applied. http://www.phytoparasitica.org posting Feb. 11, 2002.     

Impact of nitrogenous-fertilization on the population dynamics and natural control of rice leaffolders (Lep.: Pyralidae)

The effect of nitrogenous-fertilization on the population dynamics and natural control of rice leaffolders was studied in an irrigated rice area in the Philippines. Nitrogen was applied at three levels (0, 75 and 150 kg N ha^-1), and its impact on crop growth and yield, arthropod abundance, and rates of leaffolder parasitism and survival was assessed with weekly samples. Rice plants were taller and had a higher leaf nitrogen content with increasing levels of nitrogenous fertilization, but grain yield was highest at the medium nitrogen level. Herbivores, predators, and parasitoids increased in abundance with nitrogenous-fertilization level. The average density of rice leaffolder larvae at the highest nitrogen level was eight times the density at zero nitrogen level, and the peak percentage injured leaves increased from 5 to 35%. The strong increase in larval density was due to the positive effect of nitrogenous-fertilization on egg recruitment and survival of medium-sized larvae. The percentage parasitism of eggs and larvae was not affected by nitrogenous-fertilization. The increase in survival of medium-sized larvae with nitrogen levels was associated with lower predator to leaffolder ratios. The strong effect of nitrogenous-fertilization in the present small-scale experiment was attributed mainly to allowing the moths an oviposition choice between plots with different application levels of nitrogen. Therefore it is hypothesized that the effect of increasing nitrogenous-fertilization level on leaffolder larval densities will be less pronounced when implemented over a large area.     

Development of a powder formulation of Pseudomonas fluorescens for control of rice blast

Pseudomonas fluorescens strain Pf1, inhibitory to the growth of the rice blast pathogen Pyricularia oryzae in vitro , was developed as a talc-based powder formulation. When rice seeds were treated with this formulation, the bacteria spread to roots, stems and leaves of the plants and protected against leaf infection by P. oryzae . When applied as a foliar spray, the bacteria survived on the leaves. The powder formulation controlled leaf blast under greenhouse conditions. In tests as a seed treatment and foliar spray in four field trials it effectively controlled the disease and increased grain yield.     

Accumulation of pathogenesis-related proteins in the epidermis of tomato leaves infected by Cladosporium fulvum

Upon infection byCladosporium fulvum, tomato plants start to produce pathogenesis-related (PR) proteins. The PR proteins 1,3-β-glucanase, chitinase, and PR-1b accumulated near the stomata in the lower epidermis ofC. fulvum-inoculated tomato leaves as could be determined by immunolocalization with polyclonal antibodies. However, no differences in accumulation of PR proteins between a compatible and an incompatible interaction were found. Results obtained from enzyme activity measurements of 1,3-β-glucanase and chitinase on similar leaf material as used for the immunolocalization did not fully reflect the immunolocalization data. The antibodies possibly detect only the extracellular but not the intracellular enzymes. The accumulation of PR proteins near the stomata might be part of a general defence response of plants against pathogens and potential pathogens.     

水稻纹枯病菌拮抗细菌的筛选及鉴定

为获得对水稻纹枯病有生防效果的拮抗细菌,从江苏南京、徐州和常州等地采集的土样中分离细菌分离物1914株,采用平板对峙法筛选获得70株对水稻纹枯病菌有较强抑菌活性的分离物,其中11株对5种水稻病害病原菌均有抑制作用;对11株拮抗菌进行田间防效和室内促生试验,测定菌株分泌的抑菌物质和促生物质,并进行种属鉴定.结果表明,拮抗菌对水稻纹枯病的盆栽和田间小区防效在48.41％和43.03％以上;均可产生蛋白酶与嗜铁素,而不产生几丁质酶,除XF-174外其余10个菌株均可产生纤维素酶;对水稻苗株高和鲜重具有促生作用,并均可产生赤霉素(GA3);除ZF-273和XF-174外的9个菌株可产生吲哚乙酸(IAA),且细菌发酵液中IAA和GA3含量与水稻株高和鲜重的增长率呈正相关.结合各菌株形态特征、生理生化特性和16S rDNA与gyr-B序列分析结果,鉴定SF-181为枯草芽胞杆菌Bacillus subtilis,XF-174为荧光假单胞菌Pseudomonas fluorescens,其余9个菌株为解淀粉芽胞杆菌B.amyloliquefaciens.     

Induction of defence-related enzymes and resistance by the plant activator acibenzolar-S-methyl in tomato seedlings against bacterial canker caused by Clavibacter michiganensis ssp. michiganensis

Using tomato seedlings, the plant defence activator acibenzolar-S-methyl (benzo-[1,2,3]-thiadiazole-7-carbothioic acid-S-methyl ester, ASM; Bion 50 WG) was assayed for its ability to induce resistance against Clavibacter michiganensis ssp. michiganensis , the causal agent of bacterial canker of tomato. In ASM-pretreated plants, reduction in disease severity (up to 76·3%) was correlated with lower bacterial growth (up to 68·2% lower) during the time course of infection. To understand the possible mechanism of action of ASM, alterations in the activities of peroxidase (POX) and chitinase were assessed as markers of resistance. The enhanced resistance of ASM-treated plants was associated with significant increases in the activities of POX and chitinase     

Cytomolecular aspects of rice sheath blight caused by <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

Sheath blight, caused by anastomosis group 1-IA of Rhizoctonia solani Kühn (teleomorph Thanatephorus cucumeris (Frank) Donk), is one of the most destructive rice diseases worldwide. The pathogen is able to infect plants belonging to more than 27 families, including many economically important monocots and dicots such as rice, wheat, alfalfa, bean, peanut, soybean, cucumber, papaya, corn, potato, tomato and sugar beet. It is a soil borne necrotrophic fungus that survives in plant debris as sclerotia, which are small brown-to-black, rocklike reproductive structures. The sclerotia can survive in the soil for several years and infect rice plants at the water-plant interface in the flooded field by producing mycelia. Management of rice sheath blight requires an integrated approach based on the knowledge of each stage of the disease and cytomolecular aspects of rice defence responses against R. solani. This review summarizes current knowledge on molecular aspects of R. solani pathogenicity, genetic structure of the pathogen populations, and the rice-R. solani interaction with emphasis on cellular and molecular defence components such as signal transduction pathways, various plant hormones, host defence genes and production of defence-related proteins involved in basal and induced resistance in rice against sheath blight disease.     

五株病原真菌对桃小食心虫的致病力

为筛选桃小食心虫高致病性菌株,以在山西省自然感病的桃小食心虫越冬幼虫上分离获得的5株病原真菌为材料,研究各菌株感染桃小食心虫幼虫的症状、对寄主的致死率,以及与菌株毒力相关的胞外类枯草杆菌蛋白酶、几丁质酶和脂肪酶的活性变化,并评价其生物防治潜力。5个菌株在寄主上的感病症状和致死率有显著差异,感病后10天,球孢白僵菌BbTST05对寄主的致死率最高,为89.3%,其类枯草杆菌蛋白酶、几丁质酶和脂肪酶3种酶活性均最大;粉棒束孢IfTSL02对寄主的致死率及其蛋白酶、几丁质酶活性次之;尖孢镰孢菌的FoTSL01、FoTSL03和FoTSL04菌株对桃小食心虫的致死率均低于61%,3种胞外酶的活性也较低。因此,球孢白僵菌BbTST05和粉棒束孢IfTSL02可用于桃小食心虫的防治。     

Response of transgenic cucumber expressing a rice class I chitinase gene to two fungal pathogens with different infectivities

A transgenic cucumber line (CR32) over-expressing the rice class I chitinase gene exhibited resistance to Phytophthora rot (Phytophthora nicotianae var. parasitica) but not to Fusarium wilt (Fusarium oxysporum f. sp. cucumerinum). The infection behavior of these fungi on CR32 and nontransgenic plants was examined with an optical microscope. In zoosporangia of P. nicotianae var. parasitica, the rates of germination and penetration on leaves of both CR32 and nontransgenic plants were almost equal. After infection, however, the growth of infection hyphae was markedly suppressed in CR32 compared with their growth in the nontransgenic plants. In F. oxysporum f. sp. cucumerinum, the infection hyphae localized in petiole vessels of both CR32 and nontransgenic plants, and growth did not differ in the two plants. We investigated the antifungal activity of a high-molecular-weight fraction (HF) and a low-molecular-weight fraction (LF) of crude leaf extracts from CR32 and from the nontransgenic line. CR32 HF, which included the rice chitinase, had antifungal activity only against F. oxysporum f. sp. cucumerinum. In contrast, CR32 LF, which did not have the rice chitinase, had strong antifungal activity against the two fungi. These results suggested that a low-molecular-weight antifungal substance(s) was induced in CR32 and might function as a factor of resistance to P. nicotianae var. parasitica, which has cell walls that almost never contain chitin. Because rice chitinase has already been demonstrated not to localize in vessels of CR32, the infection localization of F. oxysporum f. sp. cucumerinum in vessels might enable the fungus to avoid antifungal substance(s), resulting in Fusarium wilt in transgenic cucumber.     

Enhancing Resistance of Tomato and Hot Pepper to Pythium Diseases by Seed Treatment with Fluorescent Pseudomonads

Twenty isolates of fluorescent pseudomonads were evaluated for their ability to control damping-off in tomato (Lycopersicon esculentum) and hot pepper (Capsicum annuum). These isolates were characterized as Pseudomonas fluorescens and Pseudomonas putida. Two isolates, PFATR and KKM 1 belonged to P. putida and the remaining 18 isolates belonged to P. fluorescens. Among these isolates, P. fluorescens isolate Pf1 showed the maximum inhibition of mycelial growth of Pythium aphanidermatum and increased plant growth promotion in tomato and hot pepper. P. fluorescens isolate Pf1 was effective in reducing the damping-off incidence in tomato and hot pepper in greenhouse and field conditions. Isolate Pf1 was further tested for its ability to induce production of defense-related enzymes and chemicals in plants. Earlier and increased activities of phenylalanine ammonia lyase (PAL), peroxidase (PO) and polyphenol oxidase (PPO) were observed in P. fluorescens Pf1 pretreated tomato and hot pepper plants challenged with Pythium aphanidermatum. Moreover, higher accumulation of phenolics was noticed in plants pretreated with P. fluorescens isolate Pf1 challenged with Pythium aphanidermatum. Thus, the present study shows that in addition to direct antagonism and plant growth-promotion, induction of defense-related enzymes involved in the phenyl propanoid pathway collectively contributed to enhance resistance against invasion of Pythium in tomato and hot pepper.     

Long-term effectiveness of grain protectants and structural treatments against Liposcelis decolor (Pearman) (Psocoptera: Liposcelididae), a pest of stored products

Numerous strains of the psocid pest, Liposcelis decolor (Pearman) were collected from farms and central storages and interbred to form three representative strains from three major grain-growing states of Australia: Queensland, South Australia and New South Wales. These were tested against the grain protectants and structural treatments currently registered for use in Australia. Recently, L decolor has become an important pest of stored grain in Australia, particularly in the eastern and southern parts. There is no published information available on management of this pest and the current pest-management strategy, based predominantly on phosphine fumigation, has failed to control infestations of this pest in numerous grain storages in Australia. Alternative methods of control such as use of contact insecticides were explored in the present work to supplement phosphine fumigation to manage this new pest. From eight grain protectants tested as admixtures, only chlorpyrifos-methyl, bioresmethrin plus piperonyl butoxide, and fenitrothion were found to provide long term (3-9 months) protection against all three strains of L decolor. Chlorpyrifos-methyl gave the best protection, providing a minimum of 7.5 to a maximum of 9 months protection, depending on the strains tested. Three structural treatments, azamethiphos, azamethiphos plus carbaryl and permethrin provided long-term control (8-9 months) of all three strains of L decolor on galvanised steel surfaces, with permethrin delivering 9 months protection against all strains. However, all of these treatments failed to provide long-term control of any strains on concrete surfaces. We conclude that chlorpyrifos-methyl as a grain admixture can be incorporated into fumigation strategies to optimise the control of L decolor infestations. Structural treatments, such as permethrin can be used to support a fumigation strategy in storages made of galvanised steel.     

外源茉莉酸甲酯诱导水稻抗瘟性相关防御酶和内源水杨酸的变化

 用外源茉莉酸甲酯(MeJA)处理抗稻瘟病近等基因系水稻CO39和C101LAC,显著减轻了稻瘟病的发生,而对稻瘟病菌孢子萌发及菌丝生长均无明显抑制作用,证实MeJA处理后稻瘟病病情指数的下降是由于MeJA提高了水稻幼苗的抗瘟性。对水稻抗瘟性重要防御酶的活性测定结果表明,苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)、过氧化氢酶(CAT)和脂氧合酶(LOX)活性均在MeJA处理早期上升,与亲和性互作水稻相比,高度非亲和性互作水稻中的诱导活性增加明显,且速度快。对病程相关蛋白β-1,3-葡聚糖酶和几丁质酶的活性测定结果表明,高度非亲和性互作水稻PR蛋白活性的高峰期出现和强度也明显要早且高于亲和性互作水稻。对内源水杨酸(SA)的测定结果表明,不同亲和性互作水稻的SA含量均没有明显的变化,表明MeJA诱导的水稻信号通路可能与SA信号通路无关。