Biocontrol of Rhizoctonia solani and Sclerotium rolfsii on tomato by delivering antagonistic bacteria through a drip irrigation system

In a 2-year assessment carried out on tomato crops of Central and Southern Italy, a high incidence of Rhizoctonia solani and Sclerotium rolfsii was found. These fungal pathogens attack horticultural crops and are responsible for severe crown and stem rot. Because of technical, economical and environmental issues their chemical control is an arduous task. To find alternative and eco-compatible control methods, the effectiveness of two new antagonistic bacterial isolates (Burkholderia cepacia, T1A-2B, and Pseudomonas sp., T4B-2A), previously selected from suppressive organic amendments, were tested on tomato plants grown under both growth chamber and field conditions. The potential antagonists were compared with two commercial biofungicides, based on Bacillus subtilis (BSF4) and Trichoderma asperellum (TV1), and four synthetic fungicides (tolclofos-methyl, azoxystrobin, fosetyl-Al and fosetyl-Al + propamocarb). In 2-year field experiments carried out on tomato plants, the biocontrol bacteria as well as the other treatments were applied to the soil, proximal to the plant crowns and main roots, by means of an effective and specific system of drip irrigation. In all the experiments the novel selected biocontrol bacteria significantly reduced both incidence and severity of the diseases caused by S. rolfsii or R. solani, with results demonstrating effectiveness equal to TV1, better than BSF4 and comparable with the synthetic fungicides, except for tolclofos-methyl which was the most effective treatment. In field experiments, carried out for two consecutive years, isolate T1A-2B reduced up to 58.33% and up to 63.8% the severity of the diseases caused by S. rolfsii and R. solani respectively; whereas isolate T4B-2A gave reduction of S. rolfsii and R. solani diseases severity up to 73.2% and up to 62.7%, respectively.     

Use of field-integrated information in GIS-based maps to evaluate Moko disease (Ralstonia solanacearum) in banana growing farms in Colombia

Ralstonia solanacearum (formerly Pseudomonas solanacearum) causes large losses in more than 200 different cultivated plant species. Banana and plantain “Moko” disease is caused by R. solanacearum race 2. Moko disease may cause 100% loss if prompt control measures are not applied. Although a number of investigations have been carried out, many ecological and epidemiological aspects of R. solanacearum race 2 are still unknown, particularly in the tropics. Bacteria dispersion occurs by infested tools, soil, water, insects, domestic animals and agricultural labourers. Alternative hosts have been described which may contribute to inoculum availability in crop fields. In banana crops, cable-ways are routinely used to transport fruit and materials inside plantations. In most humid tropical areas, drainage channels are made to evacuate excess soil water. It is thought that agricultural activities associated with drainage channels and cable-ways may play a role in bacteria dispersion but individual contributions have not yet been quantified. In this work we have used field-integrated information in geographical information systems (GIS)-based maps to evaluate Moko presence in the Urabá region of Colombia, during three different time periods. A previously described risk index (RI) and losses throughout the three periods were quantified. A regression model was developed to quantify the relationship between infested area and distance from every Moko focus to the nearest cable-way and drainage channels. Farms were grouped into four different profiles according to the RI. Parameters calculated using the regression model showed that 76% of Moko foci detected during the three time periods were associated with cable-ways used for transport of fruit and consumables. This finding has important implications for disease management since bacteria spread in these ways may be prevented by simple measures taken inside plantations.     

芝麻青枯病发生特点及药剂防治技术

青枯雷尔氏菌（Ralstonia solanacearum）引起芝麻青枯病,制约我国南方芝麻生产。为制定精准的药剂防治技术,在江西省南昌县建立4年的病害观察圃上试验5种药剂,研究适宜药剂和用药时期。结果发现,芝麻青枯病多于现蕾期前后开始发病,初花期病情指数增长率均最为急剧,因此可选择芝麻定苗时进行药剂喷淋护根,初花始期及其后10d左右再各喷施药剂1次为佳防治适期;300g.ai/hm2的20%噻菌铜对芝麻青枯病的防治效果最好,达71.11%~74.25%%,增产率33.95%~36.73%。2017-2018 年试验结果显示,20% 噻菌铜（300g.ai/hm2）、20% 噻唑锌（300g.ai/hm2）、3%中生菌素（22.5g.ai/hm2）和72%农用链霉素（216g.ai/hm2）对青枯病的防效范围为70.02%~75.23%; 芝麻增产范围为32.14%~35.63%,4种药剂的防效和增产幅度差异均不显著。      

Growth promotion and induction of resistance in tomato plant against Xanthomonas euvesicatoria and Alternaria solani by Trichoderma spp.

In tomato crop, the induction of resistance emerges as an important alternative for achieving the reduction of chemicals in disease control. This study aimed to evaluate the ability of 28 Trichoderma isolates to promote the growth of tomato seedlings and to induce systemic resistance (ISR) against Xanthomonas euvesicatoria and Alternaria solani, the causal agents of bacterial spot and early blight, respectively. Twelve isolates promoted the increase of plant dry matter mass (DMM) above 100%, showing the great potential of these strains. All isolates were able to colonize the root system of tomato plants. The plant growth-promoting isolates were further evaluated for potential elicitation of ISR. Treatment of the soil with all Trichoderma isolates provided protection in tomato plants from 24.13 to 95.94% against X. euvesicatoria and 30.69 to 95.23% against A. solani. The most efficient isolates in reducing the severity of bacterial spot and early blight were the isolates IB 28/07, IB 30/07, IB 37/01 and IB 28/07, IB 30/07 and IB 42/03, respectively. The effect of different time intervals between Trichoderma application and inoculation with pathogens in inducing systemic resistance in tomato plants was evaluated for the isolate IB 28/07. IB 28/07 conferred protection against both diseases at all time intervals, confirming the ability of the isolate to reduce the severity of these diseases up to 21 days after treatment of tomato plants. In vitro assays revealed that all isolates of Trichoderma were able to degrade cellulose. Only the isolate IB 34/08 showed antagonistic activity against X. euvesicatoria and none caused reduction in the in vitro mycelial growth of A. solani. Trichoderma isolates were identified at species level by DNA sequencing.     

Development of a formulation of Trichoderma asperellum to control black rot disease on pineapple caused by (Thielaviopsis paradoxa)

A local isolate of Trichoderma asperellum was tested for its antagonistic activity against Thielaviopsis paradoxa (telemorph = Ceratocyctis paradoxa). The highest antagonistic activity was achieved when the concentration of T. asperellum conidia was 1 × 10⁷ conidia/mL. The highest biomass and number of colony forming unit/mL of the T. asperellum peaked at 144 h after incubation in yeast waste residue medium. The minimum inhibition concentration value of the formulation was observed as 1% on growth of Th. paradoxa incubated at 28 ± 2 °C for 10 d. In the soil fungicide-screening test, the effect of concentrations 100-1600 μg/mL on mycelia growth was not significant (P < 0.05). Complete mycelial growth inhibition occurred at concentration above 52,600 μg/mL. Results of the fruit application tests clearly showed that all treated fruits were free of disease at the end of the incubation period. No significant differences (P > 0.05) in pH, total soluble solids and titratable acidity were observed between fruits treated with formulation of T. asperellum and the control formulation treated pineapples.     

The use of liquid swine manure for the control of potato cyst nematode through soil disinfestation in laboratory conditions

In this laboratory study the effect of liquid swine manure (LSM) on Globodera rostochiensis was evaluated. The LSM was applied to 500 g nematode-infested soil at 0.4, 2.0, 4.0, and 6.0% (v/w), equivalent to 31, 155, 310 and 465 kg of N ha⁻¹, in closed and open containers (microcosms sealed (S) and open (O)). The results showed that LSM in closed containers enriched the volatile fatty acids (VFAs) through partial anaerobic incubation process. Bacterial- and fungal-feeding nematodes predominated, while the least opportunistic groups had a very low occurrence. The LSM in (S) significantly decreased cyst nematode populations and percent egg hatching in contrast with opened ones (O), increasing some chemical parameters related to soil fertility. Hatching tests showed that individual VFA vary in their lethality to G. rostochiensis. Acetic and propionic acids were the most toxic, reducing hatching most effectively and irreversibly. Results suggest that soil disinfestation on laboratory tests with LSM, is an efficient way for the reduction of G. rostochiensis populations in acidic soils, due to the VFAs nematicidal action. To address management alternatives, field studies would be needed that compared these materials to the current management practices, and demonstrated that they are competitive in the field.     

Management of Sinapis alba subsp. mairei winter cover crop residues for summer weed control in southern Spain

Sinapis alba subsp. mairei (H. Lindb. fil.) Maire, a wild subspecies of S. alba L., which is distributed throughout the Mediterranean basin, has been recently introduced in southern Spain as a winter cover crop in olive groves. The reason behind using this cover crop is for the reduction of Verticillium dahliae inoculum. The effectiveness of this cover crop for weed control has not been assessed to date, despite weed flora in olive groves being highly diverse and competitive, especially in spring and summer, when rainfall is low, temperature is high and crop water needs are at their maximum. The objective of this work is to assess the ability and optimum management of S. alba subsp. mairei cover crop residues for controlling summer weeds. This work offers a more detailed study of the influence of this cover crop on the seedling emergence of Amaranthus blitoides (prostrate pigweed) and Chenopodium album (common lambsquarters) in rainfed field conditions. A factorial design was conducted during the 2002 and 2003 seasons. The studied factors were the following: (1) S. alba subsp. mairei cover crops versus bare soil; (2) two different S. alba residue management techniques after mowing and chopping (incorporation into the soil with shallow tillage versus leaving the residues as a mulch); and (3) the effect on two artificially sown-out summer weeds (prostrate pigweed versus common lambsquarters). The S. alba subsp. mairei cover crop residues reduced the weed infestation by 50 and 60%, and it delayed weed appearance by 3 and 4 weeks the first and second years, respectively, compared with bare soil. The optimum cover crop residue management for weed control was to leave mulch. This management was especially efficient for controlling prostrate pigweed, whether no differences were found for common lambsquarters control when the residues were incorporated into the soil with tillage. These results indicate the great ability of S. alba subsp. mairei cover crop residues to provide summer weed control in rainfed field conditions. Its use, therefore, can contribute to the reduction of the number of herbicide treatments in olive groves.     

Evaluation of actinomycete isolates obtained from herbal vermicompost for the biological control of Fusarium wilt of chickpea

A total of 137 actinomycetes cultures, isolated from 25 different herbal vermicomposts, were characterized for their antagonistic potential against Fusarium oxysporum f. sp. ciceri (FOC) by dual-culture assay. Of the isolates, five most promising FOC antagonistic isolates (CAI-24, CAI-121, CAI-127, KAI-32 and KAI-90) were characterized for the production of siderophore, cellulase, protease, hydrocyanic acid (HCN), indole acetic acid (IAA) and antagonistic potential against Rhizoctonia bataticola, which causes dry root rot in chickpea (three strains viz. RB-6, RB-24 and RB-115) and sorghum (one strain). All of the five FOC antagonistic isolates produced siderophore and HCN, four of them (except KAI-90) produced IAA, KAI-32 and KAI-90 produced cellulase and CAI-24 and CAI-127 produced protease. In the dual-culture assay, three of the isolates, CAI-24, KAI-32 and KAI-90, also inhibited all three strains of R. bataticola in chickpea, while two of them (KAI-32 and KAI-90) inhibited the tested strain in sorghum. When the FOC antagonistic isolates were evaluated further for their antagonistic potential in the greenhouse and wilt-sick field conditions on chickpea, 45-76% and 4-19% reduction of disease incidence were observed, respectively compared to the control. The sequences of 16S rDNA gene of the isolates CAI-24, CAI-121, CAI-127, KAI-32 and KAI-90 were matched with Streptomyces tsusimaensis, Streptomyces caviscabies, Streptomyces setonii, Streptomyces africanus and an identified species of Streptomyces, respectively using the BLAST searching. This study indicated that the selected actinomycete isolates have the potential for biological control of Fusarium wilt disease in chickpea.     

Eradication of Clavibacter michiganensis subsp. michiganensis by incorporating fresh crop debris into soil: Preliminary evaluations under controlled conditions

A method for the eradication of Clavibacter michiganensis subsp. michiganensis was tested for its efficacy in three experiments carried out in the laboratory and greenhouse. In the first experiment, peat moss and sand mix in pots was amended with fresh tomato debris which was either artificially infected with the pathogen, or was not amended. Pots were enclosed in plastic bags or left open. Two temperatures (25 °C and 45 °C) were tested over a 6-week period. The pathogen was not detected in the amended soil after 4 weeks treatment at 45 °C, but was not eradicated after treatments in open pots at 25 °C.In the second experiment, the survival of C.m. michiganensis in either artificially infested soil or in artificially infected tomato plants was studied to determine the behaviour of the pathogen under these conditions. Strains of saprophytic bacteria in the genera Bacillus, Paenibacillus and Brevibacillus were identified under the experimental conditions. In vitro antagonism between Bacillus subtilis and C.m. michiganensis was observed. Finally, the recovery of C.m. michiganensis introduced into disinfected substrate was determined. Survival of C.m. michiganensis in plates artificially inoculated with substrate was greatly reduced after a 4-week treatment at 25 °C, or after 1 week at 45 °C.C.m. michiganensis remained pathogenic on plant tissue after 4 weeks of either thermal treatment.It is important to take these results into account with regard the effect of different soil disinfection techniques or ecological alternatives such as biofumigation, solarization, and the addition of organic matter, as well as for integrated pest management systems.     

Algicidal effect of 3-(3-indolyl)butanoic acid, a control agent of the bacterial wilt pathogen, Ralstonia solanacearum

The chemical control agent 3-(3-indolyl)butanoic acid, previously reported as a control agent for the bacterial wilt pathogen Ralstonia solanacearum, was shown to suppress the growth of green algae during hydroponic culture of tomato. The algicidal activity of the compound was effective at 10 μg/ml, completely preventing generation of green algae under non-shaded greenhouse conditions. The algicidal effect was mainly due to suppression of the growth of motile unicellular algal cells tentatively identified as Chlamydomonas spp., which are commonly occurring in the hydroponic solution and vigorously multiply to form an algal mat on the sponge supports. The compound has potential as a non-phytotoxic algicide for hydroponically cultured crop plants.     

Reduced efficacy of rovral and botran to control Sclerotinia minor in lettuce production in the Salinas Valley may be related to accelerated fungicide degradation in soil

Accelerated soil degradation with repeated application was suggested to be responsible for the lack of efficacy of commercial fungicides to control lettuce drop caused by Sclerotinia minor in the Salinas Valley in California. In 2000, a study was initiated in a field artificially infested with S. minor to evaluate the efficacy and degradation potential of four selected fungicides (i.e., Rovral, Botran, Switch, and Endura). Crisphead lettuce was direct-seeded twice a year (spring and fall season) from 2000 to 2002. Fungicides were applied at recommended label rates twice during each season. Disease incidence was evaluated in fungicide treatments and nontreated control plots weekly starting immediately after thinning until harvest, and areas under the disease progress curves (AUDPC) were compared between treatments from 2000 to 2002. Soil samples (0–15 cm) were collected at regular intervals during the spring and fall 2002 lettuce seasons, and analyzed for residual fungicide concentration. Disease progress and final disease incidence were significantly affected by the fungicide treatment, season, year and interactions among these factors (p < 0.05). At the end of the three year experiment, up to 18 and 50% of the lettuce was infected by S. minor for the spring and fall seasons, respectively. Fungicide efficacy to control lettuce drop was highest for Endura, followed by Switch and Botran, and lowest for Rovral. Relative persistence in soils was lowest for Rovral and Botran, followed by Switch and Endura. In 2002, residual soil fungicide concentrations were 11% for Rovral and Switch, 23% for Botran, and 88% for Endura relative to their initial concentrations. Results of this study provide first evidence suggesting that reduced efficacy of lettuce drop control from Rovral and Botran in the Salinas Valley may be related to an accelerated degradation of these fungicides due to repeated application. However, insufficient fungicide persistence due to inherent properties of this sandy loam soil or the chemical structure of these fungicides can not be excluded. Both hypotheses deserve further investigation.     

Evaluation of non-fumigant alternatives to methyl bromide for weed control and crop yield in California strawberries (Fragaria ananassa L.)

In California, USA, agricultural fumigant use regulations hinder the complete transition from methyl bromide (MB) to alternative fumigants. Alternative fumigants such as 1,3-dichloropropene (1,3-D) and chloropicrin (Pic) are being used on approximately half of California conventional strawberry production fields. Geographic use limits and buffer zones set by the California Department of Pesticide Regulation for 1,3-D + Pic restrict a more complete replacement of MB. Due to the regulatory constraints and public resistance to fumigant use, it is necessary to develop fumigant-free strawberry production systems. Trials were conducted during the 2007/2008 and 2008/2009 strawberry growing seasons at Salinas and Watsonville, California. Non-fumigant treatments including steam, mustard seed meal (MSM), Muscodor albus, and furfural, fertilizers including Mustard Products & Technologies fertilizer and stabilised urea, and fungicide treatments including AG3(NP), fludioxonil + mefenoxam (mfx), and mfx + thiophanate-methyl were evaluated for weed control and strawberry fruit yield, and compared to MB + Pic (MBPic) standard soil fumigation and an untreated control. Steam treatment applied pre-plant to achieve soil temperature of ≥70 °C for 20 min up to 25 cm soil depth consistently provided weed control similar to the MBPic standard soil fumigation. Use of oxyfluorfen herbicide prior to fungicide applications in 2008/2009 also controlled weeds similar to the MBPic standard soil fumigation. Strawberry yields in steam-treated plots with the exception of steam alone in 2007/2008 at Salinas, were comparable to MBPic. At Watsonville in 2008/2009, treatment effect on strawberry yields was insignificant. Yields in furfural and MSM treatments were comparable to MBPic only in some years or sites. With the exception of steam, none of the treatments can be considered viable replacement to MB.     

Improved control of lettuce drop caused by Sclerotinia sclerotiorum using Contans combined with lignin or a reduced fungicide application

The use of Contans (Coniothyrium minitans) in Belgian commercial lettuce greenhouses has so far not always fulfilled the expectations of growers. With the aim of improving the efficacy of control of lettuce drop (Sclerotinia sclerotiorum), Contans applied alone and Contans combined with Radix (Trichoderma asperellum and Trichoderma gamsii), Kraft pine lignin and Rovral was compared in three successive lettuce crops in a naturally infested greenhouse. At harvest of each crop, the effect on disease incidence, and the effect on viability and fungal infection of sclerotia buried prior to each crop was evaluated. Contans applied alone negatively affected the sclerotial viability, but did not reduce lettuce drop symptoms. The combination of Contans with Kraft pine lignin, on the other hand, reduced sclerotial viability and lettuce drop compared with the untreated control in the last crop. Furthermore, when Contans was combined with Kraft pine lignin, a higher number of infected sclerotia was observed compared with Contans applied alone. When Radix was added to the combination, there was no extra benefit for the control of lettuce drop. The combination of one Rovral application and Contans resulted in a significant control of lettuce drop together with a negative effect on sclerotial survival. Two Rovral applications, by contrast, reduced the ability of C. minitans to parasitize and kill the sclerotia. To conclude, integration of Contans with a reduced Rovral application and with Kraft pine lignin has potential to improve the control of lettuce drop in commercial greenhouses.     

Effectiveness of plant extracts on suppression of damping-off and wilt diseases of lupine (Lupinus termis Forsik)

The aim of this study was to evaluate water and organic solvent of plant extracts for protection of lupine plants against damping-off and wilt diseases caused by Fusarium oxysporum f. sp. lupini, F. oxysporum f. sp. lupini Snyder & Hansen was isolated from diseased lupine roots collected from different locations of Minia, Assiut and New Valley governorates. Water leaf extracts of Calotropis procera, Nerium oleander, Eugenia jambolana, Citrullus colocynthis, Ambrosia maritime, Acacia nilotica and Ocimum basilicum and fruit extracts of C. colocynthis, C. procera and E. jambolana reduced damping-off and wilt diseases caused by F. oxysporum f. sp. lupini. Water extracts of E. jambolana leaves, C. colocynthis fruits and N. oleander leaves were the most effective ones for controlling such diseases. In contrast, A. nilotica and O. basilicum extracts were the least effective ones. Organic solvent extracts of tested plants reduced damping-off and wilt diseases than water extracts. Butanolic and ethereal extracts were highly effective in reduction of diseases than the other tested extracts. Under field conditions, during winter growing seasons 2008/09 and 2009/10, ethereal and butanolic extracts of N. oleander and E. jambolana leaves and C. colocynthis fruits significantly reduced the percentage of wilt severity as well as improved growth parameters (plant height, number of branches, pods and seeds per plant) and increased seed index, total seed yield/hectare compared with control treatment, while protein content in seeds was not effected. In conclusion, our study demonstrated that some plant extracts can be used for biocontrol of lupine damping-off and wilt diseases.     

Effect of plant growth promoting rhizobia on seed germination, growth promotion and suppression of Fusarium wilt of fenugreek (Trigonella foenum-graecum L.)

Five bacterial strains (TR1 to TR5) isolated from root nodules of fenugreek (Trigonella foenum-graecum) were tested for their plant growth promotory traits and biocontrol potential against Fusarium oxysporum. On the basis of morphological, physiological, biochemical and molecular characteristics, strains TR1 and TR3 - TR5 were identified as Ensifer meliloti, and TR2 as Rhizobium leguminosarum. All bacterial isolates utilized phosphate in vitro. Except TR5, all isolates produced IAA and none of them showed volatile cyanogens production. Except TR3, all isolates produced in vitro siderophore. Isolate TR1 and TR4 showed chitinase production while only TR2 showed β-1,3-glucanase activity. Isolates TR1, TR2 and TR5 exhibited ACC deaminase activity. Isolates TR1, TR2 and TR4 inhibited the growth of F. oxysporum, causing loss of structural integrity of the mycelium, hyphal perforation, lysis, fragmentation and degradation. The potential for nodulation and nitrogen fixation of the strains were confirmed by amplification of 500 bp nodC and 781 bp nifH fragments. The application of the TR1 + TR2 combination resulted in increased grain yield by 35% and 36% of fenugreek in two consecutive field trials, respectively as compared to control. Maximum increments in vigour index, nodule number and root and shoot biomass were recorded with seed inoculated with consortium (TR1 + TR2) followed by single inoculation as compared to control. The antibiotic resistant marker strain of E. meliloti TR1^strep+ and R. leguminosarum TR2^tet+ confirmed the efficient colonization of fenugreek roots. This study showed that these rhizobial isolates have properties of biocontrol agents and may be applied to promote the growth of fenugreek.     

The effect of three rates of cyantraniliprole on the transmission of tomato spotted wilt virus by Frankliniella occidentalis and Frankliniella fusca (Thysanoptera: Thripidae) to Capsicum annuum

Tomato spotted wilt virus (TSWV) is a thrips-transmitted virus that causes major losses in many crops worldwide. Management of TSWV is complex, requiring multiple preventive measures. Currently, there are few chemical options that control thrips populations before they feed upon and transmit TSWV to crop plants. Cyantraniliprole (Cyazypyr™) is an anthranilic diamide insecticide currently under development that exhibits anti-feedant properties. Transmission of TSWV by Frankliniella fusca (Hinds) to Capsicum annuum L. seedlings was reduced in plants treated with Cyazypyr™ applied to the soil at the rates of 1.45, 2.90 and 4.41 mg ai/plant. Mortality of F. fusca at 3 days post treatment did not differ significantly on excised foliage of Cyazypyr™ treated and control plants, but feeding injury was significantly less on treated foliage. Transmission of TSWV by Frankliniella occidentalis (Pergande) was not reduced in plants treated with 4.41 mg ai/plant.     

Biocontrol of vascular wilt and corm rot of gladiolus caused by Fusarium oxysporum f. sp. gladioli using plant growth promoting rhizobacterial mixture

Talc-based formulations of plant growth promoting rhizobacterial strain S2BC-2 (Bacillus atrophaeus) and strain mixture, S2BC-2 + TEPF-Sungal (Burkholderia cepacia), inhibitory to the growth of Fusarium oxysporum f. sp. gladioli (FOG), were developed for corm dressing and soil application in gladiolus. In comparison to the individual strain, the strain mixture recorded maximum spike and corm production of 100 and 150%, respectively with less vascular wilt and corm rot incidences of 73.6 and 54.8% reduction over the pathogen control in greenhouse when inoculated with FOG. Reduction in disease incidence under greenhouse conditions occurred through induction of defence gene products such as chitinase, β-1,3-glucanase, peroxidase and polyphenol oxidase. Besides disease suppression, treatment with strain mixture promoted plant growth in terms of enhanced corm and cormel production and flowering. In field experiments, the strain mixture recorded less vascular wilt and corm rot incidences of 48.6 and 46.1% mean reduction over the non-bacterised control, and was almost comparable with that of fungicide (51.5 and 47.1%, respectively). The treatment also recorded increased spike and corm yield with average increases of 58.3 and 27.4%, respectively, relative to the control.     

Combined application of antagonist Bacillus amyloliquefaciens and essential oils for the control of peach postharvest diseases

Bacillus amyloliquefaciens PPCB004 was selected as a potential antagonist to control Botrytis cinerea, Penicillium expansum and Rhizopus stolonifer on peach fruit. The HPLC data of PPCB004 indicated the lipopeptides iturin A, fengycin and surfactin as secondary metabolites. The GC/MS analysis of PPCB004 showed 3-hydroxy-2-butanone as the dominant compound (97.52% of relative peak area). Thyme (TO) and lemongrass (LO) oils showed over 50% and 25% inhibition of radial mycelial growth respectively with 8 μl oil per plate for all pathogens. Combination treatment with both oils failed to increase the percentage inhibition of radial mycelial growth of the pathogens. Combined application of PPCB004 with TO or LO was tested to assess the effectiveness in the control of these pathogens during postharvest storage. The biofilm formation of PPCB004 was significantly higher in LO than TO. LO (6 μl plate⁻¹) and PPCB004 completely inhibited the mycelial growth of the pathogens. Fruit inoculation trials with PPCB004 + LO in NatureFlex™ modified atmosphere packaging (MAP), showed lower disease incidence and severity at 25 °C for 5 d than treatments with PPCB004 + MAP, PPCB004 + TO + MAP, LO + MAP, TO + MAP or stand-alone MAP. On naturally infected fruit, PPCB004 + LO + MAP and LO + MAP treatments retained the total soluble solids/titratable acidity ratio and flesh firmness but failed to stimulate the levels of total phenolic content, phenylalanine ammonia-lyase, β-1,3-glucanase and chitinase activities. Combination of PPCB004 (spray treatment) + LO (in pad delivery system) in NatureFlex™ MAP showed absence of disease and off-flavour development, retained the overall appearance and increased the overall acceptance at market shelf conditions (20 °C for 2 d) after cold storage at 4 °C for 14 d.     

Evaluation of fungicides to control circular leaf spot of persimmon caused by Mycosphaerella nawae

The efficacy of ten fungicides against Mycosphaerella nawae, the causal agent of circular leaf spot (CLS) of persimmon, was evaluated in vitro and in field experiments. Field trials were conducted in 2009 and 2010 to investigate the comparative efficacy of the fungicides alone or combined using alternating sequences in spray programmes based on two, three or four applications. Disease incidence was assessed by estimating the percentage of affected leaves, which included leaves showing at least one necrotic spot and defoliation. Fenpropimorph, pyraclostrobin, tebuconazole and thiophanate-methyl were the most effective fungicides in inhibiting mycelial growth of M. nawae isolates (EC₅₀ < 2 ppm). In field experiments, the most effective fungicides using two spray applications were captan, chlorothalonil, mancozeb and pyraclostrobin which significantly reduced disease incidence compared with untreated plots. Regarding the number of spray applications, two applications of captan and mancozeb were less efficient than three to control the disease. However, the percentage of affected leaves provided by three applications of captan and mancozeb alone or combined with pyraclostrobin using alternating sequences in spray programmes, was not significantly different from that provided by four applications. Experimental results demonstrated that spray programmes based on three applications of these fungicides could effectively control CLS of persimmon. The advantages of spray programmes based on alternated use of strobilurins and protective fungicides are discussed.     

Biological control of Macrophomina phaseolina by chemotactic fluorescent Pseudomonas aeruginosa PN1 and its plant growth promotory activity in chir-pine

Ten strains of Pseudomonas aeruginosa (PN1 ˜ PN10) isolated from rhizosphere of chir-pine were tested for their plant growth promontory properties and antagonistic activities against Macrophomina phaseolina in vitro and in vivo. P. aeruginosa PN1 produced siderophore, IAA, cyanogen and solubilized phosphorus, besides producing chitinase and β-1,3-glucanase. In dual culture, P. aeruginosa PN1 caused 69% colony growth inhibition. However, cell free culture filtrate also posed inhibitory effect but to a lesser extent. After 90 days, P. aeruginosa PN1 increased plant growth and biomass in pots trial containing M. phaseolina-infested soil. PN1 showed the strong chemotaxis toward root exudates resulting in effective root colonization. Moreover, increased population in rhizosphere of these bacteria was also recorded after 90 days of treatment. Thus, chemotactic fluorescent P. aeruginosa PN1 exhibited strong antagonistic property against M. phaseolina, suppressed the disease and improved plant growth of the seedlings of chir-pine proving potential biocontrol agent.