Isolation of entomopathogenic Bacillus from a biodynamic olive farm and their pathogenicity to lepidopteran and coleopteran insect pests

The occurrence of Bacillus entomopathogenic bacteria on a Tunisian biodynamic farm was determined by examining 75 samples from olive tree (Olea europaea L.) habitats. A total of 40 Bacillus isolates were characterized according to their phenotypic, physiological and biochemical parameters. Isolates of the species Bacillus subtilis, Bacillus mycoides, Brevibacillus brevis, Paenibacillus polymyxa, Bacillus licheniformis, Bacillus sp. (1), Bacillus sp. (2) and a standard strain Btk HD-1 were used separately in feeding bioassays on fresh artificial diet against larvae of lepidopterans Prays oleae (Bernard) and Palpita unionalis (Hübner) and coleopterans Hylesinus oleiperda (F.) and Phloeotribus scarabaeoides (Bernard), which are olive tree pests. Larvae were successfully reared on an artificial diet with 25 g powdered olive tree leaves. Compared to the control data, only Btk and the isolates of B. licheniformis, P. polymyxa and B. brevis were entomopathogenic. Larval mortality assessed 7 days post-treatment showed high mortality rates with Btk to lepidopteran larvae (86.6% for P. oleae and 80.9% for P. unionalis) and low mortality against coleopteran pests. B. brevis isolates showed high mortality rates against P. oleae (up to 67.9%). B. licheniformis isolates caused up to 59.2% larval mortality for P. oleae and 43.6% for P. unionalis. Highest coleopteran mortality was achieved by P. polymyxa isolates (up to 55%). According to the 16S rDNA results, isolates of each of the three entomopathogenic strains were similar. Proteins in the strain supernatants were toxic to P. oleae larvae with LC50 values of 10.0 (B. brevis), 12.5 (B. licheniformis) and 37.6 μg/ml (P. polymyxa). Also, P. polymyxa showed an LC50 of 12.4 mg/l against P. scarabaeoides. Our results suggest that entomopathogenic Bacillus present locally in the biodynamic farm could be used in biological control programmes of olive tree pests.     

Assessment of rhizobacteria from grapevine for their suppressive effect on the parasitic nematode Xiphinema index

The capacity of rhizobacteria isolated from grape plants to protect grapevine roots from damage by the nematode Xiphinema index was assessed in trials carried out in the growing season 2008–2009 and 2009–2010. A total of two groups of bacteria were evaluated, one comprising 49 isolates, of which 37 were assessed in previous studies, and one comprising 90 isolates not previously evaluated. Two assays were performed on the first group, one per season, and one on the second group, lasting two growing seasons. Pots of field soil (3 L) planted with two-month old Thompson Seedless vines were inoculated with 200 or 400 specimens of X. index after inoculation with 10⁶ cfu/ml of test rhizobacteria; the posts were maintained in a shaded greenhouse. Seven isolates from the first group of bacteria (Bacillus brevis, Bacillus megaterium, Cytophaga johnsonae, Pseudomonas fluorescens, Rahnella aquatilis and Stenotrophomonas maltophilia) were effective in decreasing damage in both of the assays (P < 0.05). Ten isolates from the second group (Variovorax paradoxus, Pseudomonas pseudoalcaligenes, Bacillus mycoides, Bacillus sphaericus, Bacillus thuringiensis, Curtobacterium flaccumfaciens, Pseudomonas putida, Pseudomonas alcaligenes and Pseudomonas viridiflava) resulted in the lowest number of galls per gram root. B. brevis and B. megaterium also demonstrated good activity in previous studies on grape plants and are thus promising candidates for further research.     

Diversity and Antimicrobial Activity of Vietnamese Sponge-Associated Bacteria

This study aimed to assess the diversity and antimicrobial activity of cultivable bacteria associated with Vietnamese sponges. In total, 460 bacterial isolates were obtained from 18 marine sponges. Of these, 58.3% belonged to Proteobacteria, 16.5% to Actinobacteria, 18.0% to Firmicutes, and 7.2% to Bacteroidetes. At the genus level, isolated strains belonged to 55 genera, of which several genera, such as Bacillus, Pseudovibrio, Ruegeria, Vibrio, and Streptomyces, were the most predominant. Culture media influenced the cultivable bacterial composition, whereas, from different sponge species, similar cultivable bacteria were recovered. Interestingly, there was little overlap of bacterial composition associated with sponges when the taxa isolated were compared to cultivation-independent data. Subsequent antimicrobial assays showed that 90 isolated strains exhibited antimicrobial activity against at least one of seven indicator microorganisms. From the culture broth of the isolated strain with the strongest activity (Bacillus sp. M1_CRV_171), four secondary metabolites were isolated and identified, including cyclo(L-Pro-L-Tyr) (1), macrolactin A (2), macrolactin H (3), and 15,17-epoxy-16-hydroxy macrolactin A (4). Of these, compounds 2-4 exhibited antimicrobial activity against a broad spectrum of reference microorganisms.     

Effect of efficient pectinolytic bacterial isolates on retting and fibre quality of jute

Four bacterial strains with high balanced polygalacturonase (PG) and pectin lyase (PNL), and xylanase with minimal cellulase activity were used in jute retting. The 16S rDNA analysis revealed that the organisms were: Bacillus sp. L6 (GQ891097), Bacillus pumilus strain EK-17 (GQ891098), B. pumilus strain Geo-03-422 (GQ891103) and B. pumilus strain IK-MB12-518F (GQ891105). The microorganisms in different combinations of consortia showed synergistic effect resulting in increased PG (35.52-46.61 IU/g cell wet weight), PNL (39.79-72.12 U/ml), xylanase (0.705-0.840 μmol/ml/min) and little cellulase (0-0.153 μmol/ml/min) activities. The organisms in different combinations of consortia reduced the retting period from 11 to 13 days as compared to 19 days in the control. Microbial inoculation produced remarkable improvement in jute fibre strength (26.62-28.91 g/tex) and fineness (2.76-2.92 tex) over control. The pH of the post-retting water samples became acidic, and the electrical conductivity (Ec), chemical oxygen demand (COD) and hardness increased. The organisms could be adopted in industrial application for extraction of jute fibre.     

Enhanced selenium content in wheat grain by co-inoculation of selenobacteria and arbuscular mycorrhizal fungi: A preliminary study as a potential Se biofortification strategy

Cereal crops grown in southern Chilean Andisol provide suboptimal levels of this metalloid for human diet. Certain rhizosphere microorganisms, such as rhizobacteria and arbuscular mycorrhizal fungi can increase the selenium uptake in plants. The purpose of this study was to evaluate selenium acquisition by wheat plants through the co-inoculation of native selenobacteria strains (Stenotrophomonas sp. B19, Enterobacter sp. B16, Bacillus sp. R12 and Pseudomonas sp. R8), both individually and in mixture, as selenonanosphere source with one arbuscular mycorrhizal fungus (Glomus claroideum). Total selenium content in plant tissues and substrate was analyzed. According to our results, significant higher selenium content was found in inoculated plants in comparison to uninoculated controls (P ≤ 0.05). Independently of fungal presence, selenium content in grain from plants inoculated with Enterobacter sp. B16 (236 mg kg⁻¹) was higher than the rest of the strains (116–164 mg kg⁻¹). However, when plants were co-inoculated with a mixture of selenobacteria strains and G. claroideum, selenium content in grain was 23.5% higher (725 mg kg⁻¹) than non-mycorrhizal plants (587 mg kg⁻¹). The results suggest a synergistic effect between the selenobacteria mixture and G. claroideum associated to major biodiversity and demonstrate a great potential of these rhizosphere microorganisms for biofortification of cereals and its derivates.     

Effect of the insect growth regulator novaluron on diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae), and its indigenous parasitoids

Research to evaluate effective and selective insecticides through periodic screening is needed to strengthen diamondback moth (DBM) (Plutella xylostella L.) management. The efficacy of the insect growth regulator novaluron on DBM and effects on the parasitoids Diadegma sp. (Hym.: Ichneumonidae), Apanteles sp. (Hym.: Braconidae) and Oomyzus sokolowskii Kurdjumov (Hym.: Eulophidae) were evaluated. Novaluron was compared with the microbial insecticide Bacillus thuringiensis (Bt) var. aizawai, the pyrethroid insecticide λ- cyhalothrin and the organophosphate insecticide profenofos. Two replicated experiments were conducted on head cabbage, Brassica oleracea var. capitata, planted in March and May 2007 at Wonji in the Central Rift Valley of Ethiopia. Infestation by DBM was less with novaluron and Bt compared with other treatments for both planting dates. Infestation with λ-cyhalothrin and profenofos was comparable with the untreated control. Yields were greatest with novaluron and were 80 and 32.5% (12.4 and 7.4 t ha⁻¹) greater than the untreated treatment for the first and second plantings, respectively. Diadegma sp. accounted for 91% of the parasitoid complex observed. Parasitism of DBM by Diadegma sp. ranged between 10 and 43% among treatments. Parasitism was less with λ-cyhalothrin and profenofos compared with other treatments. Owing to its efficacy against DBM and relative safety to its parasitoid, Diadegma sp., novaluron can be used in integrated DBM management for low elevation brassica production in the Central Rift Valley of Ethiopia.     

Virulence of in vivo and in vitro produced conidia of Metarhizium brunneum strains for control of wireworms

Wireworms are the soil inhabiting larvae of click beetles and can cause severe damage to arable crops such as potatoes (Solanum tuberosum, L.). Several strains of the entomopathogenic fungus Metarhizium brunneum (Petch) are pathogenic to wireworms. In this study, three European strains of M. brunneum were tested in the laboratory against the most damaging wireworm species in Europe, Agriotes lineatus (L.), Agriotes obscurus (L.) and Agriotes sputator (L.). A Swiss strain, isolated from an A. obscurus cadaver, proved to be most effective, killing up to 73% of A. lineatus and 83% A. obscurus individuals, respectively. The median lethal time (LT₅₀) was 21 days post inoculation (dpi) for A. lineatus and 14 dpi for A. obscurus. The strain did not lose virulence through subsequent cultivation on artificial medium and thus seems to be suitable for mass production as a biocontrol agent for wireworm control.     

Selection and evaluation of potential biocontrol rhizobacteria from a raised bog environment

Understanding the interactions of bacteria with plants and the role of microorganisms in the lifecycle of plants is becoming more and more important due to the social claim for a clean agriculture. Thus the significance of studying plant associated bacteria is increasing. The main aim of our study was to isolate and characterize bacteria associated with bryophytes from a natural raised bog ecosystem. Characterization of the isolates by their beneficial properties was realized with microbiological and molecular tools. Bacterial strains were identified taxonomically, isolates belonging to the genera Pseudomonas, Bacillus, Serratia, Stenotrophomonas, Delftia, Cedecea, Lysinibacillus and Viridibacillus have been found. Pseudomonas, Serratia and Bacillus were the dominant bacterial genera associated with bryophytes. In vitro study of antagonism showed that a high number of the isolates inhibited the growth of fungal and bacterial plant pathogens – Fusarium oxysporum, Alternaria alternata, Erwinia carotovora – or produced secondary metabolic substances with biocontrol properties. Bacterial strains identified as Serratia fonticola BB17 and Pseudomonas fluorescens BE8 proved to be the most efficient against plant pathogens, with biocontrol effectiveness up to 48.28% and 55.17% respectively, suggesting the potential use of these strains in biotechnological applications.     

Effects of Bacillus subtilis strain QST 713 and storage temperatures on post-harvest disease development on greenhouse tomatoes

Tomato plants in two commercial greenhouses were treated with Rhapsody (Bacillus subtilis strain QST 713, rate of 1.45%) once every 4 weeks during 2012–2013 to determine effects on post-harvest fruit infection. Populations of Bacillus and disease incidence were monitored weekly from harvested fruit over an 18-week period. Population levels of Bacillus ranged from 75 to 110 × 10⁴ colony forming units (cfu) cm⁻² of fruit surface area one week after application to 25–30 × 10⁴ cfu cm⁻² of fruit surface area 4 weeks after application. Disease incidence on harvested fruit incubated at 21 °C for 7–10 days was variable, due to variation in inoculum levels within the greenhouse as well as variable environmental conditions. Both disease incidence and severity were significantly reduced on Rhapsody-treated fruit, especially in the 1–2 week period following application. Post-harvest storage temperature (13 °C vs. 21 °C) and incubation time (12 vs. 16 days) had a significant effect on final disease severity. Rhapsody-treated fruit incubated at 13 °C had an average of 1–2% fruit infection compared to up to 20% infection on untreated fruit at 21 °C. The most frequent pathogens affecting fruit quality were Penicillium sp. and Rhizopus stolonifer. Rhapsody applications made every 4 weeks maintained sufficiently high populations of Bacillus on the fruit surface to prevent spread of these fungi onto the fruit, resulting in significant post-harvest disease control on fresh market tomatoes. When combined with storage at 13 °C for no more than 12 days, disease was reduced to negligible levels.     

Bacillus spp. Inhibit Edwardsiella tarda Quorum-Sensing and Fish Infection

The disruption of pathogen communication or quorum-sensing (QS) via quorum-quenching (QQ) molecules has been proposed as a promising strategy to fight bacterial infections. Bacillus spp. have recognizable biotechnology applications, namely as probiotic health-promoting agents or as a source of natural antimicrobial molecules, including QQ molecules. This study characterized the QQ potential of 200 Bacillus spp., isolated from the gut of different aquaculture fish species, to suppress fish pathogens QS. Approximately 12% of the tested Bacillus spp. fish isolates (FI). were able to interfere with synthetic QS molecules. Ten isolates were further selected as producers of extracellular QQ-molecules and their QQ capacity was evaluated against the QS of important aquaculture bacterial pathogens, namely Aeromonas spp., Vibrio spp., Photobacterium damselae, Edwardsiela tarda, and Shigella sonnei. The results revealed that A. veronii and E. tarda produce QS molecules that are detectable by the Chr. violaceum biosensor, and which were degraded when exposed to the extracellular extracts of three FI isolates. Moreover, the same isolates, identified as B. subtilis, B. vezelensis, and B. pumilus, significantly reduced the pathogenicity of E. tarda in zebrafish larvae, increasing its survival by 50%. Taken together, these results identified three Bacillus spp. capable of extracellularly quenching aquaculture pathogen communication, and thus become a promising source of bioactive molecules for use in the biocontrol of aquaculture bacterial diseases.     

In Vitro Assessment of Marine Bacillus for Use as Livestock Probiotics

Six antimicrobial-producing seaweed-derived Bacillus strains were evaluated in vitro as animal probiotics, in comparison to two Bacillus from an EU-authorized animal probiotic product. Antimicrobial activity was demonstrated on solid media against porcine Salmonella and E. coli. The marine isolates were most active against the latter, had better activity than the commercial probiotics and Bacillus pumilus WIT 588 also reduced E. coli counts in broth. All of the marine Bacillus tolerated physiological concentrations of bile, with some as tolerant as one of the probiotics. Spore counts for all isolates remained almost constant during incubation in simulated gastric and ileum juices. All of the marine Bacillus grew anaerobically and the spores of all except one isolate germinated under anaerobic conditions. All were sensitive to a panel of antibiotics and none harbored Bacillus enterotoxin genes but all, except B. pumilus WIT 588, showed some degree of β-hemolysis. However, trypan blue dye exclusion and xCELLigence assays demonstrated a lack of toxicity in comparison to two pathogens; in fact, the commercial probiotics appeared more cytotoxic than the majority of the marine Bacillus. Overall, some of the marine-derived Bacillus, in particular B. pumilus WIT 588, demonstrate potential for use as livestock probiotics.     

Assessment of the Bacteriocinogenic Potential of Marine Bacteria Reveals Lichenicidin Production by Seaweed-Derived Bacillus spp.

The objectives of this study were (1) to assess the bacteriocinogenic potential of bacteria derived mainly from seaweed, but also sand and seawater, (2) to identify at least some of the bacteriocins produced, if any and (3) to determine if they are unique to the marine environment and/or novel. Fifteen Bacillus licheniformis or pumilus isolates with antimicrobial activity against at least one of the indicator bacteria used were recovered. Some, at least, of the antimicrobials produced were bacteriocins, as they were proteinaceous and the producers displayed immunity. Screening with PCR primers for known Bacillus bacteriocins revealed that three seaweed-derived Bacillus licheniformis harbored the bli04127 gene which encodes one of the peptides of the two-peptide lantibiotic lichenicidin. Production of both lichenicidin peptides was then confirmed by mass spectrometry. This is the first definitive proof of bacteriocin production by seaweed-derived bacteria. The authors acknowledge that the bacteriocin produced has previously been discovered and is not unique to the marine environment. However, the other marine isolates likely produce novel bacteriocins, as none harboured genes for known Bacillus bacteriocins.     

Effect of carbonic anhydrase-containing endophytic bacteria on growth and physiological attributes of wheat under water-deficit conditions

Drought is one of the major limitations to agricultural productivity, suppressing plant growth and yield of food crops throughout the world particularly in arid and semiarid regions. Drought-tolerant carbonic anhydrase (CA; EC 4.2.1.1)-containing endophytic bacteria may improve plant growth under stressed conditions. In the present study, effect of drought-tolerant CA-containing endophytic bacteria on growth and physiology of wheat under water-deficit conditions was studied. One hundred and fifty isolates were isolated from wheat plants and screened for their ability to tolerate polyethylene glycol (PEG) 6000-induced water-deficit stress (?0.31 to ?3.20 MPa). Fifty isolates exhibiting intrinsic ability to tolerate stress were further screened for CA activity. Ten drought-tolerant isolates with higher CA activity were evaluated for improving wheat growth under water-deficit conditions (?0.04, ?1.09, ?1.23 MPa). Results showed that PEG-mediated water-deficit stress significantly reduced growth of wheat. However, inoculation with isolates WR2, WS11 and WL19 significantly enhanced seedling growth by improving maximum root length, shoot length, root and shoot dry weight under non-stressed as well as stressed conditions. These isolates were identified by 16S rRNA as Bacillus marisflavi (WR2) Bacillus thuringiensis (WS11) and Bacillus subtilis (WL19). Isolate WL19 also improved chlorophyll content, photosynthetic rate, CA activity and relative water content compared to uninoculated control plants. Overall, our findings suggest that endophytic bacterial isolates WR2, WS11 and WL19 with CA activity can enhance photosynthesis and biomass of wheat seedlings under water-deficit conditions.

Abbreviations: CA: Carbonic anhydrase; PEG: Polyethylene glycol; CO₂: Carbon dioxide; HCO^3–: Bicarbonate; TSA: Tryptic Soy Agar; LB: Luria Bertani; A: CO₂ assimilation rate; E: Transpiration rate; g_s: Stomatal conductance; C_i: Substomatal CO₂ concentration; RWC: Relative water content; EL: Electrolyte leakage     

Management of complex weed flora in dry-seeded rice

Dry-seeded rice has been introduced as an alternative to puddled hand-transplanted rice in the north Indian states of Punjab and Haryana. In dry-seeded rice, weed flora tends to be more diverse and weeds emerge in several flushes during the crop growth cycle and substantial yield reductions due to weed competition are quite common. The efficacy and compatibility of tank mixtures of different herbicides for the control of diverse weed flora in dry-seeded rice was evaluated in field experiments during the summer seasons of 2012 and 2013. The tank mixture of fenoxaprop with ethoxysulfuron improved the control of Echinochloa crus-galli and Echinochloa colona by 43–69% as compared to fenoxaprop alone while the tank-mix of azimsulfuron with fenoxaprop was antagonistic and reduced the control of Leptochloa chinensis by 86% as compared to fenoxaprop alone. Addition of azimsulfuron or ethoxysulfuron to bispyribac did not improve the control of grass weeds as compared to bispyribac alone. Weed control with the mixture of bispyribac and fenoxaprop varied over the two years. In 2012, bispyribac and fenoxaprop mixture was antagonistic for the control of Dactyloctenum aegyptium, Acrachne racemose, and L. chinensis but in 2013, there was no apparent antagonism and the addition of bispyribac to fenoxaprop reduced grass weed biomass as compared to fenoxaprop alone. In 2013, there was a strong negative correlation (r = −0.95, P < 0.001) between weed dry matter at 45 days after sowing and rice grain yield. According to the linear regression, rice crop is likely to produce no grain yield when weed dry matter exceeds 400 g m⁻². Over the two seasons, fenoxaprop-ethoxysulfuron tank-mix produced similar grain yields (5.6–6.2 t ha⁻¹) to the weed-free check (5.6–7.1 t ha⁻¹). At the farmer fields, rice grain yield in the plots treated with pendimethalin followed by post-emergence bispyribac or a tank-mix of fenoxaprop + ethoxysulfuron ranged from 6.2 to 7.7 t ha⁻¹ as compared to 5.3–5.6 t ha⁻¹ in the plots treated with pendimethalin alone. The tank mixture of fenoxaprop with bispyribac needs further evaluation as this mixture has the potential to effectively control aerobic and aquatic grasses in dry-seeded rice. Single hand weeding prevented crop yield loss from weeds that escaped herbicide treatments only when it was performed within six weeks of sowing.     

Microbial control of the European skipper, Thymelicus lineola Ochs.

A research programme was initiated to develop biological control methods for the European skipper, Thymelicus lineola Ochs. (Lepidoptera: Hesperiidae) after its appearance in the Province of Quebec. A baculovirus and Bacillus thuringiensis could both be used to control T. lineola in the field. With B. thuringiensis a dose of 2·5 × 10⁹ International Units/ha resulted in 100% mortality of second- to fourth-instar larvae in 6–15 days; dosages of 5 and 10 × 10⁹ I.U./ha caused 100% mortality within 3–6 days. Field application of the nuclear polyhedrosis virus caused 50, 80 and 100% larval mortality after 15, 19 and 27 days respectively. B. thuringiensis provided a curative method of control, whereas the virus can be used either as a curative or as a preventive method of controlling T. lineola because it is transmitted to the progeny. It is suggested that the best way to control T. lineola is by the combined use of both micro-organisms.     

Antioxidant and Antimicrobial Potential of the Bifurcaria bifurcata Epiphytic Bacteria

Surface-associated marine bacteria are an interesting source of new secondary metabolites. The aim of this study was the isolation and identification of epiphytic bacteria from the marine brown alga, Bifurcaria bifurcata, and the evaluation of the antioxidant and antimicrobial activity of bacteria extracts. The identification of epiphytic bacteria was determined by 16S rRNA gene sequencing. Bacteria extracts were obtained with methanol and dichloromethane (1:1) extraction. The antioxidant activity of extracts was performed by quantification of total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and oxygen radical absorbance capacity (ORAC). Antimicrobial activities were evaluated against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Salmonella enteritidis, Staphylococcus aureus, Saccharomyces cerevisiae and Candida albicans. A total of 39 Bifurcaria bifurcata-associated bacteria were isolated and 33 were identified as Vibrio sp. (48.72%), Alteromonas sp. (12.82%), Shewanella sp. (12.26%), Serratia sp. (2.56%), Citricoccus sp. (2.56%), Cellulophaga sp. (2.56%), Ruegeria sp. (2.56%) and Staphylococcus sp. (2.56%). Six (15.38%) of the 39 bacteria Bifurcaria bifurcata-associated bacteria presented less than a 90% Basic Local Alignment Search Tool (BLAST) match, and some of those could be new. The highest antioxidant activity and antimicrobial activity (against B. subtilis) was exhibited by strain 16 (Shewanella sp.). Several strains also presented high antimicrobial activity against S. aureus, mainly belonging to Alteromonas sp. and Vibrio sp. There were no positive results against fungi and Gram-negative bacteria. Bifurcaria bifurcata epiphytic bacteria were revealed to be excellent sources of natural antioxidant and antimicrobial compounds.     

Antibacterial Activities of a New Brominated Diterpene from Borneon Laurencia spp

In our continuous interest to study the diversity of halogenated metabolites of Malaysian species of the red algal genus Laurencia, we examined the chemical composition of five populations of unrecorded Laurencia sp. A new brominated diterpene, 10-acetoxyangasiol (1), and four other known metabolites, aplysidiol (2), cupalaurenol (3), 1-methyl-2,3,5-tribromoindole (4), and chamigrane epoxide (5), were isolated and identified. Isolated metabolites exhibited potent antibacterial activities against clinical bacteria, Staphylococcus aureus, Staphylococcus sp., Streptococcus pyogenes, Salmonella sp. and Vibrio cholerae.     

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.     

Biorational versus conventional insecticides – Comparative field study for managing red spider mite and fruit borer on tomato

Tomato, Lycopersicum esculentum L. (Solanaceae), is an important crop worldwide that is grown both outdoors and under protected structures, for fresh market consumption and for processing. In the Mariana Islands, tomato is grown as an outdoor crop throughout the year. Tomatoes are attacked by a variety of pests, including the tomato fruitworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), and, in Pacific islands, the red spider mite Tetranychus marianae McGregor. These pests cause scarring, tissue damage, and aberrations in fruit shape or color, making the tomatoes undesirable for fresh market. Also, insect bodies, excretia or parts in fruits reduce their market suitability. Field trials aimed at improving management of these pests were undertaken at two locations in Guam (Yigo and Inarajan), USA in 2012 and 2013, assessing the efficacy of different biorational and conventional insecticides against T. marianae and H. armigera on tomato. At both locations, the mean percentage of mite-infested leaves and the population density of T. marianae were higher in control than in treated plots. An integrated pest management (IPM) program comprising sprays of selective insecticides (Petroleum spray oil, Beauveria bassiana, azadirachtin, and Bacillus thuringiensis), evaluated at 15, 30, 45 and 60 days after transplantation of tomato seedlings, significantly reduced the number of T. marianae-infested leaves and the density of T. marianae over plots treated with carbaryl, malathion, six applications of B. bassiana or B. thuringiensis and over both controls at both locations. Similarly, significantly lower fruit damage by H. armigera was recorded in the plots treated with the IPM program than in plots treated with carbaryl, malathion, or the control treatments at both locations. Marketable tomato yields from the plots which received with the IPM program were significantly greater at both locations than were those in the other treatments.     

Evaluation of bacterial biological control agents for control of root-knot nematode disease on tomato

Among 19 bacterial strains isolated in Yunnan from rhizosphere soils and plant tissues, Bacillus methylotrophicus strain R2-2 and Lysobacter antibioticus strain 13-6 exhibited the highest antagonistic activity against the tomato root-knot nematode Meloidogyne incognita in plate and greenhouse pot experiments. The two strains, when applied as soil drenches or seed treatments in greenhouse experiments, reduced root-knot severity and incidence on tomato compared to no-bacteria controls. In tomato field trials conducted in separate years, soil drench treatment with either strain reduced root-knot disease levels and increased yields compared to the control. Levels of disease control and yield enhancement provided by the strains were higher than those using the chemicals abamectin and carbofuran. This is the first report of B. methylotrophicus being used as a biocontrol agent against a plant parasitic nematode and the first demonstration that B. methylotrophicus and L. antibioticus can suppress disease caused by root-knot nematodes in the field.