Ethanol production from alkali- and ozone-treated cotton stalks using thermotolerant Pichia kudriavzevii HOP-1

In the present study, milled cotton stalks were subjected to alkali pretreatment with NaOH at 1-4% (w/v) concentrations at 121 °C for time ranging from 30 to 90 min. Ozone pretreatment was performed by passing 45 mg/L of ozone gas over 2 mm cotton stalks for 150 min at a flow rate of 0.37 L/min. The residual biomass from 4% alkali pretreatment for 60 min showed 46.6% lignin degradation accompanied by 83.2% increase in glucan content, compared with the untreated biomass. Hydrolysis of 4% alkali-treated and ozone-treated cotton stalks was conducted using enzyme combination of 20 filter paper cellulase units/gram dried substrate (FPU/g-ds), 45 IU/g-ds β-glucosidase and 15 IU/g-ds pectinase. Enzymatic hydrolysis of alkali-treated and ozone-treated biomass after 48 h resulted in 42.29 g/L glucose, 6.82 g/L xylose and 24.13 g/L glucose, 8.3 g/L xylose, respectively. About 99% of glucose was consumed in 24 h by Pichia kudriavzevii HOP-1 cells resulting in 19.82 g/L of ethanol from alkali-treated cotton stalks and 10.96 g/L of ethanol from ozone-treated cotton stalks. Simultaneous saccharification and fermentation of the alkali-treated cotton stalks after 12-h pre-hydrolysis resulted in ethanol concentration, ethanol yield on dry biomass basis and ethanol productivity of 19.48 g/L, 0.21 g/g and 0.41 g/L/h, respectively which holds promise for further scale-up studies. To the best of our knowledge, this is the first study employing SSF for ethanol production from cotton stalks.     

Mathematical modeling for extraction of essential oil from Baccharis spp. by steam distillation

Baccharis (Compositae-Asteraceae) is a large genus of plants distributed from the USA to Argentina, of which 90% are located in South America. In recent decades, species of the genus have been studied, due to their importance as sources of novel active components with possible applications in the pharmaceutical and chemical industries. This work studied three species of Baccharis: Baccharis uncinella DC, Baccharis anomala DC and Baccharis dentata (Vell) G.M. Barroso collected in São Francisco de Paula in southern Brazil. The aim of this work is the evaluation of the aromatic potential of the essential oil obtained by steam distillation method and the simulation of the extraction data by means of a mathematical model. A mathematical model based on mass transfer fundamentals is developed and applied to correlate the experimental data. This model is based on the existence of the broken and intact cells, in which the oil located in the broken cells is rapidly extracted and the oil from intact cells diffuses slowly to the surface of the vegetal material. The analysis of the essential oil was carried through by GC-MS and the major compounds identified to both processes were α-pinene, β-pinene, and spathulenol (B. uncinella), spathulenol, β-caryophyllene, and β-selinene (B. anomala), and germacrene-D, caryophyllene oxide, and spathulenol (B. dentata).     

Biorefinery of Cannabis sativa using one- and two-step steam treatments for the production of high quality fibres

Among the sources of lignocellulosic biomass that could be considered as a feedstock for a biorefinery, hemp (Cannabis sativa) is known for the quality and relative abundance of its fibre. In this work, two variations of the Feedstock Impregnation Rapid and Sequential Steam Treatment (FIRSST) process are compared for the production of both fibres and biomolecules (including liquid fuels) as the valorization of the entire carbon content of biomass is needed to reach economical viability. In the first variation, the fibres were isolated via two specific and sequential steam treatments while in the second variation, fibres were isolated using only one steam treatment. Both processes allowed the isolation of a fibre that was comparable to kraft pulp while reducing the amounts of ashes along the process. Compositions of the residual solid fibre at different steps of the treatment were evaluated using ASTM and TAPPI standard methods. Carbohydrates were identified and quantified using HPLC with an anion exchange stationary phase. The qualities of the FIRSST and Kraft pulp produced were compared using standard ATPPC methods. Both pulps showed similar indexes. The two-steps FIRSST process has leaded to direct isolation of cellulose, hemicelluloses and lignin that could be suitable for the production of energy (biofuels) and high value chemicals, with small investments in terms of chemicals and energy.     

Optimization of formic/acetic acid delignification of Miscanthus × giganteus for enzymatic hydrolysis using response surface methodology

A Box-Behnken experimental design and response surface methodology were employed to optimize the pretreatment parameters of a formic/acetic acid delignification treatment of Miscanthus × giganteus for enzymatic hydrolysis. The effects of three independent variables, namely cooking time (1, 2 and 3 h), formic acid/acetic acid/water ratio (20/60/20, 30/50/20 and 40/40/20) and temperature (80, 90 and 107 °C) on pulp yield, residual Klason lignin content, concentration of degradation products (furfural and hydroxymethylfurfural) in the black liquor, and enzymatic digestibility of the pulps were investigated. The major parameter influencing was the temperature for pulp yield, delignification degree, furfural production and enzymatic digestibility. According to the response surface analysis the optimum conditions predicted for a maximum enzymatic digestibility of the glucan (75.3%) would be obtained using a cooking time of 3 h, at 107 °C and with a formic acid/acetic acid/water ratio of 40/40/20%. Glucan digestibility was highly dependent on the delignification degree.     

Intercropping reduces Mycosphaerella pinodes severity and delays upward progress on the pea plant

Mycosphaerella pinodes is a serious pea disease of worldwide distribution. The increasing interest of sustainable tools for disease control, together with the lack of sufficient levels of genetic resistance has brought our interest in the use of intercropping as a tool for management of this disease. Effect of intercropping on M. pinodes severity was studied in field experiments performed in Spain and Tunisia, in which a susceptible pea cultivar was grown as monocrop and as two species mixed intercrop with either faba bean, barley, oat, triticale or wheat. Disease was significantly reduced in terms of both percent of diseased tissue per plant and vertical progress of lesions when pea was intercropped. Faba bean and triticale intercropped with pea showed the highest suppressive ability with above 60% of disease reduction. Oat, barley and wheat showed low to moderate M. pinodes suppressive effects. Suppressive effects can be ascribed to a combined reduction of host biomass, altered microclimate and physical barrier to spore dispersal.     

Endophytic Lecanicillium lecanii and Beauveria bassiana reduce the survival and fecundity of Aphis gossypii following contact with conidia and secondary metabolites

Lecanicillium lecanii and Beauveria bassiana are important entomopathogens of Aphis gossypii. Their capacity to colonize crop plants is also becoming widely recognized. Their presence in crop plants indicates the possibility of a much greater potential for contact between insect and fungus than previously recognized. The present experiment aimed to study the effects of endophytic strains of these fungi on the survival, and reproduction of A. gossypii. Contact with conidia of both fungi significantly reduced the rate and period of reproduction of A. gossypii. The culture filtrates of L. lecanii and B. bassiana significantly increased mortality and feeding-choice experiments indicate that insects may be able to detect metabolites of the fungi. The culture filtrate of L. lecanii also significantly reduced the reproduction of the aphid. The ethyl acetate and methanolic fractions of the culture filtrate and of mycelia of L. lecanii also caused significant mortality and reduced fecundity of A. gossypii. The methanolic fractions of mycelia of B. bassiana caused significant mortality of A. gossypii. The present investigations indicated that A. gossypii is affected by contact with both conidia and fungal metabolites. This broad influence indicates that these fungi may have a role in regulating insect pest populations.     

Comparative susceptibility of Ostrinia furnacalis, Ostrinia nubilalis and Diatraea saccharalis (Lepidoptera: Crambidae) to Bacillus thuringiensis Cry1 toxins

Transgenic corn hybrids that express toxins from Bacillus thuringiensis (Bt) are highly effective against the European corn borer, Ostrinia nubilalis (Hübner), and the closely related Asian corn borer, Ostrinia furnacalis (Guenée). Since the registration of Bt corn hybrids in the U.S. in 1996, there has been a great deal of information generated on O. nubilalis. However, relatively little information exists for O. furnacalis. To help determine whether the information generated for O. nubilalis can be leveraged for decisions regarding the use of transgenic Bt corn against O. furnacalis, experiments were designed to determine whether the pattern of sensitivity to various Bt Cry1 toxins is similar between the two species. Test insects included laboratory-reared O. furnacalis originating from Malaysia, a Bt-susceptible laboratory colony of O. nubilalis maintained at the University of Nebraska-Lincoln (UNL) and an out-group consisting of the sugarcane borer, Diatraea saccharalis (F.), from Louisiana which represents a different genus from the same family. O. furnacalis and O. nubilalis exhibited a similar pattern of susceptibility to all the Cry1 toxins and were highly susceptible to the range of Bt toxins tested including Cry1Aa, Cry1Ab, Cry1Ac and Cry1F. Both of the Ostrinia species were more tolerant to Cry1Ba compared with D. saccharalis, although sensitivity of O. furnacalis was intermediate and did not differ significantly from that of O. nubilalis and D. saccharalis. D. saccharalis was also susceptible to the range of toxins tested but unlike the two Ostrinia species, was more tolerant to Cry1F and more susceptible to Cry1Ba. These results indicate that both of the Ostrinia corn borer species are similar in sensitivity to the Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ba and Cry1F toxins, thus suggesting shared toxin receptors and mechanisms of toxicity for the two species.     

Effects of crop rotation of soybean with corn on severity of sudden death syndrome and population densities of Heterodera glycines in naturally infested soil

Sudden death syndrome, caused by Fusarium virguliforme, and the soybean cyst nematode, Heterodera glycines, combined cause the highest yield losses in soybean. The objective of this study was to determine the effectiveness of corn rotated annually with soybean on reducing severity of sudden death syndrome (SDS) and if such crop rotation is beneficial to soybean root health and thus improves disease management strategies. Experiments were conducted from 2003 to 2006 through two cycles of a corn–soybean rotation on two commercial fields in Indiana. With one exception, the rotation of soybean with corn did not provide yield benefits compared to monoculture of soybean. Severity of foliar and root symptoms of SDS in rotation plots were never less than in soybean monoculture plots. At one location, soybean monoculture resulted in suppression of SDS compared to the corn–soybean rotation, while H. glycines reproduced freely. At the other location, monoculture of soybean resulted in suppressiveness against H. glycines, while SDS was limited in all treatments. The data suggest that soil suppressiveness can independently impact the pathogens that are important in SDS development. Because H. glycines can increase SDS symptoms, its suppression may also reduce severity of SDS. Current production systems consisting of yearly rotation of soybean with corn are highly vulnerable to the development of severe soil-borne disease complexes. The simple year-to-year rotation of corn and soybean is not considered sustainable. While monoculture of soybean resulted in some disease suppression in these trials, reliance on monoculture may be detrimental due to other environmental considerations beyond the scope of these trials. Including other crops may be beneficial in improving the sustainability of soybean and corn production systems.     

Microwave-enhanced alkali treatment of Pinus yunnanensis: Physiochemical characterization of the dissolved lignins

Pinus yunnanensis was subjected to water-bath and microwave treatments in 1% NaOH aqueous solutions at 100 °C with various ratios of bath heating time to microwave heating time (0/120, 20/100, 40/80, 60/60, 80/40, 100/20 and 120/0 min). The lignins dissolved in the alkali liquors were separated and purified, and their physicochemical features were comparatively characterized by sugar analysis, GPC, FT-IR, ¹³C and HSQC NMR, as well as thermogravimetric analysis (TGA). The results showed that the lignin fractions extracted with microwave heating (20-120 min) had high molecular weights and polydispersities (M_w 3150-5710 g/mol, M_n 2130-3020 g/mol, M_w/M_n 1.48-2.00) as compared to those prepared without microwave heating (M_w 3080 g/mol, M_n 2080 g/mol, M_w/M_n 1.48). The most striking characteristic of all lignin fractions was the almost absence of associated sugars (0.16-3.25%). The TGA results indicated that the thermal stability of the lignin fraction increased with the increment of the molecular weight. FT-IR and NMR spectra suggested that the lignin fractions showed similar structures which were mainly composed of guaiacyl (G) and minor amounts of p-hydroxyphenyl (H) units. Moreover, HSQC NMR spectrum of a typical lignin fraction (prepared with microwave heating for 120 min) revealed that it contained dominant amounts of β-O-4′ linkages (64.6%) and phenylcoumaran (β-5′) substructures (25.8%) together with small amounts of resinol (β-β′) substructures (6.7%) and coniferyl alcohol end groups (2.9%).     

High level of resistance to Sclerotinia sclerotiorum in introgression lines derived from hybridization between wild crucifers and the crop Brassica species B. napus and B. juncea

Sclerotinia rot caused by the fungus Sclerotinia sclerotiorum is one of the most serious and damaging diseases of oilseed rape and there is keen worldwide interest to identify Brassica genotypes with resistance to this pathogen. Complete resistance against this pathogen has not been reported in the field, with only partial resistance being observed in some Brassica genotypes. Introgression lines were developed following hybridization of three wild crucifers (viz. Erucastrum cardaminoides, Diplotaxis tenuisiliqua and E. abyssinicum) with B. napus or B. juncea. Their resistance responses were characterized by using a stem inoculation test. Seed of 54 lines of B. napus and B. juncea obtained from Australia, India and China through an Australian Centre for International Agricultural Research (ACIAR) collaboration programme were used as susceptible check comparisons. Introgression lines derived from E. cardaminoides, D. tenuisiliqua and E. abyssinicum had much higher levels (P < 0.001) of resistance compared with the ACIAR germplasm. Median values of stem lesion length of introgression lines derived from the wild species were 1.2, 1.7 and 2.0 cm, respectively, as compared with the ACIAR germplasm where the median value for stem lesion length was 8.7 cm. This is the first report of high levels of resistance against S. sclerotiorum in introgression lines derived from E. cardaminoides, D. tenuisiliqua and E. abyssinicum. The novel sources of resistance identified in this study are a highly valuable resource that can be used in oilseed Brassica breeding programmes to enhance resistance in future B. napus and B. juncea cultivars against Sclerotinia stem rot.     

Mycoparasitic Trichoderma viride as a biocontrol agent against Fusarium oxysporum f. sp. adzuki and Pythium arrhenomanes and as a growth promoter of soybean

Trichoderma viride was proved as an effective biocontrol agent against two fungal pathogens, Fusarium oxysporum f. sp. adzuki and Pythium arrhenomanes, infecting soybean. During an in vitro biocontrol test, Trichoderma showed mycoparasitism and destructive control against the tested fungal pathogens. Both the pathogens significantly influence the germination and P. arrhenomanes had a severe effect (only 5% germination). The root system of the soybean plant was poorly developed due to the infection and it exerted a negative influence on the nodulation and further growth phases of the plant. During pot assay along with biocontrol activity, Trichoderma showed growth promoting action on the soybean plant. Trichoderma enhanced growth of shoot and root systems and fruit yield after 12 weeks of growth. Pythium and Fusarium infected plants treated with Trichoderma had ∼194% and 141% more height than pathogens alone. The fruit yield treated with Trichoderma was ∼66 per plant whereas the yield was only 41 for a control plant. The plants infected with Pythium and Fusarium and treated with Trichoderma had fruit yields of 43 and 53 respectively and those were 5 and 1.6 times higher than plants infected with pathogens.     

Mild peroxyformic acid fractionation of Miscanthus × giganteus bark. Behaviour and structural characterization of lignin

Miscanthus × giganteus bark was subjected to mild fractionation with peroxyformic acid by a two stage process. A factorial experimental design was used to study and quantify the effect of the variables (formic acid concentration (80-90%), hydrogen peroxide concentration (0.2-0.4%), temperature of the first stage (60-80 °C), and treatment time of the second stage (60-120 min)) on the main parameters of fractionation: pulp yield, remaining lignin and total polysaccharides in pulp. The dependence of lignin precipitation rate on hydrogen peroxide concentration in liquor was also studied. Hydrogen peroxide concentrations inferior to 0.5% seems to be suitable to recover high percentages of lignin. The isolated lignin was analysed by 2D-HSQC, ¹³C- and ³¹P NMR spectroscopy, FTIR spectroscopy, size-exclusion chromatography and chemical analysis. The most important chemical modifications taken place in the lignin during the fractionation were identified: β-O-4′ cleavage and hydrolysis of LC-bond structures. The C9-formula was also determined: C₉H_6.81O_2.90(OCH₃)_0.68(COOH)_0.07(OH^Ph)_0.38(OH^Al)_0.33.     

Differential response of pea (Pisum sativum) to Orobanche crenata, Orobanche foetida and Phelipanche aegyptiaca

Several broomrape species including Orobanche crenata, Orobanche foetida and Phelipanche aegyptiaca are reported to infect various grain and forage legumes in the Mediterranean and West Asia. Pea (Pisum sativum) is severely damaged by O. crenata, but there are no reports on O. foetida or P. aegyptiaca infection. We report here that pea can induce high germination of seeds of O. crenata, O. foetida and P. aegyptiaca but only O. crenata success infecting pea roots and developing further. Some differences in levels of infection by O. crenata were observed among pea accessions what can be exploited in pea resistance breeding programmes. On the contrary, all pea accessions studied were highly resistant to infection by both O. foetida and P. aegyptiaca, preventing any tubercle attachment and development. This makes pea a promising candidate as trap crop for O. foetida and P. aegyptiaca seed bank demise in infested soils.     

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.     

Both incidence and severity of white rust disease reflect host resistance in Brassica juncea germplasm from Australia,China and India

White rust (Albugo candida) is a highly destructive disease of oilseed Brassicas such as Brassica juncea and B. rapa, and has caused serious yield losses in Australia, China and India on both species. The first commercial B. juncea varieties are now being deployed in Australia, but their response to Australian strains of A. candida is yet to be defined under Australian field conditions. To identify useful sources of host resistance for Australia, China and India, in B. juncea, three field trials were undertaken in Western Australia. Forty-four B. juncea genotypes, viz. 22 from India, 12 from Australia and 10 from China, were tested. Varying levels of host resistance to Australian strains of A. candida (race 2) were identified among the genotypes from the three countries. Genotypes CBJ-001, CBJ-003 and CBJ-004 from China consistently showed high levels of resistance to A. candida on leaves across the three trials. Overall, the genotypes from China showed the best resistance, followed by the genotypes from Australia, with those from India being the most susceptible. The most susceptible genotypes were RL1359, RH30 and Seetha from India. It is noteworthy that both the incidence and severity of disease reflected varying levels of host resistance in the germplasm from the three countries, irrespective of whether screening was undertaken in the field using natural or artificial inoculation. Differentiation of resistance among these genotypes was similar to that we reported previously for artificially-inoculated seedlings or adult plants under glasshouse conditions, indicating that a choice of options is available to plant breeders to reliably differentiate host resistance among genotypes to white rust in B. juncea.     

Characterization of Solanum chomatophilum resistance to 2 aphid potato pests, Macrosiphum euphorbiae (Thomas) and Myzus persicae (Sulzer)

The aphids Macrosiphum euphorbiae (Thomas) and Myzus persicae (Sulzer) (Hemiptera: Aphididae) are responsible for yield reduction in potato (Solanum tuberosum) production by direct phloem feeding and by spreading viruses. Breeding resistant traits from Solanum chomatophilum into the potato germplasm provides alternative means to control aphid infestations. Integrated pest management strategy, using plant resistance, benefits from the characterization of the resistance and of its impact on aphid biology. Our objective was to characterize the resistance of S. chomatophilum by assessing the effects of accessions, plant parts on aphid performance, and by assessing the impact of the resistance factors on different aphid developmental stages and on alate morph production. Detailed aphid performance was obtained by measuring fecundity, survival, percentage of nymphs that reached adult moult, and population growth using whole plant and clip cage experimental designs. Accession and plant physiological age, but not aphid developmental stage, influenced all life-history parameters, except for alate morph production which was not induced on the resistant accessions. Plant part influence was independent of plant species and accession. Both experimental designs resulted in congruent resistance levels at the accession level for each of the two aphid species, supporting the use of any of them in S. chomatophilum resistance screening. PI243340 was resistant to both aphid species, while PI365324 and PI310990 were also resistant to M. euphorbiae and M. persicae, respectively.     

Variability in essential oil content and composition of Origanum hirtum L., Origanum onites L., Coridothymus capitatus (L.) and Satureja thymbra L. populations from the Greek island Ikaria

Plants belonging to four “oregano” plant species (Origanum hirtum L., Origanum onites L., Coridothymus capitatus L., and Satureja thymbra L.) were collected during flowering from 33 sites located in the eastern part of the Greek island of Ikaria in the Eastern Aegean during April, May and July 2008. C. capitatus and O. hirtum were mostly observed in higher altitudes, whereas O. onites and S. thymbra in lower ones. The spatial distribution of all species was depicted on a GIS map. All four species exhibited essential oil concentrations higher than those reported in earlier literature, namely O. onites 3-4.3%, S. thymbra 4-6.5%, C. capitatus 3.7-5.6% and O. hirtum 5.5-10.0% (v/w). Carvacrol was the main constituent of the essential oils of all species, followed by γ-terpinene, p-cymene and caryophyllene, while thymol was not detected. All constituents varied remarkably among the four species, with carvacrol exhibiting the lowest variation. Carvacrol content varied between 72.3 and 89.2% in O. onites; 46.5 and 58.0% in S. thymbra; 82.9 and 90.9% in C. capitatus; and 84.4 and 93.8% in O. hirtum. By applying hierarchical cluster analysis on the basis of the essential oil constituents two main groups, divided into four subgroups of the taxa were evident. The first group consisted of O. onites and S. thymbra, while the second one of C. capitatus and O. hirtum. The results are discussed in terms of topography and climatic variation.     

Characterization and In Vitro digestibility of rice protein prepared by enzyme-assisted microfluidization: Comparison to alkaline extraction

Microfluidization followed by density-based separation was employed to extract protein from broken rice by disrupting protein-starch agglomerates. Follow-up enzyme treatments (amylase and glucoamylase) were performed to further improve the purity of the protein-rich fraction. High protein recovery (81.87%) and purity (87.89%) were obtained. The protein composition, solubility, structural properties, and in vitro digestibility of rice proteins prepared by enzyme-assisted microfluidization (EM-RP) and alkaline extraction (AE-RP) were compared. EM-RP was mainly composed of glutelin, which had low solubility and native structure. By contrast, large quantities of prolamin and globulin appeared in the AE-RP except glutelin, leading to the richness of glutamic acid/glutamine, leucine, aromatic and charged amino acids in the AE-RP. Compared to AE-RP, EM-RP showed higher digestibility due to the richness of glutelin (an easy-to-digest protein), as evidenced by higher nitrogen release during pepsin-trypsin digestion. The presence of prolamin (an indigestible protein) in AE-RP decreased protein digestibility although alkaline extraction improved its hydrolysis. These results suggest that enzyme-assisted microfluidization could be an effective technique to non-destructively and selectively extract rice glutelin.     

Effects of crude seed extracts of Annona atemoya and Annona squamosa L. against the cabbage looper, Trichoplusia ni in the laboratory and greenhouse

Antifeedant, growth inhibitory and toxic effects of crude seed extracts of Annona squamosa and Annona atemoya from Fazenda Viveiro Bona, Parasisópolis – Minas Gerais, Brazil, were evaluated against the cabbage looper, Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) using different bioassays. Crude methanolic seed extracts deterred feeding of third instar T. ni larvae in a leaf disc choice bioassay. A. squamosa was ∼10 times more active as a feeding deterrent than A. atemoya (DC₅₀ = 2.3 mg/ml vs. 20.1 mg/ml). A. squamosa was ∼three times more active as a growth inhibitor than A. atemoya (EC₅₀ = 38.0 ppm vs. 117.0 ppm). Methanolic seed extracts of A. squamosa and A. atemoya were toxic to third instar T. ni larvae both through topical and oral application. A. squamosa was more toxic through feeding (LC₅₀ = 167.5 ppm vs. 382.4 ppm) whereas, A. atemoya exerted greater toxicity via topical application (LC₅₀ = 301.3 μg/larva vs. 197.7 μg/larva). Both A. squamosa and A. atemoya extracts reduced leaf area consumption and larval growth in a greenhouse experiment. Our results indicate that both A. squamosa and A. atemoya have potential for development as botanical insecticides, especially for local use in Brazil.     

Wilt disease management and enhancement of growth and yield of Cajanus cajan (L) var. Manak by bacterial combinations amended with chemical fertilizer

Root nodulating Sinorhizobium fredii KCC5 and Pseudomonas fluorescens LPK2 were isolated from nodules of Cajanus cajan and disease suppressive soil of tomato rhizosphere, respectively. Both strains produced IAA, siderophore, solubilized insoluble phosphate, showed chitinase and β-1,3-glucanase activities, and strongly inhibited the growth of Fusarium udum. It also caused degradation and digestion of cell wall components, resulting in hyphal perforations, empty cell (halo) formation, shrinking and lysis of fungal mycelia along with significant degeneration of conidia. LPK2 produced volatile cyanogen (HCN). Combinations of S. fredii KCC5 and P. fluorescens LPK2 with half dose of chemical fertilizers showed a significant increase in seed germination (94%) while seed germination with co-inoculated strains (KCC5 + LPK2), KCC5 and LPK2 alone was 90, 84 and 82% respectively as compared to control 77%. After 120 days of sowing, per plant number of pods, nodules, shoot length, root length, shoot weight and root weight were greater for the combination with half dose of chemical fertilizers compared to the control. Combinations of S. fredii KCC5 and P. fluorescens LPK2 with half dose of chemical fertilizers resulted in an 82% increase in grain yield per hectare compared to the control. Both strains KCC5 and LPK2 led to proto-cooperation as evidenced by synergism, aggressive colonization of the roots, and enhanced growth, suggesting potential biocontrol efficacy against Fusarium wilt in C. cajan.