Fungicidal activity of fluopicolide for suppression of Phytophthora capsici on squash

The efficacy of a new fungicide fluopicolide in suppression of Phytophthora blight caused by Phytophthora capsici was evaluated under laboratory and field conditions. Studies with 51 P. capsici isolates from vegetable crops in Georgia, USA, indicated that 5.9% of the isolates were resistant, 19.6% were intermediately sensitive, and 74.5% were sensitive to 100 μg ml⁻¹ of mefenoxam based on in vitro mycelial growth. EC₅₀ values of fluopicolide in inhibiting mycelial growth of 25 isolates, representing resistant, intermediately sensitive, and sensitive to mefenoxam, ranged from 0.05 to 0.35 μg ml⁻¹ with an average of 0.2 μg ml⁻¹ EC₅₀ values of fluopicolide in suppressing zoospore germination and sporangium production of the 25 isolates ranged from 1.1 to 4.5 μg ml⁻¹ and 0.3–9.0 μg ml⁻¹, respectively. Evaluation of a collection of 150 P. capsici isolates from vegetables and irrigation ponds found none of the isolates were resistant to 10 μg ml⁻¹ of fluopicolide. Field experiments were conducted to determine the efficacy and application methods of fluopicolide for control of P. capsici on squash in spring 2007 and 2009. Fluopicolide applied through drip irrigation or as a foliar spray at 86.6 or 115.4 g ha⁻¹ consistently provided significant disease reduction and increased squash yield. Results with fluopicolide were similar or slightly superior to mefenoxam applied at recommended rate. Fluopicolide applied at 57.7 g ha⁻¹ did not provide consistent satisfactory disease suppression. The results indicated that fluopicolide was effective in suppression of different stages of the life cycle of P. capsici and could be a viable alternative to mefenoxam for managing Phytophthora blight in squash production.     

Actinomycetes mediated biochemical responses in tomato (Solanum lycopersicum) enhances bioprotection against Rhizoctonia solani

Six actinomycetes isolates, namely Streptomyces toxytricini vh6, Streptomyces flavotricini vh8, S. toxytricini vh22, Streptomyces avidinii vh32, Streptomyces tricolor vh85 and vh41, an isolate of an unknown species of Actinomycetales, were tested for their efficacy in protecting tomato (Solanum lycopersicum) against Rhizoctonia solani under green house conditions. Actinomycetes treated plants showed better growth in terms of high chlorophyll content, higher phenylalanine ammonia lyase (PAL) activity and high total phenolic content. Qualitative and quantitative estimation of phenolic compounds from tomato leaves showed significant accumulation of six phenolic acids, gallic (29.02 μg g⁻¹ fresh leaf wt), ferulic (11.44 μg g⁻¹ fresh wt), cinnamic (56.84 μg g⁻¹ fresh wt), gentisic (24.19 μg g⁻¹ fresh wt), chlorogenic acid (1.72 μg g⁻¹ fresh wt) and salicylic (0.39 μg g⁻¹ fresh wt) acid, in actinomycetes treated plants. Biochemical profiling, when correlated with plant mortality in actinomycetes treated and untreated plants, indicated that isolates vh6 and vh8 offered 44.55% and 40.14% disease reductions, respectively compared to the control. These results established that these organisms have the potential to act as biocontrol agents.     

Efficacy and effects on yield of different fungicides for control of wet bubble disease of mushroom caused by the mycoparasite Mycogone perniciosa

Carbendazim, iprodione, prochloraz-Mn, thiabendazole and thiophanate-methyl were tested in vitro and in vivo for their effect on Mycogone perniciosa, the mycoparasite that causes wet bubble disease of white button mushroom. In vitro experiments showed that prochloraz-Mn (ED₅₀ = 0.006–0.064 μg ml⁻¹) and carbendazim (ED₅₀ = 0.031–0.097 μg ml⁻¹) were the most effective fungicides for inhibiting the mycelial growth of M. perniciosa, while iprodione (ED₅₀ = 1.90–3.80 μg ml⁻¹) was the least effective. The resistance factors calculated for the five fungicides were between 1.4 and 2. The results obtained suggest that there is very little risk that M. perniciosa will develop resistance to the fungicides assayed. The in vivo efficacy of fungicides for control of wet bubble was studied in two mushroom cropping experiments, which were artificially infected with two doses of M. perniciosa, 10⁶ and 10⁷ spores m⁻², respectively. There was, in the low dose inoculum experiment, a very high degree of effectiveness (96.5–100.0%) with all the fungicides assayed. However, iprodione performed poorly (20.5–24.4%) compared with the other fungicides (88.7–100.0%) in the high concentration inoculum experiment. The most effective treatments for controlling wet bubble did not improve the biological efficiency of Agaricus bisporus.     

Highly efficient in vitro regeneration of the industrial oilseed crop Crambe abyssinica

A highly efficient regeneration protocol for oilseed crop Crambe abyssinica has been developed using hypocotyls as explants in this study. Crambe is a potential engineering oilseed crop for industrial purposes as it contains 55-60% erucic acid in its oil and, more importantly, it does not outcross with any food oil seed crops. However, the low regeneration frequency with the currently available protocols is still a limiting factor for genetic modification of Crambe. In this study, we investigated the effects of N-source, C-source, AgNO₃, cultural conditions as well as the concentration and combination of plant growth regulators (PGR) on the regeneration frequency of C. abyssinica. The results showed that all these factors, especially the N-source and PGR concentrations and combinations, played an important role in shoot regeneration. Among all the factors tested, the combination of using hypocotyls from C. abyssinica cv. galactica, the Lepiovre basal medium supplemented with 16 g l⁻¹ glucose, 0.5 g l⁻¹ AgNO₃, 2.2 mg l⁻¹ thidiazuron (TDZ), 0.5 mg l⁻¹ α-naphthaleneacetic acid (NAA), 2.5 g l⁻¹ Gelrite, seeds germinated in dark for 3 days and explants cultured in light, gave the best regeneration frequency (over 95%). The results also suggest that reducing the content of NH₄⁺ or keeping a suitable NO₃⁻/NH₄⁺ ratio in the regeneration medium would be crucial to Crambe shoot regeneration.     

Nematicidal efficacy of isothiocyanates against root-knot nematode Meloidogyne javanica in cucumber

Isothiocyanates (ITCs), a series of new nematicides of the -NCS group, were evaluated for their efficacy against root-knot nematode Meloidogyne javanica. Of the compounds tested, AllylITC, AcITC, EtITC, BzTC, BzITC, 1-PEITC and 2-PEITC showed in vitro irreversible nematicidal activity against second-stage juveniles of M. javanica, following exposure for 72 h at concentrations as low as 5 μg mL⁻¹. When exposed to AllylITC, AcITC and EtITC at lower concentrations, motile juveniles also became irreversibly immobile in 3 days, with a LC₅₀ value at 2.76, 2.53 and 3.05 μg mL⁻¹, respectively. In the pot experiments, 1.0 ml AllylITC and 1.1 ml AcITC per kg of soil controlled M. javanica, similarly to or better than metam sodium at its recommended dose. Similar results were obtained in the field experiments using 1.0 kg AllylITC or 1.0 kg AcITC ha⁻¹. Based on the results of this study, AllylITC and AcITC have potential to be used as new bio-fumigant nematicides.     

Efficacy of a hydroxytyrosol-rich preparation from olive mill wastewater for control of olive psyllid, Euphyllura olivina, infestations

Olive mill wastewater (OMW) is a problematic by-product in Mediterranean countries. Despite this, it is a raw material that is an unfailing source of bioactive molecules. A hydroxytyrosol-rich preparation (HRP) (49.6% weight:dry weight) was extracted from fresh OMW using a hydrolysis and post-hydrolysis purification process. The field efficacy of HRP as a spray treatment (2500 l ha⁻¹) against olive psyllid, Euphyllura olivina (Hemiptera: Psyllidae), was evaluated in 2008 and 2009 in a drip-irrigated olive orchard.The HRP showed strong insecticidal activity against E. olivina at a concentration of 2 g l⁻¹ hydroxytyrosol. Application of HRP resulted in 41.1 and 72.1% control of nymphs and adults, respectively. However, HRP application did not reduce egg hatch. Neither phytotoxicity nor toxicity to auxiliary-fauna was recorded with concentrations of 1.25 g l⁻¹ or 2 g l⁻¹ HRP. But, the 2.5 g l⁻¹ concentration was slightly phytotoxic, especially at the E and F floral phenological stages of the grapes. HRP offers a natural and effective extract for control of olive psyllid and opens a new opportunity for the reuse of OMW.     

Effect of boron on the development of brown rot (Monilinia laxa) on peaches

Requirements of consumers for products with low residues of pesticides have increased the need for alternative disease management practices. The concentration of boron in fruit affects its quality, shelf life and the development of physiological disorders. However, the effect of boron on the susceptibility of peach to fruit rots has not been reported. This study investigated the effect of boron (Power B and Borax) on the development of Monilinia laxa on peaches (cv Andross). Mycelial growth of M. laxa was inhibited on potato dextrose agar supplemented with 750 μg ml⁻¹ of Borax or 1000 μg ml⁻¹ of Power B. The EC 50 values were 107.9 and 522.4 for Borax and Power B respectively. Field investigations showed that the incidence of peach infections by M. laxa was negatively correlated with the content of Boron in the leaves. Post-harvest dipping of peaches in Power B or Borax solution, at concentrations recommended by manufacturer (2 μg ml⁻¹ for Power B and 1 mg ml⁻¹ for Borax), significantly reduced the development of M. laxa. Power B, at rates of 6 μg ml⁻¹, and Borax at rates of 3 mg ml⁻¹ were the most effective in reducing infections by M. laxa. Finally, post-harvest dipping of fruit in Power B or Borax reduced losses of fruit weight and improved fruit firmness one month after storage, showing that boron increased the maintainability of peaches in cold storage. Peaches treated with 6 μg ml⁻¹ Power B or 3 mg ml⁻¹ Borax had the highest flesh firmness and the lowest water losses, while untreated control peaches were the least firm. Generally, Borax was significantly less effective than Power B, but better than the control treatment.     

Potential of extracts of the tropical plant Balanites aegyptiaca (L) Del. (Balanitaceae) to control the mealy bug, Maconellicoccus hirsutus (Homoptera: Pseudococcidae)

The effects of extracts of different parts of the perennial tropical plant Balanites aegyptiaca (L) Del., including various solvent extracts of roots, methanol extracts from leaves, fruits, flowers and roots, partially purified saponins obtained from its roots and a standard saponin were studied on the life cycle (adult longevity, number of eggs, crawlers, adults, weight of adults and % wax content) of a laboratory-reared parthenogenic line of the mealy bug, Maconellicoccus hirsutus (Homoptera: Pseudococcidae). Extracts derived from various parts of B. aegyptiaca (leaves, fruits, flowers, and roots in methanol) affected the life cycle of M. hirsutus with a methanol root extract being the most effective at a concentration of 500 μg ml⁻¹. Partially purified saponin of B. aegyptiaca and the commercial bark saponin extract (Sigma) from Quillaja saponaria at a concentration of 500 μg ml⁻¹ were effective in reducing the longevity of M. hirsutus. Significant reductions in oviposition by M. hirsutus were found for all the extracts at a concentration of 500 μg ml⁻¹. Extracts also affected the number of emerging crawlers, number of adults as well as the weight and wax content of emerging adults. These studies suggest that B. aegyptiaca plant extracts and saponins can be useful botanical insecticides for the protection of crops from mealy bugs.     

Identification and antimicrobial activity of two alkaloids from traditional Chinese medicinal plant Tinospora capillipes

Two antimicrobial alkaloids, palmatine and jatrorrhizine, were isolated from tubers of traditional Chinese medicinal plant Tinospora capillipes using activity-guided isolation method and chromatography. Their antimicrobial activity was determined in vitro. The results showed that palmatine and jatrorrhizine had inhibitory activity against plant pathogens Colletotrichum gloeosporioides, Fusarium oxysporum f. sp. niveum, Mycosphaerella sentina, Pestalotia mangiferae, Cercospora kaki, Gymnosporangium haraeanum, Rhizoctonia solani and Colletotrichum graminicola, with the EC₅₀ values of 0.0348-0.8356 g L⁻¹ and 0.0240-0.8649 g L⁻¹, respectively. Palmatine and jatrorrhizine also exhibited inhibition against animal pathogens Bacillus cereus, Bacillus megaterium, Bacillus subtilis, Staphyloccocus aureus, Staphylococcus epidermidi, Micrococcus lysodeikticus, Proteus vulgaris, Salmonella typhi and Escherichia coli, with the MIC values of 0.1-0.8 g L⁻¹ and 0.1-0.6 g L⁻¹, respectively. These results suggested that palmatine and jatrorrhizine showed relatively broad spectrum antimicrobial activity against plant and animal pathogens.     

Evaluation of post-emergence herbicides for the control of wild oat (Avena fatua L.) in wheat and barley in Argentina

Wild oat (Avena fatua L.) is the most troublesome weed in cereal crops in Argentina. With the aim of studying the effects of different herbicides, doses, and wild oat growth stage at application on weed control and crop yield, field experiments were conducted in wheat and barley crops during three growing seasons in the south of Buenos Aires Province, Argentina. Treatments were post-emergence applications of new herbicide, pinoxaden + cloquintocet mexyl (5%-1.25%), at doses that ranged from 20 g to 60 g a.i. pinoxaden ha⁻¹, applied at two to three leaves and the beginning of tillering of wild oat. In addition, standard treatments were included and applied at the same wild oat growth stages. Diclofop methyl at 511 g a.i. ha⁻¹ and fenoxaprop-p-ethyl at 55 g a.i. ha⁻¹ were applied in barley. In wheat, diclofop methyl was replaced by clodinafop-propargyl + cloquintocet mexyl (24%-6%) at 36 g a.i. clodinafop-propargyl + 9 g cloquintocet mexyl ha⁻¹ and in 2008/09 wheat experiments, iodosulfuron plus metsulfuron methyl (5%-60%) at 3.75 g a.i. ha⁻¹ + 3 g a.i. ha⁻¹ also was included. In both crops, pinoxaden at 30 g a.i. ha⁻¹ and at higher rates, fenoxaprop-p-ethyl and clodinafop-propargyl gave the best control of wild oat. In 2006/07 wheat crops, treatments applied at tiller initiation provided better control than the early timing averaged across herbicides. However, wheat yield generally was greater with early application. In barley, wild oat control and crop yield were similar regarding time of application. Variations in crop yield were correlated with grain number m⁻² both in wheat and barley, but relationships between both grain number and spikes m⁻² and with grains per spike were identified only in wheat.     

Improvement of growth and gymnemic acid production by altering the macro elements concentration and nitrogen source supply in cell suspension cultures of Gymnema sylvestre R. Br

Gymnema sylvestre is an important medicinal plant which bears bioactive compound namely gymnemic acids. The present work deals with optimization of cell suspension culture system of G. sylvestre for the production of biomass and gymnemic acid and we investigated effects of macro elements (NH₄NO₃, KNO₃, CaCl₂, MgSO₄ and KH₂PO₄ - 0.0, 0.5, 1.0, 1.5 and 2.0× strength) and nitrogen source [NH₄⁺/NO₃⁻ ratio of: 0.00/18.80, 7.19/18.80, 14.38/18.80, 21.57/18.80, 28.75/18.80, 14.38/0.00, 14.38/9.40, 14.38/18.80, 14.38/28.20 and 14.38/37.60 (mM)] of Murashige and Skoog medium on accumulation of biomass and gymnemic acid content. The highest accumulation of biomass (165.00 g l⁻¹ FW and 15.42 g l⁻¹ DW) was recorded in the medium with 0.5× concentration of NH₄NO₃ and the highest production of gymnemic acid content was recorded in the medium with 2.0× KH₂PO₄ (11.32 mg g⁻¹ DW). The NH₄⁺/NO₃⁻ ratio also influenced cell growth and gymnemic acid production; both parameters were greater when the NO₃⁻ concentration was higher than that of NH₄⁺. Maximum biomass growth (159.72 g l⁻¹ of FW and 14.95 g l⁻¹ of DW) was achieved at an NH₄⁺/NO₃⁻ ratio of 7.19/18.80, and gymnemic acid production was also greatest at the same concentration of NH₄⁺/NO₃⁻ ratio (11.35 mg g⁻¹ DW).     

Insecticidal and repellent activity of selected essential oils against of the pollen beetle, Meligethes aeneus (Fabricius) adults

The essential oils from 9 aromatic plants were tested on repellency and mortality of Meligethes aeneus adults. All the tested essential oils caused high mortality of M. aeneus adults in the tarsal tests. The lethal doses after 6 h exposure were ranged between 197 and 1508 μg cm⁻². Essential oils obtained from Carum carvi and Thymus vulgaris were most efficient where LD₅₀ was estimated as 197 and 250 μg cm⁻², respectively.Repellency declined in all the essential oils as a function of time. The longest persistence time was determined for essences obtained from C. carvi and T. vulgaris where significantly the highest repellent index of 65.6% and 63.8%, respectively, was determined. Repellent index lower than 15% was determined for the remaining essential oils.     

Regeneration in Jatropha curcas: Factors affecting the efficiency of in vitro regeneration

Factors influencing in vitro regeneration through direct shoot bud induction from hypocotyl explants of Jatropha curcas were studied in the present investigation. Regeneration in J. curcas was found to be genotype dependent and out of four toxic and one non-toxic genotype studied, non-toxic was least responsive. The best results irrespective of genotype were obtained on the medium containing 0.5 mg L⁻¹ TDZ (Thidiazuron) and in vitro hypocotyl explants were observed to have higher regeneration efficiency as compared to ex vitro explant in both toxic and non-toxic genotypes. Adventitious shoot buds could be induced from the distal end of explants in all the genotypes. The number of shoot buds formed and not the number of explants responding to TDZ treatment were significantly affected by the position of the explant on the seedling axis. Explants from younger seedlings (≤15 days) were still juvenile and formed callus easily, whereas the regeneration response declined with increase in age of seedlings after 30 days. Transient reduction of Ca²⁺ concentrations to 0.22 g L⁻¹ in the germination medium increased the number of responding explants.Induced shoot buds, upon transfer to MS medium containing 2 mg L⁻¹ Kn (Kinetin) and 1 mg L⁻¹ BAP (6-benzylamino purine) elongated. These elongated shoots were further proliferated on MS medium supplemented with 1.5 mg L⁻¹ IAA (indole-3-acetic acid) and 0.5 mg L⁻¹ BAP and 3.01-3.91 cm elongation was achieved after 6 weeks. No genotype specific variance in shoot elongation was observed among the toxic genotypes except the CSMCRI-JC2, which showed reduced response. And for proliferation among the toxic genotypes, CSMCRI-JC4 showed highest number of shoots formed. Among the rest, no significant differences were observed. The elongated shoot could be rooted by pulse treatment on half-strength MS medium supplemented with 2% sucrose, 3 mg L⁻¹ IBA (indole-3-butyric acid), 1 mg L⁻¹ IAA, 1 mg L⁻¹ NAA (α-naphthalene acetic acid) and subsequent transfer on 0.25 mg L⁻¹ activated charcoal medium. The rooted plants could be established in soil with more than 90% success. No significant differences were observed in rooting of shoots in the different toxic genotypes. However, rooting response was reduced in non-toxic genotype as compared to toxic genotypes.     

Timing and efficacy of insecticides for control of mussel scale, Lepidosaphes ulmi, on apple using predictive models

The timing and pattern of mussel scale (Lepidosaphes ulmi (L.)) crawler emergence was monitored in relation to air and bark-surface temperatures using sticky band traps around branches in apple orchards in Kent, UK in three successive years, 2007-2009. The emergence and migration of the crawlers lasted for over 4 weeks at a high level, much longer than had been previously reported. In all three years, there were clearly two peaks of emergence of the crawlers, possibly because there is a diapausing and non-diapausing form of the insect. A temperature sum model developed in the 1990s in The Netherlands using air temperatures predicted the peak emergence of crawlers to within 5-16 days; however, the model was less accurate when tree bark temperatures for the north or south of the tree were used. We have developed two models to predict the emergence of the crawlers by revising the original Dutch model. The observed emergence period (5-95%) was longer than the predicted in all three years but the two revised models performed better than the original Dutch model.In two experiments, in 2007 and 2008, the efficacy of a wide range of insecticide products at different timings during crawler emergence was evaluated. Of the 7 products compared as single or two sprays (spray volume 500 l ha⁻¹) at different emergence timings, or with the addition of adjuvants or as winter treatments, thiacloprid 480 g/l SC (375 ml ha⁻¹) and acetamiprid 20% w/w SP (375 g ha⁻¹) gave the best and similar control, reducing the percentage of fruits infested by 94%. Two sprays of fenoxycarb 25% w/w WG (600 g ha⁻¹) gave the poorest control (28%). Two sprays of spirodiclofen 240 g/l SC (600 g ha⁻¹), chlorpyrifos 75% w/w WG (960 g ha⁻¹) or flonicamid 50% w/w WG (140 g ha⁻¹) gave intermediate results, reducing the percentage infestation by 72% on average. Adjuvants gave no additional value to the treatments when applied in admixture and winter applications of mineral oil (spray volume 1000 l ha⁻¹), alone or in admixture with chlorpyrifos or thiacloprid were not effective at controlling mussel scale the following growing season.Overall, acetamiprid and thiacloprid were the most efficacious insecticides and this study indicates that sprays are best applied in the latter part of the migration (after 50% emergence) with an interval between sprays of at least two weeks to span the long emergence period of the pest.     

Chitosan enhances withanolides production in adventitious root cultures of Withania somnifera (L.) Dunal

Calli were obtained from leaf, cotyledon and internode explants of in vitro-grown plants of Indian cultivar of Withania somnifera in MS medium supplemented with 2, 4-D (2.0 mg l⁻¹) and Kinetin (0.2 mg l⁻¹). The brown, semi-friable callus (500 mg FW) derived from leaf explants produced higher number of primary adventitious roots (9 roots/callus) in half strength MS medium fortified with IBA (0.5 mg l⁻¹) and NAA (0.1 mg l⁻¹). The primary adventitious roots with an inoculum mass of 15 g FW were cultured for 6 weeks in the same medium for secondary adventitious root proliferation. Elicitation of abiotic elicitor, aluminium chloride at 10 mg l⁻¹ at the end of 4 weeks culture with 4 h exposure time enhanced withanolides productivity. Under similar culture conditions, the biotic elicitor, chitosan at 100 mg l⁻¹ stimulated higher production of all withanolides when compared to aluminium chloride treatment. This is the first report on the use of callus-derived adventitious root culture for the enhanced production of withanolides upon chitosan elicitation.     

Effect of electrolyzed oxidizing water and continuous ozone exposure on the control of Penicillium digitatum on tangerine cv. ‘Sai Nam Pung’ during storage

The effect of electrolyzed oxidizing (EO) water in combination with ozone to control postharvest decay of tangerine cv. “Sai Num Pung” was investigated. The spore suspension containing 10⁵ conidia ml⁻¹ of Penicillium digitatum was prepared. EO water was generated by electrolysis of various concentrations of NaCl solution (5, 25, 50% and saturated NaCl). The spore suspension was inoculated into EO water and incubated at 27 °C for 1, 2, 4, 8 and 32 min. It was found that the EO water with saturated NaCl completely inhibited the spore germination of the fungus within 1 min. When the fruits inoculated with P. digitatum were washed in EO water at the same concentrations as previous experiment for 4, 8 and 16 min and stored at 5 °C for 18 days, it was found that immersion of the fruit in EO water for 8 min was the most effective to reduce disease incidences. Moreover, washing fruit in EO water and kept in a refrigerated chamber at 5 °C with continuous ozone exposure at a concentration of 200 mg l⁻¹ for 2 h day⁻¹ to extend storage life suppressed the disease incidence until 28 days. However, none of the treatments had any effect on the quality of fruit such as total soluble solids, titratable acidity, percent weight loss and peel color. Therefore EO water may be useful for surface sanitation and ozone has potential to control the recontamination of postharvest diseases in tangerine fruit in storage room.     

In vitro plant regeneration from decapitated embryonic axes of Clitoria ternatea L.—An important medicinal plant

In vitro clonal propagation of Clitoria ternatea has been achieved by employing decapitated embryonic axes (DEAs) explants. The explants induced multiple shoots on cytokinin-containing medium. Several cytokinins [6-benzylaminopurine (BAP), 6-furfuryl aminopurine (KIN) and thidiazuron (TDZ)] were assayed. The best response was achieved with 2 mg l⁻¹ BAP in which 100% of cultures produced 6.0 ± 0.14 shoots per explant. MS + 1 mg l⁻¹ gibberellic acid (GA₃) was the most suitable for shoot elongation. Regenerated shoots were rooted in half-strength Murashige and Skoog (MS) medium with 0.2 mg l⁻¹ indole-3-butyric acid (IBA). Plantlets were successfully acclimatized and established in soil, and they were morphologically indistinguishable from the source plant. The plantlets attained maturity and flowered normally. The efficient regeneration protocol reported here provides an important method of micropropagation of this plant. Furthermore, this protocol may be used for genetic transformation of this valuable medicinal plant for its further improvement.     

Potassium phosphite for control of downy mildew of soybean

Downy mildew of soybean, caused by Peronospora manshurica, is widely spread throughout Brazil. The objective of this study was to evaluate the use of potassium phosphite to control this disease. Field experiments were conducted during the growing season of 2006/2007 and 2007/2008 in the state of Parana in southern Brazil. The experimental design consisted of completely randomized blocks in a factorial arrangement (4 × 2) with four replications. Four rates of potassium phosphite (0, 375, 750 and 1500 g P₂O₅ + K₂O ha⁻¹) were applied at two growth stages, V6 (fifth trifoliolate leaf) and R2 (full flowering), followed by one or two applications of pyraclostrobin and epoxiconazole (66.5 + 25 g a.i. ha⁻¹) at R3 (pod development) or R2 and R5.1 (10% of pod filling), mainly for the control of Asian soybean rust (Phakopsora pachyrhizi) and powdery mildew (Microsphaera diffusa). Field experiments were conducted to quantify the severity of downy mildew on leaves, nutrient content in leaf tissue (N, P and K), leaf area index (LAI), yield and seed weight. The maximum severity of downy mildew was observed at a growth stage of R5.3 (50% of pods were ripe), with 14% and 46% of the leaf area affected in 2006/2007 and 2007/2008, respectively. Also it was detected some effect of phosphite on Asian rust control but it was mostly in the trial of 2007/08 when the epidemic was very low (9.7-21.8% of severity). There was a linear reduction in the severity of downy mildew and a significant improvement in the LAI with an increase in the rate of phosphite applied. During the 2006/2007 growing season, a significant yield improvement was observed due to the application of the highest rate of phosphite. Two fungicide applications following phosphite application significantly improved the control of Asian soybean rust and powdery mildew, yield and seed weight when compared to a single fungicide application.     

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.     

Occurrence and characterization of fungi and oomycetes transmitted via potting mixtures and organic manures

A study was conducted to investigate the most common fungal and oomycete pathogens introduced into farms in Oman via potting mixtures and organic manures. A total of 37 commercial types of potting mixtures (2 local and 35 imported from overseas), 4 commercial types of organic manures and 11 non-commercial types of organic manures were included in the study. Identification of the isolated species was based on morphological characteristics, except for the most common species which were further identified using sequences of the internal transcribed spacer region of the ribosomal DNA (ITS rDNA). Fusarium spp. (14%), Pythium aphanidermatum (3%), Alternaria spp. (5%), Helminthosporium spp. (5%) and Cladosporium spp. (3%) were recovered at different frequencies from samples of potting mixtures. Fusarium solani (40%) and Fusarium equiseti (47%) were recovered at high frequencies from samples of organic manures. Isolations from organic manures also yielded Pythium periplocum (7%), Rhizoctonia solani (7%), Fusarium lichenicola (7%), Helminthosporium spp. (27%) and Alternaria spp. (27%). Trichoderma spp., Penicillium spp., Aspergillus spp. and Rhizopus spp. were found to be common in samples of potting mixtures and organic manures. Investigating sensitivity to hymexazol among 9 isolates of F. equiseti and 13 isolates of F. solani revealed variations among different isolates. The EC₅₀ values ranged from 1 to over 1200 (avg. 192 μg ml⁻¹) for F. equiseti isolates and from 135 to 789 (avg. 324 μg ml⁻¹) for F. solani isolates, indicating presence of resistance to this important fungicide among some Fusarium isolates. This appears to be the first report of contamination with R. solani, P. periplocum, F. solani, F. equiseti and F. lichenicola of organic manures. This study appears to report for the first time F. lichenicola in Oman and appears to be the first report of occurrence of resistance to hymexazol among F. equiseti and F. solani isolates.