Direct plant regeneration from leaf explants of Plumbago species and its genetic fidelity through RAPD markers

In vitro plant regeneration was achieved from leaf explants of Plumbago rosea and Plumbago zeylanica on Murashige & Skoog (1962) medium supplemented with 1.5 mg litre^?1 6‐benzylaminopurine, 0.25 mg litre^?1 indole‐3‐acetic acid, 50 mg litre^?1 adenine sulfate and 3% (w/v) sucrose. The shoot initials developed within 2–3 wk on the leaf margin as well as from the wounds of the leaf. High frequency shoot‐bud regeneration was achieved on similar medium in subsequent subcultures. The semi‐mature leaves produced more shoot‐buds as compared to the younger leaves. Mature leaves did not show any response for shoot bud initiation. More than 85% of the semi‐mature explants produced shoot‐buds per leaf explant within 4 wk of culture. Shoots rooted easily on medium having half‐strength basal Murashige & Skoog (1962) medium supplemented with 0.25 mg litre^?1 indole‐3‐butyric acid and 2% (w/v) sucrose; 84–92% of the in vitro rooted plantlets survived in the greenhouse. The regenerated plantlets appeared morphologically similar to the mother plants. No variation was detected among the regenerated plants by the use of Randomly Amplified Polymorphic DNA (RAPD) markers. This method might be useful for assessing plant improvement programmes.     

Determination of biokinetic parameters for ingestion of radionuclides of zirconium in animals using stable tracers

Exposure to the radioactive isotope ⁹⁵Zr, as in nuclear accidents, and to stable zirconium, due to its use in industry, has increased the interest in the biokinetics of this element. Information has been derived mainly from tests performed on animals by means of radioactive tracers. Due to the fact that extrapolation from animals to humans is always open to question, there is an increasing need of a methodology which allows data to be obtained directly from humans. The use of stable tracers, being ethically justifiable, is a powerful tool for providing this information. As two tracers of the same element must be utilized in order to evaluate gut absorption, an analytical technique which is capable of distinguishing and measuring simultaneously different isotopes of zirconium in biological samples is required. Preliminary tests on laboratory animals were performed in order to assess the feasibility of the double tracer technique combined with proton activation analysis. Received: 7 April 1999 / Accepted in revised form: 4 November 1999     

Effects of carbonyl sulfide, methyl iodide, and sulfuryl fluoride on fruit phytotoxicity and insect mortality

Three potential chemical fumigants: carbonyl sulfide (COS), methyl iodide (MI) and sulfuryl fluoride (SF) were tested at selected dosages on lemons against California red scale (Aonidiella aurantii) and MI and COS were tested on nectarines against codling moth (Cydia pomonella). In nectarines, COS was tested at 0, 20, 40, 60 and 80 mg litre^?1, MI at 0, 10, 15, 20 and 25 mg litre^?1. Both fumigants intensified nectarine peel color, delayed fruit softening, but did not alter overall fruit quality. COS at 80 mg litre^?1 resulted in 87% codling moth mortality, but the fumigant dosage was insufficient to reach the desired probits 9 level (99.9968%). MI gave 100% codling moth mortality at 25 mg litre^?1. Lemons were treated with MI at 0,10,20,40,60 mg litre^?1, SF at 0,10,20,40, 80 mg litre^?1 and COS at 0,20,40, 60 and 80 mg litre^?1. MI gave 100% red scale mortality at ≥40 mg litre^?1 but caused significant fruit injury. Conditioning lemons at 15°C for 3 days before MI fumigation lessened lemon phytotoxicity. Forced aeration at 3.5 standard litres per minute of lemons for 24 h following MI fumigation at 20 mg litre^?1 significantly reduced phytotoxicity compared to 2 h postfumigation aeration after MI treatment. SF at ≥40 mg litre^?1 gave 100% red scale mortality but resulted in commodity phytotoxicity. Lemons treated with the highest selected dose of 80 mg litre^?1 COS gave only 87% kill of red scale, but failed to reach the desired probit 9 level.     

Stable isotopes of minerals as metabolic tracers in human nutrition research

Enriched stable isotopes used as tracers have proven to be valuable in studies of the absorption and metabolism of minerals. Unlike radioisotopes, they can be used in high-risk population groups such as infants, children, and pregnant or lactating women. Estimates of mineral absorption can be made from the oral administration of a single tracer or from two tracers, one given orally and the other intravenously (IV). It is possible to determine the metabolism of the mineral with modeling based on the amount of the tracer or tracers in different biological samples. One of the key decisions in studies of this type is determining which enriched isotope and what amount to use. An example is given of calculations to estimate and compare the amounts of tracers needed for an absorption study. Methods for calculating the amounts of tracer in oral and IV doses are presented, and limits of detection and quantitation are discussed in terms of percent of enrichment and related to isotope ratio measurement precision. A general review of the use of mass spectrometric instruments for quantifying various stable isotopes is given.     

Efficacy of alternatives to antibiotic chemicals for the control of fire blight of pears

The efficacy of various chemicals as alternatives to antibiotics for the control of fireblight (Erwinia amylovora) on pear trees was tested. The chemicals were applied in two ways. In 1999 and 2000, preselected pear twigs (80–90% bloom stage) were sprayed once either preventively 1 day before inoculation or curatively one or three days after artificial inoculation with pathogen concentrations of 10⁵ and 10⁷cfu ml^?1. In 2000 and 2001, whole trees were sprayed 2 and 4 days before artificial inoculation of the flowers. From the incidence of diseased flowers it appeared that Bion (50% benzothiadiazole) at 0.2 g litre^?1 H₂O and Aliette (80% fosetyl‐Al) at 2.5 g litre^?1 H₂O showed considerable preventive action by eliciting systemic acquired resistance mostly when they were applied in the whole trees. However the best control was achieved with the antibiotic Agrept (20% streptomycin) at 0.5 g litre^?1 H₂O. This showed both preventive and curative action. Kocide (77% copper hydroxide) at 0.9 g litre^?1 H₂O, Dentamet (citric acid in chelate) at 1.5 ml litre^?1 H₂O, Bactosan (an extract from the plant Pongamia pinnatd) at 3.0 g litre^?1 H₂O and Bion at 0.1 g litre^?1 H₂O, showed preventive action, but only when the inoculum concentration was low.     

Accumulation of Trace Metals by Mangrove Plants in Indian Sundarban Wetland: Prospects for Phytoremediation

The work investigates on the potential of ten mangrove species for absorption, accumulation and partitioning of trace metal(loid)s in individual plant tissues (leaves, bark and root/pneumatophore) at two study sites of Indian Sundarban Wetland. The metal(loid) concentration in host sediments and their geochemical characteristics were also considered. Mangrove sediments showed unique potential in many- fold increase for most metal(loid)s than plant tissues due to their inherent physicochemical properties. The ranges of concentration of trace metal(loid)s for As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb and Zn in plant tissue were 0.006–0.31, 0.02–2.97, 0.10–4.80, 0.13–6.49, 4.46–48.30, 9.2–938.1, 0.02–0.13, 9.8–1726, 11–5.41, 0.04–7.64, 3.81–52.20 μg g ^?1respectively. The bio- concentration factor (BCF) showed its maximum value (15.5) in Excoecaria agallocha for Cd, suggesting that it can be considered as a high-efficient plant for heavy metal bioaccumulation. Among all metals, Cd and Zn were highly bioaccumulated in E. agallocha (2.97 and 52.2 μg g ^?1 respectively. Our findings suggest that the species may be classified as efficient metal trap for Cd in aerial parts, as indicated by higher metal accumulation in the leaves combined with BCF and translocation factor (TF) values.     

Synthesis of CpFe(CO)(L) complexes of hydantoin anions (Cp = eta5-C5H5, L = CO, PPh3), and the use of the 5,5-diphenylhydantoin anion complexes as tracers in the nonisotopic immunoassay CMIA of this antiepileptic drug.

As part of our ongoing development of the CMIA nonisotopic immunoassay method, in which the tracers are metal carbonyl complexes and detection is by Fourier transform infrared spectroscopy, we examined the potential use as tracers of the complexes CpFe(CO)2(5,5-diphenylhydantoin) 2d and CpFe(CO)(PPh3)(5, 5-diphenylhydantoin) 3. The present study involved the synthesis of a series of hydantoin complexes (2a-2d), in particular that of the derivative of 5,5-diphenylhydantoin 2d. The structure of 2d was confirmed by X-ray crystallography. The infrared analysis, establishing the position and intensity of the characteristic metal-carbonyl peaks of complexes 2d and 3 in the 1850-2200 cm-1 region, shows that measurement of the absorbance values of these characteristic peaks will permit quantitative analysis in the picomole range, the norm for routine use in immunoassay and thus suitable for use as CMIA tracers. Cross-reaction rates of these tracers with anti-DPH specific antibodies show that 2d and 3 are both recognized by anti-DPH antibodies (cross-reaction rates 43 and 20%, respectively). In developing a CMIA of DPH with these tracers, it was found that 3, with a single, intense band at 1977 cm-1, had very promising IR characteristics for use in multiassay CMIA, but probably owing to its relatively weak affinity for the antibodies, it was not possible to develop a CMIA for DPH using this tracer. Complex 2d, however, showed better recognition by the antibodies, and using this complex as a tracer, it was possible to develop a particularly sensitive monoassay of DPH by the CMIA method.     

Feeding deterrent effect of carvone, a compound from caraway seeds, on the slug Arion lusitanicus

The feeding deterrent effect of carvone on the slug Arion lusitanicus was investigated. Carvone, a natural compound from caraway seeds, was incorporated into mulch to reduce its inherent volatility. In a laboratory choice experiment, boxes were filled on one side with carvone‐treated mulch and on the other side with untreated mulch. At carvone concentrations ranging from 0.03–0.75 ml litre^?1 mulch, slugs ate significantly more lettuce on the untreated side. In a laboratory based no‐choice experiment, carvone concentrations of 0.25 and 0.75 ml litre^?1 mulch significantly reduced slug feeding in comparison with the untreated control. Moreover at the highest concentration of carvone (0.75 ml litre^?1 mulch) 50% mortality was recorded over a period of 5 days, indicating a clear molluscicidal effect. Due to its volatility carvone did not decrease plant defoliation by A. lusitanicus when applied directly onto lettuce. Subsequent field evaluation showed 0.75 ml litre^?1 mulch to partially reduce slug feeding damage. However, this effect was not sufficient to significantly increase lettuce yield. The incorporation of a higher carvone concentration into mulch is still to be tested to confirm whether carvone‐treated mulch can be recommended as an effective alternative approach to chemical slug control.     

Continuous production of L-tert-leucine in series of two enzyme membrane reactors

The L-tert-leucine synthesis was performed continuously in series of two enzyme-membrane reactors by reductive amination of trimethylpyruvate with leucine dehydrogenase. The necessary “native” cofactor NADH is regenerated with the aid of a second enzyme, formate dehydrogenase. Considering detailed kinetic studies of initial reaction rates under conditions relevant to the process a kinetic model was developed. The model shows that the overall reaction rate is strongly inhibited by the reaction product. The reactor's models combine the mass balances and proposed kinetic equations. The model adequacy was verified by using it to simulate the experiments and by comparing experimental and computed conversion, space-time yield and enzyme consumption. The calculations for the three reactor's types (batch, single CSTR and a cascade of two CSTRs in series) were compared. The results showed that a single CSTR is no favourable reactor configuration due to the very strong product inhibition. Space-time yield drops from 560 g litre^?1 day^?1 in a batch reactor to 110 g litre^?1 day^?1 in a single CSTR at the highest conversion of 98%. At the conversion of 95% the difference in biocatalyst costs between batch and two CSTR in series is negligible. Therefore the use of two enzyme membrane reactors in series was proposed. The modelling in this work shows that the optimisation of the quantity of the enzyme used results in a minimisation of the biocatalyst costs.     

STUDIES ON STRIGA SENEGALENSIS II. TRANSLOCATION OF C14-LABELLED PHOTOSYNTHATE,UREA-C14 AND SULPHUR-35 BETWEEN HOST AND PARASITE

Using a tracer technique, the nutritional relationship between Striga senegalensis and its host (Sorghum vulgare) has been studied. Preliminary trials with aqueous eosin dye indicated that a mechanism exists for the passage of watery solutions from host to parasite but not vice versa. Use of tracers like C¹⁴O₂, urea-C¹⁴ and sulphur-35 confirmed that minerals as well as organic compounds are translocated from host to parasite. However, when these tracers were applied to the parasites, translocation of labelled products into the host was insignificant. When C¹⁴O₂ was used to label the photosynthate in Sorghum, the bulk of activity appeared in sucrose, glucose and fructose, part of which was presumably obtained by the parasite. The photosynthetic ability of the green tissues of Striga was confirmed. Thus the damaging effect of Striga on the host appears to be due to removal of considerable quantities of water, minerals and organic compounds from the latter. The pattern of translocation of photosynthates and minerals from host to parasite suggests a possible application of selective, systemic weed-killers by aerial spray on the host leaves for control of the parasite.     

Effects of 1-methylcyclopropene alone and in combination with polyethylene bags on the postharvest life of mango fruit

Experiments were conducted to determine how 1‐methylcyclopropene (1‐MCP) treatments influence ethylene‐stimulated ripening of harvested mango cv. Zihua fruit at 20°C. The ripening response of fungicide (prochloraz) treated fruit was characterised following various 1‐MCP treatments in sealed jars followed by storage in polyethylene bags and/or subsequent ethephon (ethylene) exposure. Exposure of fruit to increasing concentrations of 1‐MCP for 12 h resulted in the reduced softening of produce when subsequently held in air for 7 days after ethephon treatment. Application levels of between 1 and 100 μl litre^?1 1‐MCP had increasing impact, while 200 μl litre^?1 1‐MCP apparently began to approach response saturation. Exposure of fruit to 50 or 100 μl litre^?1 concentrations of 1‐MCP for periods from 1 to 24 h subsequently resulted in reduced softening of produce when held in air for 7 days after ethephon treatment. Increasing periods of exposure from 1 to 12 h had increasing impact, while exposure times greater that 12 h appeared to reach saturation. In the absence of ethephon‐stimulation, the natural ripening of mangoes held in polyethylene bags was delayed by prior exposure to 100 μl litre^?1 1‐MCP for 12 h. Extended holding of 1‐MCP treated and non‐1‐MCP treated control fruit in polyethyene bags encouraged physiological and pathological deterioration. Following exposure to 100 μl litre^?1 1‐MCP for 12 h, mango fruit held for 10 days in polyethylene bags showed a delay in the onset of ripening relative to bagged but non‐1‐MCP treated control fruit. Treatment with 1‐MCP allowed storage of mango fruit in plastic bags at 20°C for 30 days. Observations suggest that 1‐MCP treatments do not adversely influence the quality of the post‐storage ethephon‐ripened fruit. Thus, application of 1‐MCP in combination with the use of polyethylene bags can extend the postharvest life of mango fruit at ambient temperature. Treatments that extend postharvest life are important in developing countries, such as China, where the cold chain infrastructure is often lacking.     

A Liquid Chromatography–Mass Spectrometry Method to Measure Stable Isotopic Tracer Enrichments of Glycerol and Glucose in Human Serum

Stable isotopes are commonly used as tracers for the measurement of glycerol and glucose kinetics in metabolic studies. Traditionally, the analysis of these isotopes has been performed using gas chromatography-mass spectrometry, which requires that the analytes first be derivatized. The derivatization process adds considerable complexity to the method. Liquid chromatography-mass spectrometry (LCMS) can measure many metabolites directly with limited sample preparation. We present a novel analytical method for the measurement of [1,1,2,3,3-(2)H(5)]glycerol (d(5)-glycerol) and [6,6-(2)H(2)]glucose (d(2)-glucose) isotopic tracer enrichments in human serum in a single run by LCMS. After a simple extraction step, the sample is separated isocratically by HPLC, and the isotopes are measured using positive electrospray ionization with selected ion monitoring of the sodium-adduct ions. The method is linear over a wide range of d(2)-glucose and d(5)-glycerol enrichments. The within-day standard deviation of measurement of serum samples was 0.05 mole% excess (MPE) for d(2)-glucose and 0.25 MPE for d(5)-glycerol. The variation of tracer enrichment among days was about double that measured within 1 day.     

Use of two formulations and two application techniques to deliver Bacillus thuringiensis var. israelensis for the control of Musca domestica (Diptera: Muscidae) larvae and adults in poultry houses

A field study was carried out for 6 wks to assess, from both an efficiency and economic perspective, the effect of individual and integrated success of feeding and topical applications of two formulations of Bacillus thuringiensis var. israelensis (Bti) in controlling house fly (Musca domestica L.) larvae and adults in poultry houses. There was no significant difference between the 1 g and 2 g L^?1 spray applications of Bti. In the absence of spray applications, no significant differences in larval mortalities were observed between the 250 mg and 500 mg kg^?1 feed applications. The percentage mortality of larvae accomplished as a result of using a combination of 250 mg kg^?1Bti feed and 2 g L^?1 spray applications was equivalent to that obtained as a result of combining 500 mg kg^?1Bti and 1g L^?1 spray applications. Treatment with Bti caused significant reductions in the emergence (up to 74%) of house fly adults compared to the control. The fact that the emergence of adult house flies was affected by Bti treatments implies that Bti has sublethal effects on house fly larvae. The cost–benefit analysis (expressed in terms of mortality of larvae growing) indicated that the most effective combination for house fly larvae and adult house fly emergence control was the 500 mg kg^?1 of feed and 2 g L^?1 spray application combination that resulted in 67% larval mortality and a 74% decrease in adult house fly emergence. This study presents commercial users with various alternatives for possible combinations of the two Bti formulations.     

Absorption of Iron by Enzymically Isolated Leaf Cells

Iron uptake was studied using cells enzymically isolated from green tobacco leaves. Absorption was increased both by light and succinate as probable energy sources. Bicarbonate in the incubation mixture was inhibitory, and citrate also reduced absorption presumably by chelation with the metal. Absorption of iron was temperature sensitive and optimal at 25°C. Temperature coefficients and activation energies suggested that absorption was energy mediated. NaN₃ and DNP inhibited uptake at concentrations of 10-³M and 10^?4M, respectively. The inhibition caused by DNP was not negated by an external supply of ATP. The results suggest that iron absorption is an active metabolic process in cells enzymically isolated from green tobacco leaves. Cells from Fe-chlorotic leaves of PI 54619–5–1 soybean absorbed less iron than those derived from healthy leaves of the same variety, while leaf cells from the variety Hawkeye showed no such differences.     

Use of carbon-11 in Populus shows that exogenous jasmonic acid increases biosynthesis of isoprene from recently fixed carbon

A new approach for pulse labelling of plants using the short-lived positron emitting radioisotope carbon-11 (half-life: 20.4 min) as ¹¹CO₂ is reported together with its application to measuring [¹¹C]isoprene emissions from intact leaves capturing information associated with: (1) rate of emission; (2) the relative contribution of recently fixed carbon to isoprene biosynthesis; and (3) the transit time for tracer movement through the leaf and biochemical pathways associated with isoprene biosynthesis. This approach was applied to study the response of certain Populus species to exogenous treatments of jasmonic acid (JA), a plant hormone implicated in signal transduction linked to defence response against herbivory. Twelve hours after treatment of single intact leaves of aspen (Populus tremuloides) with a 1 m m JA spray, isoprene emissions from those leaves increased 1.5 times the controls from 35.4 ± 2.2 to 53.1 ± 4.8 nmol m⁻² s⁻¹. [¹¹C]Isoprene emissions from the same leaves, reflecting the isoprene that was derived from recently fixed carbon, increased much more, to 2.2 times the controls. This increase coincided with a change in emitted [¹¹C]isoprene from 0.31 to 0.68% of ¹¹C fixed in the leaf tissue, while the tracer transit time remained constant at 12.5 min. These results suggest that JA had no effect on enzyme kinetics involved in isoprene biosynthesis, but did impact the source of recent carbon feeding that pathway. Studies with poplar (Populus nigra clone NC 5271) showed similar trends in systemic emissions (from an untreated leaf on the same plant).     

Field Evaluation of Willow Under Short Rotation Coppice for Phytomanagement of Metal-Polluted Agricultural Soils

Short rotation coppice (SRC) of willow and poplar might be a promising phytoremediation option since it uses fast growing, high biomass producing tree species with often a sufficient metal uptake. This study evaluates growth, metal uptake and extraction potentials of eight willow clones (Belders, Belgisch Rood, Christina, Inger, Jorr, Loden, Tora and Zwarte Driebast) on a metal-contaminated agricultural soil, with total cadmium (Cd) and zinc (Zn) concentrations of 6.5 ± 0.8 and 377 ± 69 mg kg^?1 soil, respectively. Although, during the first cycle, on average generally low productivity levels (3.7 ton DM (dry matter) ha^?1 y^?1) were obtained on this sandy soil, certain clones exhibited quite acceptable productivity levels (e.g. Zwarte Driebast 12.5 ton DM ha^?1 y^?1). Even at low biomass productivity levels, SRC of willow showed promising removal potentials of 72 g Cd and 2.0 kg Zn ha^?1 y^?1, which is much higher than e.g. energy maize or rapeseed grown on the same soil. Cd and Zn removal can be increased by 40% if leaves are harvested as well. Nevertheless, nowadays the wood price remains the most critical factor in order to implement SRC as an acceptable, economically feasible alternative crop on metal-contaminated agricultural soils.     

Inter-species differences in polychlorinated biphenyls patterns from five sympatric species of odontocetes: Can PCBs be used as tracers of feeding ecology?

Concentrations of thirty two polychlorinated biphenyls (PCBs) were determined in the blubber of five sympatric species of odontocetes stranded or by-caught along the Northwest coast of the Iberian Peninsula: common dolphin (Delphinus delphis), long-finned pilot whale (Globicephala melas), harbour porpoise (Phocoena phocoena), striped dolphin (Stenella coeruleoalba) and bottlenose dolphin (Tursiops truncatus). Multivariate analyses were applied to evaluate the ability of PCB patterns to discriminate these sympatric species and to determine which eco-biological factors influence these patterns, thus evaluating the relevance of PCB concentrations as biogeochemical tracers of feeding ecology. The five species could be separated according to their PCB patterns. Different exposure to these contaminants, a consequence of their different dietary preferences or habitats, together with potentially dissimilar metabolic capacities, likely explain these results; sex, age, habitat and the type of prey eaten were the most important eco-biological parameters of those tested. Although, no single congener has been specifically identified as a tracer of feeding ecology, 4 congeners from the 22 analysed seemed to be the most useful and around 12 congeners appear to be enough to achieve good discrimination of the cetaceans studied. Therefore, this study suggests that PCB patterns can be used as tracers for studying the feeding ecology, sources of contamination or even population structure of cetacean species from the Northwest Iberian Peninsula.     

PRELIMINARY RESULTS ON THE UPTAKE AND RELEASE OF 32P BY POTAMOGETON PECTINATUS

SUMMARY

The absorption and release of ³²PO₄, by Potamogeton pectinatus L during winter was investigated using a partitioned container in which the roots were separated from the stems and leaves. ³²PO₄ is absorbed by the roots as well as the stems and leaves under both light and dark conditions and transported to all parts of the plant. Transport rates (μg P plant^?1 24 hr^?1) show that more phosphorus is released by the roots than by the stems and leaves, indicating a nett movement of phosphorus from the surrounding water to the substrate. Foliar absorption of ³²P exceeds root absorption under both light and dark conditions.     

Tumor uptake study of 18F-labeled N-acetylneuraminic acids

In order to develop positron-emitting tracers for imaging metabolic functions of tumors with positron emission tomography, tumor uptake of N-acetyl-3-[¹⁸F]fluoroneuraminic acid and N-acetyl-2-deoxy-2,3-di-[¹⁸F fluoroneuraminic acid was investigated in mice or rats. The two tracers showed similar tissue distribution patterns. After i.v. injection of each tracer into mice with an FM3A tumor, the radioactivity was very rapidly cleared from normal and tumor tissues. Only tumor-to-brain and tumor-to-muscle uptake ratios were greater than 1.0 for 2 h. In 7 types of tumor models, no selective tumor uptake of tracers was observed 30 min after injection. The metabolic alteration rate of N-acetyl-3-[¹⁸F]fluoroneuraminic acid in FM3A, liver and kidney was very slow. Neither tracer may be suitable for tumor imaging in vivo.     

Effect of spraying adjuvants with the biocontrol fungus Microsphaeropsis ochracea at different water volumes on the colonization of apple leaves

In a previous study, it was observed that Microsphaeropsis ochracea was less efficacious as a biofungicide for apple scab when applied under field conditions as an unformulated spore suspension with an airblast spray using low water volumes as compared to a ground cover spray using high water volumes. The effect of spore concentration (10⁹ to 10¹² spores ha^?1), water volume (250–1500 L ha^?1), and 20 adjuvants were studied to improve colonization of apple leaves by M. ochracea. Using commercial-type spray equipment, the fungus was applied to excised apple leaves. Higher spores concentrations resulted in greater colonization. Water volume did not have a strong impact on colonization. Only a few adjuvants improved germination (glycerine) and mycelial growth (K-90, Agrimer, Ekol, and Tween 80) in some trials. When tested on apple trees, the adjuvant Assist provided the most consistent improvement in colonization in five out of eight trials. The use of oils, humectants, and surfactants could broaden the opportunities for using this biofungicide.