Determination of Homalodisca coagulata (Hemiptera: Cicadellidae) egg ages suitable for oviposition by Gonatocerus ashmeadi, Gonatocerus triguttatus, and Gonatocerus fasciatus (Hymenoptera: Mymaridae)

Homalodisca coagulata (Say) (Hemiptera: Cicadellidae) eggs 1–10 days of age were exposed to Gonatocerus ashmeadi Girault, Gonatocerus triguttatus Girault, and Gonatocerus fasciatus Girault (all Hymenoptera: Mymaridae) in no choice laboratory trials to investigate egg age utilization and to determine which egg ages are vulnerable to attack by these three parasitoids. The H. coagulata egg ages that were most suitable for oviposition by G. ashmeadi, G. triguttatus, and G. fasciatus were eggs 3, 4, and 2 days of age, respectively. Egg ages least suitable for parasitoid development were 6–10 days for G. ashmeadi (resulting in <50% parasitism), 1–2 and 7–10 days for G. triguttatus (resulting in <25% parasitism), and 3–10 days for G. fasciatus (resulting in <11% parasitism). Pooling parasitism data across all egg ages showed that parasitism by G. ashmeadi was 12.9 and 28.5% higher compared with G. triguttatus and G. fasciatus, respectively, and G. triguttatus resulted in 15.6% higher percentage parasitism compared with G. fasciatus. Egg age had a significant effect on the percentage of female G. ashmeadi offspring produced, but this was not significant for G. triguttatus, and low G. fasciatus parasitism prevented statistical analyses for comparisons. Results from tests where females were offered a choice for oviposition between eggs 1, 3, and 5 days of age demonstrated that G. ashmeadi and G. triguttatus showed no significant oviposition preference, while percentage parasitism by G. fasciatus was 29.4 and 7.4% higher when females were presented eggs 1 and 3 days of age, respectively, compared with eggs 5 days of age. Choice tests indicated that an overlap in egg age suitability for oviposition exists between G. ashmeadi, G. triguttatus, and G. fasciatus, and that interspecific competition for eggs 1, 2, and 3 days of age may occur in the field environment.     

Use of life table statistics and degree-day values to predict the invasion success of Gonatocerus ashmeadi (Hymenoptera: Mymaridae), an egg parasitoid of Homalodisca coagulata (Hemiptera: Cicadellidae), in California

Life table statistics and degree-day requirements for Gonatocerus ashmeadi Girault, a parasitoid of the glassy-winged sharpshooter Homalodisca coagulata (Say), were used to estimate the number of expected parasitoid generations in California (USA). Between two to 51 and one to 37 generations per year were estimated across different climatic regions in California, using life table and degree-day models, respectively. Temperature-based values for net reproductive rate, R_o, were estimated in California using a laboratory-derived equation and ranged from zero to approximately 48 and analyses indicate that a minimum of eight generations are required each year to sustain a population increase of G. ashmeadi. Long-term weather data from 381 weather stations across California were used with an Inverse-Distance Weighting algorithm to map temperature-based estimations for the entire state of California. This Geographic Information Systems model was used to determine number of G. ashmeadi generations based on day-degree accumulation, T_c, and R_o. GIS mapping indicated that Californian counties in the north, central west coast, central west and Sierra Nevada regions may be climatic conditions unfavorable for supporting the permanent establishment of invading populations of G. ashmeadi should H. coagulata successfully establish year-round populations in these areas. Southern counties in California that experience warmer year round temperatures and support year round populations of H. coagulata, appear to be conducive to the establishment of permanent populations of G. ashmeadi. The mechanisms facilitating G. ashmeadi invasion and the implications of these temperature-based estimates for biological control of H. coagulata are discussed.     

Amino acids as determinants of host preference for the xylem feeding leafhopper,Homalodisca coagulata (Homoptera: Cicadellidae)

Summary Homalodisca coagulata is a highly polyphagous xylem feeder with distinct seasonal patterns in it's selection of host plants. These patterns were examined in relation to the amino acid content of the xylem for four common host species; Lagerstroemia indica, Baccharis halimifolia, Prunus persica, and Prunus salicina. Xylem fluid was collected from each host species at times when numbers of feeding leafhoppers were both low and high. In each case, concentrations of amino acids were greatest when numbers were high. Similarly, comparisons between host species at given times showed that concentrations of amino acids were positively correlated with host selection. In a second study, amino acids of xylem were manipulated by budding scions of a non-preferred host (P. persica) on rootstocks of preferred (P. salinica) and non-preferred (P. persica) hosts. Morphology and phenology of the budded trees were similar to that of the scion species yet the xylem composition of amino acids was primarily dependent on the rootstock. Concentrations of amino acids and the preference of leafhoppers were roughly two-fold greater for scions of the preferred than the non-preferred rootstock. In both studies, amides (glutamine plus asparagine) were the amino acids most highly correlated with host selection. These compounds are the predominant amino acids in xylem fluid, have high nitrogen to carbon ratios, and account for a high percentage of the caloric value in xylem fluid. Many of the less abundant amino acids were positively correlated with host preference, but the correlations were less consistent and correlation coefficients were generally lower.Florida Agricultural Experiment Station Journal Series No. 9672     

Utilization of primary nutrients by the polyphagous xylophage,Homalodisca coagulata,reared on single host species

Feeding experiments were conducted to assess development of the polyphagous leafhopper, Homalodisca coagulata, on single host species. Insects were reared from eggs on two cultivars of Euonymus japonica, two cultivars of Lagerstroemia indica, and two species of Prunus. Only insects on nonvariegated E. japonica and L. indica cv. Osage survived to maturity, although a substantial proportion of those on P. salicinia developed to the fourth instar. Chemical profiles of food source (xylem fluid), insect excreta and body composition, and consumption rates were used to estimate assimilation efficiencies, daily assimilation rates, and efficiency of conversion of food into biomass for all primary nutrients identified in the xylem fluid. Rates of successful maturation were greatest on the highest nitrogen and carbon source (nonvariegated E. japonica) and were associated with low consumption rates, high assimilation efficiencies, and prolonged instar durations. Developmental period on L. indica was shorter and consumption rates were higher, yet assimilation efficiencies were reduced and young nymphs had lower survivorship. Low consumption rates, coupled with the low nutritional value of P. salicinia, precluded sufficient accumulation of nutrients to complete development. Insects fed on hosts with xylem fluid containing high ratios of amides to total organic compounds (indicative of low carbon to nitrogen ratios) had higher consumption rates but less efficient assimilation of primary nutrients. Carcass analyses of newly enclosed adults suggest that regulation of consumption rates and assimilation efficiencies to provide adequate accumulation of essential amino acids may be pivotal for successful development. © 1996 Wiley-Liss, Inc.     

Short-distance dispersal behavior and establishment of the parasitoid Gonatocerus ashmeadi (Hymenoptera: Mymaridae) in Tahiti: Implications for its use as a biological control agent against Homalodisca vitripennis (Hemiptera: Cicadellidae)

The egg parasitoid Gonatocerus ashmeadi Girault (Hymenoptera: Mymaridae), was introduced into French Polynesia as a biological control agent to control the invasive plant feeding pest Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae). The short-distance dispersal of G. ashmeadi was monitored as part of the biological control program. G. ashmeadi showed exponential dispersal capacity with 47 m/day being a minimum estimate of its natural rate of spread at high host densities (>150 nymphs per minute of sweep net sampling) in urbanized areas at sea level, which were characterized by a high diversity of exotic ornamental plants. This rate of spread contrasted starkly with almost nonexistent establishment and dispersal where host densities were very low (<2 nymphs per minute of sweep net sampling) at high elevation (800 m) with relatively undisturbed native vegetation. Survey results across different altitudes revealed an effect of vegetative diversity and host density on the measurable mobility and establishment of G. ashmeadi. In contrast, no significant influence of wind direction was found on G. ashmeadi dispersal rate or direction. Survey results for G. ashmeadi from French Polynesia suggest that the best release establishment strategies for classical biological control of H. vitripennis are: (1) many small releases where host density is high, or (2) larger and fewer releases where host densities are low.     

Production of Anagrus epos Girault (Hymenoptera: Mymaridae) on Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae) eggs

Anagrus epos Girault (Hymenoptera: Mymaridae) is a natural enemy candidate for a classical biological control program targeting the glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae), in California. Little is known about the biology or ecology of A. epos when it utilizes GWSS eggs as a host. Here, we report the results of laboratory studies that describe the host age preference for oviposition, longevity of A. epos adults provided with different food sources, and developmental rates at six different constant temperature regimes. Anagrus epos is a gregarious parasitoid in GWSS eggs with up to 14 adults emerging from each GWSS egg. In choice and no-choice tests for oviposition, A. epos females successfully parasitized all developmental ages of GWSS eggs (1–8 days old). In choice tests, parasitism rates were significantly higher in 1-, 3-, 4-, and 5-day-old GWSS eggs than in 2-, 6-, 7-, and 8-day-old eggs. If provided with honey and water, honey only, water only, or no food or water, A. epos females lived on average 8.2, 4.7, 2.6, and 1.6 days, respectively. Anagrus epos required 294.1 degree-days above a lower temperature threshold of 12.4 °C to develop from egg to adult (eclosion). Our results provide baseline information useful in the development of an efficient parasitoid mass rearing program for A. epos release and evaluation in California.     

Metabolism of amino acids, organic acids and sugars extracted from the xylem fluid of four host plants by adult Homalodisca coagulata

The efficiency of amino acid, organic acid and sugar metabolism was quantified for adult Homalodisca coagulata (Say) (Homoptera: Cicadellidae) by comparing chemical profiles of xylem fluid (food source) and insect exudate. Leafhoppers were confined in Parafilm^® sachets to stems of 4 host plants: [Baccharis halimifolia (L.), Lagerstroemia indica (L.), Prunus salicina (Lindl.), Prunus persica (L.), Batsch]. Insect feeding rates (0.09–0.27 ml h^–1), exudate osmolarity (7.8–12.8 mM) and exudate composition (mainly inorganic entities) were characteristic of a xylem feeder. Total organic solute concentration in the xylem fluid of B. halimifolia, L. indica, P. salicina and P. persica was ca. 9.4, 13.8, 5.5 and 1.8 mM, respectively. Nineteen protein amino acids, 7 organic acids and 3 or 4 sugars were identifid in the xylem fluid. Total amino acids, organic acids and sugars were metabolized with ca. 99% efficiency. Glutamine, asparagine, arginine and citric, malic and succinic acids, the predominant organic compounds in the xylem fluid of all four plant species, were metabolized with greater than 99% efficiency. Cysteine (51%), methionine (74%) and oxalic acid (77%) were metabolized with the lowest efficiency. The primary nitrogenous waste was NH _inf4 ^sup+ ; uric acid or urea were not detected. Nitrogen retention was generally less than 60% of dietary nitrogen. High feeding rates, ammonotelism and an extremely high metabolic efficiency of organic compounds permit H. coagulata to subsist on the dilute and skewed chemical profile of xylem fluid.

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Résumé L'efficacité du métabolisme des amino-acides, des acides organiques et des sucres a été quantifiée chez des H. coagulata Say (Hom. Cicadellidae) adultes, en comparant la composition chimique de la sève du xylème et du miellat des insectes. Les cicadelles ont été maintenues dans des sachets de parafilm avec des tiges de 4 plantes hôtes: Baccharis halimifolia L., Lagerstroemia indica L., Prunus salicina Lindl. et Prunus persica Batsch. Le taux de consommation (0,09 à 0,27 ml hr^–1), l'osmolarité du miellat (7,8 à 12,8 mM) et la composition du miellat (principalement des éléments inorganiques) sont caractéristiques des consommateurs de xylème. Les concentrations organiques totales en soluté de la sève du xylème étaient respectivement: 9,4; 13,8; 5,5; et 1,8 mM chez B. halimifolia, L. indica, P. salicina et P. persica. 19 amino acides protéiques, 7 acides organiques et 3 ou 4 sucres ont été identifiés dans la sève du xylème. Les acides aminés, les acides organiques et les sucres ont été métabolisés dans leur ensemble avec une efficacité de 99%. La glutamine, l'asparagine, l'arginine et les acides citrique, malique et succinique,-les principaux composés organiques de la sève du xylème de ces 4 plantes,-ont été métabolisés avec plus de 99% d'efficacité. La cystéine (51%), la méthionine (74%) et l'acide oxalique (77%) ont été métabolisés avec une plus faible efficacité. Le déchet azoté primarie était NH _inf4 ^sup+ ; l'acide urique et lurée n'ont pas été décelés. La fixation d'azote a été généralement inférieure à 60% de l'azote consommé. Des taux de consommation élevés, l'ammonotélisme et une efficacité extrêmement élevée du métabolisme des composés organiques permettent à H. coagulata de survivre malgré la composition chimique biaisée et la dilution de la sève du xylème.

     

Species identity of geographically distinct populations of the glassy-winged sharpshooter parasitoid Gonatocerus ashmeadi: morphology, DNA sequences, and reproductive compatibility

Gonatocerus ashmeadi is a common and seemingly widespread egg parasitoid of Homalodisca coagulata, the glassy-winged sharpshooter (GWSS). Location records for G. ashmeadi indicate its natural range to be the southeastern USA and northeastern Mexico (which coincides with the presumed native range of GWSS), and possibly southern and central California (CA) (the adventive range of GWSS). The purpose of our work was to determine whether G. ashmeadi in the USA and northeastern Mexico is one species or a complex of reproductively incompatible sibling species. We used three approaches to determine the species identity of different G. ashmeadi populations: (1) reassessment of key morphological features using scanning electron microscopy (SEM) to determine if subtle morphological differences exist between G. ashmeadi populations which could indicate species differences; (2) to determine if molecular differences exist between G. ashmeadi populations collected from different regions by comparing mitochondrial and ribosomal DNA sequences; (3) mating compatibility studies to determine if different populations of G. ashmeadi are reproductively isolated, or if mating occurs, whether offspring are viable thereby defining species groups on the basis of successful interbreeding. Results from these three areas (morphology, DNA sequences, and reproductive compatibility) have been evaluated collectively; leading us to the conclusion that G. ashmeadi as it is currently viewed is a valid species and not an aggregate of morphologically indistinguishable cryptic species.     

Changes in fatty acids,amino acids and carbon/nitrogen biomass during nitrogen starvation of ammonium- and nitrate-grownIsochrysis galbana

Growth of cells ofIsochrysis galbana with either nitrate or ammonium as the N-source, and the effects of subsequent N-starvation of these cells, were compared. During exponential N-sufficient growth nitrate-grown cells had double the fatty acid content of the ammonium-grown cells but lower concentrations of a few amino acids. Following resuspension in N-free medium the fatty acid content of the ammonium-grown cells increased to that of the nitrate-grown cells, but there was no further increase in fatty acid content on a C-biomass or cellular basis during the following 4 days for either culture. Fatty acid synthesis was continuous during N-starvation, while it occurred during the light-phase only in exponential growth. The proportion of 18:1n9 fatty acid increased from 10 to 25% total fatty acids during N-starvation. Intracellular free amino acid content decreased in a similar manner in both cultures on N-starvation, the ratio of intracellular free amino-N/cell-C falling more rapidly than overall cellular N/C. It was concluded that optimal amino acid and fatty acid content would be attained by growth in the presence of excess nitrate. Measurements of chlorophyll and carotenoid content and ofin vivo fluorescence indicated that these parameters had potential for monitoring the C and N biomass in cultures grown under relatively constant (not necessarily continuous) illumination.     

Population and individual polyphagy in the grasshopperTaeniopoda eques during natural foraging

Dietary patterns of free-foraging individuals of the polyphagous grasshopperTaeniopoda eques Burmeister (Romaleidae) were studied at three desert grassland sites in southern Arizona. At the population level this species was highly polyphagous at all sites, but showed evidence of selectivity in terms of frequency of feeding relative to frequency of contacts with resources. Most feeding bouts were very short, suggesting that most plants were relatively unpalatable. Both diet diversity and the mean length of feeding bouts varied among the study sites, primarily because highly preferred resources and plant tissues were not encountered with equal frequency at all sites. Individual insects were highly polyphagous. Dietary overlap calculations showed that insects at a given site generally consumed diets less similar than the resources they contacted. This result does not support the idea that all insects preferred the same subset of resources. Most differences in diet among individuals were probably due to environmental heterogeneity, but factors such as sequence of encounter, compensatory feeding on complementary resources, and intrinsic differences in preference may also have contributed to variation in diets.     

Improvement of xylose utilization in Clostridium acetobutylicum via expression of the talA gene encoding transaldolase from Escherichia coli

Clostridium acetobutylicum ATCC 824 was metabolically engineered for improved xylose utilization. The gene talA, which encodes transaldolase from Escherichia coli K-12, was cloned and overexpressed in C. acetobutylicum ATCC 824. Compared with C. acetobutylicum ATCC 824 (824-WT), the transformant bearing the E. coli talA gene (824-TAL) showed improved ability on xylose utilization and solvents production using xylose as the sole carbon source. During the fermentation of xylose and glucose mixtures with three xylose/glucose ratios (approximately 1:2, 1:1 and 2:1), the rate of xylose consumption and final solvents titers of 824-TAL were all higher than those of 824-WT, despite glucose repression on xylose uptake still existing. These results suggest that the insufficiency of transaldolase in the pentose phosphate pathway (PPP) of C. acetobutylicum is one of the bottlenecks for xylose metabolism and therefore, overexpressing the gene encoding transaldolase is able to improve xylose utilization and solvent production.     

Food consumption and utilization by 6th-instar larvae of spruce budworm, Choristoneura fumiferana: a comparison on three Picea (spruce) species

The effects of tree species, shoot age, and sex of larvae on food consumption and utilization were investigated in a factorial experiment. Diets prepared from lyophilized and powdered current-year shoots, from two age classes (sample dates) of Picea glauca (Moench) Voss, Picea rubra, Sarg., and Picea mariana (Mill.) B.S.P., were evaluated for eight performance criteria. All factors had significant effects on all criteria except for tree species on assimilation efficiency, shoot age on development time, and sex on net growth efficiency. Eight of the possible 24 two-factor interactions and one of the possible eight three-factor interactions were significant. Food utilization efficiencies and relative growth rate were in the low and moderate characterization limits for immature arthropods; relative consumption rates were high. The higher performance values on Picea glauca were related to this species greater vulnerability to spruce budworm. Performance criteria of similar larvae on an artificial diet were compared with those on the foliage-diets.

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Résumé L'auteur a déterminé la quantité d'aliments utilisées, à 22±0,5 °C, par des larves de Tordeuse des bourgeons de l'épinette parvenues à leur sixième stade de développement qui s'étaient nourries d'un régime artificiel et d'aiguilles de l'année en cours lyophilisées et réduites en poudre, prélevées à deux dates différentes, à 10 jours d'intervalle, et provenant d'épinettes blanchés (Picea glauca [Moench] Voss), d'épinettes rouges (Picea rubra Sarg.) et d'épinettes noires (Picea mariana [Mill.] B.S.P.). Les femelles ont invariablement ingéré une quantité significativement plus importante de tous les régimes que les mâles, consommant 24% de plus d'aiguilles d'épinette noire et 73% de plus d'aiguilles d'épinette blanche.Les femelles prenaient en général plus de temps pour parvenir au stade de chrysalide et avaient un taux d'utilisation des aliments significativement plus élevé. L'efficacité nette de la croissance chez les deux sexes ne présentait toutefois aucune différence significative. La performance des larves ayant ingéré des aiguilles provenant de deux classes d'âge différait de façon significative mais non pas uniforme d'un régime à l'autre: les aiguilles d'épinette blanche plus récentes donnaient une performance généralement plus élevée contrairement aux aiguilles plus jeunes d'épinette rouge et d'épinette noire. Les larves ingéraient une quantité significativement plus élevée, avaient un taux de consommation et de croissance supérieur et augmentaient davantage leur biomasse avec des aiguilles d'épinette blanche. L'efficacité d'utilisation des aliments et le taux de croissance relatif étaient de faibles à modérément faibles pour un lépidoptère arboricole phyllophage; la vitesse d'ingestion relative était élevée. La qualité des aiguilles des trois essences d'épinettes était faible. Les aiguilles d'épinette blanche permettaient aux larves d'avoir une performance significativement plus élevée, performance mesurée à partir de plusieurs critères, alors que les aiguilles d'épinette rouge et d'épinette noire donnaient une performance égale. La supériorité des performances obtenues avec l'épinette blanche explique la plus grande vulnérabilité de cette essence à la Tordeuse des bourgeons de l'épinette.

     

Morphology and development of Nigrosabulum globosum, a cleistothecial coprophile in the Bionectriaceae (Hypocreales)

Recent DNA sequence analyses indicated that Nigrosabulum globosum is a cleistothecial representative of the Bionectriaceae in the Hypocreales, but morphological characters supporting this relationship are unknown. Using light and electron microscopy we followed the development of the ascomata of this species, from the formation of gametangia through to the development of mature ascospores, and observed a series of characters that confirmed its hypocrealean affinities. These included the formation of a gel-filled centrum during early stages of ascoma development, the subsequent appearance of hyaline peridial tissue enclosed within a layer we interpret as representing a melanized uniloculate stroma, apically derived paraphyses, and an ascogenous system that gives rise to asci that were both cylindrical to clavate and globose. Ascospores, previously reported to be smooth, were ornamented with a honeycomb-like reticulum and were able to germinate within the ascoma. The carbonaceous outer (stromatic) walls of the mature, grit-like cleistothecia indicate possible resistance to UV radiation and desiccation. Furthermore, the complement of germinated ascospores would enable mature ascomata to function as propagules that could quickly initiate new growth when transferred to fresh substrate. Our reexamination of N. globosum also provides data that support the hypothesized close relationship with other bionectriaceous, cleistothecial coprophiles, i.e., species of Hapsidospora, and Bulbithecium in particular.     

Resistance in the wild crop relatives Avena macrostachya and Hordeum bogdani to the aphid Rhopalosiphum padi

In previous screening tests the two wild crop relatives Avena macrostachya (Bal., ex Coss. et Dur.) and Hordeum bogdani (Wil.) demonstrated a high degree of resistance to the aphid Rhopalosiphum padi (L.). In a choice situation using wild and cultivated oats and barley, alate aphids settled in lower numbers on the wild species. The results were, however, variable in the Avena combination. Nymph production was significantly higher, development time shorter and adult weight higher on the cultivated varieties. From the third instar and onwards the excretion of honeydew was significantly lower on the resistant plants. In general the honeydew contained less than 1% free amino acids although excreta from H. vulgare contained 3.5%. The percentage of free amino acids found in the honeydew was similar for all plant species (5.2–7.6%) except for H. vulgare, on which the aphids excreted 22% of the amounts ingested. Amino acids excreted in high proportions on all plants included asparagine, -aminobutyric acid, glutamic acid, and glycine. Tissue sectioning did not reveal any obvious mechanical barriers to stylet penetration. The potential use of these wild species as sources for aphid resistance breeding in oats and barley is considered.

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Résumè Lors d'examens systématiques antérieurs, Avena macrostachya (Bal. ex Coss. & Dur.) et Hordeum bogdani (Wil.) ont présenté une résistance élevée au puceron Rhopalosiphum padi (L.). Lorsqu'ils avaient un choix comprenant de l'avoine et de l'orge cultivés, les pucerons ailés ont atterri en nombres moins importants sur les espèces sauvages. Les résultats étaient cependant variables dans le complexe avoine. La production de nymphes et le poids des adultes étaient plus élevés sur espèces cultivées, ainsi que la durée du développement était plus longue sur les espèces sauvages. A partir du troisième stade, l'excrétion de miellat a été significativement plus faible sur les espèces résistantes. En général, le miellat y contenait moins de 1% d'acides aminés bien que sur H. vulgare il en contînt 3,5%. Les pourcentages d'acides aminés libres du miellat étaient semblables sur toutes les plantes (5,2–7,6%), à l'exception de H. vulgare sur lequel les pucerons excrétaient 22% des taux ingérés. Les acides aminés excrétés en fortes quantités sur les différentes plantes, comprenaient l'asparagine, l'acide -aminobutyrique, l'acide glutamique et la glycine. Des coupes de tissus n'ont révélé aucun obstacle mécanique clair à la pénétration des stylets. Les possibilités d'utiliser ces espèces sauvages comme source de résistance aux pucerons dans la sélection de l'avoine et de l'orge ont été examinées.

     

Hydroxylation of steroids by Fusarium oxysporum, Exophiala jeanselmei and Ceratocystis paradoxa

The potential of Fusarium oxysporum var. cubense UAMH 9013 to perform steroid biotransformations was reinvestigated using single phase and pulse feed conditions. The following natural steroids served as substrates: dehydroepiandrosterone (1), pregnenolone (2), testosterone (3), progesterone (4), cortisone (5), prednisone (6), estrone (7) and sarsasapogenin (8). The results showed the possible presence of C-7 and C-15 hydroxylase enzymes. This hypothesis was explored using three synthetic androstanes: androstane-3,17-dione (9), androsta-4,6-diene-3,17-dione (10) and 3α,5α-cycloandrost-6-en-17-one (11). These fermentations of non-natural steroids showed that C-7 hydroxylation was as a result of that position being allylic. The evidence also pointed towards the presence of a C-15 hydroxylase enzyme.The eleven steroids were also fed to Exophialajeanselmei var. lecanii-corni UAMH 8783. The results showed that the fungus appears to have very active 5α and 14α-hydroxylase enzymes, and is also capable of carrying out allylic oxidations.Ceratocystis paradoxa UAMH 8784 was grown in the presence of the above-mentioned steroids. The results showed that monooxygenases which effect allylic hydroxylation and Baeyer–Villiger rearrangement were active. However, redox reactions predominated.