Fitness consequences of infection of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> with its natural bacterial pathogen <Emphasis Type="Italic">Pseudomonas viridiflava</Emphasis>

Variation in plant resistance to pathogen infection is commonly observed in interactions between wild plants and their foliar pathogens. Models of host–pathogen interactions indicate that a large cost of infection is generally necessary to maintain this variation, yet there is limited evidence that foliar pathogens cause detectable fitness reductions in wild host plants. Most published work has focused on fungal pathogens. Pseudomonas viridiflava, a common bacterial pathogen of the annual weed Arabidopsis thaliana across its range, comprises two distinct genetic clades that cause disease symptoms of different severity. Here we measured the extent of infection of wild A. thaliana populations in the Midwest, USA, and examined the effect on seed production, in field and growth-chamber experiments, of experimental inoculation with isolates from the two clades. We found infection with P. viridiflava varied from 0 to 56% in Midwest A. thaliana populations, with the possibility of several leaves per plant infected later in the growing season. In the growth chambers, experimental inoculation reduced seed set by averages of 15 and 11% for clades A and B, respectively. In the field experiment, only clade A affected plant fitness significantly, reducing seed set by an average of 38%. Underlying these average effects we observed both negative and positive effects of infection, and variation in both fitness among plant genotypes and sensitivity to environmental conditions.     

Injured and uninjured leaf photosynthetic responses after mechanical injury on <Emphasis Type="Italic">Nerium oleander</Emphasis> leaves,and <Emphasis Type="Italic">Danaus plexippus</Emphasis> herbivory on <Emphasis Type="Italic">Asclepias curassavica</Emphasis> leaves

Insect herbivory has variable effects on plant physiology; so greater understanding is needed about how injury alters photosynthesis on individual injured and uninjured leaves. Gas exchange and light-adapted leaf chlorophyll fluorescence measurements were collected from uninjured and mechanical partial leaf defoliation in two experiments with Nerium oleander (Apocynaceae) leaves, and one experiment with Danaus plexippus herbivory on Asclepias curassavica (Asclepiadaceae) leaves. Gas exchange impairment (lower photosynthetic rate (P _n ), stomatal conductance (g _s)) indicates water stress in a leaf, suggests stomatal limitations causing injury P _n impairment. The same pattern of gas exchange impairment also occurred on uninjured leaves opposite from injured leaves in both N. oleander experiments. This is an interesting result because photosynthetic impairment is rarely reported on injured leaves near injured leaves. No photosynthetic changes occurred in uninjured A. curassavica leaves opposite from D. plexippus-fed leaves. Partially defoliated leaves that had P _n and g _s reductions lacked any significant changes in intercellular leaf [CO₂], C _i. These results neither support, nor are sufficient to reject, stomatal limitations to photosynthesis. Manually imposed midrib vein severance in N. oleander experiment #1 significantly increased leaf C _i, indicating mesophyll limitations to photosynthesis. Maximal light-adapted leaf photochemical efficiency () and also non-photochemical quenching (q _N) were reduced by mechanical or insect herbivory to both study species, suggesting leaf trouble handling excess light energy not used for photochemistry. Midrib injury on N. oleander leaves and D. plexippus herbivory on A. curassavica leaves also reduced effective quantum yield (ΦPSII) and photochemical quenching (q _P); so reduced plastoquinone pools could lead to additional PSII reaction center closure.     

Reproductive success in the Mexican rewardless <Emphasis Type="Italic">Oncidium cosymbephorum</Emphasis> (Orchidaceae) facilitated by the oil-rewarding <Emphasis Type="Italic">Malpighia glabra</Emphasis> (Malpighiaceae)

The pollination of one plant species can be facilitated by the presence of one or more neighboring plant species and evidence has been found in some rewardless species of orchid that benefit from the presence of rewarding plant species in the neighborhood. There are two pollination mechanisms by which a non-rewarding orchid attracts pollinators and increases its reproductive success: (1) A magnetic species effect that occurs even though the flowers do not resemble those of the other species, and (2) floral mimicry where the mimic’s flower resembles that of the model plant species. Oncidium cosymbephorum is a Mexican rewardless epiphytic orchid whose flowers look like those of the rewarding shrub Malpighia glabra (Malpighiaceae). The resemblance of O. cosymbephorum to the oil-offering flowers of M. glabra attracts the same pollinators, and the fitness of the orchid is higher when M. glabra is present than when it is absent. We evaluated the facilitation by M. glabra of the orchid’s pollination for natural and artificial clumps of O. cosymbephorum close to and far from M. glabra over 4 years. Two experiments were performed at five different study sites to evaluate the effect of the presence and absence of M. glabra on the reproductive success of O. cosymbephorum. In experiment 1, we recorded fruit set production in natural and artificial monospecific clumps of the orchid, and in natural and artificial heterospecific clumps of O. cosymbephorum and M. glabra. In experiment 2, we recorded the fruit set of O. cosymbephorum at different sites where individuals grow in monospecific clumps, both before and after cultivated individuals of oil-producing M. glabra had been planted in their vicinity. Both experiments showed that the reproductive success of O. cosymbephorum was greater in the presence of M. glabra than it was in its absence. This study provides experimental evidence for the magnetic species effect. Floral similarity between O. cosymbephorum and M. glabra, should be experimentally tested to determine whether it is adaptive.     

Influence of invasive <Emphasis Type="Italic">Spartina</Emphasis> growth stages on associated macrofaunal communities

In coastal wetlands, invasive plants often act as ecosystem engineers altering flow, light and sediments which, in turn, can affect benthic animal communities. However, the degree of influence of the engineer will vary significantly as it grows, matures and senesces, and surprisingly little is known about how the influence of an ecosystem engineer varies with ontogeny. We address this issue on the tidal flats of San Francisco Bay where hybrid Spartina (foliosa × alterniflora) invaded 30 years ago. The invasion has altered the physico-chemical properties of the sediment habitat, which we predicted should cause changes in macrofaunal community structure and function. Through mensurative and manipulative approaches we investigated the influence of different growth stages of hybrid Spartina on macrobenthos and the underlying mechanisms. Cross-elevation sampling transects were established covering 5 zones (or stages) of the invasion, running from the tidal flat (pre-invasion) to an unvegetated dieback zone. Additionally, we experimentally removed aboveground plant structure in the mature (inner) marsh to mimic the ’unvegetated areas’. Our results revealed four distinct faunal assemblages, which reflected Spartina-induced changes in the corresponding habitat properties along an elevation gradient: a pre-invaded tidal flat, a leading edge of immature invasion, a center of mature invasion, and a senescing dieback area. These stages of hybrid Spartina invasion were accompanied by a substantial reduction in macrofaunal species richness and an increase in dominance, as well as a strong shift in feeding modes, from surface microalgal feeders to subsurface detritus/Spartina feeders (mainly tubificid oligochaetes and capitellid polychaetes). Knowledge of the varying influence of plant invaders on the sediment ecosystem during different phases of invasion is critical for management of coastal wetlands.     

<Emphasis Type="Italic">Xanthomonas gardneri</Emphasis> exoenzymatic activity towards plant tissue

Bacterial spot caused by Xanthomonas spp. is an important tomato and pepper disease worldwide. Recent outbreaks of bacterial spot disease in Central Brazil and Canada have been attributed to Xanthomonas gardneri, which is also recognized as group D of Xanthomonas campestris pv. vesicatoria. Carotenoid-like pigments called xanthomonadins, which are diagnostic for yellow Xanthomonas spp., were extracted from X. gardneri. It was shown that the model plant Arabidopsis thaliana, member of the Brassicaceae family, can develop disease symptoms in response to different isolates of X. gardneri. Secretion of enzymes has been shown to play an important role in pathogenicity for different pathogens, and to begin to understand the interaction of X. gardneri and A. thaliana, a biochemical analysis of secreted proteins in the presence of A. thaliana leaves was performed. Different enzymatic activities such as for cellulase, α-arabinofuranosidase, pectinase, invertase and xylanase were assayed. In the presence of leaves, cellulase activity was highest after 60 and 72 h of growth and α-arabinofuranosidase activity was detected between 12 and 72 h of growth. Pectinase, invertase and xylanase activities were not detected. Cellulase and α-arabinofuranosidase activities may be important for X. gardneri acquisition of plant nutrients through degradation of cellulose fibers and hemicellulose of the cell wall, respectively, to the invasion of the host tissue and/or may generate signal molecules that are recognized by the plant. This is the first study to address how X. gardneri responds to host plant tissue.     

Systemic response to aphid infestation by <Emphasis Type="Italic">Myzus persicae</Emphasis> in the phloem of <Emphasis Type="Italic">Apium graveolens</Emphasis>

Little is known about the molecular processes involved in the phloem response to aphid feeding. We investigated molecular responses to aphid feeding on celery (Apium graveolenscv. Dulce) plants infested with the aphid Myzus persicae, as a means of identifying changes in phloem function. We used celery as our model species as it is easy to separate the phloem from the surrounding tissues in the petioles of mature leaves of this species. We generated a total of 1187 expressed sequence tags (ESTs), corresponding to 891 non-redundant genes. We analysed these ESTs in silico after cDNA macroarray hybridisation. Aphid feeding led to significant increase in RNA accumulation for 126 different genes. Different patterns of deregulation were observed, including transitory or stable induction 3 or 7days after infestation. The genes affected belonged to various functional categories and were induced systemically in the phloem after infestation. In particular, genes involved in cell wall modification, water transport, vitamin biosynthesis, photosynthesis, carbon assimilation and nitrogen and carbon mobilisation were up-regulated in the phloem. Further analysis of the response in the phloem or xylem suggested that a component of the response was developed more specifically in the phloem. However, this component was different from the stress responses in the phloem driven by pathogen infection. Our results indicate that the phloem is actively involved in multiple adjustments, recruiting metabolic pathways and in structural changes far from aphid feeding sites. However, they also suggest that the phloem displays specific mechanisms that may not be induced in other tissues.EST, macroarray and clustering data are available from our website [http://www-biocel.versailles.inra.fr/phloem]. Data deposition: The sequences reported in this paper have been deposited in the Genbank database (Accession nos.: AY607692-AY607700, AY611007, CN253939-CN255151, CV512445-CV512447 and CV651120-CV651121).     

Arbuscular mycorrhizal symbiosis increases host plant acceptance and population growth rates of the two-spotted spider mite <Emphasis Type="Italic">Tetranychus urticae</Emphasis>

Most terrestrial plants live in symbiosis with arbuscular mycorrhizal (AM) fungi. Studies on the direct interaction between plants and mycorrhizal fungi are numerous whereas studies on the indirect interaction between such fungi and herbivores feeding on aboveground plant parts are scarce. We studied the impact of AM symbiosis on host plant choice and life history of an acarine surface piercing-sucking herbivore, the polyphagous two-spotted spider mite Tetranychus urticae. Experiments were performed on detached leaflets taken from common bean plants (Phaseolus vulgaris) colonized or not colonized by the AM fungus Glomus mosseae. T. urticae females were subjected to choice tests between leaves from mycorrhizal and non-mycorrhizal plants. Juvenile survival and development, adult female survival, oviposition rate and offspring sex ratio were measured in order to estimate the population growth parameters of T. urticae on either substrate. Moreover, we analyzed the macro- and micronutrient concentration of the aboveground plant parts. Adult T. urticae females preferentially resided and oviposited on mycorrhizal versus non-mycorrhizal leaflets. AM symbiosis significantly decreased embryonic development time and increased the overall oviposition rate as well as the proportion of female offspring produced during peak oviposition. Altogether, the improved life history parameters resulted in significant changes in net reproductive rate, intrinsic rate of increase, doubling time and finite rate of increase. Aboveground parts of colonized plants showed higher concentrations of P and K whereas Mn and Zn were both found at lower levels. This is the first study documenting the effect of AM symbiosis on the population growth rates of a herbivore, tracking the changes in life history characteristics throughout the life cycle. We discuss the AM-plant-herbivore interaction in relation to plant quality, herbivore feeding type and site and the evolutionary implications in a multi-trophic context.     

Effects of browsing in relation to vegetation cover on common yew (<Emphasis Type="Italic">Taxus baccata</Emphasis> L.) recruitment in Mediterranean environments

Common yew (Taxus baccata L.) stands are recognized as prioritary habitats for biodiversity conservation within the European Union. The effects of browsing on the regeneration capacity and spatial dispersal of T. baccata recruits at the European southern limit of the species in the Mediterranean Basin have been herein studied. The efficacy of T. baccata recruitment has been evaluated at six localities in the Northern Sardinia mountains, which have similar altitude, climate, soil, and vegetation but have different types of uses (three were grazed by livestock and three were not). At each site, five habitats have been identified for T. baccata seed dispersal: reproductive female T. baccata canopy, reproductive female Ilex aquifolium canopy, non-fleshy-fruited tree canopy, fleshy-fruited shrubs, and open areas. The density of seedlings was found to be greater under fleshy-fruited trees (reproductive female T. baccata and I. aquifolium) than under shrubs, whereas the sapling density was higher in shrubby habitats, especially at grazed sites due to the mechanical protection afforded by the spiny shrubs against herbivores. Land use (LU) has been found to be the most important factor in determining the spatial distribution of seedlings and saplings in relation to forest habitats. Although browsers had an ephemeral but positive effect on seed germination through their trampling and the resultant scarification, this process eventually became ineffective as was shown by the occurrence of the lowest density of saplings in those habitats where the density of seedlings was the highest. The ultimate and most important effect of browsing was the sharp decrease in the density of saplings, and their almost complete extinction, in non-shrubby habitats. This study highlights the result that, in Mediterranean ecosystems, browsing constitutes the main negative factor on T. baccata seedling-sapling transition and furthermore confirms the necessity to preserve shrubby patches in the vicinity of reproductive female T. baccata and I. aquifolium to permit the regeneration of T. baccata in the presence of livestock. Moreover, at ungrazed sites, T. baccata is able to colonize non-shrubby shady habitats. The application of different management strategies to ungrazed and grazed sites should therefore be the main direction in the management and preservation of T. baccata stands in the Mediterranean region.     

Impact of volcanic activity on a plant-pollinator module in an island ecosystem: the example of the association of <Emphasis Type="Italic">Camellia japonica</Emphasis> and <Emphasis Type="Italic">Zosterops japonica</Emphasis>

Volcanic activity provides an indispensable opportunity to study the ecological responses of organisms to environmental devastation. We examined the reproductive success of Camellia trees to identify how volcanic activity affects the processes of leaf survival, flowering activity, fruit and seed production, pollinator abundance, pollinator visitation frequency, pollination rate, and fruit and seed maturation at different damage sites on Miyake-jima, which experienced an eruption in the summer of 2000. Volcanic gases negatively affect leaf survival and reduce flowering activity in heavily damaged areas. Pollen transfer was sufficient to ensure that higher pollination rates (83%) occurred in heavily damaged areas than in less damaged areas (26–45%), but pollinator densities were lower in response to reduced flower resources. Fruit abortion rates were greater in heavily damaged sites (78%) than in less-damaged sites (53–63%). Consequently, fruit-set rates (16–29%) did not differ significantly among sites. Seed set rates tended to increase with increasing volcanic damage. The negative correlation between seed-set rates and seed mass suggests that the decreased seed mass in severely damaged sites was attributable to the better pollination rates observed there. These results indicate that compensation mechanisms ensure better reproductive success at sites that are more strongly affected by volcanic activity.     

<Emphasis Type="Italic">In vitro</Emphasis> plant regeneration of <Emphasis Type="Italic">Alnus acuminata</Emphasis> H.B.K. ssp. <Emphasis Type="Italic">acuminata</Emphasis> and its root nodulation by <Emphasis Type="Italic">Frankia</Emphasis>

A procedure for in vitro plant regeneration of Alnus acuminata from epicotyls with cotyledonary buds was developed using different media formulations with different growth regulators and carbon sources. The development of multiple buds on explants at the initiation step was obtained with MS at 1/2 strength with either 1 or 2M of BAP but not without it. Multiplication gave up to 15 elongating shoots by explant, the best medium being MS supplemented with vitamins from B5 medium, 1M of BAP and 87mM sucrose. Rooting of about 88% occurred in the medium MS with 83 mM sucrose and 1M IBA. Alnus acuminata did not developed well on WPM. Roots of in vitro propagated plants were nodulated by Alnus-infective Frankia. The root nodules show a typical alder root nodule anatomy and differentiation pattern and effectively fixed nitrogen. Rhamnaceae-infective Frankia did not nodulate in vitro cultivated Alnus acuminata suggesting that symbiotic recognition was not altered by in vitro regeneration of the plant.     

Low soil water and nutrient availability below New Zealand kauri (<Emphasis Type="Italic">Agathis australis</Emphasis> (D. Don) Lindl.) trees increase the relative fitness of kauri seedlings

Tree species can affect the soil they are growing on and this might influence their fitness. The New Zealand gymnosperm tree species kauri (Agathis australis (D. Don) Lindl.) which grows in mixed angiosperm–gymnosperm forests has a substantial effect upon the soil. We studied the hypotheses that: (1) low soil moisture availability below mature kauri trees hampers growth of kauri seedlings and angiosperm seedlings, (2) low nutrient availability below kauri trees hampers only angiosperm seedlings, and (3) angiosperm seedlings are hampered more than kauri seedlings by the conditions below kauri trees. We tested these hypotheses by planting seedlings of kauri and mapau (Myrsine australis (A. Rich) Allan) under kauri trees and applying the following treatments: removal of herbs, removal of litter, removal of nutrient limitation, and elimination of root competition of mature kauri trees. The results indicate that low soil moisture availability, or the combination of low soil moisture availability and low nutrient fertility, hampers the growth of kauri as well as mapau seedlings below kauri trees. The mapau seedlings are hampered relatively more than the kauri seedlings which might result in an increased relative fitness of the latter.     

Effects of ozone in <Emphasis Type="Italic">Plantago lanceolata</Emphasis> and <Emphasis Type="Italic">Salix atrocinerea</Emphasis> pollen

Pollen can be modified by the increasing environmental pollution which can lead to a rising trend in pollen-induced respiratory allergies. The aim of this work was to study the fertility, protein profile and allergenicity of Plantago lanceolata and Salix atrocinerea pollen after exposure to ozone. The pollen collected directly from the anthers of both species was exposed to O₃ in an environmental chamber, for 6 h at different concentrations. After exposing the pollen, viability tests and in vitro germination were performed. Total protein content was determined by the Bradford method, the polypeptides were separated by SDS-PAGE and the antigenic properties were analyzed by immunological techniques using patients’ sera. The concentrations of O₃ tested affected the pollen fertility and the total protein content, but the protein profiles did not show any differences. Furthermore, the immunological tests revealed an increase in IgE binding reactivity when pollen was exposed to the pollutant.     

<Emphasis Type="Italic">Sporidiobolus johnsonii</Emphasis> and <Emphasis Type="Italic">Sporidiobolus salmonicolor</Emphasis> revisited

The relationship between Sporidiobolus johnsonii and S. salmonicolor was investigated using rDNA sequence data. Two statistically well-supported clades were obtained. One clade included the type strain of S. johnsonii and the other included the type strain of S. salmonicolor. However, some mating strains of S. salmonicolor were found in the S. johnsonii group. These strains belonged to mating type A2 and were sexually compatible with mating type A1 strains from the S. salmonicolor group. DNA–DNA reassociation values were high within each clade and moderate between the two clades. In the re-investigation of teliospore germination, we observed that the basidia of S. salmonicolor were two-celled. In S. johnsonii, basidia were not formed and teliospore germination resulted in direct formation of yeast cells. We hypothesize that the S. johnsonii clade is becoming genetically isolated from the S. salmonicolor group and that a speciation process is presently going on. We suspect that the observed sexual compatibility between strains of the S. johnsonii and S. salmonicolor groups and the possible genetic flow between the two species has little biological relevance because distinct phenotypes have been fixed in the two taxa and intermediate (hybrid) sequences for LSU and ITS rDNAs have not been detected. An erratum to this article can be found at     

Evolution of flower shape in <Emphasis Type="Italic">Plantago lanceolata</Emphasis>

Plantago lanceolata produces small actinomorphic (radially symmetric), wind-pollinated flowers that have evolved from a zygomorphic, biotically pollinated ancestral state. To understand the developmental mechanisms that might underlie this change in flower shape, and associated change in pollination syndrome, we analyzed the role of CYC-like genes in P. lanceolata. Related zygomorphic species have two CYC-like genes that are expressed asymmetrically in the dorsal region of young floral meristems and in developing flowers, where they affect the rate of development of dorsal petals and stamens. Plantago has a single CYC-like gene (PlCYC) that is not expressed in early floral meristems and there is no apparent asymmetry in the pattern of PlCYC expression during later flower development. Thus, the evolution of actinomorphy in Plantago correlates with loss of dorsal-specific CYC-like gene function. PlCYC is expressed in the inflorescence stem, in pedicels, and relatively late in stamen development, suggesting a novel role for PlCYC in compacting the inflorescence and retarding stamen elongation in this wind pollinated species.     

Methods for syntheses of <Emphasis Type="Italic">N</Emphasis>-methyl-<Emphasis Type="Italic">DL</Emphasis>-aspartic acid derivatives

Summary. A novel practical method for the synthesis of N-methyl-DL-aspartic acid 1 (NMA) and new syntheses for N-methyl-aspartic acid derivatives are described. NMA 1, the natural amino acid was synthesized by Michael addition of methylamine to dimethyl fumarate 5. Fumaric or maleic acid mono-ester and -amide were regioselectively transformed into beta-substituted aspartic acid derivatives. In the cases of maleamic 11a or fumaramic esters 11b, the α-amide derivative 13 was formed, but hydrolysis of the product provided N-methyl-DL-asparagine 9 via base catalyzed ring closure to DL-α-methylamino-succinimide 4, followed by selective ring opening. Efficient methods were developed for the preparation of NMA-α-amide 13 from unprotected NMA via sulphinamide anhydride 15 and aspartic anhydride 3 intermediate products. NMA diamide 16 was prepared from NMA dimethyl ester 6 and methylamino-succinimide 4 by ammonolysis. Temperature-dependent side reactions of methylamino-succinimide 4 led to diazocinone 18, resulted from self-condensation of methylamino-succinimide via nucleophyl ring opening and the subsequent ring-transformation.     

Flavonoids in the leaves of <Emphasis Type="Italic">Oxalis corniculata </Emphasis>and sequestration of the flavonoids in the wing scales of the pale grass blue butterfly, <Emphasis Type="Italic">Pseudozizeeria </Emphasis><Emphasis Type="Italic">maha</Emphasis>

Three C-glycosylflavones in the leaves of Oxalis corniculata, the host plant of the lycaenid butterfly pale grass blue (Pseudozizeeria maha), were identified as 6-C-glucosylluteolin (isoorientin), 6-C-glucosylapigenin (isovitexin) and isovitexin 7-methyl ether (swertisin). Comparative spectral and HPLC analyses between the leaf extract of the host plants and the wings of P. maha showed selective uptake of the host-plant flavonoid isovitexin to the wings of the butterfly.     

Temporal variation in the effects of habitat fragmentation on reproduction of the Mediterranean shrub <Emphasis Type="Italic">Colutea hispanica</Emphasis>

Habitat fragmentation poses a major threat to the viability of plant populations. However, the intensity of fragmentation effects may vary among years. We studied two possible effects of habitat fragmentation (patch size and isolation) on the reproduction and proportion of damaged fruits in 24 patches of the self-compatible shrub Colutea hispanica for three consecutive years with different climate conditions. We also studied the effect of fragmentation on the incidence of two main pre-dispersal seed predators, the butterflies Iolana iolas and Lampides boeticus. High between-year variability was found in number of viable seeds per fruit, number of fruits per plant, total number of viable seeds per plant and proportion of damaged fruits. In 2003, small, isolated patches had a higher fruit set and number of fruits per plant. The proportion of damaged fruits was significantly lower in isolated populations in 2003, while it was very high in all patches in 2004 and 2005. High between-year variability was also found in the proportion of fruits per plant with I. iolas eggs. In 2003 isolated patches had a lower proportion of fruits with I. iolas eggs, but no significant effect of patch size and isolation was found in 2004 or 2005. The proportion of fruits with L. boeticus eggs was similar in the three years of study, although it was slightly higher in large, non-isolated patches in 2003. Thus, the effects of fragmentation on plant reproduction cannot be generalized from one single-year survey. In contrast to the generally accepted idea that fragmentation reduces plant reproduction, plant fitness may increase in isolated patches in years with high fruit production and low seed predation.     

Arabino-Galactan Proteins from <Emphasis Type="Italic">Pistacia lentiscus</Emphasis> var. <Emphasis Type="Italic">chia</Emphasis>: isolation,characterization and biological function

Summary. Arabino-Galactan Proteins (AGPs) were isolated from Chios mastic gum (CMG) by using a buffer containing 0.1 M NaCl, 20 mM Tris–HCl, pH 7.5. Protein analytical methods, combined with specific procedures for carbohydrate characterization, indicated the presence of highly glycosylated protein backbone. In particular, staining by Yariv reagent of the electrophoretically separated molecules revealed the existence of arabinose and galactose and such a modification is characteristic for AGPs. After experiments involving extensive dialysis of the isolated extracts against water and atomic absorption, there was evidence of the existence of zinc ions that are probably covalently bound to the AGPs. By using anion-exchange chromatography, capillary electrophoresis, colorimetric methods and GC-MS, it was found that the extracts were separated into three major populations (A, B, and C), which were consistent with their respective negative charge content namely, uronic acid. The characterization of neutral sugars that was investigated with GC-MS showed the existence of arabinose and galactose in different amounts for each group. Experiments concerning the inhibition of growth of Helicobacter pylori in the presence of AGPs, as is shown for other CMG constituents, showed that the extracts of at least 1.4 g CMG affected the viability of the bacterium. There is no evidence as to whether the AGPs provoke abnormal morphologies of H. pylori, as is reported for the total CMG, or for O-glycans that possess terminal α1, 4-linked N-acetylglucosamine and are expressed in the human gastric mucosa; this has to be further investigated. Authors’ address: Theodora Choli-Papadopoulou, Laboratory of Biochemistry, School of Chemistry, Aristotle University of Thessaloniki, TK 54124 Thessaloniki, Greece     

Characterization of directly transformed weedy <Emphasis Type="Italic">Brassica rapa</Emphasis> and introgressed <Emphasis Type="Italic">B. rapa</Emphasis> with Bt <Emphasis Type="Italic">cry1Ac</Emphasis> and <Emphasis Type="Italic">gfp</Emphasis> genes

Crop to weed transgene flow, which could result in more competitive weed populations, is an agricultural biosafety concern. Crop Brassica napus to weedy Brassica rapa hybridization has been extensively characterized to better understand the transgene flow and its consequences. In this study, weedy accessions of B. rapa were transformed with Bacillus thuringiensis (Bt) cry1Ac- and green fluorescence protein (gfp)-coding transgenes using Agrobacterium to assess ecological performance of the wild biotype relative to introgressed hybrids in which the transgenic parent was the crop. Regenerated transgenic B. rapa events were characterized by progeny analysis, Bt protein enzyme-linked immunosorbent assay (ELISA), Southern blot analysis, and GFP expression assay. GFP expression level and Bt protein concentration were significantly different between independent transgenic B. rapa events. Similar reproductive productivity was observed in comparison between transgenic B. rapa events and B. rapa × B. napus introgressed hybrids in greenhouse and field experiments. In the greenhouse, Bt transgenic plants experienced significantly less herbivory damage from the diamondback moth (Plutella xylostella). No differences were found in the field experiment under ambient, low, herbivore pressure. Directly transformed transgenic B. rapa plants should be a helpful experimental control to better understand crop genetic load in introgressed transgenic weeds.