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Enniatin production by fusarium strains and its effect on potato tuber tissue.

Several Fusarium strains produce the cyclohexadepsipeptide enniatin, a host-nonspecific phytotoxin. Enniatins are synthesized by the 347-kDa multifunctional enzyme enniatin synthetase. In the present study, 36 Fusarium strains derived from a wide range of host plants were characterized with respect to enniatin production in different media. Thirteen of these strains produced enniatins on one or more of these media. To determine whether enniatin production affected virulence, an assay on potato tuber tissue was performed. Seven enniatin-producing and 16 nonproducing strains induced necrosis of potato tuber tissue, so that enniatin synthesis is not essential for the infection of potato tuber tissue. The application of a mixture of enniatins to slices of potato tuber, however, caused necrosis of the tissue. Therefore, enniatin production by the enniatin-synthesizing strains may affect their pathogenicity. The enniatin synthetase gene (esyn1) of Fusarium scirpi ETH 1536 was used as a probe to determine if similar sequences were present in the strains examined. In Southern blot analyses, DNA sequences hybridizing with the esyn1 probe were present in all but two of the strains examined. In some cases, enniatin-nonproducing strains had the same hybridization pattern as enniatin producers.

Journal Article↗

Reevaluation of the life cycle of Tuber magnatum.

Tuber spp. are ectomycorrhizal ascomycetes that produce ascocarps known as truffles. Basic aspects of Tuber biology have yet to be fully elucidated. In particular, there are conflicting hypotheses concerning the mating system and the ploidy level of the mycorrhizal and truffle hyphae. We used polymorphic microsatellites to compare the allelic configurations of asci with those from the network of the surrounding hyphae in single Tuber magnatum truffles. We then used these truffles to inoculate host plants and evaluated the microsatellite configurations of the resulting mycorrhizal root tips. These analyses provide direct evidence that T. magnatum outcrosses and that its life cycle is predominantly haploid. In addition to its scientific significance, this basic understanding of the T. magnatum life cycle may have practical importance in developing strategies to obtain and select nursery-produced mycorrhizal plants as well as in the management of artificial plantations of this and other Tuber spp.

Ascomycota↗

Adhesion to hyphal matrix and antifungal activity of Pseudomonas strains isolated from Tuber borchii ascocarps.

Pseudomonas spp. isolates from Tuber borchii ascocarps, known to be able to produce phytoregulatory and biocontrol substances in pure culture, were used to perform studies on their possible physiological role in nature. Antimycotic activity was confirmed against fungal contaminants isolated from the ascocarps, suggesting that populations associated with Tuber borchii fruit bodies may play a role in the maintenance of ascocarp health. Fifty-five percent of strains tested were also able to release metabolites which affected T. borchii mycelial growth and morphogenesis in culture. On the contrary, growth of the arbuscular mycorrhizal fungus Glomus mosseae and the ectomycorrhizal fungus Laccaria bicolor, putative competitors of Tuber for mycorrhizal infection sites on roots, was not influenced by the presence of any bacterial strain. The possibility that these bacteria, which show antifungal activity and fungal growth modulation activities, might be incorporated in the developing ascocarp by means of their preferential adhesion to Tuber mycelium is discussed.

Ascomycota↗

Topographic organization of the corticonuclear fibers from the tuber vermis and paramedian lobule in the albino rat.

The topographic organization of the corticonuclear fibers from the tuber vermis and paramedian lobule in the albino rat was investigated by autoradiographic anterograde tracing method. The medial portion of the tuber vermis projects to the dorsal part of the caudomedial subdivision of the medial cerebellar nucleus (MNcm), whereas the lateral portion of the tuber vermis projects to the dorsal part of the MNcm and the caudal part of the middle subdivision of the medial nucleus. The intermediate cortex of the paramedian lobule can be subdivided mediolaterally into three portions which project to the dorsolateral protuberance of the medial cerebellar nucleus, the rostrodorsal part of the posterior interpositus nucleus, and the caudodorsal part of the lateral anterior interpositus nucleus, respectively. The lateral cortex of the paramedian lobule can also be subdivided mediolaterally into two portions: the medial portion projects to the dorsolateral hump, and the lateral one to the lateral cerebellar nucleus. These results indicate that the cortical efferent fibers from the tuber vermis and paramedian lobule are clearly organized in the mediolateral direction in the albino rat.

Animals↗

Augmentation of the pharmacological action of corydalis tuber by saussurea root in isolated mouse ileum.

To understand the meaning of blending crude drugs in Chinese medicinal prescriptions, the influence of Saussurea root on the pharmacological action of Corydalis tuber was examined. Saussurea root increased the depression of acetylcholine-induced contraction caused by the hot water extract solution of Corydalis tuber in mouse ileum at low dosage, which showed no direct influence on acetylcholine. Dehydrocostuslactone in Saussurea root was characterized as the component having increasing activity and the relationship between the concentration of acetylcholine and the variation in the contraction depressed by Corydalis tuber alone or a mixture of the Corydalis tuber and dehydrocostuslactone was investigated for clarification of the mode of action.

Acetylcholine↗

Change in content of sugars and free amino acids in potato tubers under short-term storage at low temperature and the effect on acrylamide level after frying.

Changes in the sugar and amino acid contents of potato tubers during short-term storage and the effect on the acrylamide level in chips after frying were investigated. The acrylamide content in chips began to increase after 3 days of storage at 2 degrees C in response to the increase of glucose and fructose contents in the tubers. There was strong correlation between the reducing sugar content and acrylamide level, R(2)=0.873 for fructose and R(2)=0.836 for glucose. The sucrose content had less correlation with the acrylamide content because of its decrease after 4 weeks of storage at 2 degrees C, while the reducing sugar in potato tubers and the acrylamide in chips continued to increase. The contents of the four amino acids, i.e., asparatic acid, asparagine, glutamic acid and glutamine, showed no significant correlation with the acrylamide level. These results suggest that the content of reducing sugars in potato tubers determined the degree of acrylamide formation in chips. The chip color, as evaluated by L* (lightness), was correlated well with the acrylamide content.

Acrylamide↗

Wound-induced respiration and pyrophosphate:fructose-6-phosphate phosphotransferase in potato tubers.

A seven fold increase in the rate of respiratory O2 uptake was observed 24 h after slicing of potato tuber disks. The maximum activity of pyrophosphate:fructose-6-phosphate phosphotransferase (PFP) was 5-7 times greater than that of ATP-dependent phosphofructokinase (PFK) in fresh or aged potato slices. Thus, PFP may participate in glycolysis which supplies respiratory substrate in potato tubers. The PFP activity of desalted extracts determined in the absence of fructose-2,6-bisphosphate (F2,6BP) increased by 4.5 fold 24 h after slicing. However, maximal PFP activity determined with saturating (1 microM) F2,6BP was not changed. The Ka values of PFP for F2,6BP was lowered from 33 to 7 nM after 24 h of aging treatment. This increased susceptibility of the PFP activity to its allosteric activator, F2,6BP, may be involved in the increased respiration in wounded disks of potato tubers. Immunoblotting experiments indicated that both the alpha (66 kDa) and the beta (60 kDa) subunits of PFP were present in fresh or 24 h aged tuber slices.

Kinetics↗

Effects of light on in vitro tuberization of the potato cultivar Desiree and its relatives.

Effects of different combinations of short days and dark treatment and different light intensities during the short days on in vitro tuberization of three potato cultivars were examined. Tuberization in Desiree, Cleopatra and Gracia was induced by short-day-treatment after 4 weeks culture under long days. To preserve natural endogenous hormonal balance of in vitro plantlets growth regulators were not added to the medium. The shorter the duration of short-days period the higher light intensity necessary for earlier tuber initiation. There was also a synchronizing effect of the high light intensity on tuber initiation but it depended on photoperiod-treatments and genotypes. The higher the light intensity during induction period the less favourable the effect of light applied after the induction period.

In Vitro Techniques↗

Influence of soft rot bacteria on growth of Listeria monocytogenes on potato tuber slices.

Growth of Listeria monocytogenes on potato tuber slices and its interaction with four representative species of soft rot bacteria (Pseudomonas fluorescens, P. viridiflava, Erwinia carotovora subsp. carotovora, and Xanthomonas campestris) were investigated. When potato tuber slices were inoculated with one of two L. monocytogenes strains (Scott A and ATCC 15313), an increase in numbers of 3 to 4 logs per gram of tissue was observed with samples that were stored at 20 degrees C for 6 days. However, an increase of about 2 logs was observed with samples that were stored at 8 degrees C for 12 days. When potato slices were simultaneously inoculated with L. monocytogenes and one of the four soft rot bacteria, the growth of L. monocytogenes was inhibited in the presence of P. fluorescens or P. viridiflava but was not significantly affected in the presence of E. carotovora or X. campestris. The antagonism of the two pseudomonads to L. monocytogenes was also observed in potato tuber extract and in culture media. Formation of inhibition zones was observed only in iron-deficient media but not in the medium supplemented with FeCl3. In addition, production of fluorescent siderophore (pyoverdin) by these two pseudomonads was demonstrated. L. monocytogenes was unable to colonize macerated plant tissue induced by soft-rotting bacteria 2 days before inoculation of the pathogen. These results indicate that growth of L. monocytogenes on potato tuber slices is differentially affected by soft rot bacteria and that antagonism of fluorescent pseudomonads to L. monocytogenes is possibly caused by the production of iron-chelating siderophore by these pseudomonads.

Colony Count, Microbial↗

[Effect of cultivating measures on the tuber yield of Curcuma longa L].

The effect of cultivating measures on the tuber yield of Curcuma longa was observed. dibble planting has been proved move effective in raising the tuber yield than drill culture, and the dibble planting quantity has no effect on the tuber. The rational producing area, soil, seed tuber and so on were selected according to the experiments.

Methods↗

Size of tuber propagule influences injury of 'Kennebec' potato plants by constant light.

Chlorosis and necrotic spotting develop on the foliage of particular cultivars of potato (Solanum tuberosum L.) when grown under constant light. 'Kennebec', a cultivar severely injured by constant light when propagated from tissue-cultured plantlets, also was injured when plants were propagated from small tuber pieces (approximately 1 g). However, plants did not develop injury when propagated from large tuber pieces (approximately 100 g). Plants from large tuber pieces grew more rapidly than plants from small tuber pieces. The role of plant vigor and carbohydrate translocation in controlling injury development is discussed.

Culture Techniques↗

[Effect of elicitors on accumulation of protease inhibitors in injured potato tubers].

The time course of accumulation and the composition of proteinase-inhibiting proteins in diffusates from potato tubers treated with elicitors such as salicylic, jasmonic, and arachidonic acids were studied. The 40-kDa reserve protein patatin and the chymotrypsin inhibitors, among which proteins of 24.6, 22.0, and 16.0 kDa were prevalent, accumulated in diffusates from potato tubers. Jasmonic and arachidonic acids activated the accumulation of the chymotrypsin inhibitors in tubers in response to the injury stress, whereas salicylic acid inhibited this process. The effects of jasmonic and arachidonic acids increased when their concentrations decreased to 10(-6) M. The data suggest an important role of the lipoxygenase metabolism in signal transduction of the anti-injury defense system in the dormant potato tubers.

Arachidonic Acid↗

[Development, during ontogenetic development of gymnotids, of substance p expression in tuberous organs (electroreceptors)].

The evolution of the neuropeptidic expression of Substance P has been investigated with immunohistochemistry in the cutaneous electroreceptors, tuberous organs, during ontogenetic development of Apteronotus leptorhynchus. In the present data, antiSP antiserum has been applied to serial sections of Apteronotus leptorhynchus larvae obtained from several egg layings. Larvae were taken during development from hatching up to one hundred days old. SP immunoreactivity appeared just after hatching, in the epidermal zones which give rise to cutaneous sense organs. Four days after hatching, the tuberous organs are differentiated and immunoreactivity was observed in these organs, in both sensory cells and accessory cells. From day 30 after hatching, there was a regular decrease in the number of tuberous organs showing labelled accessory cells, and one hundred days later only 8% of tuberous organs had immunoreactive accessory cells. The adult accessory cells were no longer labelled with anti-SP antiserum. The results showed that in Apteronotus leptorhynchus, the epidermal structures which give rise to the cutaneous sensory organs were immunoreactive at a very early stage of development; this suggests that SP could have an effect upon their differentiation.

Animals↗

Focal cortical dysplasia of Taylor's balloon cell type: mutational analysis of the TSC1 gene indicates a pathogenic relationship to tuberous sclerosis.

Focal cortical dysplasia (FCD) is characterized by a localized malformation of the neocortex and underlying white matter. Balloon cells, similar to those observed in tuberous sclerosis, are present in many cases (FCD(bc)). In these patients, a hyperintense funnel-shaped subcortical lesion tapering toward the lateral ventricle was the characteristic finding on fluid-attenuated inversion recovery magnetic resonance imaging scans. Surgical lesionectomy results in complete seizure relief. Although the pathogenesis of FCD(bc) remains uncertain, histopathological similarities indicate that FCD(bc) may be related pathogenetically to tuberous sclerosis. Here, we studied alterations of the TSC1 and TSC2 genes in a cohort of patients with chronic, focal epilepsy and histologically documented FCD(bc) (n = 48). DNA was obtained after microdissection and laser-assisted isolation of balloon cells, dysplastic neurons, and nonlesional cells from adjacent normal brain tissue. Sequence alterations resulting in amino acid exchange of the TSC1 gene product affecting exons 5 and 17 and silent base exchanges in exons 14 and 22 were increased in patients with FCD(bc) compared with 200 control individuals (exon 5, 2.3% FCD(bc) vs 0% C; exon 17, 35% FCD(bc) vs 1.0% C; exon 14, 37.8% FCD(bc) vs 15% C; exon 22, 45% FCD(bc) vs 23.8% C). Sequence alterations could be detected in FCD(bc) and in adjacent normal cells. In 24 patients, DNA was suitable to study loss of heterozygosity at the TSC1 gene locus in microdissected FCD(bc) samples compared with control tissue. Eleven FCD(bc) cases exhibited loss of heterozygosity. In the TSC2 gene, only silent polymorphisms were detected at similar frequencies as in controls. Our findings indicate that FCD(bc) constitutes a clinicopathological entity with distinct neuroradiological, neuropathological, and molecular genetic features. These data also suggest a role of the TSC1 gene in the development of FCD(bc) and point toward a pathogenic relationship between FCD(bc) and the tuberous sclerosis complex.

Amino Acid Substitution↗

Expression profiling in tuberous sclerosis complex (TSC) knockout mouse astrocytes to characterize human TSC brain pathology.

Individuals with tuberous sclerosis complex (TSC) exhibit a variety of neurologic abnormalities, including mental retardation, epilepsy, and autism. Examination of human TSC brains demonstrate dysplastic astrocytes and neurons, areas of abnormal neuronal migration (tubers), and hamartomatous growths, termed subependymal nodules, which can progress to subependymal giant cell astrocytomas (SEGA). Previous studies have suggested that these neuropathologic features may result from abnormal neuroglial cell differentiation. In an effort to provide support for this hypothesis and to identify specific markers of aberrant neuroglial cell differentiation in TSC, we employed gene expression profiling on Tsc1 conditional knockout (Tsc1(GFAP)CKO) mouse astrocytes. We identified several transcripts implicated in central nervous system development that are differentially expressed in Tsc1(-/-) astrocytes compared to wild-type astrocytes. We validated the differential expression of select transcripts on the protein level both in primary cultures of Tsc1(-/-) astrocytes and in Tsc1(GFAP)CKO mouse brains. Moreover, we show that these markers are also differentially expressed within cortical tubers, but not in adjacent normal tissue from TSC patient brains. This study provides supportive evidence for a developmental defect in neuroglial cell differentiation relevant to the genesis of TSC nervous system pathology and underscores the utility of mouse modeling for understanding the molecular pathogenesis of human disease.

Animals↗

Molecular activity of sirolimus and its possible application in tuberous sclerosis treatment.

Sirolimus is one of the intensively investigated drugs with pluripotent activities. It binds to its intracellular receptor FKBP12 (FK506-binding protein 12), a member of the family of FK506-binding proteins, and inhibits the activity of mTOR, a serine/threonine kinase involved in numerous cell processes linked to cell growth control. The drug is currently registered for the prophylaxis of organ rejection and for use in coronary stents. However, unique characteristics of sirolimus make it a good candidate for anti-cancer therapy. Indeed, phase II and III clinical studies in humans with several types of neoplasms are already under way. The review describes molecular activity of sirolimus and its analogs, characteristic for specific applications, in view of very recent advances involving tuberous sclerosis complex (TSC)-mediated signaling pathways. Current studies with sirolimus performed in tuberous sclerosis animal models are presented. Possible application of sirolimus for treating tuberous sclerosis, disease caused by mutations of TSC proteins, is discussed.

Animals↗

Morphology of cerebral lesions in the Eker rat model of tuberous sclerosis.

Tuberous sclerosis (TSC) is an autosomal dominant disorder, caused by mutations of either the TSC1 or TSC2 gene. Characteristic brain pathologies (including cortical tubers and subependymal hamartomas/giant astrocytomas) are thought to cause epilepsy, as well as other neurological dysfunction. The Eker rat, which carries a spontaneous germline mutation of the TSC2 gene (TSC2+/-), provides a unique animal model in which to study the relationship between TSC cortical pathologies and epilepsy. In the present study, we have analyzed the seizure propensity and histopathological features of a modified Eker rat preparation, in which early postnatal irradiation was employed as a "second hit" stimulus in an attempt to exacerbate cortical malformations and increase seizure propensity. Irradiated Eker rats had a tendency toward lower seizure thresholds (latencies to flurothyl-induced seizures) than seen in non-irradiated Eker rats (significant difference) or irradiated wild-type rats (non-significant difference). The majority of irradiated Eker rats exhibited dysplastic cytomegalic neurons and giant astrocyte-like cells, similar to cytopathologies observed in TSC lesions of patients. The most prominent features in these brains were hamartoma-like lesions involving large eosinophilic cells, similar to giant tuber cells in human TSC. In some cells from these hamartomas, immunocytochemistry revealed features of both neuronal and glial phenotypes, suggesting an undifferentiated or immature cell population. Both normal-appearing and dysmorphic neurons, as well as cells in the hamartomas, exhibited immunopositivity for tuberin, the protein product of the TSC2 gene.

Animals↗

Non-penetrance in tuberous sclerosis.

As a result of extreme clinical variability in tuberous sclerosis, with one well-documented example of non-penetrance, phenotypically normal siblings or children of patients with tuberous sclerosis are thought to be at increased risk of having children with the disease. We report that the case of apparent non-penetrance that was previously described is the result of two independent tuberous-sclerosis mutations in the same family.

Exons↗