Promoters of auxin-induced genes from tobacco can lead to auxin-inducible and root tip-specific expression

In previous studies we have identified several mRNAs which accumulate after addition of 2,4-dichlorophenoxyacetic-acid (2,4-D) to auxin-starved tobacco cells [45, 46]. The mRNAs corresponding to cDNA clone pCNT103 were found to accumulate transiently prior to the cell division response due to auxin treatment. In this study we determined the sequences of three 103-like cDNAs and two 103-like genes, GNT1 and GNT35. To further study the regulation of the expression of these genes their 5 regions were translationally fused with the -D-glucuronidase reporter gene (GUS). The GNT1 5 region led to GUS expression only in the root tips of transgenic plants. By using transgenic hairy-root cultures and transformed cell suspension cultures it was shown that the 5 regions of both GNT1 and GNT35 lead to 2,4-D-inducible expression of GUS activity. The homology of the 103-like genes with other auxin-regulated genes is evaluated.Department of Plant Molecular Biology, Leiden University     

High-level expression of a tobacco chitinase gene in Nicotiana sylvestris. Susceptibility of transgenic plants to Cercospora nicotianae infection

Endochitinases (E.C. 3.2.14, chitinase) are believed to be important in the biochemical defense of plants against chitin-containing fungal pathogens. We introduced a gene for class I (basic) tobacco chitinase regulated by Cauliflower Mosaic Virus 35S-RNA expression signals into Nicotiana sylvestris. The gene was expressed to give mature, enzymatically active chitinase targeted to the intracellular compartment of leaves. Most transformants accumulated extremely high levels of chitinase-up to 120-fold that of non-transformed plants in comparable tissues. Unexpectedly, some transformants exhibited chitinase levels lower than in non-transformed plants suggesting that the transgene inhibited expression of the homologous host gene. Progeny tests indicate this effect is not permanent. High levels of chitinase in transformants did not substantially increase resistance to the chitin-containing fungus Cercospora nicotiana, which causes Frog Eye disease. Therefore class I chitinase does not appear to be the limiting factor in the defense reaction to this pathogen.     

In vitro clonal multiplication of the actinorhizal plant Comptonia peregrina

Micropropagation of the actinorhizal plant Comptonia peregrina of the Myricaceae was achieved successfully by the induction of root buds in excised root culture with cytokinin (1.0 M benzyladenine). Excised root segments with initiated root buds were subcultured in Woody Plant Medium (WPM) lacking growth regulators, developing extensive callus which subsequently gave rise to multiple adventitious buds. Shoot elongation was facilitated by transfer of calluses to more aerated conditions. Root initiation was induced on shoots by brief treatment with auxin (<1 M indolebutyric acid) and transfer to WPM for plantlet development. Controlled light and aeration in liquid medium were critical conditions for successful micropropagation.     

Biodiversity amongst filamentous fungi

Diversities in fungi are manifold. Fungi themselves are heterogeneous and constitute at least three unrelated major taxa. Structural diversity reflects, in most cases, adaptive and functional strategies. Diversity in nucleic acids and chemical compounds is very high in several fungal taxa. Fungi play an essential role in the function of ecosystems. The diversity of plant parasites is extremely high and species-dependent associations exist. Saprobic fungi are most important in wood and litter decay and diverse taxa comprise the main decomposers in specific successional niches. Two dominating symbiotic systems have evolved convergently in various fungal groups, notably lichens and mycorrhizas, both remarkably diverse in their heterotrophic partners.     

Initiation of acellular extrinsic fiber cementum on human teeth

Summary The development of acellular extrinsic fiber cementum (AEFC) has never before been studied in human teeth. We have therefore examined the initiation of AEFC in the form of a collagenous fiber fringe and its attachment to the underlying dentinal matrix, in precisely selected, erupting human premolars with roots developed to 50%–60% of their final length. Freshly extracted teeth were prefixed in Karnovsky's fixative, decalcified in EDTA and subdivided into about 10 blocks each, cut from the mesial and distal root surfaces, vertical to and along the root axis. The blocks were postfixed in osmium tetroxide, embedded in Epon and cut for light- and electron-microscopic investigation. Starting at the advancing edge of the root, within a region extending about 1 mm coronal to this edge, fibroblast-like cells were seen closely covering the external root surface. Along the first 100 m from the root edge, these cells extended cytoplasmic processes and contacted the dentinal collagen fibrils. Between these cells and the dentinal matrix, new collagen fibrils and very short collagen fibers gradually developed. Within the second 100 m from the root edge, this resulted in the formation of a cell-fiber fringe network. Newly formed fibers of the fringe were directly attached to the non-mineralized matrix containing dentinal collagen fibrils and could be distinguished from the latter by differences in fibril orientation. During the process of dentin mineralization, the transitional zone between the fiber-fringe base and the dentinal matrix, i.e., the future dentino-cemental junction, also mineralized. It is suggested that this fiber fringe is the base of AEFC, which later increases in thickness by fiber extension and subsequent mineralization.Abbreviations AEFC acellular extrinsic fiber cementum - AIFC acellular intrinsic fiber cementum - CIFC cellular intrinsic fiber cementum - CMSC cellular mixed stratified cementum - ARE advancing root edge - CP cytoplasmic process - D dentin - DCJ dentinocemental junction - E enamel - EBL external basal lamina - EC epithelial cell - EDTA ethylene diaminetetraacetic acid - ERM epithelial rests of Malassez - FF fiber fringe - HRS Hertwig's epithelial root sheath - IBL internal basal lamina - MD mineralized dentin - NMD non-mineralized dentin - OB odontoblast - PD predentin - PL periodontal ligament     

Neurofilament protein triplet immunoreactivity in the dorsal root ganglia of the guinea-pig

Summary Immunoreactivity for the neurofilament protein triplet was investigated in neurons of the dorsal root ganglia of the guinea-pig by using a battery of antibodies. In unfixed tissue, nearly all neurons in these ganglia demonstrated some degree of neurofilament protein triplet immunoreactivity. Large neurons generally displayed intense immunoreactivity, whereas most small to medium-sized neurons showed faint to moderate immunoreactivity. Double-labelling immunofluorescence demonstrated that most antibodies to the individual subunits of the neurofilament protein triplet had the same distribution and intensity of labelling in sensory neurons. Increasing durations of tissue fixation in aldehyde solutions selectively diminished neurofilament protein triplet immunoreactivity in small to medium-sized neurons. Double-labelling with neurofilament protein triplet antibodies in combination with antibodies to other neuronal markers, such as neuron-specific enolase, substance P and tyrosine hydroxylase, showed that tissue processing conditions affect the degree of co-localization of immunoreactivity to the neurofilament protein triplet and to these other neuronal markers. These results indicate that, with a judicious manipulation of the duration of tissue fixation, neurofilament protein triplet immunoreactivity can be used in combination with other neuronal markers to distinguish groups of neurons according to their size and chemical coding.     

Influence of Meloidogyne incognita on Resistant and Susceptible Sweet Potato Cultivars

Effects of several population densities ofMeloidogyne incognita on the sweet potato cultivars Centennial (susceptible) and Jasper (moderately resistant) were studied. Field plots were infested with initial levels (Pi) of 0, 10, 100, 1,000, 5,000, and 10,000 eggs and juveniles/500 cm³ soil in 1980 and 0, 100, 1,000, 2,000, 3,000, 4,000, and 5,000 in 1981. M. incognita population development trends were similar on both cultivars; however, at high Pi, more eggs and juveniles were recovered from Centennial than from Jasper. The highest Pi did not result in the highest mid-season (Pm) counts. Pi was negatively correlated with the number of marketable roots and root weight but positively correlated with total cracked roots, percentage of cracked roots, and cracking severity. Jasper tolerated higher Pi with greater yields and better root quality than Centennial. Cracking of fleshy roots occurred with both cultivars at low Pi.