Philippe Vergne

Thin Film Colorimetric Interferometry

Measurement technique for the study of very thin lubrication films down to one nanometer in a point contact between a steel ball and a transparent disc is used to explore the relationship between central and minimum film thickness and rolling speed at the interface between elastohydrodynamic and boundary lubrication for a series of lubricating fluids. This technique based on the colorimetric interferometry combines powerful film thickness mapping capabilities with high accuracy. It was confirmed that both hexadecane and mineral base oil obey the linear relationship between log central and minimum film thickness and log rolling speed predicted by elastohydrodynamic theory down to approximately one nanometer. Conversely, squalane and additive-treated mineral base oil showed film thickness enhancement at slow speeds caused by boundary layers formation within the lubricant film. Obtained experimental data was used for the determination of pressure-viscosity coefficients of test fluids. The measurement technique also enabled us to produce information about the influence of boundary layers on film thickness shape. Presented as a Society of Tribologists and Lubrication Engineers Paper at the ASME/STLE Tribology Conference in Seattle, Washington, October 1–4, 2000

Analysis of gene expression in rose petals using expressed sequence tags

Single‐pass sequences were obtained from the 5′‐ends of a total of 1794 rose petal cDNA clones. Cluster analysis identified 242 groups of sequences and 635 singletons indicating that the database represents a total of 877 genes. Putative functions could be assigned to 1151 of the transcripts. Expression analysis indicated that transcripts of several of the genes identified accumulated specifically in petals and stamens. The cDNA library and expressed sequence tag database described here represent a valuable resource for future research aimed at improving economically important rose characteristics such as flower form, longevity and scent.

Biosynthesis of the major scent components 3,5-dimethoxytoluene and 1,3,5-trimethoxybenzene by novel rose O-methyltransferases

In Chinese rose species and in many modern varieties, two methylated phenolic derivatives, 3,5‐dimethoxytoluene and 1,3,5‐trimethoxybenzene, are major scent components. We show that cell‐free extracts of rose petals catalyse the synthesis of 3,5‐dimethoxytoluene and 1,3,5‐trimethoxybenzene by methylation of precursor molecules. An expressed sequence tag approach was used to identify four highly similar O‐methyltransferase sequences expressed specifically in petals and anthers. Thin layer chromatography analysis showed that the activities of these enzymes with different substrates and the proportions of reaction products produced closely mimicked those observed using cell‐free petal extracts, indicating that orcinol O‐methyltransferases are responsible for the biosynthesis of 3,5‐dimethoxytoluene and 1,3,5‐trimethoxybenzene from un‐methylated precursors in this organ.

Expression of heat shock factor and heat shock protein 70 genes during maize pollen development

We have analysed the expression of heat shock protein 70 (HSP70) and heat shock factor (HSF) gene during maize pollen development, HSFs being the transcriptional activators of hsp genes. In order to eliminate the sporophytic tissues of anthers, we have isolated homogeneous cell populations corresponding to five stages of maize pollen development from microspores to mature pollen. We show that in the absence of heat stress, hsp 70 genes are highly expressed late-bicellular pollen as compared to other stages. HSP70 transcripts are significantly accumulated in response to a heat shock at the late microspore stage but to a much lower extent than in vegetative tissues. The latest stages of pollen development, i.e. mid-tricellular and mature pollen, do not exhibit heat-induced accumulation of HSP70 transcripts. Therefore, we analysed the expression of hsf genes throughout pollen development. We demonstrate that at least three hsf genes are expressed in maize and that transcripts corresponding to one hsf gene, whose expression is independent of temperature in somatic as well as in microgametophytic tissues, are present at similar levels throughout pollen development. In addition, we show that the expression of the two other hsf genes is heat-inducible in maize vegetative tissues and is not significantly increased after heat shock at any stage of pollen development. These results indicate that the loss of hsp gene expression at late stages of pollen development is not due to a modification of hsf gene expression at the mRNA level and that hsf gene expression is differentially regulated in vegetative and microgametophytic tissues.

Genes normally expressed in the endosperm are expressed at early stages of microspore embryogenesis in maize

Reproduction in flowering plants is characterized by double fertilization and the resulting formation of both the zygotic embryo and the associated endosperm. In many species it is possible to experimentally deviate pollen development towards an embryogenic pathway. This developmental switch, referred to as microspore embryogenesis or androgenesis, leads to the formation of embryos similar to zygotic embryos. In a screen for genes specifically expressed during early androgenesis, two maize genes were isolated by mRNA differential display. Both genes represent new molecular markers expressed at a very young stage of androgenic embryogenesis. When their expression pattern was studied during normal reproductive development, both showed early endosperm-specific expression. Investigation of the cytological features of young androgenic embryos revealed that they present a partially coenocytic organization similar to that of early endosperm. These findings suggest that maize androgenesis may possibly involve both embryogenesis and the establishment of endosperm-like components.

ZMOCL1, AN HDGL2 FAMILY HOMEOBOX GENE, IS EXPRESSED IN THE OUTER CELL LAYER THROUGHOUT MAIZE DEVELOPMENT

The formation of a morphologically distinct outer cell layer or protoderm is one of the first and probably one of the most important steps in patterning of the plant embryo. Here we report the isolation of ZmOCL1 (OCL for outer cell layer), a member of the HDGL2 (also known as HD-ZIP IV) subclass of plant-specific HD-ZIP homeodomain proteins from maize. ZmOCL1 transcripts are detected very early in embryo development, before a morphologically distinct protoderm is visible, and expression then becomes localised to the protoderm of the embryo as it develops. Subsequently, expression is observed in the L1 cell layer of both the developing primary root and shoot meristems, and is maintained in developing leaves and floral organs. We propose that ZMOCL1 may play a role in the specification of protoderm identity within the embryo, the organisation of the primary root primordium or in the maintenance of the L1 cell layer in the shoot apical meristem. We also show that the expression of ZmOCL1 is different from that of another epidermal marker gene, LTP2 (lipid transfer protein) and, in meristems, is complementary to that of Kn1 (Knotted) which is transcribed only in underlying cell layers.

Role of Petal-Specific Orcinol O-Methyltransferases in the Evolution of Rose Scent

Orcinol O-methyltransferase (OOMT) 1 and 2 catalyze the last two steps of the biosynthetic pathway leading to the phenolic methyl ether 3,5-dimethoxytoluene (DMT), the major scent compound of many rose (Rosa x hybrida) varieties. Modern roses are descended from both European and Chinese species, the latter being producers of phenolic methyl ethers but not the former. Here we investigated why phenolic methyl ether production occurs in some but not all rose varieties. In DMT-producing varieties, OOMTs were shown to be localized specifically in the petal, predominanty in the adaxial epidermal cells. In these cells, OOMTs become increasingly associated with membranes during petal development, suggesting that the scent biosynthesis pathway catalyzed by these enzymes may be directly linked to the cells secretory machinery. OOMT gene sequences were detected in two non-DMT-producing rose species of European origin, but no mRNA transcripts were detected, and these varieties lacked both OOMT protein and enzyme activity. These data indicate that up-regulation of OOMT gene expression may have been a critical step in the evolution of scent production in roses.

Effects of Grease Composition and Structure on Film Thickness in Rolling Contact

The aim of this work is to show the correlation between the tribological behavior of grease and its composition and structure. A tribological investigation was conducted on various lubricants. The following parameters were varied: base oil, soap and presence of additives. To ensure efficient control of grease composition, greases containing the same type of soap were manufactured from the same concentrated soap sample. Film thickness measurements showed that the thickener microstructure (revealed by TEM observations) is not the determining factor for the formation of a thick lubricant film, i.e. a film following EHL equations. Nevertheless, the soap - base oil interaction is an essential parameter. The composition of a grease influences oil bleeding, mechanical stability, and rheological behavior. The elastic modulus G seems to be the only parameter directly linked to tribological behavior. Greases with low G have a greater capacity to form a thick EHD film compared to greases with large G.

Complexity and Genetic Variability of Heat-Shock Protein Expression in Isolated Maize Microspores

The expression of heat-shock proteins (HSPs) in isolated maize (Zea mays L.) microspores has been investigated using high-resolution two-dimensional electrophoresis coupled to immunodetection and fluorography of in vivo synthesized proteins. To this end, homogeneous and viable populations of microspores have been purified in sufficient amounts for molecular analysis from plants grown in controlled conditions. Appropriate conditions for thermal stress application have been defined. The analysis revealed that isolated microspores from maize display a classical heat-shock response characterized by the repression of the normal protein synthesis and the expression of a set of HSPs. A high complexity of the response was demonstrated, with numerous different HSPs being resolved in each known major HSP molecular weight class. However, the extent of this heat-shock response is limited in that some of these HSPs do not accumulate at high levels following temperature elevation. Comparative analysis of the heat-shock responses of microspores isolated from five genotypes demonstrated high levels of genetic variability. Furthermore, many HSPs were detected in microspores at control temperature, indicating a possible involvement of these proteins in pollen development at stages close to first pollen mitosis.

Genomic approach to study floral development genes in Rosa sp.

Cultivated for centuries, the varieties of rose have been selected based on a number of flower traits. Understanding the genetic and molecular basis that contributes to these traits will impact on future improvements for this economically important ornamental plant. In this study, we used scanning electron microscopy and sections of meristems and flowers to establish a precise morphological calendar from early rose flower development stages to senescing flowers. Global gene expression was investigated from floral meristem initiation up to flower senescence in three rose genotypes exhibiting contrasted floral traits including continuous versus once flowering and simple versus double flower architecture, using a newly developed Affymetrix microarray (Rosa1_Affyarray) tool containing sequences representing 4765 unigenes expressed during flower development. Data analyses permitted the identification of genes associated with floral transition, floral organs initiation up to flower senescence. Quantitative real time PCR analyses validated the mRNA accumulation changes observed in microarray hybridizations for a selection of 24 genes expressed at either high or low levels. Our data describe the early flower development stages in Rosa sp, the production of a rose microarray and demonstrate its usefulness and reliability to study gene expression during extensive development phases, from the vegetative meristem to the senescent flower.