Stress Abiotiques et Différenciation des Végétaux Cultivés

A method for the simultaneous determination of free chlorogenic acids (CGA) and sesquiterpene lactones (STL) in chicory root and its dried (flour) and roasted (grain) forms is described. The method uses one extraction and one analysis for all chicory root products. Various solvents with low to high polarity, such as methanol, chloroform, or n-hexane, were tested alone, in combination in different proportions or with acidified or neutral aqueous solvent. The water/chloroform/methanol (30/30/40, v/v/v) mixture generated the best extraction yield, 21% higher than alcohol mixtures. The profiling of CGA and STL content was performed through a conventional HPLC-DAD method using a PFP core shell column in a fast single run. Good retention time and area repeatability (RDD mean % 0.46 and 5.6, resp.) and linearity (R 2 ≥ 0.96) were obtained. The STL and chlorogenic acids levels determined were 254.7 and 100.2 μg/g of dry matter in the root, 792.5 and 1,547 μg/g in flour, and 160.4 and 822.5 μg/g in the roasted grains, respectively. With an average recovery of 106% and precision of 90%, this method is rapid, reproducible, and straightforward way to quantify the chlorogenic acids and STL in chicory raw material and end products.

The present paper reports the various obstacles cleared during the past decade with the view of generating red blood cells in vitro from various sources of stem cells, for transfusion purposes. We also consider the next developments to be performed for achieving this goal. Starting from the natural source, the haematopoietic stem cells, the major advance resides in the establishment of the proof of principle for transfusion in human, by showing a normal life span of cultured Red blood cells compared to their native counterpart. The best available source of highly proliferative adult stem cells is cord blood, with the capacity of generating the equivalent of 100 units of packed RBC from one average unit. It is, however, a limited source in terms of haematopoietic stem cells and remains dependent on donations as observed from conventional blood supply. Critical advances have allowed the in vitro production of functional RBC from pluripotent human stem cells, embryonic and induced pluripotent stem cells, in the past 5 years. Because induced pluripotent stem cells (iPS) can proliferate indefinitely and be selected for a phenotype of interest, they appear the most favourable source of stem cells. Proof of concept of the generation of RBC from iPS has been made, but still needs to be optimized. We also discuss the key points that remain to be resolved to achieve an application for clinical transfusion. Several crucial points remain to be resolved notably to ensure the safety of iPS of clinical grade, the optimization of the erythrocyte differentiation and cellular amplification, and finally, the definition of GMP conditions for industrial production.

A simple, efficient protocol for direct in vitro shoot organogenesis and regeneration was established for three species of Miscanthus including two clones of Miscanthus x giganteus, one clone of M. sinensis and one clone of M. sacchariflorus. Shoots were induced from the axillary nodes of both M. x giganteus and M. sacchariflorus and from apical meristems of both M. sinensis and M. sacchariflorus. A tillering method was used to accelerate shoot proliferation. Shoots were rooted in a wet perlite substrate in pots in the greenhouse. Subsequently, rooted plants were transferred to the field. The genetic uniformity of regenerated plants was evaluated using amplified fragment length polymorphism analysis and compared to that of rhizome-propagated plants. A total of 33,443 fragments were generated, representing 869 markers. There were 21 fragments (0.06 % of the fragments) or 19 markers (2.19 % of the markers) that were polymorphic, and almost all of these were singletons. The three species showed similar polymorphisms. Genetic variability was also found in the rhizome-propagated plants, sometimes at a higher rate than in the in vitro culture, indicating that the genetic uniformity was not altered by the protocol. This protocol may help breeders produce new clones of Miscanthus in the future.

To increase yield in pea (Pisum sativum L.), autumn sowing would be preferable. Hence, frost tolerance of pea became a major trait of interest for breeders. In order to better understand the cold acclimation in pea, Champagne a frost tolerant line and Terese, a frost sensitive line, and their recombinant inbred lines (RIL) were studied. RIL frost tolerance was evaluated by a frost damage scale under field as well as controlled conditions. A quantitative trait loci (QTL) approach was used to identify chromosomal regions linked to frost tolerance. The detected QTL explained from 6.5 to 46.5% of the phenotypic variance. Amongst them, those located on linkage groups 5 and 6 were consistent with over all experiments, in field as well as in controlled environments. In order to improve the understanding of the frost tolerance mechanisms, several cold acclimation key characters such as concentration of sugars, electrolyte leakage, osmotic pressure, and activity of RuBisCO were assessed. Some of these physiological QTL colocalised with QTL for frost damage, in particular two raffinose QTL on LG5 and LG6 and one RuBisCO activity QTL on LG6, explaining 8.8 to 27.0% of the phenotypic variance. In addition, protein quantitative loci were mapped; some of them colocalised with frost damage and physiological QTL on LG5 and LG6, explaining 16.0-43.6% of the phenotypic variance. Raffinose metabolism and RuBisCO activity and its effect on photosynthesis might play a major role in cold acclimation of pea.

ABSTRACTEstimating the proportion of N remobilization and postsilking N uptake allocated to kernels may help to improve N‐use efficiency in maize (Zea mays L.). In this study, we show theoretically and experimentally, that 15N labeling at the beginning of stem elongation can be used in the field to estimate the proportion of N remobilized from the vegetative parts to the kernels of maize by measuring 15N distribution only at maturity. In the same way, 15N labeling at silking allows a determination of the proportion of postsilking N uptake allocated to the kernels by measuring 15N distribution only at maturity. Two 1‐yr experiments with three and four genotypes and one 2‐yr experiment with testcross progenies from 66 recombinant inbred lines were developed. Nitrogen‐15 labeling during vegetative growth provided an estimate of the proportion of N remobilized with a greater accuracy compared with the “balance” method, which computes the difference between the amount of N present in the stover at silking and the amount of N present in the stover at maturity. The validity of our 15N method is mainly based on the assumption that less than 15 to 20% of 15N is taken up after silking. The determination of the proportion of postsilking N uptake allocated to kernels requires assumptions that are more difficult to fulfill. Nitrogen‐15 labeling in the field at the beginning of rapid stem growth appears to be a useful tool for studying the genetic variability of N remobilization using a large number of genotypes.

In sugarbeet (Beta vulgaris subsp. vulgaris), many linkage maps have been constructed, but the availability of markers continues to limit utility of genetic maps in public domain programs. Here a framework genetic map is presented that is expandable and transferable to research programs interested in locating their markers on a consensus map. In its current framework, the primary markers used were amplified fragment length polymorphisms (AFLPs) that were anchored to Butterfass chromosome‐nomenclature linkage groups using linkage group specific markers validated in other populations. Thus, a common framework has been established that anchors 331 markers, including 23 newly mapped simple sequence repeat (SSR) markers, having a combined total of 526.3 cM among the nine beet linkage groups. The source of the mapping population was a sugarbeet × table beet population, and this is the first report of a map constructed with a relatively wide cross in B. vulgaris Segregation distortion was common (22% of loci), particularly extreme for Butterfass Chromosome 5, and predominantly favored the sugarbeet (seed parent) allele. Physical segments of the beet genome that carry mapped markers have been identified, demonstrating that physical and genetic mapping are facile and complementary applications for beet improvement.

The improvement of energy efficiency in the greenhouse production of cut rose and pot rose can be achieved through the use of rose cultivars having vigorous growth. A better understanding of the inheritance of vigour and its related traits will assist the breeding activities. Quantitative trait locus (QTL) analyses were performed with the help of an integrated linkage map of a diploid rose population originating from a cross between Rosa multiflora-derived genotypes. The underlying datasets for ten vigour-related traits were collected in an evaluation study of this population in two greenhouse experiments with suboptimal temperatures for growth. We identified ten chromosomal regions, scattered over the seven linkage groups, containing QTLs for these traits. Considering each trait separately, we detected a total of 42 QTLs. Among these QTLs, 24 were found in both of the experiments, eight and ten were specific to either of the two experiments. The number of QTLs for individual traits varied from three to five with a respective contribution to the phenotypic variation from 12 to 35%. QTLs for highly correlated traits frequently co-localized, indicating a common genetic basis. Clustering of QTLs for different traits was noted in some chromosome regions, for instance, one on chromosome 2 included major QTLs for eight of ten traits under study, suggesting co-localization of several separate genes or/and the occurrence of various genes having pleiotropic effects. The discovery of markers associated to QTL regions is in roses the first step towards marker-assisted selection for vigour improvement enabling the transfer of useful QTL-alleles of R. multiflora to pot and cut roses.

All plants contain an unusual class of hemoglobins that display bis-histidyl coordination yet are able to bind exogenous ligands such as oxygen. Structurally homologous hexacoordinate hemoglobins (hxHbs) are also found in animals (neuroglobin and cytoglobin) and some cyanobacteria, where they are thought to play a role in free radical scavenging or ligand sensing. The plant hxHbs can be distinguished from the others because they are only weakly hexcacoordinate in the ferrous state, yet no structural mechanism for regulating hexacoordination has been articulated to account for this behavior. Plant hxHbs contain a conserved Phe at position B10 (Phe(B10)), which is near the reversibly coordinated distal His(E7). We have investigated the effects of Phe(B10) mutation on kinetic and equilibrium constants for hexacoordination and exogenous ligand binding in the ferrous and ferric oxidation states. Kinetic and equilibrium constants for hexacoordination and ligand binding along with CO-FTIR spectroscopy, midpoint reduction potentials, and the crystal structures of two key mutant proteins (F40W and F40L) reveal that Phe(B10) is an important regulatory element in hexacoordination. We show that Phe at this position is the only amino acid that facilitates stable oxygen binding to the ferrous Hb and the only one that promotes ligand binding in the ferric oxidation states. This work presents a structural mechanism for regulating reversible intramolecular coordination in plant hxHbs.

Asparagus officinalis L. is a perennial monocotyledon, a member of the Liliaceae family. It grows in temperate climates and under subtropical conditions. Growth of spears normally takes place in sandy soils. There are two culture types: the white asparagus, in which the spears are harvested from earthed up plants, where spears are cut at the crown level just as they emerge through the soil, and the green asparagus, in which the spears are harvested at or about ground level when they reach a fixed height above the soil. The main producers are the USA, Spain, France, and Taiwan. White asparagus production predominates in Europe (France and Spain) and Taiwan, while green asparagus is essential in North America and Australasia. In European countries, white asparagus is produced mainly for the fresh market, whereas canned white asparagus is produced in Taiwan.

Leaves from 2-month-old in vitro grown plantlets of a clone ofCichorium placed in agitated liquid induction medium at 35°C in the dark produce embryoids after 5 days of culture, without synchronization. Vascular sheath parenchyma cells react first, but every mesophyll cell is potentially embryogenic. Single cells show an early patchy callosic wall and undergo dedifferentiation. With SEM the cells of those proembryoids just emerging through the epidermis are seen to be linked by a fibrillar network, the nature of which is discussed. Four FITC-labelled lectins were tested; only DBA shows embryogenic specificity.