A plant parasite uses light cues to detect differences in host-plant proximity and architecture.

Sunlight filtered by green plant tissue becomes diminished in its ratio of red to far-red wavelengths (R:FR). Some parasitic plants exploit this change by growing towards regions of low R:FR to locate host plants. In principle, variation in R:FR can also convey ecologically relevant information about host proximity or architecture. Here, we demonstrate that the parasitic vine Cuscuta epilinum Weihe (Convolvulaceae) can distinguish fine-scale differences in R:FR associated with differences in the proximity and shape of potential host plants. We conducted dual-choice experiments by placing parasite seedlings between targets, including low R:FR fields manipulated via LED lighting and pairs of model plants exhibiting realistic R and FR reflectance but differing in proximity or shape. Seedlings consistently distinguished between low-R:FR fields of differing intensity. Furthermore, they exhibited preferences for nearer plant models versus identical models placed 4 cm further away and between same-sized models exhibiting shape differences. Our results indicate that parasites can discriminate minute differences in R:FR signatures corresponding to host factors (proximity and shape) that impact seedling survival. This keen sensory ability underpins the parasite's sophisticated foraging behaviour and highlights the broader importance of light cues in plant ecology.

Genome-wide identification and expression analysis of R2R3, 3R- and 4R-MYB transcription factors during lignin biosynthesis in flax (Linum usitatissimum).

MYB transcription factors (TFs) have vital roles in regulating lignin or fiber development. Flax (Linum usitatissimum) is known as one of the plants with high fiber production capacity. However, no studies have been conducted to identify and characterize MYB TFs in the flax genome. Results showed that flax genome harbours 167 R2R3, seven 3R, and one 4R-type MYB TFs. 22 MYB genes (%13) were estimated to be tandem duplicated dated around 13.3-86.98 Mya. 130 flax MYB members have apparent orthologous with Arabidopsis, in which 17 R2R3 MYBs are associated with lignin biosynthesis. MYB062, MYB072, MYB096, MYB141, and MYB146 genes were up-regulated in tissues having higher lignin production capacity. In opposite, MYB012 and MYB113 genes were down-regulated which points out the involvement of those genes in the lignin biosynthesis mechanism. This comprehensive study can provide a basis for understanding the role of MYBs in fiber or lignin production in flax.

Consensus genetic linkage map construction and QTL mapping for plant height-related traits in linseed flax (Linum usitatissimum L.).

BACKGROUND: Flax is an important field crop that can be used for either oilseed or fiber production. Plant height and technical length are important characters for flax. For linseed flax, plants usually have a short technical length and plant height than those for fiber flax. As an important agronomical character for fiber and linseed flax, plant height is usually a selection target for breeding. However, because of limited technologies and methods available, there has been little research focused on discovering the molecular mechanism controlling plant height. RESULTS: In this study, two related recombinant inbred line (RIL) populations developed from crosses of linseed and fiber parents were developed and phenotyped for plant height and technical length in four environments. A consensus linkage map based on two RIL populations was constructed using SNP markers generated by genotyping by sequencing (GBS) technology. A total of 4497 single nucleotide polymorphism (SNP) markers were included on 15 linkage groups with an average marker density of one marker every 2.71 cM. Quantitative trait locus (QTL) mapping analysis was performed for plant height and technical length using the two populations. A total of 19 QTLs were identified for plant height and technical length. For the MH population, eight plant height QTLs and seven technical length QTLs were identified, five of which were common QTLs for both traits. For the PH population, six plant height and three technical length QTLs were identified. By comparing the QTLs and candidate gene information in the two population, two common QTLs and three candidate genes were discovered. CONCLUSIONS: This study provides a foundation for map-based cloning of QTLs and marker-assisted selection for plant height-related traits in linseed and fiber flax.

Flax stems: from a specific architecture to an instructive model for bioinspired composite structures.

The present paper proposes to carefully study and describe the reinforcement mechanisms within a flax stem, which is an exceptional natural model of composite structure. Thanks to accurate microscopic investigations, with both optical and SEM method, we finely depicted the flax stem architecture, which can be view as a composite structure with an outer protection, a unidirectional ply on the periphery and a porous core; each component has a specific function, such as mechanical reinforcement for the unidirectional ply and the porous core. The significant mechanical role of fibres was underlined, as well as their local organisation in cohesive bundles, obtained because of an intrusive growth and evidenced in this work through nanomechanical AFM measurement and 3D reconstruction. Following a biomimetic approach, these data provide a source of inspiration for the composite materials of tomorrow.

Style polymorphism in Linum (Linaceae): a case of Mediterranean parallel evolution?

Heterostyly is a sex polymorphism that has challenged evolutionary biologists ever since Darwin. One of the lineages where heterostyly and related stylar conditions appear more frequently is Linum (Linaceae). This group is particularly suitable for testing competing hypotheses about ancestral and transitional stages on the evolutionary building up of heterostyly. We generated a phylogeny of Linum based on extensive sampling and plastid and nuclear DNA sequences, and used it to trace the evolution of character states of style polymorphism. We also revised available data on pollination, breeding systems, and polyploidy to analyse their associations. Our results supported former phylogenetic hypotheses: the paraphyly of Linum and the non-monophyly of current taxonomic sections. Heterostyly was common in the genus, but appeared concentrated in the Mediterranean Basin and the South African Cape. Ancestral character state reconstruction failed to determine a unique state as the most probable condition for style polymorphism in the genus. In contrast, approach herkogamy was resolved as ancestral state in some clades, giving support to recent hypotheses. Some traits putatively related to heterostyly, such as life history and polyploidy, did show marginal or non-significant phylogenetic correlation, respectively. Although pollinator data are limited, we suggest that beeflies are associated with specific cases of heterostyly. The consistent association between style polymorphism and heteromorphic incompatibility points to ecological factors as drivers of the multiple evolution of style polymorphism in Linum. Albeit based on limited evidence, we hypothesised that specialised pollinators and lack of mating opportunities drive evolution of style polymorphism and loss of the polymorphism, respectively.

Microtomography imaging of an isolated plant fiber: a digital holographic approach.

This paper describes a method for optical projection tomography for the 3D in situ characterization of micrometric plant fibers. The proposed approach is based on digital holographic microscopy, the holographic capability being convenient to compensate for the runout of the fiber during rotations. The setup requires a telecentric alignment to prevent from the changes in the optical magnification, and calibration results show the very good experimental adjustment. Amplitude images are obtained from the set of recorded and digitally processed holograms. Refocusing of blurred images and correction of both runout and jitter are carried out to get appropriate amplitude images. The 3D data related to the plant fiber are computed from the set of images using a dedicated numerical processing. Experimental results exhibit the internal and external shapes of the plant fiber. These experimental results constitute the first attempt to obtain 3D data of flax fiber, about 12 µm×17 µm in apparent diameter, with a full-field optical tomography approach using light in the visible range.

Preparation, processing and properties of lignosulfonate-flax composite boards.

Hemp, hay, straw for animal litters, raffia and sisal stems, abaca and jute bleached pulp fibres, miscanthus stems and flax fibres were mixed to lignosulfonate at 70% filler concentration and compressed in the form of 5 cm-thick boards. Flax was found to give the best mechanical properties measured in bending mode and used for all tests. Several methods able to improve adhesion between matrix and flax fibres were studied. A treatment of flax fibres with NaOH-water was found to decrease the mechanical properties of composites. Ethanol or dichloromethane solvents that are known to dewax flax fibre surfaces improve the mechanical properties of final board. The addition of pectin to the lignosulfonate matrix was found to improve the mechanical properties in the same order of magnitude as with the ethanol treatment. Both methods improve the flexural strength by 60% while keeping the elastic modulus constant. Mechanical improvement shows that these two methods are increasing the lignosulfonate/flax fibre interfacial adhesion. The best compositions have mechanical properties above the normalized minimum required for wood-based boards.

Contrasting seasonal overlaps between primary and secondary growth are linked to wood anatomy in Mediterranean sub-shrubs.

Whole-plant approaches allow quantification of the temporal overlap between primary and secondary growth. If the amount of time available to grow is short, there may be a high temporal overlap between shoot growth and wood formation. We hypothesise that such overlap depends on the duration of the growing season and relates to wood anatomy. We evaluated wood anatomy, shoot longitudinal and radial growth rates, fine root production and the concentrations of non-structural carbohydrates (NSC) in the wood of six sub-shrub species growing in sites with contrasting climatic conditions (Lepidium subulatum, Linum suffruticosum, Salvia lavandulifolia, Satureja montana, Ononis fruticosa, Echinospartum horridum). Sub-shrub species living in sites with a short growing season displayed a high overlap between aboveground primary and secondary growth and formed wide vessels, whereas species from the warmest and driest sites presented the reverse characteristics. The highest overlap was linked to a rapid shoot extension and thickening through the enhanced hydraulic conductivity provided by wide vessels. The reductions in NSC concentrations when growth peaked were low or moderate, indicating that sub-shrubs accumulate NSC in excess, as do trees. The temporal overlap among primary and secondary growth in woody plants may be connected to the duration and rates of shoot and wood growth, which in turn depend on the vessel lumen area.

Effects of cadmium and mycorrhizal fungi on growth, fitness, and cadmium accumulation in flax (Linum usitatissimum; Linaceae).

PREMISE OF THE STUDY: Agricultural soils have become contaminated with a variety of heavy metals, including cadmium. The degree to which soil contaminants affect plants may depend on symbiotic relationships between plant roots and soil microorganisms. We examined (1) whether mycorrhizal fungi counteract the potentially negative effects of cadmium on the growth and fitness of flax (Linum usitatissimum) and (2) whether mycorrhizal fungi affect the accumulation of cadmium within plant parts. METHODS: Two flax cultivars (Linott and Omega) were grown in three soil cadmium environments (0, 5, and 15 ppm). Within each cadmium environment, plants were grown in either the presence or absence of mycorrhizal fungi. Upon senescence, we measured growth and fitness and quantified the concentration of cadmium within plants. KEY RESULTS: Soil cadmium significantly decreased plant fitness, but did not affect plant growth. Mycorrhizal fungi, which were able to colonize roots of plants growing in all cadmium levels, significantly increased plant growth and fitness. Although mycorrhizal fungi counteracted the negative effects of cadmium on fruit and seed production, they also enhanced the concentration of cadmium within roots, fruits, and seeds. CONCLUSIONS: The degree to which soil cadmium affects plant fitness and the accumulation of cadmium within plants depended on the ability of plants to form symbiotic relationships with mycorrhizal fungi. The use of mycorrhizal fungi in contaminated agricultural soils may offset the negative effects of metals on the quantity of seeds produced, but exacerbate the accumulation of these metals in our food supply.

Isolation of the floral morph-related genes in heterostylous flax (Linum grandiflorum): the genetic polymorphism and the transcriptional and post-transcriptional regulations of the S locus.

Heterostylous species have two types of flowers, thrum and pin morphs, and these are controlled by a single diallelic locus designated the 'S locus'; fertilization between these two types of flowers is successful. The S gene and the molecular mechanism by which it operates remain to be uncovered, although heterostyly has been studied since the time of Darwin. We compared transcripts and proteins of the thrum and pin flowers of heterostylous flax (Linum grandiflorum) to characterize the molecular differences between them and to elucidate the molecular machinery of heterostyly. Twelve floral morph-related genes were eventually isolated by an integrated study of subtraction and 2D-PAGE analyses, and four genes, TSS1, LgAP1, LgMYB21 and LgSKS1, were predicted to be related to heterostyly. TSS1, a thrum style-specific gene, showed some features suitable for the S gene. Although its biological function is unclear, TSS1 was expressed only in the thrum style and is probably linked to the S locus. LgMYB21, another thrum style gene, would be involved in floral morphogenesis. LgMYB21 was highly expressed in the thrum style, which is shorter than the pin style, and its overexpression in Arabidopsis reduced pistil length. Furthermore, a comparison of transcript and protein accumulations showed no differences in the mRNA accumulation of some thrum-specific proteins, including LgSKS1, suggesting that these are regulated by floral morph-specific post-transcriptional regulation. The Linum S locus regulates not only S specificity but also many floral phenotypes. Dynamic regulation of transcripts and proteins would be necessary for the pleiotropic function of the Linum S locus.

[Combining ability for the traits of stem branching and plant height in linseed lines].

Combining ability for the traits of stem branching and plant height has been studied in ten pure lines of flax under complete and incomplete diallel crosses. High heritability of the traits "plant height", "the number of lateral stems" and "the number of lateral shoots" and essential role of genes with dominant effects of interaction in genetic control of the traits of stem branching and plant height have been shown. On the basis of combining ability indexes the ways for usage of certain genotypes and crossing combinations in flax breeding were defined. As a result of individual selection from hybrid combinations some new complexes of habit traits and agriculturally valuable plant characteristics were obtained.

Ontogeny of floral organs in flax (Linum usitatissimum; Linaceae).

PREMISE OF THE STUDY: Flax (Linum usitatissimum) is an important crop worldwide; however, a detailed study on flower development of this species is lacking. Here we describe the pattern of initiation and a program of key developmental events in flax flower ontogeny. This study provides important fundamental information for future research in various aspects of flax biology and biotechnology. METHODS: Floral buds and organs were measured throughout development and examined using scanning electron microscopy. KEY RESULTS: Floral organs were initiated in the following sequence: sepals, stamens and petals, gynoecium, and nectaries. The five sepals originated in a helical pattern, followed evidently by simultaneous initiation of five stamens and five petals, the former opposite of the sepals and the latter alternate to them. The gynoecium, with five carpels, was produced from the remaining, central region of the floral apex. Stamens at early stages were dominated by anther growth but filaments elongated rapidly shortly before anthesis. Early gynoecium development occurred predominantly in the ovary, and ovule initiation began prior to enclosure of carpels. A characteristic feature was the twisted growth of styles, accompanied by the differentiation of papillate stigmas. Petal growth lagged behind that of other floral organs, but petals eventually grew rapidly to enclose the inner whorls after style elongation. Flask-shaped nectaries bearing stomata developed on the external surface of the filament bases. CONCLUSIONS: This is the first detailed study on flax floral organ development and has established a key of 12 developmental stages, which should be useful to flax researchers.

Haustorially expressed secreted proteins from flax rust are highly enriched for avirulence elicitors.

Rust fungi, obligate biotrophs that cause disease and yield losses in crops such as cereals and soybean (Glycine max), obtain nutrients from the host through haustoria, which are specialized structures that develop within host cells. Resistance of flax (Linum usitatissimum) to flax rust (Melampsora lini) involves the induction of a hypersensitive cell death response at haustoria formation sites, governed by gene-for-gene recognition between host resistance and pathogen avirulence genes. We identified genes encoding haustorially expressed secreted proteins (HESPs) by screening a flax rust haustorium-specific cDNA library. Among 429 unigenes, 21 HESPs were identified, one corresponding to the AvrL567 gene. Three other HESPs cosegregated with the independent AvrM, AvrP4, and AvrP123 loci. Expression of these genes in flax induced resistance gene-mediated cell death with the appropriate specificity, confirming their avirulence activity. AvrP4 and AvrP123 are Cys-rich proteins, and AvrP123 contains a Kazal Ser protease inhibitor signature, whereas AvrM contains no Cys residues. AvrP4 and AvrM induce cell death when expressed intracellularly, suggesting their translocation into plant cells during infection. However, secreted AvrM and AvrP4 also induce necrotic responses, with secreted AvrP4 more active than intracellular AvrP4, possibly as a result of enhanced formation of endoplasmic reticulum-dependent disulfide bonds. Addition of an endoplasmic reticulum retention signal inhibited AvrM-induced necrosis, suggesting that both AvrM and AvrP4 can reenter the plant cell after secretion in the absence of the pathogen.

Polyhydroxybutyrate synthesis in transgenic flax.

Flax (Linum usitatissimum L.) is an annual plant species widely cultivated in temperate climates for bast fibres and linseed oil. Apart from traditional textile use, the fibres are fast becoming an integral part of new composite materials utilized in automobile and constructive industry. Especially attractive for environmental safety demands are biodegradable and renewable biocomposities based on polyhydroxybutyrate (PHB) polymer as a matrix and reinforced with the flax fibres. Manufacturing of PHB by bacteria fermentation is however substantially more expansive as compared to technologies producing conventional plastics. We report for the first time generation of transgenic plants which produce both components of flax/PHB composites, i.e. the fibres and the thermoplastic matrix in the same plant organ of a crop. The flax (cv. Nike) plants were transformed using constructs bearing either single cDNA, encoding the beta-ketothiolase enzyme (C plants), or all three of the genes necessary for poly-beta-hydroxybutyrate (PHB) synthesis (M plants). Both constructs contained a plastidial targeting sequence. The amount of PHB produced by the transgenic plants was up to over 70-fold higher than in wild-type plants, when analysed using the gas chromatography/mass spectrometry (GC-MS method). The PHB accumulation in plastids caused change both in their shape and size. The use of a stem-specific promoter for transgene expression protected the transgenic plant from growth retardation and also provided higher PHB synthesis than in the case of constructs governed by the 35S CaMV constitutive promoter. None toxic effects that could lead to stunted growth or the loss of fertility were observed, when 14-3-3 promoter was used as the stem-specific. Significant modifications in stem mechanical properties were accompanied to the PHB accumulation in growing cell of fibres in the transgenic plants. The Young's modulus E, the average measure of stem tissues resistance to tensile loads increased up to twice in M plants as compared to a single gene transformed ones. However, a wide range of E values, from 24.1 to 54.4 MPa, was observed in dependence of tested strain. Potential commercial significance of the genetic manipulation approach enabling synthesis of thermoplastic in crops cultivated for fibres is discussed.