Ecological constraints increase the climatic debt in forests.

Biodiversity changes are lagging behind current climate warming. The underlying determinants of this climatic debt are unknown and yet critical to understand the impacts of climate change on the present biota and improve forecasts of biodiversity changes. Here we assess determinants of climatic debt accumulated in French forest herbaceous plant communities between 1987 and 2008 (that is, a 1.05 °C mean difference between the observed and bioindicated temperatures). We show that warmer baseline conditions predispose plant communities to larger climatic debts, and that climate warming exacerbates this response. Forest plant communities, however, are absorbing part of the temperature increase mainly through the species' ability to tolerate changing climate. As climate warming is expected to accelerate during the twenty-first century, plant migration and tolerance to climatic stresses probably will be insufficient to absorb this impact posing threats to the sustainability of forest plant communities.

NtRING1, putative RING-finger E3 ligase protein, is a positive regulator of the early stages of elicitin-induced HR in tobacco.

KEY MESSAGE: NtRING1 is a RING-finger protein with a putative E3 ligase activity. NtRING1 regulates HR establishment against different pathogens. Loss-/gain-of-function of NtRING1 altered early stages of HR phenotype establishment. Plant defence responses against pathogens often involve the restriction of pathogens by inducing a hypersensitive response (HR). cDNA clones DD11-39, DD38-11 and DD34-26 were previously obtained from a differential screen aimed at characterising tobacco genes with an elicitin-induced HR-specific pattern of expression. Our precedent observations suggested that DD11-39, DD38-11 and DD34-26 might play roles in the HR establishment. Only for DD11-39 a full-length cDNA sequence was obtained and the corresponding protein encoded for a type-HC RING-finger/putative E3 ligase protein which we termed NtRING1. The expression of NtRING1 was upregulated upon HR induction by elicitin, Ralstonia solanacearum, or tobacco mosaic virus (TMV) in tobacco. Silencing of NtRING1 remarkably delayed the establishment of elicitin-induced HR in tobacco as well as the expression of different early induction genes in tissues undergoing HR. Accordingly, transient overexpression of NtRING1 accelerated the HR launching upon elicitin treatment. Taking together, our data suggests that NtRING1 plays a functional role in the early establishment of HR.

[Probable flora: an expression mean of ecological gradients in France]. / La flore probable : un mode d'expression des gradients écologiques en France.

The application of the criterion of fidelity of plants to plants over four millions botanical observations in France is considered to characterize the ecology of 215,000 phytosociological surveys. Among those discriminant plants, some are missing of the surveys, but they can have a certain probability of occurrence: these plants are called "probable plants" and they represent the "probable flora" of a territory. The study of their geographical distribution shows ecological gradients of flora across France in a better way than only considering the botanical observations. In fact, this method mitigates the discontinuities of taxa observations whose absence may be due to historical and/or anthropogenic factors.

Changes in plant community composition lag behind climate warming in lowland forests.

Climate change is driving latitudinal and altitudinal shifts in species distribution worldwide, leading to novel species assemblages. Lags between these biotic responses and contemporary climate changes have been reported for plants and animals. Theoretically, the magnitude of these lags should be greatest in lowland areas, where the velocity of climate change is expected to be much greater than that in highland areas. We compared temperature trends to temperatures reconstructed from plant assemblages (observed in 76,634 surveys) over a 44-year period in France (1965-2008). Here we report that forest plant communities had responded to 0.54 °C of the effective increase of 1.07 °C in highland areas (500-2,600 m above sea level), while they had responded to only 0.02 °C of the 1.11 °C warming trend in lowland areas. There was a larger temperature lag (by 3.1 times) between the climate and plant community composition in lowland forests than in highland forests. The explanation of such disparity lies in the following properties of lowland, as compared to highland, forests: the higher proportion of species with greater ability for local persistence as the climate warms, the reduced opportunity for short-distance escapes, and the greater habitat fragmentation. Although mountains are currently considered to be among the ecosystems most threatened by climate change (owing to mountaintop extinction), the current inertia of plant communities in lowland forests should also be noted, as it could lead to lowland biotic attrition.

[The phytoclimates of France: probabilistic classification of 1874 bio-indicators of the climate]. / Les phytoclimats de France : classification probabiliste de 1874 bio-indicateurs du climat.

This article presents a synthesis of the relationships between plants and climates at the scale of France, based on a probabilistic classification of 1874 bio-indicators. This classification defines plants groups that indicate the climate, named phytoclimates, expressing the climatic gradients in France. This classification shows 210 phytoclimatic groups distributed into ten cluster levels. The analysis of the various hierarchical levels shows two main phytoclimates testifying the importance of the marine masses and the altitude. The analysis of the third hierarchical level underlines particular phytoclimates which would not be easily recognizable by only analysing the overlapping of floristic and climatic territories. This classification allows one to select taxa that are indicators of the climate. The distribution monitoring or modeling of these taxa should show the effects of the global change on the ecosystems.

[Relationships between plants and climate in France: calibration of 1874 bio-indicators]. / Relations entre plantes et climats en France: étalonnage de 1874 bio-indicateurs.

The influence of climate on plants geography is studied through a probabilistic calibration between a botanical database, containing 12 000 plots, and a meteorological database composed of 574 climatic stations. The calibration measures the climatical optimum (position) and the indicator power (concentration) of 1874 plants for six climatic variables. The validation of these relations is based upon the comparison of the estimation of climate by plants and the values measured by climatic stations near the plots. This validation underlines that plants are accurate (accuracy=88.5%) and stable (stability=96.5%) bio-indicators of climate variables.

NtLRP1, a tobacco leucine-rich repeat gene with a possible role as a modulator of the hypersensitive response.

Plant defense responses against pathogens often involve the restriction of the pathogen to its site of penetration achieved through the combined effects of the hypersensitive response (HR) and its tightly connected localized acquired resistance (LAR). The tobacco DD9-3 expressed sequence tag was previously isolated from a screen designed to isolate genes induced early during the HR, thus potentially involved in the induction/regulation of the HR or LAR. Translation of the open reading frame of DD9-3 revealed a leucine-rich repeat (LRR) domain highly homologous with the receptor domain of a receptor kinase, suggesting a potential function in signaling pathways. The full-length cDNA was cloned. It encodes a small (232 amino acids) LRR protein, designated Nicotiana tabacum leucine-rich protein 1 (NtLRP1), containing a signal peptide, four leucine zipper repeats, five LRR repeats, and a C-terminal domain rich in proline. NtLRP1 expression is induced early during the HR initiated by elicitins, Ralstonia solanacearum, or Tobacco mosaic virus. NtLRP1 coupled with the green fluorescent protein localizes to the endoplasmic reticulum (ER). Loss-of-function through virus-induced gene silencing or through RNA interference did not modify the elicitin-induced HR or LAR. Gain-of-function experiments through transient Agrobacterium tumefaciens-mediated NtLRP1 expression in tobacco leaves caused the suppression of the HR induced by 2 nM elicitin and delayed the HR when the elicitin was applied at higher concentrations. The results suggest that NtLRP1 acts as a modulator of the HR and that retention in the ER is essential for its function.

Defense and resistance-inducing activities in tobacco of the sulfated beta-1,3 glucan PS3 and its synergistic activities with the unsulfated molecule.

Laminarin, a beta-1,3 glucan with single beta-glucose branches at position 6, was chemically sulfated to produce PS3 with a degree of sulfation of 2.4. PS3 has previously been shown to activate the salicylic acid (SA) signaling pathway in infiltrated tobacco and Arabidopsis thaliana leaf tissues. Here, we investigated whether PS3 induces systemic defense and resistance responses in tobacco. Using a radiolabeled compound, it was first demonstrated that PS3 remains strictly localized to the infiltrated tissues. PS3 is also resistant to beta-glucanase degradation. In transgenic PR1-beta-glucuronidase (GUS) tobacco plants, PS3 causes a strong increase in GUS activity in treated tissues but none in untreated leaves. PS3-infiltrated tissues challenged with tobacco mosaic virus (TMV) 8 d after elicitor application show a decrease in both the lesion number and the lesion size, whereas treatment with laminarin, the unsulfated native glucan, affected only the lesion number. PS3 does not induce systemic acquired resistance to TMV. PS3 and laminarin show synergistic effects in promoting the oxidative burst in tobacco cell suspensions and in increasing the expression of genes encoding O-methyltransferases of the phenylpropanoid pathway in tobacco plants. No synergistic effect was observed on the expression of either the SA-dependent acidic PR1 gene or the ethylene-dependent basic PR5 gene in tobacco plants.

Identification of tobacco ESTs with a hypersensitive response (HR)-specific pattern of expression and likely involved in the induction of the HR and/or localized acquired resistance (LAR).

Plant defense responses against pathogens often involve the restriction of the pathogen to its site of penetration. Restriction is achieved through the combined effects of the hypersensitive response (HR) and its tightly connected localized acquired resistance (LAR). As LAR is induced by unknown signals released by the cells undergoing the HR, LAR inducing/regulating genes must show a HR-specific pattern of expression. Here, we describe a differential display reverse-transcript polymerase chain reaction (DDRT-PCR) strategy to isolate tobacco expressed sequence tags (ESTs) characterized by such an expression profile, which also characterizes genes involved in the induction/execution of the HR. We compared the DDRT-PCR profile of tobacco cell suspensions treated with beta-megaspermin inducing the HR with that of untreated cells and cells treated with alpha-megaspermin inducing a Defense No Death (DND) phenotype. The expression profile of the selected ESTs was analyzed in tobacco plants expressing a beta-megaspermin-induced HR or a DND phenotype, including LAR, induced by three different elicitors. This comprehensive analysis allowed to identify 24 HR-specific ESTs, half of them shows no or non-significant homology with ESTs and genes in the databases. The other half exhibits homology with genes encoding a receptor-like kinase protein, proteins involved in the regulation of plasma membrane structure, proteins of the ubiquitin/26S proteasome proteolytic system, RNA binding proteins, and a protein hypothesized to be a true regulator of the HR.

Beta-1,3 glucan sulfate, but not beta-1,3 glucan, induces the salicylic acid signaling pathway in tobacco and Arabidopsis.

Sulfate substituents naturally occurring in biomolecules, such as oligosaccharides and polysaccharides, can play a critical role in major physiological functions in plants and animals. We show that laminarin, a beta-1,3 glucan with elicitor activity in tobacco (Nicotiana tabacum), becomes, after chemical sulfation, an inducer of the salicylic acid (SA) signaling pathway in tobacco and Arabidopsis thaliana. In tobacco cell suspensions, the oxidative burst induced by the laminarin sulfate PS3 was Ca2+ dependent but partially kinase independent, whereas laminarin triggered a strickly kinase-dependent oxidative burst. Cells treated with PS3 or laminarin remained fully responsive to a second application of laminarin or PS3, respectively, suggesting two distinct perception systems. In tobacco leaves, PS3, but not laminarin, caused electrolyte leakage and triggered scopoletin and SA accumulation. Expression of different families of Pathogenesis-Related (PR) proteins was analyzed in wild-type and mutant tobacco as well as in Arabidopsis. Laminarin induced expression of ethylene-dependent PR proteins, whereas PS3 triggered expression of ethylene- and SA-dependent PR proteins. In Arabidopsis, PS3-induced PR1 expression was also NPR1 (for nonexpressor of PR genes1) dependent. Structure-activity analysis revealed that (1) a minimum chain length is essential for biological activity of unsulfated as well as sulfated laminarin, (2) the sulfate residues are essential and cannot be replaced by other anionic groups, and (3) moderately sulfated beta-1,3 glucans are active. In tobacco, PS3 and curdlan sulfate induced immunity against Tobacco mosaic virus infection, whereas laminarin induced only a weak resistance. The results open new routes to work out new molecules suitable for crop protection.

Biological and molecular comparison between localized and systemic acquired resistance induced in tobacco by a Phytophthora megasperma glycoprotein elicitin.

We have compared localized (LAR) and systemic (SAR) acquired resistance induced in tobacco by a hypersensitive response (HR) inducing Phytophthora megasperma glycoprotein elicitin. Three different zones were taken into account: LAR, SAR(T) and SAR(S). The LAR zone was 5-10 mm wide and surrounded the HR lesion. SAR(T) was the tissue of the elicitor-treated leaf immediately beyond the LAR zone. The systemic leaf was called SAR(S). Glycoprotein-treated plants showed enhanced resistance to challenge infection by tobacco mosaic virus (TMV). Disease resistance was similar in SAR(T) and SAR(S), and higher in LAR. The expression pattern, in glycoprotein-treated plants, of acidic and basic PR1, PR2, PR3 and PR5 proteins and of O-methyltransferases (OMT), enzymes of the phenylpropanoid pathway, was similar to that in TMV-infected plants. OMT was stimulated in LAR but not in SAR(T) and SAR(S). The four classes of acidic and basic PR proteins accumulated strongly in LAR. Reduced amounts of acidic PR1, PR2, PR3 and only minute amounts of basic PR2 and PR3 accumulated in SAR(T) and SAR(S). In glycoprotein-treated plants, expression of the acidic and basic PR proteins in LAR and SAR of transgenic NahG and ETR tobacco plants and in LAR of plants treated with inhibitors of salicylic acid accumulation and of ethylene biosynthesis indicated a salicylic acid-dependent signalling pathway for acidic isoform activation and an ethylene-dependent signalling pathway for basic isoform activation.

Molecular cloning and biological activity of alpha-, beta-, and gamma-megaspermin, three elicitins secreted by Phytophthora megasperma H20.

We report on the molecular cloning of the Phytophthora megasperma H20 (PmH20) glycoprotein shown previously as an inducer of the hypersensitive response, of localized acquired resistance and of systemic acquired resistance in tobacco (Nicotiana tabacum), and of the PmH20 alpha- and beta-megaspermin, two elicitins of class I-A and I-B, respectively. The structure of the glycoprotein shows a signal peptide of 20 amino acids followed by the typical elicitin 98-amino acid-long domain and a 77-amino acid-long C-terminal domain carrying an O-glycosylated moiety. The molecular mass deduced from the translated cDNA sequence is 14,920 and 18,676 D as determined by mass spectrometry. This structure together with multiple sequence alignments and phylogenetic analyses indicate that the glycoprotein belongs to class III elicitins. It is the first class III elicitin protein characterized, which we named gamma-megaspermin. We compared the biological activity of the three PmH20 elicitins when applied to tobacco cv Samsun NN plants. Although alpha- and gamma-megaspermin were similarly active, beta-megaspermin was the most active in inducing the hypersensitive response and localized acquired resistance, which was assessed by measuring the levels of acidic and basic pathogenesis-related proteins and of the antioxidant phytoalexin scopoletin. The three elicitins induced similar levels of systemic acquired resistance measured as the expression of acidic PR proteins and is increased resistance to challenge tobacco mosaic virus infection.