Multiple Technology Approach Based on Stable Isotope Ratio Analysis, Fourier Transform Infrared Spectrometry and Thermogravimetric Analysis to Ensure the Fungal Origin of the Chitosan.

Chitosan is a natural polysaccharide which has been authorized for oenological practices for the treatment of musts and wines. This authorization is limited to chitosan of fungal origin while that of crustacean origin is prohibited. To guarantee its origin, a method based on the measurement of the stable isotope ratios (SIR) of carbon δ13C, nitrogen δ15N, oxygen δ18O and hydrogen δ2H of chitosan has been recently proposed without indicating the threshold authenticity limits of these parameters which, for the first time, were estimated in this paper. In addition, on part of the samples analysed through SIR, Fourier transform infrared spectrometry (FTIR) and thermogravimetric analysis (TGA) were performed as simple and rapid discrimination methods due to limited technological resources. Samples having δ13C values above -14.2‱ and below -125.1‱ can be considered as authentic fungal chitosan without needing to analyse other parameters. If the δ13C value falls between -25.1‱ and -24.9‱, it is necessary to proceed further with the evaluation of the parameter δ15N, which must be above +2.7‱. Samples having δ18O values lower than +25.3‱ can be considered as authentic fungal chitosan. The combination of maximum degradation temperatures (obtained using TGA) and peak areas of Amide I and NH2/Amide II (obtained using FTIR) also allows the discrimination between the two origins of the polysaccharide. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) based on TGA, FTIR and SIR data successfully distributed the tested samples into informative clusters. Therefore, we present the technologies described as part of a robust analytical strategy for the correct identification of chitosan samples from crustaceans or fungi.

Bio-Inspired Rhamnolipids, Cyclic Lipopeptides and a Chito-Oligosaccharide Confer Protection against Wheat Powdery Mildew and Inhibit Conidia Germination.

Plant protection is mainly based on the application of synthetic pesticides to limit yield losses resulting from diseases. However, the use of more eco-friendly strategies for sustainable plant protection has become a necessity that could contribute to controlling pathogens through a direct antimicrobial effect and/or an induction of plant resistance. Three different families of natural or bioinspired compounds originated from bacterial or fungal strains have been evaluated to protect wheat against powdery mildew, caused by the biotrophic Blumeria graminis f.sp. tritici (Bgt). Thus, three bio-inspired mono-rhamnolipids (smRLs), three cyclic lipopeptides (CLPs, mycosubtilin (M), fengycin (F), surfactin (S)) applied individually and in mixtures (M + F and M + F + S), as well as a chitosan oligosaccharide (COS) BioA187 were tested against Bgt, in planta and in vitro. Only the three smRLs (Rh-Eth-C12, Rh-Est-C12 and Rh-Succ-C12), the two CLP mixtures and the BioA187 led to a partial protection of wheat against Bgt. The higher inhibitor effects on the germination of Bgt spores in vitro were observed from smRLs Rh-Eth-C12 and Rh-Succ-C12, mycosubtilin and the two CLP mixtures. Taking together, these results revealed that such molecules could constitute promising tools for a more eco-friendly agriculture.

Increased contribution of wheat nocturnal transpiration to daily water use under drought.

Increasing evidence suggests that in crops, nocturnal water use could represent 30% of daytime water consumption, particularly in semi-arid and arid areas. This raises the questions of whether nocturnal transpiration rates (TRN ) are (1) less influenced by drought than daytime TR (TRD ), (2) increased by higher nocturnal vapor pressure deficit (VPDN ), which prevails in such environments and (3) involved in crop drought tolerance. In this investigation, we addressed those questions by subjecting two wheat genotypes differing in drought tolerance to progressive soil drying under two long-term VPDN regimes imposed under naturally fluctuating conditions. A first goal was to characterize the response curves of whole-plant TRN and TRN /TRD ratios to progressive soil drying. A second goal was to examine the effect of VPDN increase on TRN response to soil drying and on 13 other developmental traits. The study revealed that under drought, TRN was not responsive to progressive soil drying and - intriguingly - that TRN seemingly increased with drought under high VPDN consistently for the drought-sensitive genotype. Because TRD was concomitantly decreasing with progressive drought, this resulted in TRN representing up to 70% of TRD at the end of the drydown. In addition, under drought, VPDN increase was found not to influence traits such as leaf area or stomata density. Overall, those findings indicate that TRN contribution to daily water use under drought might be much higher than previously thought, that it is controlled by specific mechanisms and that decreasing TRN under drought might be a valuable trait for improving drought tolerance.

Nighttime evaporative demand induces plasticity in leaf and root hydraulic traits.

Increasing evidence suggests that nocturnal transpiration rate (TRN ) is a non-negligible contributor to global water cycles. Short-term variation in nocturnal vapor pressure deficit (VPDN ) has been suggested to be a key environmental variable influencing TRN . However, the long-term effects of VPDN on plant growth and development remain unknown, despite recent evidence documenting long-term effects of daytime VPD on plant anatomy, growth and productivity. Here we hypothesized that plant anatomical and functional traits influencing leaf and root hydraulics could be influenced by long-term exposure to VPDN . A total of 23 leaf and root traits were examined on four wheat (Triticum aestivum) genotypes, which were subjected to two long-term (30 day long) growth experiments where daytime VPD and daytime/nighttime temperature regimes were kept identical, with variation only stemming from VPDN , imposed at two levels (0.4 and 1.4 kPa). The VPDN treatment did not influence phenology, leaf areas, dry weights, number of tillers or their dry weights, consistently with a drought and temperature-independent treatment. In contrast, vein densities, adaxial stomata densities, TRN and cuticular TR, were strongly increased following exposure to high VPDN . Simultaneously, whole-root system xylem sap exudation and seminal root endodermis thickness were decreased, hypothetically indicating a change in root hydraulic properties. Overall these results suggest that plants 'sense' and adapt to variations in VPDN conditions over developmental scales by optimizing both leaf and root hydraulics.

High resolution mapping of traits related to whole-plant transpiration under increasing evaporative demand in wheat.

Atmospheric vapor pressure deficit (VPD) is a key component of drought and has a strong influence on yields. Whole-plant transpiration rate (TR) response to increasing VPD has been linked to drought tolerance in wheat, but because of its challenging phenotyping, its genetic basis remains unexplored. Further, the genetic control of other key traits linked to daytime TR such as leaf area, stomata densities and - more recently - nocturnal transpiration remains unknown. Considering the presence of wheat phenology genes that can interfere with drought tolerance, the aim of this investigation was to identify at an enhanced resolution the genetic basis of the above traits while investigating the effects of phenology genes Ppd-D1 and Ppd-B1 Virtually all traits were highly heritable (heritabilities from 0.61 to 0.91) and a total of mostly trait-specific 68 QTL were detected. Six QTL were identified for TR response to VPD, with one QTL (QSLP.ucl-5A) individually explaining 25.4% of the genetic variance. This QTL harbored several genes previously reported to be involved in ABA signaling, interaction with DREB2A and root hydraulics. Surprisingly, nocturnal TR and stomata densities on both leaf sides were characterized by highly specific and robust QTL. In addition, negative correlations were found between TR and leaf area suggesting trade-offs between these traits. Further, Ppd-D1 had strong but opposite effects on these traits, suggesting an involvement in this trade-off. Overall, these findings revealed novel genetic resources while suggesting a more direct role of phenology genes in enhancing wheat drought tolerance.

Genotype-dependent influence of night-time vapour pressure deficit on night-time transpiration and daytime gas exchange in wheat.

In crop plants, accumulating evidence indicates non-marginal night-time transpiration (TRNight) that is responsive to environmental conditions, especially in semiarid areas. However, the agronomical advantages resulting from such phenomenon remain obscure. Recently, drought-tolerance strategies directly stemming from daytime TR (TRDay) responses to daytime vapour pressure deficit VPD (VPDDay) were identified in wheat (Triticum spp.), but the existence of similar strategies resulting from TRNight response to night-time VPD (VPDNight) remains to be investigated, especially that preliminary evidence on this species indicates that TRNight might be responsive to VPDNight. Our study aims at investigating such strategies among a group of diverse lines including drought-tolerant genotypes. The study revealed that: (i) TRNight can be as high as 55% that of the maximal TRDay; (ii) VPDNight is the major driver of TRNight in a genotype-dependent fashion and has an impact on following daytime gas exchange; and (iii) a strong correlation exists between TR sensitivities to VPD under night-time and daytime conditions, revealing that tolerance strategies such as conservative water use do also exist under night-time environments. Overall, this report opens the way to further phenotyping and modelling work aiming at assessing the potential of using TRNight as a trait in breeding new drought-tolerant germplasm.