Impact of Paraburkholderia phytofirmans PsJN on Grapevine Phenolic Metabolism

Phenolic compounds are implied in plant-microorganisms interaction and may be induced in response to plant growth-promoting rhizobacteria (PGPRs). Among PGPR, the beneficial bacterium Paraburkholderia phytofirmans PsJN was previously described to stimulate the growth of plants and to induce a better adaptation to both abiotic and biotic stresses. This study aimed to investigate the impact of PsJN on grapevine secondary metabolism. For this purpose, gene expression (qRT-PCR) and profiling of plant secondary metabolites (UHPLC-UV/DAD-MS QTOF) from both grapevine root and leaves were compared between non-bacterized and PsJN-bacterized grapevine plantlets. Our results showed that PsJN induced locally (roots) and systemically (leaves) an overexpression of PAL and STS and specifically in leaves the overexpression of all the genes implied in phenylpropanoid and flavonoid pathways. Moreover, the metabolomic approach revealed that relative amounts of 32 and 17 compounds in roots and leaves, respectively, were significantly modified by PsJN. Once identified to be accumulated in response to PsJN by the metabolomic approach, antifungal properties of purified molecules were validated in vitro for their antifungal effect on Botrytis cinerea spore germination. Taking together, our findings on the impact of PsJN on phenolic metabolism allowed us to identify a supplementary biocontrol mechanism developed by this PGPR to induce plant resistance against pathogens

Paraburkholderia phytofirmans PsJN-Plants Interaction: From Perception to the Induced Mechanisms

The use of plant-associated bacteria has received many scientific and economic attention as an effective and alternative method to reduce the chemical pesticides use in agriculture. The genus Burkholderia includes at least 90 species including pathogenic strains, plant pathogens, as well as plant beneficial species as those related to Paraburkholderia, which has been reported to be associated with plants and exerts a positive effect on plant growth and fitness. Paraburkholderia phytofirmans PsJN, a beneficial endophyte able to colonize a wide range of plants, is an established model for plant-associated endophytic bacteria. Indeed, in addition to its plant growth promoting ability, it can also induce plant resistance against biotic as well as abiotic stresses. Here, we summarized an inventory of knowledge on PsJN-plant interaction, from the perception to the resistance mechanisms induced in the plant by a way of the atypical colonization mode of this endophyte. We also have carried out an extensive genome analysis to identify all gene clusters which contribute to the adaptive mechanisms under different environments and partly explaining the high ecological competence of P. phytofirmans PsJN

Embryogenèse pollinique chez l orge (Hordeum vulgare L.) : importance du prétraitement

L'embryogenèse pollinique est une technique d'amélioration des espèces qui permet d'obtenir des plantes haploïdes doublées. Les microspores initialement programmées pour former des grains de pollen sont réorientées afin de produire des plantes haploïdes. Cette technique voit son utilisation réduite chez les céréales à cause de la régénération de plantes albinos. Nous avons cherché à optimiser le protocole afin d'améliorer les rendements en plantules chlorophylliennes chez des cultivars récalcitrants et à identifier les mécanismes moléculaires impliqués dans la réorientation de la microspore en nous focalisant sur les défenses. Notre travail a porté sur la culture d'anthères chez l'orge (Hordeum vulgare L.). Afin d'améliorer le protocole, nous avons utilisé le cv. d'hiver Igri régénérant majoritairement des plantules chlorophylliennes et cinq cvs. de printemps produisant exclusivement des plantules albinos. L'apport de sulfate de cuivre dans les milieux de prétraitement et de culture a permis d'augmenter le nombre d'anthères formant des embryons, le nombre de plantules régénérées et le pourcentage de plantules chlorophylliennes régénérées chez l'ensemble des cvs. testés. L'étude de la saison à laquelle les plantes mères sont cultivées a fait apparaitre que les rendements de la culture d'anthères étaient meilleurs pendant les mois les plus longs de l'année. Nous avons également constaté que les résultats obtenus dépendent de la position de l'épi sur le plateau de tallage. Enfin le suivi de la production d'O2- et le suivi d'expression de différents gènes de réponse aux stress nous a permis de mettre en évidence l'activation des défenses dans l'anthère au cours du prétraitementThe production of doubled haploid through pollen embryogenesis represents a modern tool for the improvement of cultivated species enabling plant breeders to produce homozygous lines in a few months. Pollen embryogenesis consists in the regeneration of haploid plantlets from microspores, which are initially destined to develop into pollen grains. This technique allows to decrease the time and the cost of classical selection, and to study the mechanisms involved in the embryo formation. Albinism restrains the use of pollen embryogenesis in cereals. This study is based on applied and fundamental aspects in order to 1- optimize the anther culture protocol and to improve green plantlet regeneration in recalcitrant cvs. and 2- identify molecular mechanisms involved in the switch of microspore, especially defenses. In order to optimize the protocole of anther culture in barley (Hordeum vulgare L.) we used the winter cv. Igri which regenerates chlorophyllous plantlets at 90%, and 5 spring cvs. giving rise mostly to albino plantlets. The copper sulphate addition in media improved the anthers response, the quantity of regenerated plantlets and the percentage of regenerated chlorophyllous plantlets, with all tested cvs. Spring and summer are the best seasons for pollen embryogenesis in cvs used. Finally, we noticed that results also depended on spike position on the tillering plate. Our results indicate that the pretreatment induces defence mechanisms in anthers, as revealed by an oxidative burst and expression gene induction. Copper sulphate addition in the media induced more progressive and less important induction of gene expressionREIMS-BU Sciences (514542101) / SudocSudocFranceF

Analyses of Lysin-motif Receptor-like Kinase (<i>LysM-RLK</i>) Gene Family in Allotetraploid <i>Brassica napus</i> L. and Its Progenitor Species: An In Silico Study

The LysM receptor-like kinases (LysM-RLKs) play a crucial role in plant symbiosis and response to environmental stresses. Brassica napus, B. rapa, and B. oleracea are utilized as valuable vegetables. Different biotic and abiotic stressors affect these crops, resulting in yield losses. Therefore, genome-wide analysis of the LysM-RLK gene family was conducted. From the genome of the examined species, 33 LysM-RLK have been found. The conserved domains of Brassica LysM-RLKs were divided into three groups: LYK, LYP, and LysMn. In the BrassicaLysM-RLK gene family, only segmental duplication has occurred. The Ka/Ks ratio for the duplicated pair of genes was less than one indicating that the genes’ function had not changed over time. The BrassicaLysM-RLKs contain 70 cis-elements, indicating that they are involved in stress response. 39 miRNA molecules were responsible for the post-transcriptional regulation of 12 Brassica LysM-RLKs. A total of 22 SSR loci were discovered in 16 Brassica LysM-RLKs. According to RNA-seq data, the highest expression in response to biotic stresses was related to BnLYP6. According to the docking simulations, several residues in the active sites of BnLYP6 are in direct contact with the docked chitin and could be useful in future studies to develop pathogen-resistant B. napus. This research reveals comprehensive information that could lead to the identification of potential genes for Brassica species genetic manipulation

In Silico Analyses of Autophagy-Related Genes in Rapeseed (Brassica napus L.) under Different Abiotic Stresses and in Various Tissues

The autophagy-related genes (ATGs) play important roles in plant growth and response to environmental stresses. Brassica napus (B. napus) is among the most important oilseed crops, but ATGs are largely unknown in this species. Therefore, a genome-wide analysis of the B. napus ATG gene family (BnATGs) was performed. One hundred and twenty-seven ATGs were determined due to the B. napus genome, which belongs to 20 main groups. Segmental duplication occurred more than the tandem duplication in BnATGs. Ka/Ks for the most duplicated pair genes were less than one, which indicated that the negative selection occurred to maintain their function during the evolution of B. napus plants. Based on the results, BnATGs are involved in various developmental processes and respond to biotic and abiotic stresses. One hundred and seven miRNA molecules are involved in the post-transcriptional regulation of 41 BnATGs. In general, 127 simple sequence repeat marker (SSR) loci were also detected in BnATGs. Based on the RNA-seq data, the highest expression in root and silique was related to BnVTI12e, while in shoot and seed, it was BnATG8p. The expression patterns of the most BnATGs were significantly up-regulated or down-regulated responding to dehydration, salinity, abscisic acid, and cold. This research provides information that can detect candidate genes for genetic manipulation in B. napus

Adaptation of grapevine flowers to cold involves different mechanisms depending on stress intensity.

Grapevine flower development and fruit set are influenced by cold nights in the vineyard. To investigate the impact of cold stress on carbon metabolism in the inflorescence, we exposed the inflorescences of fruiting cuttings to chilling and freezing temperatures overnight and measured fluctuations in photosynthesis and sugar content. Whatever the temperature, after the stress treatment photosynthesis was modified in the inflorescence, but the nature of the alteration depended on the intensity of the cold stress. At 4°C, photosynthesis in the inflorescence was impaired through non-stomatal limitations, whereas at 0°C it was affected through stomatal limitations. A freezing night (-3°C) severely deregulated photosynthesis in the inflorescence, acting primarily on photosystem II. Cold nights also induced accumulation of sugars. Soluble carbohydrates increased in inflorescences exposed to -3°C, 0°C and 4°C, but starch accumulated only in inflorescences of plants treated at 0 and -3°C. These results suggest that inflorescences are able to cope with cold temperatures by adapting their carbohydrate metabolism using mechanisms that are differentially induced according to stress intensity

Kluyveromyces marxianus, an Attractive Yeast for Ethanolic Fermentation in the Presence of Imidazolium Ionic Liquids

Imidazolium ionic liquids (ILs) are promising solvents for lignocellulosic biomass (LCB) pretreatment and allow the achievement of higher ethanolic yields after enzymatic hydrolysis and ethanolic fermentation. However, residual ILs entrapped in pretreated biomass are often toxic for fermentative microorganisms, but interaction mechanisms between ILs and cells are still unknown. Here we studied the effects of 1-ethyl-3-methylimidazolium acetate [Emim][OAc] and 1-ethyl-3-methylimidazolium methylphosphonate [Emim][MeO(H)PO2] on Kluyveromyces marxianus, a thermotolerant ethanologenic yeast. Morphological impacts induced by ILs on K. marxianus were characterized by Scanning Electron Microscopy analysis and showed wrinkled, softened, and holed shapes. In Yeast-Malt-Dextrose (YMD) medium, K. marxianus tolerated IL additions up to 2% for [Emim][OAc] and 6% for [Emim][MeO(H)PO2]. Below these thresholds, some IL concentrations enhanced ethanolic yields up to +34% by switching the metabolic status from respiratory to fermentative. Finally, K. marxianus fermentation was applied on several substrates pretreated with [Emim][OAc] or [Emim][MeO(H)PO2] and enzymatically hydrolyzed: a model long fiber cellulose and two industrial LCBs, softwood (spruce) and hardwood (oak) sawdusts. The maximum ethanolic yields obtained were 1.8 to 3.9 times higher when substrates were pretreated with imidazolium ILs. Therefore K. marxianus is an interesting fermentative yeast in a second-generation bioethanol process implying IL pretreatment