Increased Ascorbate Biosynthesis Does Not Improve Nitrogen Fixation Nor Alleviate the Effect of Drought Stress in Nodulated Medicago truncatula Plants.

Legume plants are able to establish nitrogen-fixing symbiotic relations with Rhizobium bacteria. This symbiosis is, however, affected by a number of abiotic constraints, particularly drought. One of the consequences of drought stress is the overproduction of reactive oxygen (ROS) and nitrogen species (RNS), leading to cellular damage and, ultimately, cell death. Ascorbic acid (AsA), also known as vitamin C, is one of the antioxidant compounds that plants synthesize to counteract this oxidative damage. One promising strategy for the improvement of plant growth and symbiotic performance under drought stress is the overproduction of AsA via the overexpression of enzymes in the Smirnoff-Wheeler biosynthesis pathway. In the current work, we generated Medicago truncatula plants with increased AsA biosynthesis by overexpressing MtVTC2, a gene coding for GDP-L-galactose phosphorylase. We characterized the growth and physiological responses of symbiotic plants both under well-watered conditions and during a progressive water deficit. Results show that increased AsA availability did not provide an advantage in terms of plant growth or symbiotic performance either under well-watered conditions or in response to drought.

Robust Cytonuclear Coordination of Transcription in Nascent Arabidopsis thaliana Autopolyploids.

Polyploidy is hypothesized to cause dosage imbalances between the nucleus and the other genome-containing organelles (mitochondria and plastids), but the evidence for this is limited. We performed RNA-seq on Arabidopsis thaliana diploids and their derived autopolyploids to quantify the degree of inter-genome coordination of transcriptional responses to nuclear whole genome duplication in two different organs (sepals and rosette leaves). We show that nuclear and organellar genomes exhibit highly coordinated responses in both organs. First, organelle genome copy number increased in response to nuclear whole genome duplication (WGD), at least partially compensating for altered nuclear genome dosage. Second, transcriptional output of the different cellular compartments is tuned to maintain diploid-like levels of relative expression among interacting genes. In particular, plastid genes and nuclear genes whose products are plastid-targeted show coordinated down-regulation, such that their expression levels relative to each other remain constant across ploidy levels. Conversely, mitochondrial genes and nuclear genes with mitochondrial targeting show either constant or coordinated up-regulation of expression relative to other nuclear genes. Thus, cytonuclear coordination is robust to changes in nuclear ploidy level, with diploid-like balance in transcript abundances achieved within three generations after nuclear whole genome duplication.

Effect of Different Agitation Techniques on the Penetration of Irrigant and Sealer into Dentinal Tubules.

OBJECTIVE: The aim of this study was to compare the effectiveness of five intracanal agitation techniques on the penetration of irrigant and sealer into dentinal tubules. BACKGROUND DATA: Intracanal agitation techniques could promote chemomechanical debridement and the sealing of root canals during endodontic treatment. However, there is limited evidence for the agitation effect of Nd:YAP laser. MATERIALS AND METHODS: Human maxillary premolars with single straight canals (N = 60) were prepared with ProTaper Next® NiTi rotary files (Dentsply Maillefer) up to X4 (ISO 40 size). Rhodamine B-labeled sodium hypochlorite was used for final irrigation together with the conventional syringe (control), sonic, ultrasonic, Nd:YAP laser, or V-Clean™ endodontic agitation system. All canals were obturated with gutta-percha and fluorescein isothiocyanate-labeled AH Plus sealer. Transverse sections were obtained at 2, 5, and 8 mm from the apex and observed under confocal laser scanning microscopy. Maximum penetration depth and penetration percentage of both irrigant and sealer were recorded. Kruskal-Wallis and Mann-Whitney tests were performed for multiple comparisons. The Spearman coefficient was calculated to confirm correlations between irrigant and sealer penetration. RESULTS: Laser agitation attained the most irrigant and sealer penetration depth and penetration percentage (p < 0.05). Sonic and ultrasonic agitation attained significantly more penetration percentage than the control group at 2 mm from the apex (p < 0.05). Patterns of irrigant and sealer penetration correlated significantly for all agitation techniques (p < 0.001). CONCLUSIONS: Nd:YAP laser was superior to other agitation techniques in dentinal tubule penetration of irrigant and sealer at one or more sectioned levels from the apex.