Histidine kinase two-component response regulators Ssk1, Skn7 and Rim15 differentially control growth, developmental and volatile organic compounds emissions as stress responses in Trichoderma atroviride

The Skn7, Ssk1 and Rim15 proteins are response regulators involved in osmotic, oxidative and nutritional stress in fungi. In order to verify the involvement of these genes in Trichoderma atroviride IMI206040’s growth, conidiation, direct antagonism against plant pathogens (Rhizoctonia solani and Sclerotinia sclerotiorum), production of volatile organic compounds (VOCs) with fungistatic effect, and interaction with plants (growth promotion), single mutants were generated, and the phenotypic patterns were analysed in comparison to the wild-type (wt) strain. The mutants were submitted to osmotic, oxidative, membrane and cell wall stress conditions in vitro. The Δskn7 and Δrim15 mutants did not show either significant differences at morphological level, or marked decreases in mycelial growth and conidiation in relation to wt, whereas Δssk1 had altered phenotypes in most conditions tested. The plant-growth promotion of Arabidopsis thaliana seedlings induced by VOCs was not quantitatively modified by any of the mutants in relation to the wt strain, although possible differences in secondary root hairs was noticed for Δrim15. The fungistatic activity was significantly altered for Δssk1 and Δrim15. Overall, the Δssk1 strain showed remarkable morphological differences, with decrease in mycelial growth and conidiation, being also affected in the antagonistic capacity against plant pathogens. The impacts demonstrated by the deletion of ssk1 suggest this gene has a relevant participation in the signalling response to different stresses in T. atroviride and in the interactive metabolism with phytopathogens and plants. On the other hand, unlike other fungal models, Skn7 did not appear to have a critical participation in the above-mentioned processes; Rim15 seemed to confirm its involvement in modulating cellular responses to nutritional status, although with a possible cross-talk with other cellular processes. Our results suggest that Ssk1 likely plays a key regulatory role, not only in basic metabolisms of T. atroviride, but also in biocontrol-related characteristics

Species-Specific Expansion and Molecular Evolution of the 3-hydroxy-3-methylglutaryl Coenzyme A Reductase (HMGR) Gene Family in Plants

Kazakh dandelion (Taraxacum kok-saghyz, Tk) is a rubber-producing plant currently being investigated as a source of natural rubber for industrial applications. Like many other isoprenoids, rubber is a downstream product of the mevalonate pathway. The 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) enzyme catalyzes the conversion of 3-hydroxy-3-methylglutaryl-CoA to mevalonic acid, a key regulatory step in the MVA pathway. Such regulated steps provide targets for increases in isoprenoid and rubber contents via genetic engineering to increase enzyme activities. In this study, we identify a TkHMGR1 gene that is highly expressed in the roots of Kazakh dandelion, the main tissue where rubber is synthesized and stored. This finding paves the way for further molecular and genetic studies of the TkHMGR1 gene, and its role in rubber biosynthesis in Tk and other rubber-producing plants

Does nature learn? Information integration and rare events in systems of increasing complexity

The environment is a continuous source of matter and energy, which dynamizes the adaptive processes of biological systems, so that these systems emerge, persist or are extinguished as a consequence of their reactions to the environment. This perspective, forged from classical physics, gives way to multiple ecological theories, with evolution being the most prominent one. In all these cases, information would be both dependent and subsequent to matter and energy. Thus, the emergence and dynamics of genetic material or ecological attributes such as abundance, richness or diversity depend mainly on the interaction of these two fundamental states. However, recent approaches from quantum physics and complexity views put forward the notion that information can be independent and prior to matter and energy, which allows us to see ecological processes from another perspective, i.e., as including complex biological systems as capable of showing emergent properties such as cognition. We proposed here a set of postulates and ideas that suggests how the ability to manipulate (internalize, integrate, store and generate) information can be developed by those systems, which would directly and non-randomly influence ecological attributes and their dynamics; i.e., how this property can possibly help replacing the notion of the environment as the ultimate cause of changes. Besides fully detailing the sources of knowledge and our rationale in this sense, we have also discussed how these thoughts and possibilities can be employed in devising better and more comprehensive approaches for biological conservation strategies

Six-month Follow-up of Cervical Composite Restorations Placed With a New Universal Adhesive System: A Randomized Clinical Trial

SUMMARY Purpose: The objective of this double-blind, randomized clinical trial was to evaluate the six-month clinical performance of a new universal adhesive (Xeno Select, Dentsply) in noncarious cervical lesions (NCCLs) using two evaluation criteria: World Dental Federation (FDI) and the US Public Health Service (USPHS). Methods and Materials: A total of 124 restorations were randomly placed in 31 patients according to the following groups: ER-D = etch-and-rinse/dry dentin; ER-M = etch-and-rinse/moist dentin; SE-et = selective enamel etching; and SET = self-etch. The composite resin EVOLUX (Dentsply) was placed incrementally. The restorations were evaluated after one week (baseline) and at six months using the FDI and USPHS criteria. Statistical analyses were performed using appropriate tests (α=0.05). Results: Fifteen restorations were lost or fractured at six months (one for ER-D, three for ER-M, five for SE-et, and six for SET) (p&amp;gt;0.05 at six-month recall). When ER (ER-D and ER-M) was compared with SE (SE-et and SET) there was a significant difference in the retention rate after six months (p=0.001). Marginal staining and postoperative sensitivity to air were only observed in three (one for ER-M and two for SET) and two restorations (two for ER-D) in both evaluation criteria (p&amp;gt;0.05), respectively. Forty-seven restorations were considered to have minor discrepancies in marginal adaptation at the six-month recall using the FDI criteria (13 for ER-D, 10 for ER-M, 11 for SE-et, and 13 for SET; p&amp;gt;0.05 between groups). However, for all groups, a significant difference was detected when baseline and six-month data were compared (p&amp;lt;0.05). Conclusions: The six-month clinical behavior of Xeno Select Universal Adhesive depends on the bonding strategy used. The universal adhesive did not fulfill the American Dental Association criteria for full approval when used in the self-etch mode.</jats:p

Gene transfer utilizing pollen-tubes of Albuca nelsonii and Tulbaghia violacea

Developing a tissue culture-independent genetic transformation system would be an interesting technique for gene transfer in valuable medicinal and horticultural plants. Efficient gene delivery (Agrobacterium tumefaciens strain LBA 4404: harbouring PBI121 plasmid) was achieved with Km-resistant pollen grains (pollen tube technique) and were found to be GUS-positive for Albuca nelsonii (31.3%) and Tulbaghia violacea (32.6%). The Km-resistance (95.6% for A. nelsonii and 86.7% for T. violacea) and GUS-positive (100% for A. nelsonii and 97.5% for T. violacea) putative transgenic seedlings in vitro were obtained with 200 mg L-1 Km. The in vitro plants were obtained from leaf explants of putative transgenic seedlings and were confirmed to be Km-resistant and GUSpositive (T. violacea, 73.7% and A. nelsonii, 80.5%). The plants were successfully acclimatized in the greenhouse. We describe a tissue culture-independent gene transfer technique with high efficiency clonal transgenic plant production for A. nelsonii and T. violacea. This can also be applied to biotechnological crop improvement of the same species and potentially to other plants

Genomic sequencing identifies novel Bacillus thuringiensis Vip1/Vip2 binary and Cry8 toxins that have high toxicity to Scarabaeoidea larvae

The Bacillus thuringiensis strain HBF-18 (CGMCC 2070), which has previously been shown to encode the cry8Ga toxin gene, is active against both Holotrichia oblita and Holotrichia parallela. Recombinant Cry8Ga however is only weakly toxic to these insect pests suggesting the involvement of additional toxins in the native strain. We report that through the use of Illumina sequencing three additional, and novel, genes, namely vip1Ad1, vip2Ag1, and cry8-like, were identified in this strain. Although no protein corresponding to these genes could be identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the HBF-18 proteome, reverse transcription (RT)-PCR indicated that all three genes were transcribed in the native strain. The two vip genes were cloned and expressed and, as with other Vip1/2 toxins, appeared to function as a binary toxin and showed strong activity against H. oblita, H. parallela and Anomala corpulenta. This is the first report to demonstrate that the Vip1/Vip2 binary toxin is active against these Scarabaeoidea larvae. The cry8-like gene appeared to be a C-terminally truncated form of a typical cry8 gene and was not expressed in our usual recombinant Bt expression system. When however the missing C-terminal region was replaced with the corresponding sequence from cry8Ea, the resulting hybrid expressed well and the toxin was active against the three test insects