Light intensity affects RNA silencing of a transgene in Nicotiana benthamiana plants

Abstract Background Expression of exogenous sequences in plants is often suppressed through one of the earliest described RNA silencing pathways, sense post-transcriptional gene silencing (S-PTGS). This type of suppression has made significant contributions to our knowledge of the biology of RNA silencing pathways and has important consequences in plant transgenesis applications. Although significant progress has been made in recent years, factors affecting the stability of transgene expression are still not well understood. It has been shown before that the efficiency of RNA silencing in plants is influenced by various environmental factors. Results Here we report that a major environmental factor, light intensity, significantly affects the induction and systemic spread of S-PTGS. Moreover, we show that photoadaptation to high or low light intensity conditions differentially affects mRNA levels of major components of the RNA silencing machinery. Conclusions Light intensity is one of the previously unknown factors that affect transgene stability at the post-transcriptional level. Our findings demonstrate an example of how environmental conditions could affect RNA silencing.</p

MIWI N-terminal arginines orchestrate generation of functional pachytene piRNAs and spermiogenesis

N-terminal arginine (NTR) methylation is a conserved feature of PIWI proteins, which are central components of the PIWI-interacting RNA (piRNA) pathway. The significance and precise function of PIWI NTR methylation in mammals remains unknown. In mice, PIWI NTRs bind Tudor domain containing proteins (TDRDs) that have essential roles in piRNA biogenesis and the formation of the chromatoid body. Using mouse MIWI (PIWIL1) as paradigm, we demonstrate that the NTRs are essential for spermatogenesis through the regulation of transposons and gene expression. The loss of TDRD5 and TDRKH interaction with MIWI results in attenuation of piRNA amplification. We find that piRNA amplification is necessary for transposon control and for sustaining piRNA levels including select, nonconserved, pachytene piRNAs that target specific mRNAs required for spermatogenesis. Our findings support the notion that the vast majority of pachytene piRNAs are dispensable, acting as self-serving genetic elements that rely for propagation on MIWI piRNA amplification. MIWI-NTRs also mediate interactions with TDRD6 that are necessary for chromatoid body compaction. Furthermore, MIWI-NTRs promote stabilization of spermiogenic transcripts that drive nuclear compaction, which is essential for sperm formation. In summary, the NTRs underpin the diversification of MIWI protein function

Modulation of Aub-TDRD interactions elucidates piRNA amplification and germplasm formation.

Aub guided by piRNAs ensures genome integrity by cleaving retrotransposons, and genome propagation by trapping mRNAs to form the germplasm that instructs germ cell formation. Arginines at the N-terminus of Aub (Aub-NTRs) interact with Tudor and other Tudor domain-containing proteins (TDRDs). Aub-TDRD interactions suppress active retrotransposons via piRNA amplification and form germplasm via generation of Aub-Tudor ribonucleoproteins. Here, we show that Aub-NTRs are dispensable for primary piRNA biogenesis but essential for piRNA amplification and that their symmetric dimethylation is required for germplasm formation and germ cell specification but largely redundant for piRNA amplification

ANNULUS CLOSURE TECHNOLOGY DEVELOPMENT INSPECTION/SALT DEPOSIT CLEANING MAGNETIC WALL CRAWLER

The Liquid Waste Technology Development organization is investigating technologies to support closure of radioactive waste tanks at the Savannah River Site (SRS). Tank closure includes removal of the wastes that have propagated to the tank annulus. Although amounts and types of residual waste materials in the annuli of SRS tanks vary, simple salt deposits are predominant on tanks with known leak sites. This task focused on developing and demonstrating a technology to inspect and spot clean salt deposits from the outer primary tank wall located in the annulus of an SRS Type I tank. The Robotics, Remote and Specialty Equipment (RRSE) and Materials Science and Technology (MS&amp;T) Sections of the Savannah River National Laboratory (SRNL) collaborated to modify and equip a Force Institute magnetic wall crawler with the tools necessary to demonstrate the inspection and spot cleaning in a mock-up of a Type I tank annulus. A remote control camera arm and cleaning head were developed, fabricated and mounted on the crawler. The crawler was then tested and demonstrated on a salt simulant also developed in this task. The demonstration showed that the camera is capable of being deployed in all specified locations and provided the views needed for the planned inspection. It also showed that the salt simulant readily dissolves with water. The crawler features two different techniques for delivering water to dissolve the salt deposits. Both water spay nozzles were able to dissolve the simulated salt, one is more controllable and the other delivers a larger water volume. The cleaning head also includes a rotary brush to mechanically remove the simulated salt nodules in the event insoluble material is encountered. The rotary brush proved to be effective in removing the salt nodules, although some fine tuning may be required to achieve the best results. This report describes the design process for developing technology to add features to a commercial wall crawler and the results of the demonstration testing performed on the integrated system. The crawler was modified to address the two primary objectives of the task (inspection and spot cleaning). SRNL recommends this technology as a viable option for annulus inspection and salt removal in tanks with minimal salt deposits (such as Tanks 5 and 6.) This report further recommends that the technology be prepared for field deployment by: (1) developing an improved mounting system for the magnetic idler wheel, (2) improving the robustness of the cleaning tool mounting, (3) resolving the nozzle selection valve connections, (4) determining alternatives for the brush and bristle assembly, and (5) adding a protective housing around the motors to shield them from water splash. In addition, SRNL suggests further technology development to address annulus cleaning issues that are apparent on other tanks that will also require salt removal in the future such as: (1) Developing a duct drilling device to facilitate dissolving salt inside ventilation ducts and draining the solution out the bottom of the ducts. (2) Investigating technologies to inspect inside the vertical annulus ventilation duct

DCL3 and DCL4 are likely involved in the light intensity-RNA silencing cross talk in Nicotiana benthamiana

Plants have substantially invested in RNA silencing as the central defense mechanism to combat nucleotide “invaders” such as viruses, trasposable elements and transgenes. The quantity and quality of light perceived by a plant as a constant environmental stimulus refining cell homeostasis and RNA silencing mechanism seems not to be an exception. In our recent paper in BMC Plant Biology we documented that light intensity, in physiological ranges, positively affects silencing initiation and spread.1 Here, we show that virus induced gene silecing under high light conditions results in more frequent systemic silencing events of a transgene and is acompanied by elevated DCL3 and DCL4 mRNA levels. In addition, our results show that DCL3 holds a vital role in systemic silencing spread and the positive effect of light intensity on RNA silencing requires DCL4 function

Investigating the Martian soil at the InSight landing site

International audienceThe InSight mission is a geophysical mission aimed at better understanding the structure of Mars and of the other rocky planets of the solar system. To do so, a lander accommodating two cameras, a very sensitive seismometer, and a dynamic self-penetrating heat probe nicknamed the mole were placed on the Mars surface by the Instrument Deployment Arm (IDA). Besides geophysical data (which definitely enriched the existing knowledge on the structure of Mars), the InSight instruments significantly increased the knowledge of the geological and geotechnical characteristics of the surface material at the InSight site. Small strain (elastic) parameters were derived from wave velocity measurements during the hammering sessions between the self-penetrating probe and the seismometer. A detailed observation of the soil profile along a depth of 37 cm was made possible thanks to the photos taken by the cameras, and to a detailed analysis of the mole penetration process. Further information was provided by an intense campaign of scraping and piling conducted by the IDA on the surface sand/dust layer. It was shown that the soil profile was composed of a surface 1 cm thick sand/dust layer, overlaying an around 20 cm thick loose duricrust made up of a cohesive matrix containing some pebbles, located above a 12 cm layer of sand overlaying a gravel/sand deposit. It is believed that the geology and soil mechanics data provided by the InSight mission will help for further robotic exploration of Mars