Potato virus Y HC-Pro reduces the ATPase activity of NtMinD, which results in enlarged chloroplasts in HC-Pro transgenic tobacco

Potato virus Y (PVY) is an important plant virus and causes great losses every year. Viral infection often leads to abnormal chloroplasts. The first step of chloroplast division is the formation of FtsZ ring (Z-ring), and the placement of Z-ring is coordinated by the Min system in both bacteria and plants. In our lab, the helper-component proteinase (HC-Pro) of PVY was previously found to interact with the chloroplast division protein NtMinD through a yeast two-hybrid screening assay and a bimolecular fluorescence complementation (BiFC) assay in vivo. Here, we further investigated the biological significance of the NtMinD/HC-Pro interaction. We purified the NtMinD and HC-Pro proteins using a prokaryotic protein purification system and tested the effect of HC-Pro on the ATPase activity of NtMinD in vitro. We found that the ATPase activity of NtMinD was reduced in the presence of HC-Pro. In addition, another important chloroplast division related protein, NtMinE, was cloned from the cDNA of Nicotiana tabacum. And the NtMinD/NtMinE interaction site was mapped to the C-terminus of NtMinD, which overlaps the NtMinD/HC-Pro interaction site. Yeast three-hybrid assay demonstrated that HC-Pro competes with NtMinE for binding to NtMinD. HC-Pro was previously reported to accumulate in the chloroplasts of PVY-infected tobacco and we confirmed this result in our present work. The NtMinD/NtMinE interaction is very important in the regulation of chloroplast division. To demonstrate the influence of HC-Pro on chloroplast division, we generated HC-Pro transgenic tobacco with a transit peptide to retarget HC-Pro to the chloroplasts. The HC-Pro transgenic plants showed enlarged chloroplasts. Our present study demonstrated that the interaction between HC-Pro and NtMinD interfered with the function of NtMinD in chloroplast division, which results in enlarged chloroplasts in HC-Pro transgenic tobacco. The HC-Pro/NtMinD interaction may cause the formation of abnormal chloroplasts in PVY-infected plants

Medicago truncatula transporter database: a comprehensive database resource for M. truncatula transporters

<p>Abstract</p> <p>Background</p> <p><it>Medicago truncatula </it>has been chosen as a model species for genomic studies. It is closely related to an important legume, alfalfa. Transporters are a large group of membrane-spanning proteins. They deliver essential nutrients, eject waste products, and assist the cell in sensing environmental conditions by forming a complex system of pumps and channels. Although studies have effectively characterized individual <it>M. truncatula </it>transporters in several databases, until now there has been no available systematic database that includes all transporters in <it>M. truncatula</it>.</p> <p>Description</p> <p>The <it>M. truncatula </it>transporter database (MTDB) contains comprehensive information on the transporters in <it>M. truncatula</it>. Based on the TransportTP method, we have presented a novel prediction pipeline. A total of 3,665 putative transporters have been annotated based on International Medicago Genome Annotated Group (IMGAG) V3.5 V3 and the <it>M. truncatula </it>Gene Index (MTGI) V10.0 releases and assigned to 162 families according to the transporter classification system. These families were further classified into seven types according to their transport mode and energy coupling mechanism. Extensive annotations referring to each protein were generated, including basic protein function, expressed sequence tag (EST) mapping, genome locus, three-dimensional template prediction, transmembrane segment, and domain annotation. A chromosome distribution map and text-based Basic Local Alignment Search Tools were also created. In addition, we have provided a way to explore the expression of putative <it>M. truncatula </it>transporter genes under stress treatments.</p> <p>Conclusions</p> <p>In summary, the MTDB enables the exploration and comparative analysis of putative transporters in <it>M. truncatula</it>. A user-friendly web interface and regular updates make MTDB valuable to researchers in related fields. The MTDB is freely available now to all users at <url>http://bioinformatics.cau.edu.cn/MtTransporter/</url>.</p

Signal enhancement with stacked magnets for high-resolution radio frequency glow discharge mass spectrometry

A method for signal enhancement utilizing stacked magnets was introduced into high-resolution radio frequency glow discharge-mass spectrometry (rf-GD-MS) for significantly improved analysis of inorganic materials. Compared to the block magnet, the stacked magnets method was able to achieve 50−59% signal enhancement for typical elements in Y2O3, BSO, and BTN samples. The results indicated that signal was enhanced as the increase of discharge pressure from 1.3 to 8.0 mPa, the increase of rf-power from 10 to 50 W with a frequency of 13.56 MHz, the decrease of sample thickness, and the increase of number of stacked magnets. The possible mechanism for the signal enhancement was further probed using the software “Mechanical APDL (ANSYS) 14.0”. It was found that the distinct oscillated magnetic field distribution from the stacked magnets was responsible for signal enhancement, which could extend the movement trajectories of electrons and increase the collisions between the electrons and neutral particles to increase the ionization efficiency. Two NIST samples were used for the validation of the method, and the results suggested that relative errors were within 13% and detection limit for six transverse stacked magnets could reach as low as 0.0082 μg g−1. Additionally, the stability of the method was also studied. RSD within 15% of the elements in three nonconducting samples could be obtained during the sputtering process. Together, the results showed that the signal enhancement method with stacked magnets could offer great promises in providing a sensitive, stable, and facile solution for analyzing the nonconducting materials

De novo transcriptome analysis of Medicago falcata reveals novel insights about the mechanisms underlying abiotic stress-responsive pathway

BACKGROUND: The entire world is facing a deteriorating environment. Understanding the mechanisms underlying plant responses to external abiotic stresses is important for breeding stress-tolerant crops and herbages. Phytohormones play critical regulatory roles in plants in the response to external and internal cues to regulate growth and development. Medicago falcata is one of the stress-tolerant candidate leguminous species and is able to fix atmospheric nitrogen. This ability allows leguminous plants to grow in nitrogen deficient soils. METHODS: We performed Illumina sequencing of cDNA prepared from abiotic stress treated M. falcata. Sequencedreads were assembled to provide a transcriptome resource. Transcripts were annotated using BLASTsearches against the NCBI non-redundant database and gene ontology definitions were assigned. Acomparison among the three abiotic stress treated samples was carried out. The expression of transcriptswas confirmed with qRT-PCR. RESULTS: We present an abiotic stress-responsive M. falcata transcriptome using next-generation sequencing data from samples grown under standard, dehydration, high salinity, and cold conditions. We combined reads from all samples and de novo assembled 98,515 transcripts to build the M. falcata gene index. A comprehensive analysis of the transcriptome revealed abiotic stress-responsive mechanisms underlying the metabolism and core signalling components of major phytohormones. We identified nod factor signalling pathways during early symbiotic nodulation that are modified by abiotic stresses. Additionally, a global comparison of homology between the M. falcata and M. truncatula transcriptomes, along with five other leguminous species, revealed a high level of global sequence conservation within the family. CONCLUSIONS: M. falcata is shown to be a model candidate for studying abiotic stress-responsive mechanisms in legumes. This global gene expression analysis provides new insights into the biochemical and molecular mechanisms involved in the acclimation to abiotic stresses. Our data provides many gene candidates that might be used for herbage and crop breeding. Additionally, FalcataBase (http://bioinformatics.cau.edu.cn/falcata/) was built for storing these data. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-2019-x) contains supplementary material, which is available to authorized users

Assessing the Relationship Between Motor Anticipation and Cortical Excitability in Subacute Stroke Patients With Movement-Related Potentials

Background: Stroke survivors may lack the cognitive ability to anticipate the required control for palmar grasp execution. The cortical mechanisms involved in motor anticipation of palmar grasp movement and its association with post-stroke hand function remains unknown.Aims: To investigate the cognitive anticipation process during a palmar grasp task in subacute stroke survivors and to compare with healthy individuals. The association between cortical excitability and hand function was also explored.Methods: Twenty-five participants with hemiparesis within 1–6 months after first unilateral stroke were recruited. Twenty-five matched healthy individuals were recruited as control. Contingent negative variation (CNV) was measured using electroencephalography recordings (EEG). Event related potentials were elicited by cue triggered hand movement paradigm. CNV onset time and amplitude between pre-cue and before movement execution were recorded.Results: The differences in CNV onset time and peak amplitude were statistically significant between the subacute stroke and control groups, with patients showing earlier onset time with increased amplitudes. However, there was no statistically significant difference in CNV onset time and peak amplitude between lesioned and non-lesioned hemisphere in the subacute stroke group. Low to moderate linear associations were observed between cortical excitability and hand function.Conclusions: The earlier CNV onset time and higher peak amplitude observed in the subacute stroke group suggest increased brain computational demand during palmar grasp task. The lack of difference in CNV amplitude between the lesioned and non-lesioned hemisphere within the subacute stroke group may suggest that the non-lesioned hemisphere plays a role in the motor anticipatory process. The moderate correlations suggested that hand function may be associated with cortical processing of motor anticipation

Development and application of a porous cage carrier method for detecting trace elements in soils by direct current glow discharge mass spectrometry

The accurate and reliable determination of trace elements in soil still remains a big challenge for glow discharge mass spectrometry due to the poor conductive nature of soils. In the present work, a porous cage carrier was developed and used in the analysis of soils. The investigation results suggested that the carrier with a circular cross-sectional area in the range from 20 to 38 mm2, length from 15 to 17 mm and diameter of hole size from 1.5 mm to 2.0 mm could obtain good signals. Then the porous cage carrier method was systematically evaluated by analysing three types of soil reference materials. The discharge process was kept stable for more than 100 minutes, which was much longer than the boric acid method and indium sheet method. The investigations suggested that the internal precision was obtained within 16%, the external precision was better than 20% and the relative error was in the range from 0.7% to 17%. The detection limit of Tb could reach 0.014 μg g−1, which indicated that the new method qualified for the analysis of trace elements in soils. Compared to traditional tablet-pressed methods, the porous cage carrier method was not only convenient for sample preparation, but also showed good stability, reproducibility and better detection limits for trace elements. Furthermore, this method was proved to promote the potential application of GD-MS in the environmental field