Drosophila bang-sensitive paralytic mutants suffer from hyperactivity and paralysis following a mechanical shock; after recovery from paralysis, they cannot be paralyzed for a refractory period lasting up to 1 hr. Previously, we have shown that in easily shocked (eas), a typical bang-sensitive mutant, electrical shocks delivered to the brain cause seizure-like activity in the dorsal longitudinal flight motor neurons (DLMmns), and failure of giant fiber (GF) stimulation to evoke DLM potentials via the escape response pathway (Pavlidis et al., 1994). Here, we show that seizure and failure in the GF pathway with a refractory period is common to all six members of the bang-sensitive class. This syndrome was not found in any of eight other excitability mutants, including those affecting voltage-gated sodium or potassium-channel function. We show that failure occurs at the synapse between a peripherally synapsing interneuron (PSI) and the DLMmns, while the DLMmn-DLM neuromuscular junctions remain functional. Additionally, failure occurs in all other GF pathway-activated muscles. Failures occurred without seizures in the tergotrochanteral jump muscle (TTM), as was also found in approximately 10% of DLM tests, suggesting that seizures and failures may be independent events. This hypothesis is supported by the finding that, in double mutant combination with mlenapts, which suppresses behavioral bang sensitivity, DLM failures, but not seizures, were reduced.

The basal ganglia (BG), serving as an intermediate bridge between the cerebral cortex and thalamus, are believed to play crucial roles in controlling absence seizure activities generated by the pathological corticothalamic system. Inspired by recent experiments, here we systematically investigate the contribution of a novel identified GABAergic pallido-cortical pathway, projecting from the globus pallidus externa (GPe) in the BG to the cerebral cortex, to the control of absence seizures. By computational modelling, we find that both increasing the activation of GPe neurons and enhancing the coupling strength of the inhibitory pallido-cortical pathway can suppress the bilaterally synchronous 2–4 Hz spike and wave discharges (SWDs) during absence seizures. Appropriate tuning of several GPe-related pathways may also trigger the SWD suppression, through modulating the activation level of GPe neurons. Furthermore, we show that the previously discovered bidirectional control of absence seizures due to the competition between other two BG output pathways also exists in our established model. Importantly, such bidirectional control is shaped by the coupling strength of this direct GABAergic pallido-cortical pathway. Our work suggests that the novel identified pallido-cortical pathway has a functional role in controlling absence seizures and the presented results might provide testable hypotheses for future experimental studies.

BackgroundRecent innovation in the field of genome engineering encompasses numerous levels of plant genome engineering which attract the substantial excitement of plant biologist worldwide. RNA-guided CRISPR Cas9 system has appeared a promising tool in site-directed mutagenesis due to its innovative utilization in different branches of biology. CRISPR-Cas9 nuclease system have supersedes all previously existed strategies and their associated pitfalls encountered with site-specific mutagenesis.Results Here we demonstrated an efficient sequence specific integration/mutation of FAD2–2 gene in soybean using CRISPR-Cas9 nuclease system. A single guided RNA sequence was designed with the help of a number of bioinformatics tools aimed to target distinct sites of FAD2–2 loci in soybean. The binary vector (pCas9-AtU6-sgRNA) has been successfully transformed into soybean cotyledon using Agrobacterium tumafacien. Taken together our findings complies soybean transgenic mutants subjected to targeted mutation were surprisingly detected in our target gene. Furthermore, the detection of Cas9 gene, BAR gene, and NOS terminator were carried out respectively. Southern blot analysis confirmed the stable transformation of Cas9 gene into soybean. Real time expression with qRT-PCR and Sanger sequencing analysis confirmed the efficient CRISPR-Cas9/sgRNA induced mutation within the target sequence of FAD2–2 loci. The integration of FAD2–2 target region in the form of substitution, deletions and insertions were achieved with notably high frequency and rare off-target mutagenesis.ConclusionHigh frequent mutation efficiency was recorded as 21% out of all transgenic soybean plants subjected to targeted mutagenesis. Furthermore, Near-infrared spectroscopy (NIR) indicates the entire fatty acid profiling obtained from the mutants seeds of soybean. A considerable modulation in oleic acid content up to (65.58%) whereas the least level of linoleic acid is (16.08%) were recorded. Based on these finding CRISPR-Cas9 system can possibly sum up recent development and future challenges in producing agronomically important crops.Electronic supplementary materialThe online version of this article (10.1186/s12896-019-0501-2) contains supplementary material, which is available to authorized users.

Porcine circovirus type 2 (PCV2) is the essential infectious agent of postweaning multisystemic wasting syndrome (PMWS). In the present study, we obtained sequences of 31 PCV2 isolates from different farms of 11 provinces of eastern China and analyzed the genetic characterization of 136 eastern China-derivate PCV2 isolated during 2001-2009. The results showed that these PCV2 isolates could be divided into two groups, PCV2b (108 of 1A/1B, 19 of 1C) and PCV2a (1 of 2A, 2 of 2D, 6 of 2E). Among the 9 PCV2a isolates, eight were found before the year 2005. Meanwhile, three major heterogenic regions were observed in amino acid positions 53-91, 121-151, and 190-210; a few specific substitution patterns were found in each subgroup and several variant or conserved epitopes were also observed in the Cap protein. It indicated that PCV2b predominated in PMWS occurrences in eastern China and a few recombinants of different genotypes existed in nature.

b-Amyloid precursor protein binding protein 1 (APP-BP1) was previously identified based on its binding to the carboxyl terminal of b-amyloid precursor protein. In this report, we have discovered that a mutation of dAPP-BP1 (Drosophila ortholog of APP-BP1) hinders tissue development, causes apoptosis in imaginal disc cells, and blocks the NEDD8 conjugation pathway. We show that dAPP-BP1 specifically binds the intracellular domain of APP-like protein (APPL). The dAPP-BP1 mutation partially suppresses the abnormal macrochaete phenotype of Appl d , while overexpression of dAPP-BP1 causes abnormal macrochaetes. When APPL is overexpressed, the normal bristle pattern in the fly thorax is disturbed and apoptosis is induced in wing imaginal discs. APPL overexpression phenotypes are enhanced by reducing the level of dAPP-BP1. APPL overexpression is shown to inhibit the NEDD8 conjugation pathway. APPL-induced apoptosis is rescued by overexpression of dAPP-BP1. Our data suggest that APPL and dAPP-BP1 interact antagonistically during Drosophila development.

Experiments in soil columns were conducted to evaluate the single and interactive effects of water, salt and nitrogen stress at different sunflower (Helianthus annuus L.) growth stages in Hetao Irrigation District, China. The study factors included soil salinity (S 0 : EC e =2.5-3.6 dS m-1 ; S 1 : EC e =9.6-10.7 dS m-1), soil moisture (W 0 : 35 ％-55％ of field water capacity; W 1 : 75％-100％ of field water capacity), and nitrogen application rates (N 0 : 0 kg N ha-1 ; N 1 : 135 kg N ha-1). The results indicated that the S 1 treatments increased the duration of the seedling stages by 23.91% but decreased the duration of maturity by 33.09% on average compared with the S 0 treatments. Similarly, water deficit significantly retarded anthesis and prolonged the total growth period. The comprehensive stress assessment index (CSAI) was obtained using principal component analysis (PCA) and membership function analysis (MFA). The CSAIs in different treatments showed that soil salinity was the main limiting factor for sunflower vegetative growth from seeding to bud (SS 1), whereas water stress dominated the development from bud to flowering (SS 2) and flowering to maturity (SS 3). Although statistically non-significant, nitrogen stress was intensified after bud initiation and the CSAI in W 1 S 0 N 0 treatment was 40.68% lower than W 1 S 1 N 1 treatment in SS 3. Moreover, the interactive effects of the three factors were complicated. Our experiments suggested that adequate water supply after bud initiation and the reasonable nitrogen application rate (135 kg N ha-1) can alleviate adverse effects on sunflower reproductive growth under different saline conditions.

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A facility of BaPS (Barometric Process Separation) and indoor incubation experiments were used to determine the effect of soil salinity on soil respiration and nitrogen transformation. The rates of soil respiration, gross nitrification, denitrification, ammonium and nitrate nitrogen concentrations and relevant soil parameters were measured. Results showed that soil respiration and nitrification and denitrification rates were all affected by soil salinity. Furthermore, the effect of soil salinity level on nitrification and denitrification rates had a threshold value (EC1:5 = 1.13 dS/m). When soil salinity level was smaller to this threshold value, the rates of nitrification and denitrification increased with soil salinity while they were reduced when soil salinity level was larger than the threshold value. Moreover, the changing law of soil respiration rate with soil salinity was similar with the nitrification and denitrification rates while the variation tendency was opposite. In addition, the transformation form urea to ammonium and nitrate nitrogen was also reduced with the increase of soil salinity and the reduced effect could be expressed by exponential functions.