Sporisorium scitamineum is the causative agent responsible for smut disease of sugarcane worldwide. However, lack of efficient gene manipulation system makes this fungus much behind the type model of ...the smut fungi in molecular biology. Here, we report the development of a CRISPR/Cas9 and T-DNA based dual vector system that allowed efficient knock-out or knock-in of a gene of interest in the S. scitamineum in a site-specific manner. By using Mfa2, a key player in the mating event in S. scitamineum as a tester gene, site-specific insertions of the introduced fragments were achieved both for Mfa2 knockout and complementation. Of particular advantage of this system is the simplicity of selection and identification for the desired transformants by using drug resistance coupled with PCR. This system greatly facilitates the gene function study in S. scitamineum, and could potentially be used for other basidiomycete fungi.
Marine unmanned vehicle is a novel robot widely used in ocean observation, and its accurate control is of significance to their path planning. We want to find a method to predict the velocity and ...course of this robot, which can help us realize the accurate control of it. The paper proposed a promising type of hybrid robotic fish (HRF), which can realize two kinds of motion modes on the sea surface. Firstly, the configuration and dynamic model of the HRF were analyzed elaborately. Then, to realize accurate velocity prediction under two kinds of motion modes of HRF, the influence factors are presented in a complex marine environment. Based on the influence factors to its maneuverability, such as wind or wave parameters, a velocity prediction algorithm based on back-propagation neural network (BPNN) was introduced. However, BPNN has the disadvantages of extended learning and training time, easily falling into local optimum. Then we found that genetic algorithm (GA), which is a kind of evolutionary algorithm, is suitable for our problem. Therefore, the accuracy and efficiency of the prediction algorithm were improved by adopting the genetic algorithm to optimize the weight and threshold of BPNN. Taking the experimental data from the pool test, the back-propagation neural network with genetic algorithm (GA-BPNN) forecasting model was established. Besides, the other prediction methods were compared and evaluated under the same assessment criterion to validate the proposed forecasting model. The experimental results demonstrate that the GA-BPNN model has higher accuracy and efficiency compared with other prediction algorithms, which verifies the feasibility of the velocity prediction model for hybrid robotic fish in complex ocean environments.
The basidiomycete fungus
Sporisorium scitamineum
is the causative agent of sugarcane smut disease. Mating between two strains of the opposite mating type is essential for filamentous growth and ...infection in sugarcane plants. However, the mechanisms underlying mating and pathogenicity are still not well understood. In this work we used gene disruption to investigate the role of
Ssubc2
, the gene encoding a kinase regulator in
S. scitamineum
. Deletion of
Ssubc2
did not alter the haploid cell morphology or growth rate
in vitro
or tolerance to stress, but mutants with both alleles deleted lost mating ability and infectivity. Deletion of one
Ssubc2
allele in a pair with a wild-type strain resulted in impaired mating and reduced virulence. Transcriptome profiling revealed that about a third of genes underwent reprogramming in the wild types during mating. Although gene expression reprogramming occurred in the pairing of
Ssubc2
-null mutants, their transcriptomic profile differed significantly from that of the wild types, in which 625 genes differed from those present in the wild types that seemed to be among the required genes for a successful mating. These genes include those known to regulate mating and pathogenicity, such as components of the MAPK pathway and
hgl1
. Additionally, a total of 908 genes were differentially expressed in an out-of-control manner in the mutants. We conclude that SsUbc2 functions as a key factor to coordinate the reprogramming of gene expression at the global level and is essential for the transition from monokaryotic basidial growth to dikaryotic hyphal growth through mating.
Angelica sinensis is a long-standing medicine used by Chinese medical practitioners and well-known for its blood-tonic and blood-activating effects. Ferulic acid, ligustilide, and eugenol in Angelica ...sinensis activate the blood circulation; however, the material basis of their blood-tonic effects needs to be further investigated. In this study, five homogeneous Angelica sinensis polysaccharides were isolated, and their sugar content, molecular weight, monosaccharide composition, and infrared characteristics determined. Acetylphenylhydrazine (APH) and cyclophosphamide (CTX) were used as inducers to establish a blood deficiency model in mice, and organ indices, haematological and biochemical parameters were measured in mice. Results of in vivo hematopoietic activity showed that Angelica sinensis polysaccharide (APS) could elevate erythropoietin (EPO), granulocyte colony-stimulating factor (G-CSF), and interleukin-3 (IL-3) serum levels, reduce tumor necrosis factor-α (TNF-α) level in mice, and promote hematopoiesis in the body by regulating cytokine levels. Biological potency test results of the in vitro blood supplementation indicated strongest tonic activity for APS-H 2 O, and APS-0.4 has the weakest haemopoietic activity. The structures of APS-H 2 O and APS-0.4 were characterized, and the results showed that APS-H 2 O is an arabinogalactan glycan with a main chain consisting of α-1,3,5-Ara(f), α-1,5- Ara(f), β-1,4-Gal(p), and β-1,4-Gal(p)A, and two branched chains of β-t-Gal(p) and α-t-Glc(p) connected to each other in a (1→3) linkage to α-1,3,5-Ara(f) on the main chain. APS-0.4 is an acidic polysaccharide with galacturonic acid as the main chain, consisting of α-1,4-GalA, α-1,2-GalA, α-1,4-Gal, and β-1,4-Rha. In conclusion, APS-H 2 O can be used as a potential drug for blood replenishment in patients with blood deficiency, providing a basis for APS application in clinical treatment and health foods, as well as research and development of new polysaccharide-based drugs.
In eukaryotic genomes, DNA methylation is an important type of epigenetic modification that plays crucial roles in many biological processes. To investigate the impact of a hypovirus infection on the ...methylome of
, the chestnut blight fungus, whole-genome bisulfite sequencing (WGBS) was employed to generate single-base resolution methylomes of the fungus with/without hypovirus infection. The results showed that hypovirus infection alters methylation in all three contexts (CG, CHG, and CHH), especially in gene promoters. A total of 600 differentially methylated regions (DMRs) were identified, of which 144 could be annotated to functional genes. RNA-seq analysis revealed that DNA methylation in promoter is negatively correlated with gene expression. Among DMRs, four genes were shown to be involved in conidiation, orange pigment production, and virulence. Taken together, our DNA methylomes analysis provide valuable insights into the understanding of the relationship between DNA methylation and hypovirus infection, as well as phenotypic traits in
.
Biotrophic fungi have to infect their host to obtain nutrients and must establish an interaction with the host to complete their life cycle. In this process, effectors play important roles in ...manipulating the host’s immune system to avoid being attacked. Sporisorium scitamineum is the causative agent of sugarcane smut, the most important disease in sugarcane-producing regions worldwide. In this work, we functionally characterized the conserved effector PEP1 in S. scitamineum. The mating process and the expression of genes in the MAPK signaling pathway and the a and b loci were adversely affected in Sspep1-null mutants. The requirement for SsPEP1 in pathogenicity and symptom development was allele dosage-dependent, i.e., deleting one Sspep1 allele in the mating pair turned a normal black whip with abundant teliospores into a white whip with few teliospores; however, deleting both alleles almost abolished infectivity and whip development. ΔSspep1 mutants produced significantly less mycelium mass within infected plants. Additionally, SsPEP1 was identified as a potent inhibitor of sugarcane POD-1a peroxidase activity, implying that SsPEP1 may function to relieve reactive oxygen species-related stress within the host plant. Taken together, our work demonstrated that SsPEP1 is a multifaceted effector essential for S. scitamineum growth, development, and pathogenicity.
The biotrophic basidiomycetous fungus
causing smut disease in sugarcane is characterized by a life cycle composed of a yeast-like nonpathogenic haploid basidiosporial stage outside the plant and ...filamentous pathogenic dikaryotic hyphae within the plant. Under field conditions, dikaryotic hyphae are formed after mating of two opposite mating-type strains. However, the mechanisms underlying genetic regulation of filamentation and its association with pathogenicity and development of teliospores are unclear. This study has focused on the characterization and genetic dissection of haploid filamentous mutants derived from T-DNA insertional mutagenesis. Our results support the existence of at least three genotypes among the six haploid filamentous mutants that differentially contribute to virulence and development of the whip and teliospore, providing a novel foundation for further investigation of the regulatory networks associated with pathogenicity and teliospore development in
.
Colon cancer (CC) is one of the most common gastrointestinal malignancies. Effectiveness of the existing therapies is limited. Immunotherapy is a promising complementary treatment approach for CC. ...Major histocompatibility complex class I-related protein A and B (MICA/B) are ligands for NK cells. Shedding of MICA/B from the surface of tumor cells by cleavage of MICA/B at the membrane proxial region in MICA/B α3 structural domain is one of immune evasion strategies leading to escape of cancer cells from immunosurveillance. In this study, we generated a panel of MICA/B monoclonal antibodies (mAbs) and identified one of mAbs, mAb RDM028, that had high binding affinity to MICA/B and recognized a site on MICA/B α3 structural domain that is critically important for cleavage of MICA/B. Our study has further demonstrated that RDM028 augmented the surface expression of MICA/B on HCT-116 human CC cells by inhibiting the MICA/B shedding resulting in the enhanced cyotoxicity of NK cells against HCT-116 human CC cells and mediated anti-tumor activity in nude mouse model of colon cancer. These results indicate that mAb RDM028 could be explored for developing as an effective immuno therapy against CC by targeting the MICA/B α3 domain to promot immunosurveillance mediated by MICA/B-NKG2D interaction.
A primary criterion of designing wireless sensor networks is energy efficiency. Focused on the energy problem of routing protocol in wireless sensor networks, we must mention to the clustering ...routing protocol. LEACH is one of the first clustering routing algorithms for sensor networks, and it is able to increase the network lifetime. However, it selects a cluster head based on random probability model and doesn't consider the distribution and the remaining energy of sensor nodes. This paper proposes eNergy-Efficient Cluster Head Selection (NECHS) mechanism base on fuzzy logic in clustering routing. By using fuzzy logic technique, appropriate cluster head is selected based on two factors: neighbor nodes and remaining energy. Simulation results show that the NECHS protocol achieves about 0.09 to 26.8% improvement when the rounds of first dead, tenth dead, half dead and all dead compared to the LEACH protocol.
One of fundamental issues in Wireless Sensor Networks (WSNs) is sensor deployment, which affects the performance and effectiveness of WSNs. Sensor node redeployment is an efficient way to extend the ...lifetime of the network by deploying additional sensor nodes in the sensing region. In this paper, we formulate the event occurrence probability distribution of the redeployment problem in WSNs as Binomial Distribution rather than Uniform Distribution and use Bayesian Estimation to estimate the probability of each node's sensing area. In addition, we proposed a novel algorithm called MFCSB-static to solve the problem under static routing schemes. The experimental results show that the proposed algorithm greatly outperforms the algorithm using Uniform Distribution (MFCSU-static) in the percentage of successfully predicting the earliest energy drain out node.
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