[Current progress in functions of axon guidance molecule Robo and underlying molecular mechanism].

The axon guidance molecule Robo is a transmembrane protein which is conserved during evolution. Robo and its ligand, Slit, have been implicated in regulating many developmental processes, such as axon guidance, neuronal migration, tumor metastasis, angiogenesis, lung morphogenesis, kidney morphogenesis, heart morphogenesis, ovary development and gonad development. Robo function mainly depends on the binding of its Ig1 domain to the LRR-2 domain of Slit ligand. Meanwhile, Robo function is also mediated by binding to some signaling molecules, including the heparan sulfate proteoglycans (HSPGs), GTPase-activating proteins (GAPs) and tyrosine kinase Abelson. Several transcription factors, including Hox, Midline and Nkx2.9, were shown to regulate robo expression. In addition, alternative splicing and transport regulation also affect Robo function. In this review, we summarized the studies on the molecular structure, functions and molecular mechanism of Robo, which would propose a novel strategy for the research of neural development, as well as prevention and treatment of nervous system diseases and cancers.

[Current progress in functions of axon guidance molecule Slit and underlying molecular mechanism].

The axon guidance molecule Slit is a secreted glucoprotein which is conserved during evolution. Slit has been implicated in regulating a variety of life activities, such as axon guidance, neuronal migration, neuronal morphological differentiation, tumor metastasis, angiogenesis and heart morphogenesis. Slit function mainly depends on the binding of its LRR-2 domain to the Ig1 domain of Roundabout (Robo) receptor, meanwhile Slit function is also mediated by a range of signaling molecules, including the heparan sulfate proteoglycans (HSPGs), GTPase-activating proteins (GAPs), tyrosine kinase Abelson, calcium ions, MicroRNA-218 and other axon guidance molecules. Several transcription factors, including Single-minded, Irx and Midline, were shown to regulate slit expression. In addition, multiple Slit isoforms exist as a consequence of alternative spliced transcripts. The research on guidance mechanism of Slit will facilitate the understanding of molecular mechanism underlying neural networks formation in the process of neural development and regeneration. Meanwhile, the studying of Slit guidance mechanism could promote the prevention and treatment of human neurological diseases and cancer metastasis.

Effect ofBacillus velezensis JC-K3 on Endophytic Bacterial and Fungal Diversity in Wheat Under Salt Stress.

Plant growth-promoting bacteria (PGPB) can effectively reduce salt damage in plants. Currently, there are many studies on the effects of PGPB on the microbial community structure of rhizosphere soil under salt stress, but fewer studies on the community structure of endophytic bacteria and fungi. We propose that inoculation of endophytic bacteria into the rhizosphere of plants can significantly affect the microbial community structure of the plant's above-ground and underground parts, which may be the cause of the plant's "Induced Systemic Tolerance." The isolated endophytes were re-inoculated into the rhizosphere under salinity stress. We found that, compared with the control group, inoculation with endophytic Bacillus velezensis JC-K3 not only increased the accumulation of wheat biomass, but also increased the content of soluble sugar and chlorophyll in wheat, and reduced the absorption of Na in wheat shoots and leaves. The abundance of bacterial communities in shoots and leaves increased and the abundance of fungal communities decreased after inoculation with JC-K3. The fungal community richness of wheat rhizosphere soil was significantly increased. The diversity of bacterial communities in shoots and leaves increased, and the richness of fungal communities decreased. JC-K3 strain improved wheat's biomass accumulation ability, osmotic adjustment ability, and ion selective absorption ability. In addition, JC-K3 significantly altered the diversity and abundance of endophytic and rhizosphere microorganisms in wheat. PGPB can effectively reduce plant salt damage. At present, there are many studies on the effect of PGPB on the microbial community structure in rhizosphere soil under salt stress, but there are few studies on the community structure changes of endophytic bacteria and fungi in plants.

The Effect of Salt-Tolerant Antagonistic Bacteria CZ-6 on the Rhizosphere Microbial Community of Winter Jujube (Ziziphus jujuba Mill. "Dongzao") in Saline-Alkali Land.

As the main economic crop cultivated in the Yellow River Delta, winter jujube contains various nutrients. However, soil salinization and fungal diseases have affected the yield and quality of winter jujube. In order to use plant growth-promoting rhizobacteria (PGPR) to reduce these damages, the antagonistic bacteria CZ-6 isolated from the rhizosphere of wheat in saline soil was selected for experiment. Gene sequencing analysis identified CZ-6 as Bacillus amyloliquefaciens. In order to understand the salt tolerant and disease-resistant effects of CZ-6 strain, determination of related indicators of salt tolerance, pathogen antagonistic tests, and anti-fungal mechanism analyses was carried out. A pot experiment was conducted to evaluate the effect of CZ-6 inoculation on the rhizosphere microbial community of winter jujube. The salt tolerance test showed that CZ-6 strain can survive in a medium with a NaCl concentration of 10% and produces indole acetic acid (IAA) and 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase. Studies on the inhibition mechanism of pathogenic fungi show that CZ-6 can secrete cellulase, protease, and xylanase. Gas chromatography-mass spectrometry (GC-MS) analysis showed that CZ-6 can release volatile organic compounds (VOCs), including 2-heptanone and 2-nonanone. In addition, the strain can colonize the rhizosphere and migrate to the roots, stems, and leaves of winter jujube, which is essential for plant growth or defense against pathogens. Illumina MiSeq sequencing data indicated that, compared to the control, the abundance of salt-tolerant bacteria Tausonia in the CZ-6 strain treatment group was significantly increased, while the richness of Chaetomium and Gibberella pathogens was significantly reduced. Our research shows that CZ-6 has the potential as a biological control agent in saline soil. Plant damage and economic losses caused by pathogenic fungi and salt stress are expected to be alleviated by the addition of salt-tolerant antagonistic bacteria.

BmRobo1a and BmRobo1b control axon repulsion in the silkworm Bombyx mori.

The development of the nervous system is based on the growth and connection of axons, and axon guidance molecules are the dominant regulators during this course. Robo, as the receptor of axon guidance molecule Slit, plays a key role as a conserved repellent cue for axon guidance during the development of the central nervous system. However, the function of Robo in the silkworm Bombyx mori is unknown. In this study, we cloned two novel robo genes in B. mori (Bmrobo1a and Bmrobo1b). BmRobo1a and BmRobo1b lack an Ig and a FNIII domain in the extracellular region and the CC0 and CC2 motifs in the intracellular region. BmRobo1a and BmRobo1b were colocalized with BmSlit in the neuropil. Knock-down of Bmrobo1a and Bmrobo1b by RNA interference (RNAi) resulted in abnormal development of axons. Our results suggest that BmRobo1a and BmRobo1b have repulsive function in axon guidance, even though their structures are different from Robo1 of other species.

BmRobo2/3 is required for axon guidance in the silkworm Bombyx mori.

Axon guidance is critical for proper wiring of the nervous system. During the neural development, the axon guidance molecules play a key role and direct axons to choose the correct way to reach the target. Robo, as the receptor of axon guidance molecule Slit, is evolutionarily conserved from planarians to humans. However, the function of Robo in the silkworm, Bombyx mori, remained unknown. In this study, we cloned robo2/3 from B. mori (Bmrobo2/3), a homologue of robo2/3 in Tribolium castaneum. Moreover, BmRobo2/3 was localized in the neuropil, and RNAi-mediated knockdown of Bmrobo2/3 resulted in the longitudinal connectives forming closer to the midline. These data demonstrate that BmRobo2/3 is required for axon guidance in the silkworm.

[Medium optimization for antagonistic Streptomyces S24 and its inhibition on Aspergillus flavus].

Streptomyces S24 has broad spectrum against Aspergillus spp. in food and feed, such as Aspergillus flavus, Aspergillus niger and Asperegillus alutacells. The objective of this study was to improve the production of antifungal substances produced by S24 and to test their inhibitory effects on Aspergillus flavus. By using one-factor-at-a-time experiment and orthogonal design method, we optimized the fermentation medium. The composition of an optimized medium for the production of antifungal substances contained (g/L): starch soluble, 10; Glucose, 40; yeast extract, 8; soybean powder, 24; KH2PO4 4; and CaCO3 0.8. As a result, the productivity of antifungal substances could reach to 10 235.45 microg/mL, and this value was 2.81 times higher than that of initial medium before optimization. Additionally, inhibitory effects of the products on Aspergillus flavus were analyzed. Antagonistic tests indicated that the antifungal substances greatly inhibited mycelium growth and spores germination of Aspergillus flavus. We observed through microscope that the mycelia grew abnormally, such as contorting, bulging, vacuole increasing and the cytoplasmic contents inside effusing and the spores appeared unusual, such as gathering, deforming, cytoplasmic contents inside effusing and fracturing.

[Extraction and characterization of antifungal substances produced by antagonistic Streptomyces S24].

Streptomyces S24 has broad spectrum resistance to the Aspergillus in food and feed, such as Aspergillus flavus, Aspergillus niger, Asperegillus alutacells and so on. We studied the adsorption and desorption properties of antifungal substance from Streptomyces S24 on macroporous resins, screened the best elution solution and also investigated some physical and chemical characters of antifungal substance by determining the antifugal activity using oxford plate assay system. According to the analysis results, AB-8 resin offered the best adsorption and desorption capacity for antifungal substance and its saturated absorption capacity was 7.0822 x 10(4) microg/g, the optimal elution solution was 85% acetone and the dynamic desorption rate could reach 93.82%. The antifungal substance was stable to heat and alkali, not sensitive to organic solvents, and sensitive to ultraviolet rays and acid. Based on its ultraviolet spectrometry, the antifungal substance was identified as heptaene macrolide antibiotic.