番茄匍柄霉叶斑病拮抗细菌的筛选与鉴定

应用平板对峙法从连作多年的番茄根际土样中筛选得到对番茄匍柄霉叶斑病具有较强拮抗活性的细菌菌株ZF161,该菌株对番茄匍柄霉的平板抑制率达到70.51%。离体叶片试验显示,菌株ZF161对番茄匍柄霉叶斑病防治效果达到69.83%。通过菌落形态观察、生理生化特性、Biolog测定和多基因系统发育树综合分析,鉴定菌株ZF161为枯草芽孢杆菌(Bacillussubtilis)。番茄盆栽试验结果表明,菌株ZF161对番茄匍柄霉叶斑病的防治效果达到63.27%。进一步平板对峙抑菌谱试验结果显示,菌株ZF161对其他7种病原真菌也具有较好的抑制效果。上述结果说明,菌株ZF161对番茄匍柄霉叶斑病具有良好的防治效果,且对多种病原真菌也具有抑制作用,具有开发应用的潜力。     

赤峰地区番茄灰叶斑病病原菌的鉴定

以番茄灰叶斑病为研究对象,采用形态学鉴定、致病性检测和分子生物学等方法,研究了赤峰地区番茄灰叶斑病的病原菌种类,以期为该病害的防治提供参考依据。结果表明:病原菌(ZTWYF16081515)分生孢子淡褐色,长方形至圆柱形,大小为(21.4~71.9)μm×(10.5~27.3)μm。分离菌株喷雾接种,7d后番茄叶片上形成典型的番茄灰叶斑病病斑。核糖体RNA基因内转录间隔区(ITS)和gpd区的PCR产物经测序后进行BLAST分析,均表明该菌与茄匍柄霉S.solani的ITS和gpd序列100%相同,确定该病原为茄匍柄霉S.solani。     

番茄灰霉病拮抗细菌的筛选与X-75菌株鉴定

采用琼脂平板扩散法筛选对番茄灰霉菌具有拮抗作用的细菌,经过对200株分离自土壤的细菌菌株进行初筛、复筛后,得到了1株具有较高拮抗活性的菌株X-75。对该菌株进行形态观察和生理生化特征分析,初步将其鉴定为芽孢杆菌属(Bacillus sp.),将其16SrDNA序列与GenBank中已知标准菌株的16SrDNA序列进行比对,并用Neighbor-joining方法构建X-75菌株进化树,结果表明,X-75与标准菌株AB245422 Bacillus velezensis聚于同一分支,同源性最高,达99.93%;利用番茄离体叶片对X-75菌株的防效进行初步检测,结果表明X-75菌株对番茄灰霉病菌具有明显的拮抗作用。     

番茄灰霉病拮抗细菌的分离筛选及鉴定

以番茄灰霉病菌(Botrytis cinerea)为供试菌,采用平板对峙方法,研究了不同植物根围土壤中的细菌对番茄灰霉病(Botrytis cinerea)的拮抗效果。结果表明:对分离纯化的75株细菌进行拮抗测定,其中30株对番茄灰霉病有不同程度抑制作用,分离频率为40.00%。其中3株拮抗细菌YM8、FQ10、FQ11对番茄灰霉病病原菌拮抗效果较好,抑菌圈直径在1.5~3.0cm。通过生理生化试验、形态特征观察对其进行了种属初步鉴定,鉴定结果为菌株YM8为地衣芽孢杆菌(Bacillus licheniformis),FQ10为短芽孢杆菌(Bacillus brevis),FQ11为蜡状芽孢杆菌(Bacillus cereus)。     

木霉拮抗菌的筛选与鉴定

采用涂布平板法从植物和土壤中共分离得到301株细菌,其中从羊蹄甲、芦荟和香蕉皮等植物中分离到内生细菌86株,从土壤中分离到细菌215株。以食用菌栽培中的主要污染菌木霉,包括哈茨木霉（Trichoderma harzianum）、长枝木霉（Trichoderma longibrachiatum）等4种共8株木霉菌株以及蘑菇（Agaricus bisporus）、香菇（Lentinul aedodes）等14种不同食用菌为实验菌,通过平板对峙培养法筛选出对木霉有拮抗作用而对食用菌拮抗性弱甚至无拮抗作用的细菌75株。采用传统分类法以及16S rDNA序列分析对其中作用效果最好的5株细菌进行了鉴定,结果显示,5株细菌均为枯草芽孢杆菌Bacilluss subtilis。在实验中还发现,从羊蹄甲、芦荟和香蕉皮等植物中更容易分离到木霉拮抗菌。     

枯草芽孢杆菌NBF809防治番茄棒孢叶斑病研究

采用平板对峙结合显微观察确定枯草芽孢杆菌NBF809菌株对多主棒孢的抑菌活性,采用活体盆栽试验研究其对番茄棒孢叶斑病的防治效果。结果表明,在平板对峙条件下NBF809菌株对多主棒孢具有良好的抑菌活性,显微镜下观察到菌丝发生扭曲、肿胀和变形;盆栽试验结果表明NBF809菌株对番茄棒孢叶斑病的防治效果达到73.26%。聚合酶链式反应用于检测NBF809菌株的抗菌素生物合成基因,扩增到bac D、itu C、itu D、mrs M和myc C 5个基因,涉及Bacilysin、Iturin、Mersacidin和Mycosubtilin 4种抗菌素的合成。     

枯草芽孢杆菌M6和木霉10对番茄灰病的防治效果研究

分别研究了枯草芽孢杆菌M6、木霉10和两者配成的复合菌剂W对番茄灰霉病的防治效果,测定了它们对番茄灰霉病的抑制作用.结果表明,枯草芽孢杆菌M6、木霉10、复合菌剂W对番茄灰霉病均有抑制作用,但复合菌 剂W对番茄灰霉病抑制作用更强,抑制率达到62.2％,且复合菌剂W的使用浓度不能低于105 cfu/mL.田间防治试...     

宁夏红枣叶斑病病原菌鉴定及室内药剂筛选

以宁夏红枣叶斑病典型病叶为试材,采用组织分离法、离体叶片菌块接种法和菌丝生长抑制法,研究了该地区红枣叶斑病的致病菌种类及有效防治药剂。结果表明:从宁夏红枣叶斑病典型病叶上主要分离得到匍柄霉属真菌(Stemphyliumsp.),占总分离物的85.0%,其中匍柄霉菌株Y-14具有很强的致病性,是宁夏红枣叶斑病的主要病原菌种类;7种杀菌剂对病原菌的毒力强弱依次为25%丙环唑EC25%咪鲜胺EC1 000亿/g枯草芽孢杆菌WP10%多抗霉素WP250g/L嘧菌酯SC70%代森锰锌WP1%申嗪霉素SC,尤其前5种杀菌剂的EC50值1.0g/L,可作为宁夏红枣叶斑病生物防治和化学防治的首选药剂。     

草莓病害拮抗细菌的筛选及其对草莓褐色叶斑病的防效

通过组织分离法和稀释分离法从草莓根、茎、叶和根际土壤中分离内生细菌183株及根际细菌55株,其中9株拮抗菌对草莓褐色叶斑病病原菌（Pilidium concavum）、灰葡萄孢菌（Botrytis cinerea）、尖孢镰刀菌（Fusarium oxysporum）、链格孢菌（Alternaria alternate）4种草莓病原菌具有广谱拮抗效果。结合病原菌形态学特征、生理生化特性和分子生物学鉴定结果,将拮抗菌株G3-23、J3-20、G3-29鉴定为贝莱斯芽孢杆菌（Bacillus velezensis）,J6-1、J1-4、G2-12、G3-k2和J1-11鉴定为枯草芽孢杆菌（Bacillus subtilis）,G3-20鉴定为耐盐芽孢杆菌（Bacillus halotolerans）,G3-17鉴定为蜡样芽孢杆菌（Bacillus cereus）。菌株G3-23能够分泌蛋白酶和纤维素酶,对由P. concavum引起的草莓褐色叶斑病具有良好的防治效果,并且该菌株能在离体草莓叶片上成功定殖,表现出较好的生防潜力。     

白菜黑斑病菌拮抗细菌LBC-1菌株的分离与鉴定

从湖南、湖北、河北等地区菜地采集土样,采用抑菌圈法筛选对白菜黑斑病病原菌芸薹链格孢菌（Alternaria brassicae）具有拮抗作用的芽孢杆菌菌株。通过初筛得到具有拮抗作用的菌株20株,复筛出1株拮抗活性较高的菌株LBC-1。通过生理生化试验、形态特征观察及16S rDNA序列分析对其进行了种属鉴定。鉴定结果为菌株LBC-1属于解淀粉芽孢杆菌（Bacillus amyloliquefaciens）,是1株广谱拮抗芽孢细菌菌株。     

Effect of MCPH1 on irradiation induced DNA damage in esophageal cancer cells

AIM: To discover the effect of MCPH1 on the DNA damage induced by ionizing radiation in esophageal cancer cells. METHODS: ECA109 cancer cells were radiated at dose of 8 Gy. The nuclear foci of relevant factors were detected 1 h after irradiation in the ECA109 cells after silence of MDC 1 gene. A cell line was established that was stable low expression of MCPH 1 . The nuclear foci induced by ionizing radiation after silence of MCPH 1 were determined. RESULTS: The MCPH1 gene silenced ECA109 cell line was successfully constructed. A strong relationship between MDC1, MCPH1 and γ-H2AX was observed 1 h after 8 Gy irradiation. Silence of MDC 1 did not affect the nuclear foci formation of γ-H2AX and MCPH1. The nuclear foci of MDC1 but not γ-H2AX significantly reduced after silencing of MCPH 1 . CONCLUSION: MCPH1 is located in the downstream of H2AX and upstream formation of MDC1, and regulates the nuclear foci formation of MDC1 during DNA damage response.     

Activation of repair pathways after neuronal DNA damage induced by bupivacaine

AIM To investigate the activation of related repair pathways after bupivacaine-induced neuronal DNA damage by cDNA gene screening. METHODS The bupivacaine-induced SH-SY5Y neuronal damage and DNA damage model was established. The technique of cDNA microplate array was used to screen the 21 important regulatory factors in the DNA damage repair pathway. Post-analysis of these differentially expressed repair genes for the repair pathway enrichment and distribution was performed. The data were analyzed by GraphPad Prism 6 statistical software to compare differences between groups. RESULTS The viability of SH-SY5Y cells treated with bupivacaine at different concentrations （detected by CCK-8 assay） showed that the IC₅₀ value of bupivacaine was 1.5 mmol/L. The comet assay related index （the comet tail） was increased （P<0.05）, the phosphorylation level of γH2AX protein was increased （P<0.05）, indicating that DNA damage in the SH-SY5Y cells was significantly aggravated after bupivacaine treatment. The results of cDNA microplate assay showed that compared withcontrol group, the differentially expressed genes after bupivacaine treatment were DNA-PKcs, PTEN, NTH1, RAD9, CSB, GADD45, XPD, XPC-HR23B and P53. The analysis showed that these repair genes were mainly concentrated in the following 3 repair mechanisms： base excision repair, nucleotide excision repair, and non-homologous reconstitution. CONCLUSION The repair genes differentially expressed after neuronal DNA damage caused by local anesthetics are mainly concentrated in the pathways of non-homologous end-joining, base excision repair and nucleotide excision repair.     

Involvement of extracellular signal regulated kinase in the regulation of amyloid precursor protein processing in PC12 cells by TPPB

AIM: To explore the signal transduction pathways involved in the regulation of amyloid precursor protein (APP) processing by protein kinase C (PKC) activator TPPB.METHODS: PC12 cells were treated with TPPB (PKC activator) for 3 h and various signal transduction inhibitors were added to the conditioned medium to investigate their effects on α-secretase form of soluble amyloid precursor protein (sAPPα) secretion after TPPB treatment via Western blotting. Extracellular signal regulated kinase (ERK, p42/44MAPK) and phospho-p42/44MAPK were also measured after TPPB treatment.RESULTS: TPPB (1 μmol/L) significantly increased sAPPα secretion as compared with control group. The increase in sAPPα secretion by TPPB was partially blocked by ERK inhibitor U0126, c-Jun N-terminal kinase (JNK) inhibitor SP600125 and protein tyrosine kinase (PTK) inhibitor genistein, but not by p38MAPK inhibitor SB203580. TPPB (1 μmol/L) increased the expression of phospho-p42/44MAPK without altering total p42/44MAPK levels.CONCLUSION: ERK, JNK and PTK may be involved in the regulation of APP processing by TPPB.     

GDC-0032 inhibits cell viability and induces DNA damage in U251 human glioma cells

AIM: To investigate the effect of phosphatidylinostiol 3-kinase (PI3K) inhibitor GDC-0032 on cell viability, cell cycle and DNA damage in human glioma U251 cells. METHODS: The cell viability was analyzed by MTT assay. The cell cycle distribution of U251 cells was examined by flow cytometry. The protein expression was examined by Western blot. The expression and localization of γ-H2AX were determined by laser-scanning confocal microscopy. RESULTS: GDC-0032 inhibited the cell viability in a dose-dependent manner. U251 cells showed G₁ phase arrest accompanied with upregulation of p27 protein after exposure to GDC-0032, while the expression of cell division cycle protein 2 (Cdc2) was inhibited. GDC-0032 treatment increased the formation of γ-H2AX foci and histone H2AX phosphorylation in the U251 cells. In addition, GDC-0032 upregulated the phosphorylation levels of mitogen-activated protein kinases, including extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), in the glioma cells, while the expression of survivin was inhibited. CONCLUSION: GDC-0032 inhibits U251 cell growth by inhibition of cell viability and cell cycle progression. GDC-0032 also induces DNA damage of U251 cells. The anticancer activity of GDC-0032 is associated with increasing the activity of ERK and JNK and downregulating the expression of survivin.     

Regulatory effect of miR-214-5p on myocardial injury and immune response in ischemia-reperfusion rats by targeting PAK4

AIM: To investigate the effect of microRNA-214-5p (miR-214-5p) on myocardial injury and immune response in rats with ischemia-reperfusion (I/R) by targeting p21-activated protein kinase 4 (PAK4). METHODS: The rats were divided into sham group, I/R group, Ad-Scramble group, and Ad-miR-214 group (n=9). Adenovirus was injected into 6 different sites on the anterior wall of the left ventricle of the rats. Four days later, the I/R model was constructed by suturing the left anterior descending coronary artery. The expression level of miR-214 was detected by RT-qPCR. Myocardial injury was observed by HE staining. The levels of heart damage markers (CK-MB, Mb, and cTnI) and inflammatory factors (IL-6, IL-1β and TNF-α) were measured by ELISA. The rate of cardiomyocyte apoptosis was analyzed by flow cytometry. The content of MDA and the activity of SOD were detected by commercially available kits. Target genes were predicted by genetic software and verified by dual-luciferase reporter assay. The protein levels of caspase-3, caspase-9, Bcl-2, Bax, PAK4, p-Akt and p-mTOR were determined by Western blot. RESULTS: miR-214 was down-regulated in the cardiomyocytes of I/R rats (P<0.01). Over-expression of miR-214-5p attenuated myocardial injury in the I/R rats, down-regulated the expression of CK-MB, Mb and cTnI, decreased the apoptotic rate of cardiomyocytes, up-regulated the expression of Bcl-2, down-regulated Bax, caspase-3 and caspase-9 expression, increased SOD activity, and decreased the content of MDA, IL-6, IL-1β and TNF-α (P<0.01). The binding sites of miR-214-5p and PAK4 were pre-sent in the 3’-UTR, and over-expression of miR-214-5p up-regulated the protein levels of PAK4, p-Akt and p-mTOR (P<0.01). CONCLUSION: miR-214-5p over-expression attenuates myocardial injury in I/R rats by targeting PAK4, inhibits cardiomyocyte apoptosis, oxidative stress and inflammation, and activates the PI3K/Akt/mTOR pathway.