拮抗枯草芽孢杆菌KC-5的分离鉴定及其发酵优化

为了筛选出对植物病原菌具有拮抗作用的生物防治细菌,采用平板对峙法从蔬菜根际土壤中分离获得一株对多种病原真菌具有抑制作用的枯草芽孢杆菌,并对抑制后的病原菌菌丝进行了观察,结果表明该菌株对尖孢镰刀菌、串珠镰刀菌、层出镰刀菌以及腐皮镰刀菌均具有抑菌活性。通过形态特征、生理生化特征及16S rRNA序列分析,将该菌株鉴定为枯草芽孢杆菌(Bacillus subtilis),并对其发酵培养基进行了优化。     

棉花枯萎、黄萎病拮抗芽孢杆菌的抗菌蛋白特性

从土贝母和腊肠等中药和发酵食品中筛选出对棉花枯萎、黄萎病菌有广谱拮抗作用的芽孢杆菌29株,其中有12株菌产抗菌蛋白。有5株抑菌活性较强:H110、H184、H216、B316和B382。经初步鉴定,H110和H184为枯草芽孢杆菌,H216、B316和B382为地衣芽孢杆菌。5株菌的蛋白粗提液对热稳定,对蛋白酶K、胰蛋白酶均不敏感,但H184、H216的蛋白粗提液对胃蛋白酶部分敏感。     

海水养殖系统中37株枯草芽孢杆菌生理代谢、遗传特性及抑菌效果差异分析

【背景】由水产致病菌导致的病害不断暴发,寻找安全有效的抗生素替代品是目前生产的迫切需求。人们通常过于关注益生菌效应,而对其安全性评价重视度不够。【目的】分析我国海水养殖系统中不同来源枯草芽孢杆菌菌株的表型及遗传特征,并寻找绿色安全且具有多重抑菌作用的菌株。【方法】以2009-2021年从我国海水养殖系统中分离的37株枯草芽孢杆菌为对象,利用纸片扩散法(K-B法)检测其对不同抗生素的抗性;利用培养基平板法测定淀粉酶、蛋白酶和溶血能力;通过PCR方法检测枯草芽孢杆菌溶血相关基因携带风险;采用牛津杯法测定其对副溶血弧菌、溶藻弧菌、爱德华氏菌、哈维氏弧菌、美人鱼发光杆菌和假交替单胞菌等6种病原菌的抑菌作用;并对候选益生性枯草芽孢杆菌的安全性进行评估。【结果】药敏检测结果显示,37株枯草芽孢杆菌对甲氧苄啶、吡哌酸、链霉素表现出强耐药性,对磺胺嘧啶表现出中等耐药,对头孢噻肟、环丙沙星、舒巴坦的耐药率低,对克拉霉素、诺氟沙星、氟苯尼考、氟甲喹、复方新诺明、四环素表现为完全敏感。蛋白酶、淀粉酶活性测试结果显示,37株枯草芽孢杆菌能不同程度地水解酪蛋白和淀粉。溶血性测试结果显示,37株枯草芽孢杆菌中有4株出现溶血现象,而8个溶血相关基因在37株枯草芽孢杆菌中均有检出,溶血表型与检测基因关联分析表明,产生溶血现象的菌株与其溶血基因携带间无直接相关性。抑菌试验分析表明,37株枯草芽孢杆菌均对2种及以上病原菌有抑制作用,对6种病原菌均具有良好抑菌作用的有2株(菌株Bs4和Bs7)。对凡纳滨对虾的安全试验表明,菌株Bs4对凡纳滨对虾具有高安全性,7 d对虾存活率为100%。【结论】通过对37株枯草芽孢杆菌生理代谢表型、遗传特性及病原拮抗特性进行比较分析,揭示了我国海水养殖系统中枯草芽孢杆菌具有多元化的表型及遗传特征,并筛选出一株生态安全且具有多重抑菌活性的益生性枯草芽孢杆菌,为水产养殖病害防控、开发抑菌类微生态制剂及水产养殖行业健康绿色发展提供了理论基础和技术支撑。     

拮抗菌株Kc-t99的鉴定及其抑菌活性研究

通过传统分类与分子生物学技术相结合的方法对从京郊菜园土壤中分离筛选的拮抗菌株Kc-t99进行鉴定,并采用生物测定的方法评价其抑菌活性.结果表明,菌株Kc-t99的形态学特征和生理生化特性与枯草芽孢杆菌基本一致,其16S rDNA序列与GenBank中已鉴定的枯草芽孢杆菌的16S rDNA序列同源性高达98.06%,据此初步确定其为枯草芽孢杆菌.抑菌试验表明该菌株对供试的5种病原真菌和4种细菌均具抑菌活性,其中对甘蓝枯萎病菌、黄瓜角斑病菌和桃褐腐病菌抑制作用显著.     

向日葵枯萎病拮抗芽胞杆菌的筛选与生防评价

目的筛选向日葵枯萎病拮抗芽胞杆菌菌株并研究其抗菌谱,探讨环境条件对菌株抑菌活性的影响并通过植物栽培完成生防评价。方法从向日葵根际土壤中选择性分离芽胞杆菌,通过5点对峙法确定1株高效拮抗菌株,进行鉴定后,测定其抑菌谱;单因素实验探讨环境条件对抑菌活性的影响;向日葵发芽实验完成生防评价。结果确定1株高效的枯萎病拮抗菌株WBFL-1,经鉴定为枯草芽胞杆菌,该菌对镰刀菌属具有广谱抗性,其抑菌活性的最佳条件是温度40℃,pH值7.0,接种量150μL,发酵时间48 h。生防评价表明该菌对枯萎病的拮抗效果显著。结论该菌可为农作物枯萎病的生物防治提供有效菌种储备。     

拮抗Bacillus subtilis的筛选及发酵条件对拮抗能力的影响

筛选了3株对大肠埃希菌具有拮抗活性的枯草芽胞杆菌,并对这3个菌株的拮抗能力最强的Bf菌株的发酵条件进行了初步研究。先将不同枯草芽胞杆菌菌株与株大肠埃希菌进行拮抗试验,根据抑菌率,筛选出对大肠埃希菌拮抗效果较好的枯草芽胞杆菌Bf。然后,在不同pH值、温度以及不同的培养时间下观察记录Bf对大肠埃希菌的拮抗作用。根据实验结果分析得到:筛选到的枯草芽胞杆菌在pH值为7.0,37℃下培养40 h后抑菌效果最好。     

具有分泌蛋白能力的短芽孢杆菌的筛选及鉴定

短芽孢杆菌（Bacillus　brevis）具有分泌蛋白能力强和胞外蛋白酶活性低的特性,是分泌表达外源蛋白较理想的宿主。为获得分泌表达系统较理想的宿主菌,建立了短芽孢杆菌高效筛选模型,从800余株细菌中筛得8株具有高蛋白分泌能力且没有胞外蛋白酶活性的候选菌。经多相分类学初步鉴定其中5株为短芽孢杆菌。     

一株解淀粉芽孢杆菌的分离鉴定及其抑菌条件研究

旨在从海洋环境中分离筛选抑制鲈鱼病原鳗弧菌的拮抗菌,进行菌种鉴定,并对其抑菌条件及药物敏感性进行研究。采用十字交叉划线法和平板点种法筛选鲈鱼病原鳗弧菌的拮抗菌,采用形态学、生理生化反应及16S r DNA基因序列的系统发育分析方法对分离拮抗菌进行鉴定,采用单因子变量法对拮抗菌WTD的最佳抑菌条件进行研究,采用平板药敏纸片法对其药物敏感性进行研究。结果显示,从海洋环境样品中分离出126株细菌,筛选出19株拮抗菌。其中菌株WTD的拮抗效果最强,细菌鉴定结果表明,该菌为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。最佳抑菌条件研究表明,该菌在温度35℃、p H7、盐度为1%Na Cl的时候,拮抗效果最好。药敏实验结果表明,该菌对麦迪霉素等29种抗生素敏感,而对多粘菌素具有抗性。拮抗菌解淀粉芽孢杆菌WTD对鲈鱼病原菌鳗弧菌有较强的抑制作用,具有潜在的应用价值。     

可可西里低温适生拮抗芽孢杆菌的筛选鉴定及脂肽化合物分析

极端环境特殊微生物资源研究和开发具有广阔的应用前景和研究意义.对分离筛选自青海可可西里境内植被根围的8株在4和10 ℃条件下生长良好的低温适生芽孢杆菌进行鉴定分析.结果表明: 通过生理生化特征分析、rep-PCR指纹图谱分析、16S rDNA及gyrB基因序列分析鉴定,8株供试菌株分别为莫哈韦芽孢杆菌(Bacillus mojavensis)3株,解淀粉芽孢杆菌(B. amyloliquefaciens)1株和简单芽孢杆菌(B. simplex)4株.采用平板对峙试验从中筛选到4株对油菜菌核病原真菌(Sclerotinia sclerotiorum)及水稻白叶枯病原细菌(Xanthomonas oryzae pv. oryzae)均具有显著拮抗活性的生防菌株;采用MALDI-TOF-MS质谱分析生防菌株的拮抗活性物质,结果显示菌株KKD-1 (B. mojavensis)产生脂肽类化合物泛革素和表面活性素,菌株KKD-2(B. amyloliquefaciens)产生脂肽类化合物伊枯草菌素A、泛革素和表面活性素,推断生防菌株的拮抗活性可能与脂肽化合物的合成及分泌有关.该研究为低温适生性芽孢杆菌生物肥料和生物农药的研发提供了菌株资源.     

可可西里低温适生拮抗芽孢杆菌的筛选鉴定及脂肽化合物分析

极端环境特殊微生物资源研究和开发具有广阔的应用前景和研究意义.对分离筛选自青海可可西里境内植被根围的8株在4和10℃条件下生长良好的低温适生芽孢杆菌进行鉴定分析.结果表明:通过生理生化特征分析、rep-PCR指纹图谱分析、16S rDNA及gyrB基因序列分析鉴定,8株供试菌株分别为莫哈韦芽孢杆菌(Bacillus mojavensis)3株,解淀粉芽孢杆菌(B.amyloliquefaciens)1株和简单芽孢杆菌(B.simplex)4株.采用平板对峙试验从中筛选到4株对油菜菌核病原真菌(Sclerotinia sclerotiorum)及水稻白叶枯病原细菌(Xanthomonasoryzae pv.oryzae)均具有显著拮抗活性的生防菌株;采用MALDI-TOF-MS质谱分析生防菌株的拮抗活性物质,结果显示菌株KKD1(B.mojavensis)产生脂肽类化合物泛革素和表面活性素,菌株KKD2(B.amyloliquefaciens)产生脂肽类化合物伊枯草菌素A、泛革素和表面活性素,推断生防菌株的拮抗活性可能与脂肽化合物的合成及分泌有关.该研究为低温适生性芽孢杆菌生物肥料和生物农药的研发提供了菌株资源.     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Synthesis and antimicrobial studies of hydrazone derivatives of 4-[3-(2,4-difluorophenyl)-4-formyl-1H-pyrazol-1-yl]benzoic acid and 4-[3-(3,4-difluorophenyl)-4-formyl-1H-pyrazol-1-yl]benzoic acid

Microbial resistance to antibiotics is an unresolved global concern, which needs urgent and coordinated action. One of the guidelines of the Centers for Disease Control and Preventions (CDC) to combat antibiotic resistance is the development of new antibiotics to treat drug-resistant bacteria. In our effort to find new antibiotics, we report the synthesis and antimicrobial studies of 30 new pyrazole derivatives. These novel molecules have been synthesized by using readily available starting materials and benign reaction conditions. Some of these molecules have shown activity with MIC values as low as 0.78?µg/mL against four bacterial strains; Staphylococcus aureus, methicillin-resistant S. aureus, Bacillus subtilis, and Acinetobacter baumannii. Furthermore, active molecules are non-toxic to mammalian cell line.

     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Novel compounds with potent CDK9 inhibitory activity for the treatment of myeloma

Cyclin-dependent kinases (CDKs) and Polo-like kinases (PLKs) play key role in the regulation of the cell cycle. The aim of our study was originally the further development of our recently discovered polo-like kinase 1 (PLK1) inhibitors. A series of new 2,4-disubstituted pyrimidine derivatives were synthesized around the original hit, but their PLK1 inhibitory activity was very poor. However the novel compounds showed nanomolar CDK9 inhibitory activity and very good antiproliferative effect on multiple myeloma cell lines (RPMI-8226).