大莲湖池杉林湿地土壤可培养细菌的分离与鉴定

采用牛肉膏蛋白胨培养基培养,从大莲湖池杉林土壤中共分离得到20个菌落形态不同的菌株。通过对这些菌株的形态、培养特征、生理生化特征的研究以及16S rDNA序列分析,初步确定这些菌株分别属于假单胞菌属(Pseudomonas)、芽胞杆菌属(Bacillus)、红球菌属(Rhodococcus)、北里孢菌属(Kitasatosporia)、金黄杆菌属(Chryseobacterium)、不动杆菌属(Acinetobacter)、黄杆菌属(Flavobacterium)、鞘氨醇杆菌属(Sphingobacte-rium)和丛毛单胞菌属(Comamonas)等9个属细菌。其中芽胞杆菌属和不动杆菌属细菌是优势菌,分离到的红球菌属、北里孢菌属、鞘氨醇杆菌属和丛毛单胞菌属细菌在国内湿地土壤中报道较少。     

明党参内生菌的分离鉴定及其对金黄色葡萄球菌的抑制作用

研究郑州市栽培明党参内生菌资源状况,筛选出对金黄色葡萄球菌具有拮抗作用的内生菌。采用平板培养法从明党参中分离内生细菌,通过混菌法和牛津杯双层平板扩散法筛选出对金黄色葡萄球菌具有明显抑制作用的菌株,并通过培养特征、形态特征、生化特征、全自动微生物质谱检测、16S rDNA基因序列分析和系统进化树分析对菌株进行了分类学鉴定。在明党参中共分离到29株内生细菌,通过初筛和复筛后,筛选出对金黄色葡萄球菌具有明显抑制作用的4株菌株NMY1、NMJ10、NMG10、PMG3,其中NMJ10对金黄色葡萄球菌的抑菌圈直径达20.3 mm,经鉴定菌株NMJ10和菌株PMG3为芽胞杆菌属新菌种(Bacillus spp.),菌株NMY1为枯草芽胞杆菌(B.subtilis),菌株NMG10为假蕈状芽胞杆菌(B.pseudomycoides)。明党参内生菌具有多样性,其中芽胞杆菌属为优势菌株,有拮抗金黄色葡萄球菌活性高的菌株存在,其代谢产物对金黄色葡萄球菌有较高抑制作用,为抗菌药源成分的开发提供了新的来源。     

仙草植物内生芽胞杆菌种群多样性研究

为了研究仙草植株内生芽胞杆菌种群的多样性,利用营养琼脂培养基分离仙草根、茎、叶中的内生芽胞杆菌,采用16SrRNA序列鉴定法对分离获得的芽胞杆菌进行鉴定。同一组织内,对16SrRNA序列及形态完全一样的菌株合并冗余,用ClustalX对分离得到的菌株16SrRNA进行排序,构建Neighbor-Joining系统进化树,并计算多样性指数。实验结果显示,共分离得到58株仙草内生芽胞杆菌(根29株、茎28株、叶1株),分属于芽胞杆菌属(Bacillus)、类芽胞杆菌属(Paenibacillus)和赖氨酸芽胞杆菌属(Lysinibacillus),共3个属的11个种,其中与阿氏芽胞杆菌B.aryabhattai、简单芽胞杆菌B.simplex和B.flexus 3个种最接近的内生芽胞杆菌占总分离芽胞杆菌株数的62%;根部内生芽胞杆菌数量及种属均最丰富,其多样性指数高于茎和叶。     

极端沙漠干燥炎热环境土壤中耐热和耐辐射菌的分离、培养与鉴定

目的:了解库姆塔格沙漠土壤耐热和耐辐射微生物的多样性,获得耐热及耐辐射微生物资源,为沙漠极端微生物资源的开发利用及其重要功能基因的发掘提供重要的物质基础。方法:无菌采集新疆维吾尔自治区吐鲁番盆地鄯善县库木塔格沙漠(北纬24°50′11″,东经90°12′30″,海拔440 m)土壤,利用常规平板分离方法进行菌株分离、培养,定向筛选耐高温和耐紫外线菌株并纯化,通过菌株16S r DNA基因序列测定,结合系统发育研究对分离得到的细菌进行鉴定分析。结果:共分离得到9株菌,相互间相似性较高(相似度90%),均为芽孢杆菌(Bacillus)。序列测定后为4个种,分别为枯草芽胞杆菌枯草亚种(Bacillus subtilis subsp.subtilis)、地芽胞杆菌属(Geobacillus sp.)、芽胞杆菌属(Bacillus sp.)和热噬淀粉芽胞杆菌(Bacillus thermoamylovorans)。结论:本研究分离、驯化并鉴定了9株新疆干旱沙漠环境细菌,具有一定的耐高温和耐辐射特性,为极端环境微生物资源的开发利用提供了基础理论依据。     

新疆乌尔禾地区盐渍土壤耐盐细菌多样性与群落结构研究

研究新疆北部乌尔禾地区盐渍土壤中微生物群落结构及多样性,以期发现新的高盐环境耐盐性微生物资源菌株。采用传统分离培养法获得可培养耐盐菌株并对菌株形态学、16S rRNA基因测序、耐盐特性进行研究,同时结合高通量测序技术分析新疆乌尔禾地区盐渍土壤耐盐细菌的多样性与群落结构。共分离得到耐盐细菌11株,分属6个属,均为中度耐盐菌,以芽胞杆菌属(Bacillus)为优势菌。对盐渍土壤微生物16S rRNA(V3～V4)基因测序,共获得细菌序列290 952条,分属24个门410个属,变形菌门(Proteobacteria, 60.31%)、厚壁菌门(Firmicutes, 21.52%)、拟杆菌门(Bacteroidetes, 6.9%)和放线菌门(Actinobacteria, 6%)相对丰度较高。优势属为克吕沃尔菌属(Kluyvera,21%)、Hafnia-Obesumbacterium(19.6%)和假单胞菌属(Pseudomonas,7.5%)。结果表明,新疆乌尔禾地区盐渍土壤耐盐细菌优势菌群以芽胞杆菌属(Bacillus)居多,细菌群落结构较复杂,潜在可利用微生物资源较为丰富,对高盐极端环境耐盐微生物新资源有进一步研究的意义。     

东亚飞蝗肠道细菌的研究

目的从微生态学角度研究东亚飞蝗的营养生理活动。方法从东亚飞蝗自然种群的雌、雄成虫肠道环境中分离、纯化、培养,获得细菌16个属的菌株,对菌体形态、染色反应、培养性状、生理生化反应进行系统研究。结果上述16个细菌菌株分别属于沙雷菌属(Serratia)、短状杆菌属(Brachybacterium)、预研菌属(Yokenella)、肠杆菌属(Penterobacter)、微杆菌属(Microbacterium)、柠檬酸杆菌属(Citrobacter)、类芽胞杆菌属(Paenibacillus)、沙门菌属(Salmonella)、棍状杆菌属(Clavibacter)、放线杆菌属(Actinobacillus)、米勒菌属(Moelleralla)、不动杆菌属(Acinetobacter)、埃希菌属(Escherichia)、葡萄球菌属(Staphylococcus)、克吕沃尔菌属(Kluyvera)、克雷伯菌属(Kleb-siella)。结论东亚飞蝗雌性成虫肠道环境细菌种类为12个,雄性为10个,其中6个菌株相同;数量之间存在明显差异。     

五倍子内生菌的分离与鉴定

目的分离五倍子内生菌,并进行筛选和鉴定。方法分别采用牛肉膏蛋白胨固体、液体培养基,高氏1号培养基,马丁孟加拉红培养基外加4‰的国产硫酸庆大霉素(800万单位)四种培养基对五倍子内生菌进行分离。结果得到5株菌株,经菌落形态显微观察与16S rDNA序列分析鉴定,5株菌株均为革兰阳性菌;A1、A5和B1为杆菌,同属芽胞杆菌属;A3为球菌,微球菌属;A6为球菌,葡萄球菌属;A5与B1同源性最高,达到100%。结论从五倍子中分离得到5株内生细菌,将有利于五倍子的深入开发利用。     

青海可可西里嗜碱芽胞杆菌资源调查

【目的】了解可可西里嗜碱芽胞杆菌资源多样性及产酶多样性,为芽胞杆菌功能资源挖掘和菌剂开发提供基础。【方法】采用Horikoshi I培养基,通过可培养法分离青海可可西里土壤中的嗜碱芽胞杆菌,利用16S r RNA基因序列初步鉴定分离获得的芽胞杆菌。采用透明圈法分析分离菌株的产蛋白酶、纤维素酶及木聚糖酶活性。【结果】从青海可可西里土壤中共分离获得66株嗜碱芽胞杆菌,根据16S r RNA基因序列相似性分析,发现它们隶属于6个属22个种,分别为芽胞杆菌属(Bacillus)、纤细芽胞杆菌属(Gracilibacillus)、喜盐芽胞杆菌属(Halobacillus)、咸海鲜芽胞杆菌属(Jeotgalibacillus)、类芽胞杆菌属(Paenibacillus)和嗜冷芽胞杆菌属(Psychrobacillus),其中以芽胞杆菌属(Bacillus)为优势属。2株嗜碱芽胞杆菌与它们最近匹配模式菌株的16S r RNA基因序列相似性为97.00%和98.65%,为潜在新种。三种酶活检测结果表明产酶菌株约占总分离菌株的95.00%,其中55株具有产蛋白酶活性,27株具有产纤维素酶活性,8株能够产木聚糖酶。【结论】青海可可西里蕴藏着较丰富的嗜碱芽胞杆菌资源及丰富的产酶资源,为后续嗜碱芽胞杆菌的挖掘提供理论基础。     

滇西北兰坪金顶铅锌矿矿区可培养细菌多样性初步研究

目的对兰坪金顶铅锌矿矿区样品中的可培养细菌进行分离并对其多样性进行研究。方法采集云南兰坪金顶铅锌矿矿区土样和矿石样,采用固体肉汤培养基、卯黄培养基及PYGV培养基分离该矿区环境中的可培养细菌,利用16SrRNA基因序列分析构建系统发育树,并统计不同种属细菌的数量,初步评估细菌多样性。结果兰坪金顶铅锌矿矿区环境细菌的主要种群包括放线菌门、变形菌门和厚壁菌门的不同菌属：微球菌属、节杆菌属、假单胞菌属、短波单胞菌属、芽孢杆菌属、类芽孢杆菌属、考克菌属、葡萄球菌属、芽孢八叠球菌及Skermanella属的菌株,其中抗逆性较强的优势菌群为放线菌门的细菌。结论本研究初步证实兰坪金顶铅锌矿矿区可培养细菌种类丰富。     

基于可培养方法分析云南腾冲小空山火山谷芽胞杆菌分布特征

【目的】为了解云南腾冲小空山火山谷土壤中可培养芽胞杆菌种类分布特征。【方法】采用可培养手段对小空山火山谷阳坡、谷底和阴坡土壤中的芽胞杆菌进行分离培养,根据16S rRNA基因序列同源性对分离菌株进行鉴定,并分析系统发育地位。利用Canoco5软件分析采样点芽胞杆菌种类分布特征与土壤样品理化性质的相关性。【结果】从火山谷土壤样品中共分离获得180株芽胞杆菌,16S rRNA基因测序鉴定结果表明分离菌株隶属于芽胞杆菌纲2个科(芽胞杆菌科和类芽胞杆菌科)、6个属、34个种,其中芽胞杆菌属(Bacillus) 11个种,类芽胞杆菌属(Paenibacillus) 14个种,短芽胞杆菌属(Brevibacillus)3个种,赖氨酸芽胞杆菌属(Lysinibacillus)4个种,嗜冷芽胞杆菌属(Psychrobacillus)1个种和绿芽胞杆菌属(Viridibacillus)2个种,其中7个菌株与其最近模式菌株16SrRNA相似性低于种的界定阈值(98.65%),为芽胞杆菌潜在新物种。优势属为芽胞杆菌属和类芽胞杆菌属,优势种为蕈状芽胞杆菌(Bacillusmycoides),图瓦永芽胞杆菌(Bacillustoyonensis),蜡状芽胞杆菌(Bacilluscereus),解木糖赖氨酸芽胞杆菌(Lysinibacillusxylanilyticus),蜂房类芽胞杆菌(Paenibacillusalvei)和沙地绿芽胞杆菌(Viridibacillus arenosi)。其中16个种分离自阳坡,29个种分离自阴坡,9个种分离自谷底,三者共同种类为6种。阳坡、谷底和阴坡的芽胞杆菌种群分布Bray-Curtis相似性为62.4%,多样性分析结果表明,Shannon指数(H′)大小次序为阴坡阳坡谷底。环境因子分析发现,芽胞杆菌种群分布多样性特征与其土壤的海拔高度、碳氮比和硫含量呈负相关,而和碳源和氮源含量呈正相关。【结论】从以上结果得出,云南腾冲火山谷有着较为丰富的芽胞杆菌资源,且还存在可分离培养的芽胞杆菌的潜在新物种,为利用火山微生物资源提供了保障。     

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.     

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.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.