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。     

农林生态区土壤中的可培养芽胞杆菌资源调查——以四川省和重庆市部分地区为例

调查了四川和重庆农林生态区土壤中的芽胞杆菌资源种群分布,为芽胞杆菌功能资源挖掘和菌剂开发提供基础。采用可培养法从农林生态区土壤中分离获得芽胞杆菌,利用细菌16S rRNA基因初步确定其分类地位,并通过多样性指数分析芽胞杆菌种群结构。结果表明,共分离获得芽胞杆菌95株,经16S rRNA基因鉴定,7个属38个种,其中芽胞杆菌属(Bacillus)22个种、赖氨酸芽胞杆菌属(Lysinbacillus)4个种、类芽胞杆菌属(Paenibacillus)7个种、短芽胞杆菌属(Brevibacillus)2个种、嗜冷芽胞杆菌属(Psychrobacillus)1个种、鲁梅尔芽胞杆菌属(Rummeliibacillus)1个种和虚构芽胞杆菌属(Fictibacillus)1个种。其中,6株芽胞杆菌与其最相近模式菌株的16S rRNA相似性低于98.65%,为潜在新种。农林生态区土壤芽胞杆菌多样性指数香农指数大小依次为乐山林地盐亭县农地重庆林地盐亭县林地。以芽胞杆菌种类为样本,含量为指标,当欧式距离λ=15时,芽胞杆菌种类可分为高含量分布广型和低含量分布不均匀型。综上所述,农林生态区土壤蕴藏着较丰富的芽胞杆菌种类,为芽胞杆菌资源开发和利用提供了资源保障。     

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。     

养猪微生物发酵床芽胞杆菌空间分布多样性

了解微生物发酵床大栏养猪垫料中的芽胞杆菌多样性和空间分布规律,为微生物发酵床管理、芽胞杆菌新资源挖掘及菌剂开发奠定基础。将发酵床划分为32个方格(4行×8列),采用五点取样法获得每个方格的样品。采用可培养法从32份样品中分离芽胞杆菌菌株,利用16S rRNA基因序列初步鉴定所分离获得的芽胞杆菌种类。利用聚集度指标和回归分析法,分析芽胞杆菌的样方空间分布型。通过Shannon-Wiener多样性指数、Simpson优势度指数、Hill指数及丰富度指数分析,揭示微生物发酵床中芽胞杆菌的空间分布多样性。从32份样品中共获得芽胞杆菌452株,16S rRNA基因鉴定结果表明它们分别隶属于芽胞杆菌纲的2个科、8个属、48个种。其中,种类最多的为芽胞杆菌属(Bacillus),30种;赖氨酸芽胞杆菌属(Lysinibacillus),6种;类芽胞杆菌属(Paenibacillus),5种;短芽胞杆菌属(Brevibacillus),3种;鸟氨酸芽胞杆菌属(Ornithinibacillus)、大洋芽胞杆菌属(Oceanibacillus)、少盐芽胞杆菌属(Paucisalibacillus)和纤细芽胞杆菌属(Gracilibacillus)各1个种。芽胞杆菌种类在发酵床空间分布差异很大,根据其空间出现频次,可分为广分布种类,如地衣芽胞杆菌(Bacillus licheniformis);寡分布种类,如根际芽胞杆菌(B.rhizosphaerae);少分布种类,如弯曲芽胞杆菌(B.flexus)。依据其数量,可分为高含量组优势种群,如地衣芽胞杆菌(B.licheniformis);中含量组常见种群,耐盐赖氨酸芽胞杆菌(Lysinibacillus halotolerans);寡含量组寡见种群,如根际芽胞杆菌(B.rhizosphaerae);低含量组偶见种群,如土地芽胞杆菌(B.humi)。空间分布型聚集度和回归分析测定表明,芽胞杆菌在微生物发酵床的分布类型为聚集分布。微生物发酵床垫料中芽胞杆菌种类总含量高达4.41×108个/g,其种类含量范围为0.01—94.1×106个/g(均值为8.96×106个/g),丰富度指数(D)、优势度指数(λ)、Shannon-Wiener指数(H')和均匀度指数(J')分别为0.4928、0.2634、1.3589和0.9803,其中香农指数最大的单个芽胞杆菌种类为地衣芽胞杆菌(B.licheniformis)。根据芽胞杆菌种类多样性指数聚类分析,当欧式距离λ=17时,可分为高丰富度高含量和低丰富度低含量类型。微生物发酵床的芽胞杆菌种类丰富、数量高,是一个天然的菌剂"发酵罐",有望直接作为微生物菌剂,应用于土壤改良、作物病害防控、污染治理等领域。     

新疆伊犁马铃薯根际芽胞杆菌纯培养多样性

了解马铃薯根际土壤中芽胞杆菌种类多样性,可为挖掘芽胞杆菌新资源提供基础。从新疆伊犁州9个地点采集了30份马铃薯根际土壤样品,采用可培养法从中分离芽胞杆菌,通过16S rRNA基因同源性鉴定了分离菌株的分类地位。共获得芽胞杆菌349株,基于菌落形态特征和16S rRNA基因序列确定了66个代表菌株,其中14株与近缘种模式菌株的16S rRNA基因序列相似性介于93.0–98.5%之间,为潜在芽胞杆菌新种。349株芽胞杆菌属于9属66种,分别为芽胞杆菌属(Bacillus)的41个种(41/66,62.1%),为最优势属;类芽胞杆菌属(Paenibacillus)有9种(9/66,13.6%),赖氨酸芽胞杆菌属(Lysinibacillus)共6种(6/66,9.1%),嗜冷芽胞杆菌属(Pscychrobacillus)共4种(4/66,6.1%),虚构芽胞杆菌属(Fictibacillus)有2种(2/66,3.0%),短芽胞杆菌属(Brevibacillus)、大洋芽胞杆菌属(Oceanobacillus)、鲁梅尔芽胞杆菌属(Rummelibacillus)和解硫胺素芽胞杆菌属(Aneurinibacillus)皆为1种。新疆9个地点马铃薯根际土壤中芽胞杆菌含量为2.20–8.86×104 cfu/g,其中特克斯蒙古乡马铃薯根际土壤中芽胞杆菌的菌落含量和种类最多,分别为8.86×104 cfu/g和37种;尼勒克县马场马铃薯根际土壤中的芽胞杆菌含量最少,特克斯去昭苏路上和阜康西沟村的芽胞杆菌种类最少,仅9种。但每个地点的优势种均相同,均为简单芽胞杆菌(Bacillus simplex)和阿氏芽胞杆菌(B.aryabhattai)。芽胞杆菌种类可划分为高频度分布型(简单芽胞杆菌等4种)、中频度分布型(植物内生芽胞杆菌等12种)和低频度分布型(其余51种芽胞杆菌,如蕈状芽胞杆菌)。新疆9个地点芽胞杆菌种类分布可分为高含量高丰度型(特克斯蒙古乡和农技推广园)和低含量低丰度型(如尼勒克县马场等地)。相关性分析发现,马铃薯根际芽胞杆菌种类分布与海拔高度无显著相关性。新疆马铃薯根际土壤蕴含种类丰富的芽胞杆菌资源,可为芽胞杆菌功能菌株开发提供丰富的菌种来源。     

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。     

基于可培养法分析亚热带植物内生与根际芽胞杆菌种类分布异质性

【背景】芽胞杆菌是农业上重要的微生物菌剂,对植物具有促生、防病、防虫等作用。【目的】了解亚热带植物内生和根际芽胞杆菌的种群分布,为其功能挖掘提供科学依据。【方法】采用可培养手段对甘蔗(Saccharum officinarum)、红麻(Hibiscus cannabinus L.)、黄麻(Corchorus capsularis L.)、黄秋葵(Abelmoschus esculentus)和玫瑰茄(Hibiscus sabdariffa)根际土壤和根内生芽胞杆菌进行分离,利用16S rRNA基因测序对分离菌株进行分类鉴定,并分析系统发育地位。【结果】共获得了可培养芽胞杆菌菌株144株,其中根内生芽胞杆菌82株,根际土壤62株。经16S rRNA基因测序鉴定为芽胞杆菌4个属37个种,分别为芽胞杆菌属、短芽胞杆菌属、类芽胞杆菌属和赖氨酸芽胞杆菌属。芽胞杆菌菌落数量和种类在根部及其根际土壤中差异较大,根际土壤中芽胞杆菌菌落数量远远大于根部,根际土壤中芽胞杆菌菌落含量范围为(0.2-370.0)×10~5CFU/g,而根部为(0.1-81.0)×10~3 CFU/g。根部分离获得的芽胞杆菌种类远大于根际土壤中,从作物根际土壤中共获得了芽胞杆菌19个种,从根部获得内生芽胞杆菌共32个种。阿氏芽胞杆菌(Bacillus aryabhattai)、仙草芽胞杆菌(Bacillus mesonae)和假蕈状芽胞杆菌(Bacillus pseudomycoides)同时存在于4种作物的根际土壤和根部,其他芽胞杆菌种类仅存在于1种作物的根际或者根部。【结论】亚热带作物根际土壤和根部芽胞杆菌种类和数量极为丰富,而且还存在可分离培养的芽胞杆菌潜在新物种,这为了解植物与根际微生物相互作用、根际环境生态平衡提供了理论基础和科学依据。     

芦笋老茎堆肥中嗜热细菌的分离与鉴定

利用稀释涂布法对芦笋老茎堆肥不同发酵阶段6个样品中的嗜热细菌进行分离,并采用16S rDNA序列分析方法对分离得到的菌落形态有明显区别的22株细菌进行鉴定.根据16S rDNA序列分析结果,22株细菌菌株中13株属于芽胞杆菌属(Bacillus),1株属于类芽胞杆菌属(Paenibacillus),4株属于假黄色单胞菌属(Pseudoxanthomonas),1株属于肠杆菌属(Enterobacter),1株属于副球菌属(Paracoccus),1株属于短芽胞杆菌属(Brevibacillus),菌株D-b2在GenBank数据库中未找到与其相似的已知细菌属的序列,分类地位待定.从以上鉴定结果可以看出,芦笋老茎堆肥中的优势嗜热细菌主要是芽胞杆菌(Bacillus spp.)和假黄色单胞菌(Pseudoxanthomonas spp.).     

1株抗菌植物内生菌EJH-2菌株的分离和鉴定

目的从中药植物金银花的组织中分离到1株具有抗菌活性的植物内生菌EJH-2菌株,并对其进行了分子生物学鉴定。方法通过总DNA提取,PCR扩增,得到1300bp的16S rRNA序列。PCR产物序列通过BLAST软件在NCBI网站中进行同源性比较。通过Bioedit7．0和Treedrawing软件绘制系统发育树。结果EJH-2的16SrRNA序列和数据库中的类多粘芽胞杆菌KCTC 1663菌株的序列的同源性为99．76％。在系统发育树中,EJH-2菌株和多粘类芽胞杆菌在同一分支。结论EJH-2菌株应归属于多粘类芽胞杆菌（Paenibacillus polymyxa）。     

四川海螺沟冰川土样芽胞杆菌资源分析

【背景】芽胞杆菌(Bacillus-like)是一类能形成具有强抗性芽胞且可在多种极端环境下存活的细菌,其产生的多种功能代谢产物在多种领域具有重要研究价值。由于冰川低温、寡营养的独特生态环境,其存在的芽胞杆菌可能具有特殊性,因此研究冰川芽胞杆菌有利于发掘新基因、丰富芽胞杆菌多样性。【目的】了解四川海螺沟冰川土壤芽胞杆菌资源,为挖掘芽胞杆菌新资源提供基础。【方法】采用纯培养法分离获得冰川土壤芽胞杆菌资源,利用16S r RNA基因进行系统发育分析,测定代表性菌株的生理生化特性并采用类平均法和欧氏距离模型进行聚类分析。【结果】共筛选到可培养细菌44株,经16S r RNA基因鉴定确定其中36株为芽胞杆菌,隶属于4个属的19个种,分别为芽胞杆菌属(Bacillus)11个种24株、类芽胞杆菌属(Paenibacillus)2个种3株、短芽胞杆菌属(Brevibacillus)4个种5株和赖氨酸芽胞杆菌属(Lysinibacillus)2个种4株,其中以芽胞杆菌属(Bacillus)为优势属。分离菌中仅3株芽胞杆菌可在4°C生存,7株能在50°C生长,大部分菌株在30°C下生长良好;有74%菌株能耐碱,有37%菌株能在无盐条件下生长。根据生理生化结果,采用类平均法和欧氏距离模型进行聚类分析,可分为3组,分别包含7种、4种和6种芽胞杆菌。第1组均可以水解七叶灵和利用葡萄糖,第2组均不能水解七叶灵和利用葡萄糖,第3组仅能共同利用葡萄糖。【结论】四川海螺沟冰川土壤蕴藏着较为丰富的芽胞杆菌资源,为芽胞杆菌新资源挖掘提供了资源保障。     

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.     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.     

Composition and diversity of bird communities in Swedish farmland–forest mosaic landscapes

CapsuleThe amount of forest (at local and landscape scales) and occurrence of residual habitats at the local scale are shown to be the major factors influencing bird community composition in farmland–forest landscapes in central Sweden.

Aims To investigate the importance of local habitat and landscape structure for breeding birds in farmland–forest landscapes in central Sweden.

Methods Breeding birds were censused at 292 points. A detailed habitat mapping was made within 300 m of the points. Within a 300–600 m radius only two major habitats (forests and arable fields) were identified.

Results Cluster analyses of bird communities identified three site types that also differed in habitat composition: (i) partially forested sites in forested landscapes; (ii) heterogeneous sites with residual habitats in mosaic landscapes; and (iii) field-dominated farmland sites in open landscapes. A total of 19 of 25 farmland bird species (restricted to farmland or using both farmland and forest) had the highest abundance in farmland sites with mosaics of forest and farmland, while only six farmland species had the highest abundance in field-dominated sites. The bird community changed from being dominated by farmland species to being dominated by forest species (common in forest landscapes without farmland) at small proportions (10–20%) of forest at the local scale. A major difference in habitat composition between heterogeneous and field-dominated sites was the occurrence of different residual habitats (e.g. shrubby areas and seminatural grasslands). These habitats seemed to influence bird community composition more than land-use, despite covering <10% of the area. Seminatural grasslands were important for bird community composition and species-richness, but grazing seemed to be less important. Among different land-use types, cereal crops were the least preferred fields. Set-asides with tall vegetation and short rotation coppices were positively associated with species-richness of farmland birds.

Conclusion In general, the composition of the landscape was important for bird community composition, although amount and distribution of forests, occurrence of residual habitats and land-use of fields at the local scale had the strongest influence on bird community composition. The possible implications of these patterns for managing farmland–forest landscapes are discussed.     

Chemical constituents from the fruits of Rhus typhina L. and their chemotaxonomic significance

This work describes the isolation and characterization of twenty-nine compounds from the fruits of Rhus typhina L., including eleven flavonoids (1–11), eleven phenols (12–22), two pentacyclic triterpenes (23–24), two organic acids (25–26), one lumichrome (27), one courmarin (28) and one pyrimidine (29) on the basis of their spectroscopic data. Compounds apigenin (1), daidzein (4), orobol (5), 3′, 5, 5′, 7-tetrahydroxyflavanone (6), naringenin (7), butein (8), (-)-catechin (9), quercetin-3-O-α-L-(3″-O-galloyl)-rhamnoside (11), 2-hydroxybenzoic acid (13), 4-hydroxybenzaldehyde (14), vanillin (15), methyl 3,4-dihydroxybenzoate (16), 3,5-dihydroxybenzamide (18), tyrosol (19), caffeic acid (20), 3-(2,4,6-trihydroxyphenyl)-1-(4-hydroxyphenyl)-propan-1-one (21), phlorizin (22), friedelin (23), oleanolic acid (24), 4,4-dimethyl-heptanedioic acid (25), anthranilic acid (26), lumichrome (27), scoparone (28) and uracil (29) have not been recorded before in this plant. This is the first report on the occurrence of compounds 4–7, 9, 11, 13–14, 16, 18–21, 25–29 from the genus Rhus. Moreover, the chemotaxonomic significance of these isolated compounds was also summarized.