基于高通量测序分析桃蚜体内微生物群落结构及多样性

为探明桃蚜Myzus persicae体内微生物群落结构及其种类多样性,采用Illumina HiSeq二代测序技术检测桃蚜体内细菌16S rRNA基因和真菌ITS基因序列的方法,分析取食白菜Brassica pekinensis和甘蓝Brassica oleracea的无翅孤雌桃蚜成虫体内微生物群落结构及多样性。研究结果获得桃蚜体内细菌16S rDNA和真菌ITS1优质序列分别为473 750条和472 980条,并根据序列相似性对其进行聚类分析,分别获得959个和1 424个OTUs。基于OTUs分类结果,共注释鉴定细菌类群26个门、55个纲、128个目、227个科、419属、451种,真菌类群10个门、31个纲、77个目、172个科、343属、441种。其中,在门级水平上,取食白菜和甘蓝的桃蚜体内细菌类群均以变形菌门Proteobacteria内的细菌（占73.11%,80.10%）为优势菌;真菌类群均以子囊菌门Ascomycota真菌（占51.91%,50.98%）为优势菌。在属级水平上,取食白菜和甘蓝的桃蚜体内细菌均以布赫纳氏菌属Buchnera（占60.82%,56.11%）为优势属,而其次优势细菌属则分别为未培养的叶绿体菌Chloroplas（占2.84%）和立克次氏小体属Rickettsiella（占10.04%）;取食白菜的桃蚜体内真菌优势属为青霉属Penicillium（占5.15%）,次优势菌属为被孢霉属Mortierella（占3.83%）,而取食甘蓝的桃蚜体内真菌优势属为曲霉属Aspergillus（占3.93%）,次优势菌属为被孢霉属Mortierella（占3.86%）。以上研究结果表明取食不同植物的桃蚜体内微生物群落结构和种类多样性存在一定差异。α多样性指数表明桃蚜体内微生物群落结构具有较高的丰富度和多样性。研究结果为进一步挖掘桃蚜体内关键微生物,并解析其生物学功能及其利用等方面奠定了重要基础。     

短花针茅荒漠草原土壤微生物群落组成及结构

为详细了解内蒙古短花针茅荒漠草原生态系统中土壤微生物群落组成与结构。对其土壤中微生物的总DNA提取后,采用高通量测序技术对土壤中细菌的16Sr DNA和真菌的ITS基因进行了序列测定,分析了短花针茅荒漠草原土壤中微生物群落结构特征。结果表明,共获得细菌OTUs13711个,真菌OTUs 5929个。物种分类显示,细菌种类隶属于29门57纲111目191科485属,其中优势类群为Gammaproteobacteria和Thermoleophilia,它们的相对丰度分别为32.68%和26.83%。真菌隶属于4门16纲45目78科105属,优势类群为Ascomycota和Basidiomycota,它们的相对丰度分别为35.76%和25.90%。土壤中细菌群落的多样性和丰富度均高于真菌。     

毛黄堇根腐病根际土壤微生物群落结构及多样性

目的：研究健康毛黄堇植株和患根腐病毛黄堇植株根际土壤微生物群落多样性和组成差异，探讨其与毛黄堇病害发生的关系，为针对性开展防治工作提供科学依据。方法：利用Illumina MiSeq技术，比较健康和患病根际土壤微生物群落结构特征及差异。结果：患病根际土细菌群落α多样性降低，真菌群落α多样性升高。健康和患病植株细菌、真菌门水平的组成结构及相对丰度差异均较小，细菌优势门为变形菌门和放线菌门，真菌为子囊菌门。属水平上组成结构复杂，优势细菌属为Subgroup＿6等，优势真菌属为曲霉属。患病植株差异标志物种主要为小单孢菌科细菌和核盘菌科真菌。结论：土壤微生物群落多样性和结构与毛黄堇健康状态密切相关，致病真菌相对丰度增加可能是导致病害发生的重要原因。     

宁夏南部生态移民迁出区不同恢复模式土壤微生物群落特征

为了揭示人工林对土壤微生物环境的作用机理,利用高通量测序技术,比较了宁南山区刺槐、河北杨、油松、青海云杉和自然恢复林地的土壤真菌、细菌群落组成及多样性,分析了土壤理化性质与优势菌群的关系。结果表明: 1)不同恢复模式土壤真菌优势菌门为子囊菌门、担子菌门、被孢霉门、未分类真菌,占总真菌群落的90%以上;细菌优势菌门为放线菌门、变形菌门、酸杆菌门、绿弯菌门和其他细菌门,占总细菌群落相对丰度的80%以上。2)油松林地土壤真菌多样性最高,Shannon指数和 Simpson 指数分别为3.72±0.37、0.07±0.04。自然恢复林地真菌物种数量最高,Ace和Chao1指数分别为708.19±137.25、706.26±125.34;油松林地细菌多样性和物种数量都最高,Shannon、Simpson、Ace和Chao1指数分别为6.57±0.04、0.004±0.00、3439.81±41.67和3463.14±32.16。3)不同恢复模式相对丰度差异显著的真菌属为产油菌属、枝孢菌属、链格孢属,细菌属为67-14科未定名属、红色杆菌科红色杆菌属、鞘脂单胞菌科鞘脂单胞菌属。4)微生物优势菌群与土壤理化性质的冗余分析(RDA)表明,土壤容重(BD)、碳氮比(C/N)、pH是影响真菌优势菌群的主要因子,BD、氮磷比(N/P)、全磷(TP)、全碳(TC)是影响细菌优势菌群的主要因子。总体而言,不同恢复模式间真菌丰度差异性高于细菌丰度,表明真菌群落组成及其多样性对于不同树种和土壤环境变化较细菌群落更为敏感。研究结果将为宁南山区植被恢复措施及生态系统功能稳定性的维持提供理论支持。     

马缨杜鹃根系微生物群落结构及其变化

【目的】为了解野生马缨杜鹃根系微生物的群落结构特征,比较百里杜鹃内马缨杜鹃根际土壤、根表及根内3个生态位微生物群落组成差异,探究杜鹃根系生态位之间、变化菌属间的相互关系,以期为今后杜鹃根系微生物研究提供具有参考价值的数据。【方法】对百里杜鹃内野生马缨杜鹃根系微生物16S rRNA V4区和ITS1区进行高通量测序,比较分析根际土壤、根表及根内微生物群落多样性及组成差异;并基于共变化网络分析进一步探究差异菌属间的相互关系。【结果】马缨杜鹃根系不同的生态位之间,微生物群落结构存在差异,其中以根际土壤与根表、根内差异更为显著,且细菌群落差异小于真菌群落。且从根际-根表-根内,马缨杜鹃根系细菌α-多样性显著下降。马缨杜鹃根系微生物分布于41细菌门和6个真菌门。优势细菌门为变形菌门(Proteobacteria,39.64%±0.08,69.47%±0.12,77.34%±0.07)、酸杆菌门(Acidobacteria,34.10%±0.11,11.03%±0.04,9.18%±0.04)以及放线菌门(Actinobacteria,10.19%±0.03,8.70%±0.02,7.08%±0.01),它们占整体细菌群落总丰度的80%以上。同时8个细菌菌门的相对丰度在根际土壤、根表和根内间显著变化,且它们的相对丰度占细菌群落总丰度的75%以上。真菌主要分布于接合菌门(Zygomycota)、担子菌门(Basidiomycota)和子囊菌门(Ascomycota),它们占整体菌群的99%以上。马缨杜鹃根系存在589个属的细菌,390个属的真菌,从根际-根表-根内,其中25个细菌属和10个真菌属的相对丰度发生显著变化。马缨杜鹃根系微生物群落共变化网络分析表明:在马缨杜鹃根系不同生态位间,除Waitea外,包括枝孢菌属(Cladosporium)、拟盘多毛孢属(Pestalotiopsis)等在内的8个差异真菌菌属均与细菌菌属显著相关,它们相互作用调控微生物群落结构的变化。Bryobacter、Nocardia、Rhizomicrobium和Telmatobacter等核心菌属对马缨杜鹃根系微生物群落共变化网络的变化具有十分重要的调控作用。【结论】百里杜鹃地区马缨杜鹃根际土壤、根表以及根内3个生态位间,微生物群落组成存在差异;而造成微生物组成存在差异这一结果,可能与马缨杜鹃根系密切相关。同时,共变化网络分析揭示出马缨杜鹃根系生态位之间,细菌和真菌彼此间互作。     

广东省本地油茶和引种油茶根际土壤微生物群落特征

【背景】近年来,油茶低效林面积较大,根际土壤微生物影响林木抗性和生长,对林业可持续发展具有重要意义。【目的】了解广东省本地油茶和引种油茶根际土壤微生物群落特征。【方法】利用高通量测序分析油茶根际土壤微生物群落组成。【结果】油茶根际土壤细菌有26门77纲201目377科593属676种,真菌有14门50纲121目266科502属631种。油茶根际土壤中的优势细菌为酸杆菌门和变形菌门,优势真菌为子囊菌门和担子菌门。两种油茶根际土壤微生物组成差异显著,本地油茶根际土壤的细菌多样性显著高于引种油茶。在门水平上,脱硫杆菌门细菌和罗兹菌门、被孢霉门真菌的相对丰度在两种油茶间差异显著,Amorphotheca在本地油茶根际土壤中特异性富集。两种油茶根际土壤细菌碳代谢相对丰度差异显著,真菌以腐生营养型为主,其次为病理营养型和共生营养型。本地油茶根际土壤中显著富集土壤腐生菌,而共生营养型真菌(尤其是丛枝菌根真菌)相对丰度(6.43%)显著低于引种油茶中(21.83%)。此外,有机质和养分含量是影响油茶根际土壤微生物群落的关键因子。【结论】本地油茶和引种油茶根际土壤微生物群落组成和结构差异显著,Amorp...     

银北盐渍化土壤中6种耐盐植物根际细菌群落结构及其多样性

土壤微生物对土壤肥力的形成和植物营养的转化起着积极的作用。对盐渍化土壤中植物根际微生物群落组成和多样性进行研究,有助于发现新的重要微生物功能类群或者功能潜力,对于盐碱土壤的植被恢复和生态重建都具有十分重要的意义。通过高通量测序和分离培养方法,对宁夏银北地区盐渍化土壤中的6种耐盐植物根际细菌的群落结构和多样性进行了分析。结果表明:所有土壤样品共检测到31门(亚门) 67纲253科400属细菌类群,不同植物根际微生物群落组成相似,但丰度各有差异,厚壁菌门、变形菌门、放线菌门和拟杆菌门为主要优势类群;在纲水平芽孢杆菌纲相对丰度最高,其次为α-变形菌、γ-变形菌和梭菌纲;在属水平上,芽孢杆菌属以极高的丰度(15.57%—53.85%)占据绝对优势,其次是鞘氨醇单胞菌属、不动杆菌属和节杆菌属。6种植物根际细菌的群落结构组成表现为芨芨草和柽柳相似,柳枝稷、苜蓿与枸杞相似,苦豆子则与另外5种差别最大。分离培养出的110株根际细菌多数具有较强的盐碱耐受性和一种以上生物学活性,分别隶属于芽孢杆菌属、假单胞菌属、鞘氨醇杆菌属、节杆菌属和中华根瘤菌属,在属水平上多样性单一,其中芽孢杆菌属是优势属。传统的分离培养和高通量测序结果都反映出银北地区盐渍化土壤中定殖着许多有重要生态功能的微生物类群,可用于进一步筛选和利用根际微生物改良盐碱土壤。     

微生物型育苗垫片对水稻根围土壤微生物多样性的影响

[目的]分析微生物型育苗垫片对水稻根围土壤微生物多样性和群落组成的影响,为微生物型育苗垫片的示范应用提供科学参考。[方法]采用水稻秧盘育苗试验,基于常规微生物培养和高通量测序技术测定微生物型垫片育苗对水稻根围土壤微生物种群数量、多样性及群落组成变化。[结果]与普通壮秧剂育苗相比,微生物型垫片育苗的水稻根围土壤细菌数量在出苗后第7 d增加了36. 59%,第14 d和第21 d时降低但差异不显著;放线菌数量分别显著增加了271. 11%、702. 48%和100%;真菌数量在第7 d和第14 d分别降低了127. 05%和740%,第21 d时明显增加了160. 98%。在水稻幼苗根围微生物群落组成中,变形菌门、放线菌门、酸杆菌门和螺旋体菌门为优势细菌,子囊菌门、担子菌门、接合菌门和壶菌门为优势真菌;与对照相比,微生物型育苗垫片组中变形菌门增加,螺旋体菌门和酸杆菌门减少,其中高丰度的假单胞菌属、根瘤菌属、节杆菌属、克雷伯氏菌属等根围促生菌均为水稻幼苗根围优势菌;真菌的子囊菌门增加,担子菌门、接合菌门和壶菌门减少,后期子囊菌门成为单一优势菌,该门的腐质霉属、假裸囊菌属和接合菌门的被孢霉属菌为优势菌。[结论]微生物型垫片育苗在水稻幼苗前期可提高土壤细菌的丰富度,后期降低土壤真菌的丰富度和多样性,具有改善水稻幼苗根围微生态环境的作用。     

基于高通量测序的禁牧对土壤微生物群落结构的影响

麋鹿的采食、躺卧和践踏行为均会对栖息地土壤环境造成影响,进而影响土壤微生物群落结构。利用高通量测序技术,分析江苏大丰麋鹿国家级自然保护区禁牧点和补饲点土壤细菌和真菌群落结构差异,并结合土壤理化性质探究禁牧对土壤微生物群落结构的影响。结果表明细菌群落的优势菌门为变形菌门,真菌群落的优势菌门为子囊菌门。禁牧改变了土壤微生物群落结构,在门水平上提高了变形菌门、放线菌门和担子菌门的相对丰度,降低了绿弯菌门、厚壁菌门和子囊菌门的相对丰度,禁牧点与补饲点土壤微生物群落多样性的相似性较低。冗余分析中,细菌受土壤环境因子的影响大于真菌,其中土壤pH是影响细菌和真菌群落最大的土壤环境因子。研究揭示了禁牧对土壤微生物群落结构的影响,为保护区制定麋鹿生境恢复方案提供参考。     

热带珊瑚岛植物种植对土壤改良及其微生物群落形成的影响

植被及关联的土壤微生物对于维持热带珊瑚岛的生态系统功能和稳定性具有重要作用。采集了植物种植前的种植土和种植早期珊瑚砂土,以及栽植幼苗基质中的根际土和非根际土,利用扩增子测序的手段,分析其中的真菌和细菌的多样性、群落组成及变化。分析结果显示,种植土、幼苗基质中的非根际土和根际土可能是珊瑚砂土壤中微生物的重要来源,其中最主要的类群包括以担子菌门和接合菌门为代表的真菌,以及以变形菌门、酸杆菌门为代表的细菌。珊瑚砂土的真菌丰度（864.2±41.4,为每0.25 g土的物种数）显著低于来自苗基质的根际土（1086.1±64.3,P=0.014）和苗非根际土（1251.4±48.1,P < 0.001）;珊瑚砂土的细菌丰度与苗基质根际土和非根际土之间并没有显著性差异。通过对比种植土和植物种植后的珊瑚砂土的微生物群落β-多样性,也发现植物种植对真菌和细菌类群组成产生了影响;主要类群接合菌门真菌相对丰度从0.2%增加到17.4%,而伞菌纲真菌的相对丰度从20.8%下降到0.9%,β-变形菌纲细菌的相对丰度从17.7%下降到0.1%。研究结果启示,珊瑚砂土壤微生物的组成并非是对不同来源微生物进行简单的集合,其中生活史与植物关系密切的微生物类群,目前还未在珊瑚砂土壤中表现出优势分布;相反,一些在植物根际和非根际土中较少出现的微生物,在珊瑚砂土壤中则广为存在,从而说明土壤微生物群落恢复过程的复杂性。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Physiological characteristics of various species of strains of carboxydobacteria

Seven strains of aerobic carbon monoxide-oxidizing bacteria (carboxydebacteria) when growing on CO as sole source of carbon and energy had doubling times which ranged from 12–42 h. The activity profiles obtained after discontinuous sucrose density gradient centrifugation indicated that the CO-oxidizing enzymes are soluble and the hydrogenases are membrane-bound in all strains examined. The CO-oxidizing enzymes of Pseudomonas carboxydohydrogena, Pseudomonas carboxydoflava, Comamonas compransoris, and the so far unidentified strains OM2, OM3, and OM4 had a molecular weight of 230,000; that of Achromobacter carboxydus amounted to 170,000. The molecular weights of the CO-oxidizing and H₂-oxidizing enzymes turned out to be identical. The cell sonicates were shown to catalyze the oxidation of both CO and H₂ with methylene blue, thionine, phenazine methosulfate, toluylene blue, dichlorophenolindophenol, cytochrome c or ferricyanide as electron acceptors. Methyl viologen, benzyl viologen, FAD⁺, FMN⁺, and NAD(P)⁺ were not reduced. The spectrum of electron acceptors was identical for all strains tested. Neither free formate, hydrogen nor oxygen gas were involved in the CO-oxidation reaction. Methylene blue was reduced by CO at a 1:1 molar ratio. The results indicate that CO-oxidation by carboxydobacteria is catalyzed by identical or similar enzymes and that the reaction obeys the equation CO+H₂OCO₂+2H⁺+2e^- as previously shown for Pseudomonas carboxydovorans.Dedicated to Otto Kandler remembering almost three decades of enjoyable cooperation     

Modulation of allostery of pyruvate kinase by shifting of an ensemble of microstates

Since the introduction of the concepts of allostery about four decades ago, much advancement has been made in elucidating the structure-function correlation in allostery. However, there are still a number of issues that remain unresolved. In this review we used mammalian pyruvate kinase (PK) as a model system to understand the role of protein dynamics in modulating cooperativity. PK has a triosephosphate isomerase (TIM)(α/β)₈ barrel structural motif. PK is an ideal system to address basic questions regarding regulatory mechanisms about this common (α/β)₈ structural motif. The simplest model accounting for all of the solution thermodynamic and kinetic data on ligand-enzyme interactions involves two conformational states, inactive E^T and active E^R. These conformational states are represented by domain movements. Further studies provide the first evidence for a differential effect of ligand binding on the dynamics of the structural elements, not major secondary structural changes. These data are consistent with our model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the inactive E^T and active E^R represent the two extreme end states. Sequence differences and ligands can modulate the distribution of states leading to alterations of functions. The future work includes: defining the network of functionally connected residues; elucidating the chemical principles governing the sequence differences which affect functions; and probing the nature of mutations on the stability of the secondary structural elements, which in turn modulate allostery.     

（鱼句）亚科花（鱼骨）型鱼类骨骼系统的比较

对我国花型Hemibarbuspattern鱼类作了骨骼系统比较,结果表明,此类型鱼类脑颅较长,副蝶骨平直或稍弯曲,眶蝶骨腹纵嵴发达(铜鱼Coreius septentrionalis例外),下颞窝和咽突中等大,基枕骨后突发达;脑颅中的上筛骨的后突、侧筛骨的外筛突,蝶耳骨的外突、上耳骨的后突、围眶骨和后颞窝等均有明显的差异;咽颅中的舌颌骨、尾舌骨、鳃盖骨和下咽齿的列数等又有显著的区别;附肢骨骼中的腰带骨、脊椎骨中的复合神经骨和第4椎骨腹侧的悬器等也有不同之处。据此,这些差异和区别可作为属间或种间的分类依据。