培菌白蚁菌圃和粪便微生物多样性分析

【背景】培菌白蚁是属于白蚁科的一类与鸡枞菌属真菌共生的高等白蚁,其与体内肠道微生物和体外菌圃微生物形成三维共生体系。【目的】分析培菌白蚁菌圃和粪便的微生物多样性,并与肠道微生物进行比较。【方法】通过Illumina MiSeq高通量测序方法对培菌白蚁菌圃和粪便样品进行细菌16S rRNA基因和真菌ITS测序分析。【结果】高通量测序获得培菌白蚁菌圃和粪便样品细菌和真菌的有效序列和OTU数目。5个样品细菌OTU数目在90-199之间,而真菌OTU在10-58之间,细菌的种类多样性明显大于真菌。不论是细菌还是真菌,粪便样品的OTU数目多于菌圃样品。经物种分类分析,菌圃样品主要优势细菌是变形菌门(Proteobacteria),其相对含量超过82.4%;其次是拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes);粪便样品中优势细菌为拟杆菌门,其次是变形菌门,粪便优势菌属为别样杆菌属和营发酵单胞菌属,这与培菌白蚁肠道菌多样性组成一致。培菌白蚁菌圃和粪便样品共生真菌主要为担子菌门(Basidiomycota)和子囊菌门(Ascomycota)。菌圃优势真菌为鸡枞菌属(Termitomyces),相对含量在51.83%以上,菌圃中还鉴定到炭角菌属(1%,Xylaria)。【结论】为今后培菌白蚁-体内外微生物共生关系研究以及微生物的分离培养提供了依据和参考。     

传统醋曲储存过程中的微生物多样性

【背景】醋曲是我国传统谷物醋酿造中的重要微生物来源,通常一次制备分批使用。【目的】解析传统醋曲储存过程中微生物群落结构变化规律。【方法】从山西晋南一家百年老醋坊分别采集大曲原料、新制醋曲、储存7个月和12个月的醋曲,利用高通量测序技术分析微生物多样性。【结果】从4组样品中共找到610个真菌可操作分类单元(operational taxonomic unit, OTU)和747个细菌OTU。子囊菌门(Ascomycota,占比95%)和厚壁菌门(Firmicutes,占比81%)分别为优势的真菌和细菌类群。醋曲成品中约1/3的真菌OTU和约95%的细菌OTU可在醋曲原料中找到,说明原料是醋曲的重要微生物来源。相较于新制醋曲,储存7个月和12个月醋曲中的真菌和细菌多样性均显著降低。醋曲贮存过程中微生物群落结构发生明显改变,并且相较于真菌群落结构,细菌群落结构更易波动。相较于醋曲原料,醋曲成品中显著富集扣囊复膜孢酵母(Saccharomycopsis fibuligera)、东方伊萨酵母(Issatchenkiaorientalis)等真菌,以及克罗彭斯特菌属(Kroppenstedtia)...     

南极三洋藓(Sanionia uncinata)细菌群落组成分析

【背景】南极苔藓中蕴藏着丰富的极地微生物资源,然而目前对南极苔藓细菌群落组成的研究较少,限制了对这类极端生境微生物资源的开发和应用。【目的】在南极长城站不同地点随机取样,揭示南极三洋藓(Sanioniauncinata)的细菌群落结构组成。【方法】采用IlluminaHiSeq高通量测序技术对3个苔藓样品的16S rRNA基因V4区进行测序及生物信息学分析。【结果】3个苔藓样品中共获得细菌总优化序列273 367条。基于97%序列相似度,细菌优化序列聚类为9 579个OTU。分类地位明确的细菌可归为14门27纲50属。优势类群是变形菌门(Proteobacteria)(30.70%)、拟杆菌门(Bacteroidetes)(19.67%)、疣微菌门(Verrucomicrobia)(12.43%)、蓝细菌门(Cyanobacteria)(10.55%)及放线菌门(Actinobacteria)(9.36%)。在属水平上,56.73%属于未知细菌。【结论】南极三洋藓(Sanionia uncinata)中具有丰富多样的细菌,该研究为今后极地微生物研究奠定了基础。     

基于高通量测序研究青藏高原茶卡盐湖微生物多样性

【目的】茶卡盐湖(Chaka Salt Lake,CSL)是青藏高原有名的天然结晶盐湖,具有独特的石盐盐湖矿床,盛产青盐。盐湖卤水环境中存在丰富的嗜盐菌资源和潜在的新种,细菌和古菌的群落结构特征和物种多样性尚不明确。【方法】采用Illumina高通量测序平台对茶卡盐湖水样和底泥混合物中的细菌和古菌群落进行16S r RNA基因(V3-V5区)高通量测序,检测4个样本的群落结构差异和微生物多样性。【结果】获得细菌和古菌总有效序列分别为117 192和110 571条。结果分析表明细菌和古菌的物种注释(Operational taxonomic unit,OTU)数目分别为421和317,获得分类地位明确的细菌种类为14门28纲170属,古菌为5门4纲34属。细菌的优势类群是厚壁菌门(Firmicutes),所占比例为68.37%,其次为变形菌门Proteobacteria(20.49%);优势种属依次为芽孢杆菌属Bacillus(41.94%)、海洋芽孢杆菌属Oceanobacillus(8.03%)、假单胞菌属Pseudomonas(7.67%)、盐厌氧菌属Halanaerobium(7.42%)和乳球菌属Lactococcus(7.38%);古菌的优势类群以广古菌门(Euryarchaeota)盐杆菌纲(Halobacteria)为主,优势菌是Halonotius(17.21%)和盐红菌属Halorubrum(16.23%)。【结论】揭示了茶卡盐湖中细菌和古菌的群落结构及物种多样性,为嗜盐菌的开发及后续微生物资源的挖掘提供了理论依据。     

基于高通量测序的褐飞虱肠道微生物多样性分析

【目的】探明褐飞虱Nilaparvata lugens成虫肠道微生物群落结构和多样性。【方法】分离褐飞虱成虫完整肠道并提取总DNA,利用Illumina MiSeq(PE300)技术对其肠道细菌16S rRNA的V3-V4变异区和真菌ITS2序列进行测序,统计肠道微生物的操作分类单元(operational taxonomic unit, OTU)数量,分析其物种组成、丰度及Alpha多样性。并通过qPCR技术验证随机挑选注释到的4种肠道菌的高通量测序数据的有效性。【结果】分别获得褐飞虱成虫肠道细菌16S rRNA和真菌ITS2优质序列32 395和24 986条,根据序列相似性进行聚类分析分别获得235和128个OTUs。其中,细菌共注释到7个门, 15个纲, 26个目, 45个科和73个属;真菌共鉴定到3个门, 9个纲, 12个目, 15个科和18个属。在门分类水平上,细菌以变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)为优势门类;真菌以子囊菌门(Ascomycota)为绝对优势菌门。在属分类水平上,细菌的优势属为不动杆菌属Acinetobacter以及紫单胞菌科(Porphyromonadaceae)未确定属和毛螺菌科(Lachnospiraceae)未确定属,其丰度分别为36.37%, 17.22%和15.01%;真菌的优势属为粪壳菌纲(Sordariomycetes)未确定属,丰度为95.77%。Alpha多样性分析结果显示,褐飞虱肠道细菌(真菌)的观测物种数、Chao1指数、Shannon指数和Simpson 指数分别为235(128), 262.64(165.40), 3.90(0.91)和0.62(0.75)。4种肠道菌的qPCR结果显示高通量测序数据具有较高的有效性。【结论】褐飞虱成虫肠道细菌和真菌群落整体多样性比较丰富。研究结果为从共生微生物角度解析褐飞虱的环境适应性以及开发基于微生物防治的新技术等方面提供了依据。     

新疆蟠桃叶附生酵母种群结构分析

通过Illumina MiSeq平台, 对石河子143团两个蟠桃园所采集的叶片样本附生酵母菌26S rDNA D1/D2区进行高通量测序, 并分析了其物种的组成、Alpha多样性以及菌种群落结构。结果显示,两个桃园中共检测到1470个OTU (Operational Taxonomic Units, 操作分类单元), 桃园1中含有个1217个OTU, 桃园2中含有609个OTU, 其中有356个为共有OTU。共鉴定出酵母菌2门22属, 属于担子菌门(Basidiomycota)、子囊菌门(Ascomycota), 两个桃园共有的酵母属有线黑粉酵母属(Filobasidium), 接合酵母属(Zygosaccharomyces), 红酵母属(Rhodotorula), 隐球酵母属(Cryptococcus), Papiliotrema, 毕赤酵母属(Pichia), 假丝酵母属(Candida), Kwoniella, Cystobasidium, 两个桃园优势属均为线黑粉酵母属。研究结果表明石河子地区蟠桃叶表面附生酵母菌资源丰富, 值得进一步开发与利用。     

白蜡虫蜡花微生物多样性分析

【目的】探究白蜡虫Ericerus pela(Chavannes)雄若虫分泌的蜡花中微生物的多样性,比较不同厚度蜡花(随雄若虫发育蜡质层厚度增加)中细菌及真菌的种类和变化。【方法】采用Mi Seq高通量测序方法对不同发育阶段白蜡虫雄若虫分泌的蜡花中细菌16S rRNA和真菌ITS进行测序,利用Mothur软件计算蜡花中细菌和真菌的操作分类单元(operational taxonomic unit,OTU)及其丰富度、Alpha多样性分析。【结果】细菌3个样品共获470个OTU,真菌2个样品共获264个OTU。细菌共鉴定出18门30纲92科173属;真菌共鉴定出3门15纲43科68属。其中丛毛单胞菌属Comamonas细菌在蜡花中为优势菌,分别占前、中、后期雄若虫分泌的蜡花中细菌总量的79.79%,41.61%和46.86%;枝胞属Cladosporium在前期雄若虫分泌的蜡花中占真菌总量的17.01%,隐球菌属Cryptococcus在后期雄若虫分泌的蜡花中占真菌总量的60.84%,是蜡花中真菌重要组成部分。【结论】不同发育阶段雄若虫分泌的蜡花中细菌及真菌群落存在差异,随着蜡花厚度增加,细菌多样性先增加后减少,而真菌多样性降低。     

高通量测序分析云南腾冲热海热泉微生物多样性

【背景】云南腾冲热海热泉中蕴含着丰富的极端微生物资源。【目的】揭示云南腾冲热海热泉中微生物物种多样性及群落结构差异,发掘酸性热泉中铁、硫氧化功能微生物。【方法】采用Illumina HiSeq高通量测序技术对3处热泉15个水体样品中微生物16SrRNA基因V4-V5区进行测序及生物信息学分析。【结果】3处热泉中共获得578061条有效序列,聚类为141个可操作分类单元(Operational taxonomic unit,OTU),包括19个门66个属。鼓鸣泉(GMQ)、蛤蟆嘴(HMZ)、黄瓜箐(HGQ)3处热泉均以泉古菌门(Crenarchaeota)和厚壁菌门(Firmicute)为主。从属水平分析,碱性热泉鼓鸣泉(GMQ)和中性热泉蛤蟆嘴(HMZ)分别注释到37、32个属,优势属均为芽孢杆菌属(Bacillus)和热棒菌属(Pyrobaculum)。酸性热泉黄瓜箐(HGQ)共注释到20个属,优势属为酸杆菌属(Acidibacillus)和酸硫杆状菌属(Acidithiobacillus),此外,具有铁、硫氧化潜力的菌属有喜酸菌属(Acidicaldus)、硫化芽孢杆菌属(Sulfobacillus)、硫化叶菌属(Sulfolobus)及生金球菌属(Metallosphaera)等,进一步通过硫氧化培养基分离获得了这些菌属中的纯菌株。【结论】云南腾冲热海热泉水体中蕴含丰富的微生物资源,热泉间微生物物种组成差异明显;酸性热泉中存在多种具有潜在铁、硫代谢功能的菌种;未分类类群、非培养类群丰度很高,尤其是蕴藏着可观的古菌资源。     

枯草芽胞杆菌菌肥对有机冬瓜根区土壤微生态的影响

【背景】微生物肥料已广泛应用于我国有机作物的种植,其对有机种植土壤微生态的影响尚需科学评测。【目的】高通量测序技术可用于精确分析土壤微生物群落,从细菌、真菌群落结构和多样性的角度阐释枯草芽胞杆菌菌肥对有机农田根区土壤微生物群落的影响。【方法】在有机农田轮作种植条件下,施用枯草芽胞杆菌菌肥后提取冬瓜根区土壤基因组DNA,通过PCR扩增建立文库,利用IlluminaMiSeq高通量测序技术,并结合相关生物信息学方法分析土壤细菌16SrRNA基因V3-V4区和真菌ITS1区的多样性指数及群落结构;测定根区土壤化学性质及酶活性,分析有机冬瓜果实品质,并作相关分析。【结果】从6个有机冬瓜根区土壤样本中获得14199个细菌操作分类单元(OTU)和3378个真菌OTU,细菌和真菌文库测序覆盖率分别在98%、99%以上。枯草芽胞杆菌菌肥会在一定程度上提高土壤细菌种群多样性而降低真菌种群多样性,丰富了细菌群落结构,但显著降低了真菌群落丰富度(P0.05);并减少了根区土壤特有细菌和真菌物种。变形菌门、厚壁菌门和放线菌门是优势细菌,子囊菌门是优势真菌;枯草芽胞杆菌菌肥会提高绿弯菌门和子囊菌门的相对丰度,比例分别为46.23%、10.01%;降低变形菌门和担子菌门的相对丰度,比例分别为11.14%、74.72%。枯草芽胞杆菌菌肥显著降低了土壤pH,显著提高了有机冬瓜果实总氨基酸、可溶性固形物等营养成分含量(P0.05)。【结论】施用枯草芽胞杆菌菌肥改变有机冬瓜根区土壤细菌和真菌的丰富度和多样性,降低了土壤pH,提高了有机冬瓜果实品质。     

猎豹(Acinonyx jubatus)肠道微生物多样性及其性别差异研究（英文）

【目的】探讨猎豹(Acinonyx jubatus)肠道微生物多样性特征。【方法】通过采集新鲜粪便样品,对9只健康成年野生猎豹(4只雄性,5只雌性)的肠道微生物16S rRNA基因V3–V4区进行高通量测序,对猎豹肠道微生物多样性进行研究。【结果】测序共获得肠道微生物16S rRNA基因V3–V4区有效序列599349条,序列平均长度405 bp。通过以97%的序列相似性进行分类,共获得操作分类单元(OTU)268个。经序列比对和分类鉴定,这些OTU都属于细菌域,包括10个门,21个纲,35个目,72个科,144个属。其中,丰度最高的5个细菌门是厚壁菌门(Firmicutes,平均占OTU总数的42.29%%)、放线菌门(Actinobacteria,31.54%)、梭杆菌门(Fusobacteroidetes,16.66%)、变形菌门(Proteobacteria,5.30%)和拟杆菌门(Bacteroidetes,4.19%)。拟杆菌门的丰度较低是猎豹肠道微生物的主要特征。丰度最高的5个科依次是红蝽杆菌科(Coriobacteriaceae,31.28%)、消化链球菌科(Peptostreptococcaceae,平均占17.66%),梭杆菌科(Fusobacteriaceae,15.46%)、毛螺菌科(Lachnospiraceae,12.40%)、梭菌科I(Clostridiaceae_I,6.93%)等。丰度最高的5个属依次是柯林斯氏菌属(Collinsella,30.16%)、梭杆菌属(Fusobacterium,15.46%)、艰难梭菌属(Peptoclostridium,11.46%)、Blautia属(8.28%)和狭窄梭菌属1(Clostridium_sensu_stricto_1,6.39%)。约有2.32%的OTU没有归类到属。群落alpha多样性分析结果显示,猎豹肠道微生物群落Shannon指数为2.93–4.41,Simpson指数为0.72–0.91。通过依据性别进行分组,对雌雄两组之间的alpha多样性比较表明,雄性组的物种和Shannon指数略高于雌性组。Beta多样性分析表明,雌雄两组之间的差异高于各组内部不同个体之间的差异。然而,聚类分析显示,相同性别的猎豹的肠道微生物并没有聚在一起。【结论】本文通过高通量测序技术研究了猎豹肠道微生物多样性特征和性别差异,为猎豹的保护、救护饲养和消化生理学研究提供了基础数据。     

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.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

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.     

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。     

Time of detection of recessive genes: effects of system of mating and number of examined individuals

Summary Anthers were cultured from two sets of seven lines of hexaploid wheat (Triticum aestivum L.) with different cytoplasms, the euplasmic nucleus donors, Siete Cerros 66 and Penjamo 62, as well as their six alloplasmic lines derived from wild relative species of the genera Triticum and Aegilops. Significant cytoplasmic and nuclear effects but no cytoplasmic-nuclear interaction were found for embryogenic anther response, with the best performance of Penjamo 62 in Ae. kotschyi cytoplasm. Plant regeneration was not affected significantly by the cytoplasmic background of the lines cultured. The possible genetic implications of the observed cytoplasmic and nuclear influences on the in vitro haploid induction of wheat are discussed.     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.