神农架林区和自然保护区链霉菌拮抗性的研究(Ⅱ)

用琼脂移块法对由神农架林区和自然保护区土壤中分离并鉴定了的３６种５７株链霉菌进行拮抗性试验,结果显示３６种５５株链霉菌对细菌或真菌有拮抗性,或对细菌和真菌都有拮抗性,并发现这些链霉菌以较高的百分率拮抗丝状真菌和酵母菌。     

拉鲁湿地自然保护区放线菌组成分析及生物活性测定

探究拉鲁湿地自然保护区的放线菌组成及其抑菌和酶活性,为放线菌新药物先导化合物和高活性酶的筛选提供资源。从拉鲁湿地自然保护区不同土壤类型、不同优势植被采集25份土样。用分散差速离心法分离了拉鲁湿地中温放线菌和低温放线菌。从中温放线菌中选择15株代表菌株进行了初步分类鉴定。采用打孔法检测了其对2株细菌和4株病原真菌的抑菌活性。结果表明：（1）拉鲁湿地放线菌数量从水生环境向陆地生态系统递增,中温放线菌数量显著多于低温放线菌;（2）拉鲁湿地土壤中分离到链霉菌属、小单孢菌属、诺卡氏菌属、马杜拉菌属、小链孢菌属5个放线菌属。其中以链霉菌属和小单孢菌属为优势属。链霉菌属以金色类群、白孢类群和粉红孢类群为主,小单孢菌分离到黄橙类群和黑褐类群;（3）供试菌株分解纤维素能力较强,分解蛋白质活性较低,具有抗菌活性的菌株很少,且抗菌活性较弱;（4）供试菌株耐毒性物质的能力较强。这些菌可用于毒害有机物污染物的处理。     

西双版纳、白茫雪山、波罗的海南岸地衣纯培养放线菌多样性

【背景】在地衣共生系统中除了共生真菌、藻类以外,还蕴藏着丰富的放线菌资源。【目的】采集来自云南西双版纳、白茫雪山、德国波罗的海南岸3个地区的地衣,对获得的地衣纯培养放线菌进行多样性分析。【方法】采用3种放线菌选择分离培养基,通过平板稀释涂布法分离放线菌。通过比较16S rRNA基因序列相似性以及构建系统发育树,确定纯培养放线菌的分类地位。【结果】共分离菌株1 123株,鉴定417株。其中从西双版纳17份地衣样品中分离纯化到107株放线菌,分布在7个目14个科33个属,潜在新种18株,其中链霉菌为优势菌属;从白茫雪山7份地衣样品中分离纯化到103株放线菌,分布在4个目5个科9个属,潜在新种16株。其中链霉菌为优势菌属,占比39%;从波罗的海南岸5份地衣样品中分离纯化到65株放线菌,分布在4个目8个科18个属,潜在新种5株,潜在新种菌和链霉菌为优势菌属。【结论】在本研究条件下,西双版纳可培养地衣放线菌多样性较白茫雪山和波罗的海南岸丰富。白茫雪山地衣链霉菌居多,潜在新种占比15.5%。3个地区地衣放线菌的区系组成各不相同,这与3个地区地衣所处地理环境、完全不同的气候密切相关。     

古巴土壤中放线菌的研究

本文对古巴土壤中放线菌的生态分布进行了系统研究。土壤中的放线菌绝大部分均属链霉菌属。对水平分布的不同土类中链霉菌的数量、种类和种的组成以及不同海拔高度垂直分布情况均有详细介绍。并对林地、荒地、放牧、耕作等不同利用的情况下土壤中链霉菌的数量、种类和种的组成也进行了比较研究,提出了链霉菌生态地理分布规律的有力依据。放线菌是土壤中重要的微生物类群,对有机质的分解起着重要作用,同时也是抗生素的主要生产菌。近年来还发现放线菌能产一些植物生长刺激素,有的能与高等植物共生固定大气氮素、有的则对滴滴涕和西玛津等农药起到降解的作用。因此,研究土壤中放线菌的生态分布特别具有现实意义。     

广西山口红树林内生放线菌的分离、筛选及初步鉴定

从广西山口国家红树林生态自然保护区采集8种真红树植物和5种半红树植物的根、茎、叶及胚轴等组织样品共计41份。经过表面消毒后粉碎,分别以几丁质、腐殖酸、木聚糖等作为主要营养及能量来源设计了8种分离培养基,从植物组织中分离得到118株放线菌。使用6种新药筛选模型对分离菌株进行了生理活性筛选,77株放线菌的发酵液样品在一个或多个药物筛选模型中显示为阳性,初筛阳性率为65.3%。对77株筛选阳性菌株进行的初步分类研究结果显示,其中44株属于链霉菌属,25株为小单孢属的菌株,3株属于糖丝菌属,3株为诺卡氏属菌株,1株是拟诺卡氏属菌株,1株是伦茨氏菌属的菌株。对一株在抗多重耐药检定菌HH22和降血脂模型上都显示出良好活性的菌株I07A-01824进行表型和基因型分析,确定I07A-01824为黄白链霉菌(Streptomyces albidoflavus)菌株。该研究显示,红树植物内生环境不仅仅孕育了丰富多样的内生放线菌,而且红树植物内生放线菌也是新型天然生理活性次级代谢产物的有效来源。     

塔里木盆地荒漠盐碱生境土壤放线菌的初步研究

采用常规方法研究了新疆塔里木盆地荒漠盐碱生境土壤放线菌区系、放线菌的耐盐、碱性及对植物病原真菌的拮抗性。结果表明:土壤放线菌区系较单一,仅分离到链霉菌属和拟诺卡氏菌属,其中以链霉菌属为主(98.57%),链霉菌可分10个类群,以白孢类群为优势类群。土壤含盐量愈高,链霉菌组成愈简单。放线菌的最适生长盐浓度多集中在6%~8%,最适生长pH集中在7~8。24株供试放线菌中,有4株中度嗜盐放线菌对供试靶标菌有拮抗性,占供试菌的16.67%。     

山东半岛农药污染点源放线菌多样性及抑菌活性

【目的】探究山东半岛农药污染点源放线菌多样性及非链霉菌抑菌活性,试图发现新放线菌和新抗生素。【方法】通过16S rDNA序列对已分离得到的154株纯培养放线菌进行初步分类鉴定并进行系统发育分析;采用管碟法和菌丝生长速率法检测10株非链霉菌的抑菌活性。【结果】154株放线菌覆盖7个科,8个属,有一株非链霉菌(205)可能为潜在的新种。10株非链霉菌的发酵液均对供试的植物病原真菌和细菌有不同程度的抑制作用,尤其是菌株Microbacterium oxydans JN853773和Kocuria rosea JN192402对所有供试病原菌都有强烈的抑制作用。【结论】农药污染点源可以作为开发新型抗生素产生菌的重要来源。     

瑞拉菌素产生菌的鉴定

自陕西秦岭太白山土壤中分离到 1株编号为S 5 12 0的放线菌。根据对其生物特征鉴定、生理生化特征分析 ,它与链霉菌属中委内瑞拉链霉菌最为相近 ,但菌种S 5 12 0对梨黑星病菌、苹果腐烂病菌等多种引起植物病害的病原真菌有拮抗和溶菌作用 ,故认为S 5 12 0是委内瑞拉链霉菌的一个新变种 ,定名为委内瑞拉链霉菌秦岭变种(Streptomycesvenezuelaevar .qinlingensis.n .Var)。     

宁夏白芨滩自然保护区苦豆子内生放线菌多样性及其分布

【目的】探索宁夏干旱荒漠区苦豆子内生放线菌的多样性及区系组成,为苦豆子内生放线菌的合理开发和利用提供理论依据。【方法】从宁夏白芨滩自然保护区不同植被和土壤类型的6个样区采集健康苦豆子样品30份,采用组织匀浆法从苦豆子植株的根部、茎部、叶部和种子中分离内生放线菌,根据培养性状、菌落、孢子等的形态特征和16S rRNA基因序列分析对分离菌株进行鉴定;根据苦豆子内生放线菌的相对频率、物种多样性指数、丰富度指数和相似性系数分析其区系组成特点。【结果】从30份苦豆子样品中,共分离得到内生放线菌111株,这些菌株分属于链霉菌属(Streptomyces)、拟诺卡氏菌属(Nocardiopsis)、普劳斯氏菌属(Prauserella)、Actinophytocola、微杆菌属(Microbacterium)、束丝放线菌属(Actinosynnema)、嗜热油菌属(Thermoleophilum)、糖霉菌属(Glycomyces)和糖丝菌属(Saccharothrix) 9个属,其中链霉菌属(Streptomyces)和拟诺卡氏菌属(Nocardiopsis)为优势属。苦豆子内生放线菌的分布具有一定的组织特异性,根和种子中的内生放线菌数量高于茎和叶。植被类型中,沙生植被草原(样区Ⅳ)分离的苦豆子内生放线菌物种多样性指数最高,而荒漠草原(样区Ⅴ)最低。荒漠草原(样区Ⅴ)和林地(样区Ⅵ)内生放线菌群落有密切的相似性。【结论】苦豆子体内含有丰富的内生放线菌资源,其内生放线菌具有很高的宿主特异性,而且其分布受生境影响。     

人参病原菌拮抗放线菌的分离筛选与鉴定

采用平板对峙法、生长速率测定法等对人参根际分离得到的4株放线菌的抑菌活性进行了研究,并通过形态学特征观察、生理生化测定和16S rDNA序列分析进行分类鉴定。结果表明,4株放线菌对人参常见病病原真菌的抑菌效果明显。其中,FX-10对毁灭柱孢菌、灰葡萄孢菌和FX-61对人参核盘菌的抑制率分别达到91.28%、91.8%和90.07%。FX-10、FX-61、FX-137对人参的5种病原菌和FX-139对人参的7种病原菌有抑制作用,并且抑菌效果稳定。通过形态、生理生化和分子生物学鉴定,FX-139为酒红土褐链霉菌(Streptomyces vinaceus-drappus),FX-10为黑浅灰链霉菌(Streptomyces nigrogriseolus),FX-137为酒红链霉菌(Streptomyces vinaceus),FX-61为链霉属的灰褐类群。     

越南原始森林粘细菌的分离与鉴定

为了探讨越南热带原始森林环境中可培养粘细菌的多样性,以原始森林的土壤、腐木及树皮样品为研究对象,采用大肠杆菌法和滤纸法从中分离纯化粘细菌,将形态鉴定与16S rRNA基因测序分析相结合,对分离到的纯培养物进行分类鉴定,确定其分类地位。结果显示,从越南16个土样和22个腐木、树皮样品中共分离到70株粘细菌,经纯化后获得了32株。这些菌株分属于6个属,分别是粘球菌属（Myxococcus）14株、珊瑚球菌属（Corallococcus）7株、侏囊菌属（Nanncystis）6株、蜂窝囊菌属（Melittangium）3株、原囊菌属（Archangium）1株、软骨霉状菌属（Chondromyces）1株。这表明粘细菌在越南原始森林环境中分布广泛,并存在较为罕见的粘细菌类群。     

Nitrogen-fixing bacteria communities occurring in soils under different uses in the Western Amazon Region as indicated by nodulation of siratro (Macroptilium atropurpureum)

Understanding native communities is a crucial step for the management of biological nitrogen fixation, since they may be either a source of efficient strains or a limiting factor when efficient strains need to be introduced. This work aimed to evaluate the density, diversity and efficiency of Leguminosae nodulating bacterial (LNB) communities and their component strains in soils under various land use systems (LUSs): pristine forest, agriculture, pasture, agroforestry, young secondary forest, and old secondary forest,. The LNB communities were trapped from these soils by using the promiscuous host siratro under controlled conditions. We also studied their relationships with physical and chemical attributes of the soil. Agroforestry and agriculture soil samples induced the highest number of nodules in siratro, while forest soil samples induced the lowest number of nodules. No relationship was found between LNB and Leguminosae species diversity in the LUSs. The soil chemical variables that were most related to differences in nodule number and shoot dry matter weight of plants inoculated with soil suspensions of the LUSs were, respectively: Ca²⁺, Mg²⁺, base saturation, exchangeable bases and Cu²⁺; and pH, cation exchange capacity, B, Cu²⁺ and clay. Although, LNB communities from all LUSs were efficient under controlled and similar conditions, they were found to be composed of strains with variable efficiency: inefficient, efficient, highly efficient and superior efficiency. Efficient strains occurred at the highest frequency in all LUSs. The isolated strains presented similar and new sequences that were phylogenetically related to well known LNB genera in α-and β-Proteobacteria. Unusual genera in these branches, as well as in other branches, which are probably endophytic bacteria, were also isolated from nodules. These data support siratro as a useful trap species to study the LNB biodiversity of diverse ecosystems in tropical soils. The fact that the highest diversity and nodulation were seen in managed systems such as agriculture and agroforestry suggests a high resilience of LNB communities to changes in land use after deforestation in a region where large forest areas are still preserved and can be a source of propagules.     

砒砂岩区不同立地类型人工沙棘林下草本物种多样性环境解释

砒砂岩区是黄土高原区水土流失最为严重的地区之一。砒砂岩区沙棘人工林有良好的水土保持功能,沙棘林下草本植物是该人工林生物多样性的重要组成部分。目的是探明砒砂岩区人工沙棘林下草本物种多样性与环境的关系,依据生境条件的差异,沙棘林被划分为7个类型。野外调查记录了各类型所处的地形条件,采用样方法调查了每个样地草本的生物多样性,并取土样测定其养分和水分状况。采用Patrick指数、Pielou指数、Shannon指数、Simpson指数对砒砂岩区不同立地类型人工沙棘林下草本植物进行了物种多样性分析,并通过CCA、PCCA方法研究了物种分布与环境的关系。结果表明:(1)砒砂岩区不同立地类型人工沙棘林下草本层优势种不同,阴坡混交林物种数最多为23种,阳坡混交林Pielou指数最高为0.89,阴坡纯林和混交林Shannon指数均为最高,Simpson指数没有明显差异。(2)影响砒砂岩区人工沙棘林下草本物种的分布的环境因子顺序为地形因子土壤因子灌木层因子,其中坡向、坡度、土壤含水量、土层厚度、郁闭度、混交结构所占解释量较大。本研究可为当地沙棘人工林生物多样性保护和植被建设提供基础数据。     

毛竹入侵马尾松林的土壤菌群多样性变化

为揭示毛竹入侵邻域林分过程中对土壤细菌群落多样性变化的影响,以马尾松林为研究对象,采集纯毛竹林、竹针混交林和常绿针叶(马尾松)混交林3类样地的混合土样,基于高通量测序技术分析土壤细菌群落多样性和结构变化.结果表明: 研究期间共得到细菌类群39门88纲134目160科511属;在门的分类水平上,与竹针混交林、常绿针叶混交林相比,纯毛竹林酸杆菌门所占比例显著较低,而放线菌门、拟杆菌门、TM7和衣原体门所占比例较高;在属的分类水平上,相对于纯毛竹林,竹针混交林和常绿针叶混交林均有多属表现出所占比例显著上升或下降,单独出现在竹针混交林或常绿针叶混交林且所占比例在0.005%～0.1%的非优势属有130属;α多样性指数均表现为常绿针叶混交林>竹针混交林>纯竹林,且纯竹林与两者均有显著差异,而常绿针叶混交林和竹针混交林之间差异不显著.PCoA分析表明,毛竹入侵对土壤细菌的种群多样性和群落结构产生了影响;所占比例小于0.1%的非优势菌群门分类水平的百分比特征与土壤环境梯度(水溶性有机氮和硝态氮)之间有显著相关性,两者对毛竹入侵马尾松林后土壤细菌群落的非优势种群影响巨大,具有重要的参考价值.     

The relationships between microbiological attributes and soil and litter quality in pure and mixed stands of native tree species in southeastern Bahia, Brazil

This study was conducted to link soil and litter microbial biomass and activity with soil and litter quality in the surface layer for different pure and mixed stands of native tree species in southeastern Bahia, Brazil. The purpose of the study was to see how strongly the differences among species and stands affect the microbiological attributes of the soil and to identify how microbial processes can be influenced by soil and litter quality. Soil and litter samples were collected from six pure and mixed stands of six hardwood species (Peltogyne angustifolia, Centrolobium robustum, Arapatiella psilophylla, Sclerolobium chrysophyllum, Cordia trichotoma, Macrolobium latifolium) native to the southeastern region of Bahia, Brazil. In plantations of native tree species in humid tropical regions, the immobilization efficiency of C and N by soil microbial biomass was strongly related to the chemical quality of the litter and to the organic matter quality of the soil. According to the variables analyzed, the mixed stand was similar to the natural forest and dissimilar to the pure stands. Litter microbial biomass represented a greater sink of C and N than soil microbial biomass and is an important contributor of resources to tropical soils having low C and N availability.     

宁夏罗山自然保护区3种典型林分凋落物和土壤层水源涵养能力综合评估

森林凋落物层和土壤层是森林生态水文效应中的主要贡献层，对森林生态系统水土保持功能和水源涵养能力有重要影响。对比宁夏罗山自然保护区3种典型林分类型凋落物和土壤层水文效应的变化规律和水源涵养能力大小，为该地区的森林生态水文、水土保持和森林管理提供科学依据。以该自然保护区青海云杉纯林、油松纯林和青海云杉油松混交林为研究对象，运用称量、室内浸泡、环刀法和回归分析法对凋落物和土壤层的水文效应进行测定和拟合，并使用熵权法对二者的水源涵养能力进行评估。结果表明：（1）青海云杉油松混交林凋落物的总厚度和总储量显著高于青海云杉纯林和油松纯林（P<0.05），3种林分类型的半分解层的厚度和储量高于未分解层。（2）凋落物层最大持水量范围为63.29-95.08t/hm²，最大持水率范围为335.97%-353.85%，有效拦蓄量范围为34.09-63.92t/hm²，三者均为青海云杉油松混交林>云杉纯林>油松纯林。（3）3种林分类型的凋落物持水量（Q）与浸泡时间（t）呈对数函数关系，吸水速率（V）与浸泡时间（t）呈幂函数关系。（4）从3种林分类型的土壤物理性质和持水特性得出，3种林分类型的土壤层水文效应的等级排序为云杉油松混交林>油松纯林>青海云杉纯林。（5）凋落物层和土壤层的水源涵养能力大小为青海云杉油松混交林（0.43）>油松林（0.3）>青海云杉林（0.27）。综合来看，青海云杉油松混交林的凋落物层和土壤层的水源涵养能力最优，其次是纯林，说明混交林在水土保持和水源涵养方面比纯林更具优势。     

A new cultivation independent approach to detect and monitor common Trichoderma species in soils

A set of primers was developed for the detection, identification and quantification of common Trichoderma species in soil samples. Based on a broad range master alignment primers were derived to amplify an approximate 540 bp fragment comprising the internal transcribed spacer region 1 (ITS 1), 5.8S rDNA and internal transcribed spacer region 2 (ITS 2) from all taxonomic Clades of the genus Trichoderma. The primer set was applied to test strains as well as community DNA isolated from arable and forest soil. For all tested isolates the corresponding internal transcribed spacer regions of Trichoderma spp. strains were amplified, but none of non-Trichoderma origin. PCR with community DNA from soil yielded products of the expected size. Analysis of a clone library established for an arable site showed that all amplified sequences originated exclusively from Trichoderma species mainly being representatives of the Clades Hamatum, Harzianum and Pachybasioides and comprising most of the species known for biocontrol ability. In a realtime PCR approach the primer set uTf/uTr also proved to be a suitable system to quantify DNA of Trichoderma spp. in soils.     

子午岭林区白桦-辽东栎混交林光合生理生态特征研究

对黄土高原子午岭次生林区白桦林、辽东栎林和白桦-辽东栎混交林3种林分的土壤物理特性和叶片光合特性进行了研究。结果表明:(1)白桦-辽东栎混交林地的土壤水分明显改善,其土壤容重最小、土壤孔隙度最大,且均优于纯林,即混交林地有深层次的土壤水分可供利用,并改善了土壤的物理结构;(2)辽东栎林的光合速率和气孔导度最大,其次为白桦-辽东栎混交林,水分利用率(WUE)为混交林白桦>混交林辽东栎>辽东栎林>白桦林;(3)混交林中白桦、辽东栎的Fv/Fm和Fv/Fo值均较大,与纯林差异不显著;白桦林和辽东栎林的qP和NPQ值均大于混交林。     

Characteristics of soil animal community in the subalpine/alpine forests of western Sichuan during onset of freezing

Seasonal freeze–thaw cycle is a common phenomenon in the subalpine/alpine forest region, and may have a significant influence on the structure and function of soil animal community. To understand the characteristics and dynamics of soil animal community as well as its response to repeated freeze–thaw events in this region during onset of freezing, a field experiment was conducted to investigate the composition, abundance, and diversity of soil fauna in primary fir (Abies faxoniana) forest, fir and birch (Betula albosinensis) mixed forest and secondary fir (A. faxoniana) forest, which were three representative forests in the subalpine and alpine forest region in western Sichuan. Soil samples were collected from November 3 to November 27, 2008, which was defined as onset of freezing based on the simultaneous monitoring of soil temperature. Soil macrofauna were picked up by hand in the fields. Mesofauna were separated and collected from the soil samples by Baermann and Tullgren methods, respectively. By preliminary identification, 40,942 individuals were collected, which belonged to 7 phyla, 15 classes and 25 orders in the three sampling forests. 16,557, 14,669 and 9716 individuals were found in primary forest, mixed forest and secondary forest, respectively. In comparison with the mineral soil layer, the soil organic layer had higher density and groups of soil fauna. Furthermore, following the repeated freeze–thaw events, density and groups of soil fauna experienced a distinct decrease in both soil organic layer and mineral soil layer, and this trend in soil organic layer was more significant in the primary forest, owing to higher intense and more frequent freeze–thaw cycles in soil organic layer of the primary forest. The results revealed that soil animals were sensitive bio-indicators to environmental changes, such as repeated freeze–thaw events and dry–wet cycle. Meanwhile, the results also implied that the dynamics of the structure and function of soil animal community during the onset of freezing was of ecological significance to understand the wintertime ecological process in soils.     

中亚热带几种典型森林土壤养分含量分析及综合评价

分析及综合评价不同森林植被对土壤养分因子的影响,对揭示森林植被与土壤养分的相互关系,全面衡量森林土壤养分水平具有重要的意义。以中亚热带杉木(Cunninghamia lanceolata)纯林、木荷(Schima superb)纯林、木荷+楠木(Phoebe ahenre)混交林、木荷+杉木混交林、青冈栎(Cyclobalanopsis glauca)天然次生林、毛竹(Phyllostachys edulis)林6种森林类型为研究对象,在分析不同森林类型土壤养分特征状况差异的基础上,采用结构方程模型确定土壤养分因子权重,对其土壤养分水平进行综合评价。结果表明:(1)6种森林类型0-60 cm土壤有机质、全氮、碱解氮和有效磷的含量分别为8.24-28.17 g/kg、0.67-1.31 g/kg、44.88-89.31 mg/kg和1.24-6.50mg/kg,其含量最高的都是青冈栎天然次生林,土壤有机质和碱解氮含量最低的是毛竹林,土壤全氮和有效磷的含量最低的分别为杉木纯林和木荷纯林,0-60 cm土壤全磷和速效钾的含量分别为0.15-0.21 g/kg和35.54-54.32mg/kg,其含量最高的都是木荷+楠木混交林,含量最低的分别为木荷纯林和毛竹林;(2)土壤有机质、全氮、全磷和有效磷的含量在6种森林类型中都一致表现出随土层加深而逐渐减小的规律,而碱解氮和速效钾含量在土壤垂直剖面的变化却因森林类型而异,没呈现出具体的变化规律;(3)栎类天然次生林的土壤养分等级综合评分值为3.47,评价等级属于中上水平,杉木纯林、木荷纯林、木荷+楠木混交林、木荷+杉木混交林的土壤养分等级综合评分值分别为2.45、2.76、2.83、2.68,评价等级均属于中下水平,而毛竹林的土壤养分等级综合评分值仅为1.95,评价等级属于土壤养分缺乏。