不同生境黑老虎根际和内生真菌多样性分析

为探究不同生长条件下黑老虎根际和根部内生真菌群落组成和多样性及其与土壤环境因子的相关性,该文应用Illumina高通量测序方法对贵州3个不同生境下黑老虎根际和根部内生真菌进行了研究。结果表明:(1)3种生境下,根际土壤真菌OTU数量(3 867)远多于根部内生真菌(801),其中根际土壤真菌共有的OTU为72个,共注释到5个门、49个属,大多为子囊菌门; 属水平上被孢霉属、外瓶柄霉属、 柱孢属占比较高; 根部内生真菌共有的OTU为14个,共注释到2个门、11个属,子囊菌门(13个,占比92.9%)占绝对优势,属水平上被孢霉属、外瓶柄霉属、 柱孢属和丛赤壳属占比最高; 所有样本中,共有的OTU仅为6个,注释到2个门、5个属,子囊菌门(5个,占比83.3%)为优势门; 在属水平上,占比最高的为外瓶柄霉属(2个,33.3%),其余分别为被孢霉属、柱孢属和丛赤壳属 。Alpha多样性分析表明,根际土壤的真菌群落多样性和丰富度均显著高于根部内生真菌,而野生生境的真菌多样性高于栽培生境。(2)在门水平上,3个生境下主要内生真菌类群均为子囊菌门(Ascomycota)和担子菌门(Basidiomycota),占总菌群的88.28%; 在属水平上,不同生境条件下,根际和根部内生真菌群落结构差异明显; 栽培生境下,根部内生真菌菌群具有一定的偏好性,而野生生境下,根部内生真菌菌群均匀度更高; FUNGuild真菌群落功能预测显示,栽培生境下的病理-腐生营养型(pathotroph-saprotroph)在根部内生真菌中占比较高,而野生生境下的腐生营养型(saprotroph)及共生营养型(symbiotroph)占比较高。(3)土壤环境因子对根部内生真菌和根际真菌的影响方式不同,其中土壤总钾(TK)和土壤总磷(TP)与黑老虎根部内生真菌香农指数和辛普森指数显著正相关,而土壤有机质(SOM)、总氮(TN)和速效氮(AN)与黑老虎根际土壤真菌Ace指数和Chao1指数显著正相关。综上表明,土壤有机质、总氮、速效氮是影响黑老虎根际土壤真菌群落的主要土壤环境因子。     

自然林下与田间根腐三七根际微生物群落特征及比较分析

【背景】三七根际微生物群落特征与其土传根腐病害密切相关,而针对自然林下根腐三七的相关研究鲜见报道。【目的】比较分析自然林下与田间根腐三七根际土壤微生物群落的组成特征,结合土壤理化性质与酶活性分析,为三七根腐病害防治与仿野生栽培提供科学依据。【方法】采集自然林下与田间根腐三七根际土壤,利用高通量测序技术,分析土壤细菌与真菌群落的物种组成与多样性,并测定土壤理化性质和酶活性。【结果】自然林下与田间根腐三七根际土壤中细菌和真菌群落组成具有明显差异,自然林下根腐三七根际土壤中担子菌门(Basidiomycota)、酸杆菌门(Acidobacteria)和疣微菌门(Verrucomicrobia)的相对丰度较高,而田间根腐三七根际土壤中子囊菌门(Ascomycota)、变形菌门(Proteobacteria)和绿弯菌门(Chloroflexi)的相对丰度较高。在属分类水平,镰刀菌属(Fusarium)是自然林下根腐三七根际土壤中的优势菌群,相对丰度为17.30%,而癣囊腔菌属(Plectosphaerella)是田间根腐三七根际土壤中的优势菌群,相对丰度为22.55%;Candidatus Ba...     

青枯病易感和钝感桑树品种根际土壤真菌群落结构比较

为研究青枯病易感和钝感桑树品种植株根际土壤真菌群落组成,该研究以ITS1F和ITS2R为引物,基于高通量测序技术对桑树青枯病易感品种(台湾长果桑,SM)和桑树青枯病钝感品种(桂桑12号,IM)植株根际土壤真菌群落结构进行分析。结果表明：(1)两个品种间指示真菌丰富度的ACE、Chao1指数及表征多样性的Shannon指数无显著差异,门分类水平,被孢霉门(Mortierellomycota)和球囊菌门(Glomeromycota)是青枯病钝感桑树品种植株根际土壤中特有的优势真菌门;而属分类水平,Apiotrichum、地丝菌属(Geotrichum)、足放线病菌属(Scedosporium)和腐质霉属(Humicola)等是青枯病易感桑树品种植株根际土壤中富集的特有优势真菌属。(2)青枯病易感桑树品种植株根际土壤中,缺失了被孢霉门、球囊菌门真菌,以及被孢霉属(Mortierella)、镰刀菌属(Fusarium)、曲霉菌属(Aspergillus)和毛壳菌属(Chaetomium)等具有生防功能的优势真菌门属,可能是其易感青枯病的重要原因。(3)根据真菌群落对同类环境资源的利用途径进行功...     

The brown root rot fungus Phellinus noxius affects microbial communities in different root-associated niches of Ficus trees

Brown root rot (BRR) caused by Phellinus noxius is a destructive tree disease in tropical and subtropical areas. To understand how BRR affects the composition of the plant rhizoplane-enriched microbiota, the microbiomes within five root-associated compartments (i.e., bulk soil, old/young root rhizosphere soil, old/young root tissue) of Ficus trees naturally infected by P. noxius were investigated. The level of P. noxius infection was determined by quantitative PCR. Illumina sequencing of the internal transcribed spacer and 16S rRNA revealed that P. noxius infection caused a significant reduction in fungal diversity in the bulk soil, the old root rhizosphere soil, and the old root tissue. Interestingly, Cosmospora was the only fungal genus positively correlated with P. noxius. The abundance and composition of dominant bacterial taxa such as Actinomadura, Bacillus, Rhodoplanes, and Streptomyces differed between BRR-diseased and healthy samples. Furthermore, 838 isolates belonging to 26 fungal and 35 bacterial genera were isolated and tested for interactions with P. noxius. Antagonistic activities were observed for isolates of Bacillus, Pseudomonas, Aspergillus, Penicillium, and Trichoderma. Cellophane overlay and cellulose/lignin utilization assays suggested that Cosmospora could tolerate the secretions of P. noxius and that the degradation of lignin by P. noxius may create suitable conditions for Cosmorpora growth.     

General microflora,arbuscular mycorrhizal colonization and occurrence of endophytes in the rhizosphere of two age groups of <Emphasis Type="Italic">Ginkgo biloba</Emphasis> L.of Indian Central Himalaya

The populations of the general microflora (bacteria, actinomycetes and fungi) in the rhizosphere and their corresponding non-rhizosphere soil samples of Ginkgo biloba L. of two age groups (Group A, <25 years-young trees; Group B, >60 years-old trees) growing under a temperate location of Indian Himalayan Region (IHR) have been determined. Observations were also made for the diversity, distribution and colonization of arbuscular mycorrhizal (AM) fungi and occurrence of endophytes in roots of G. biloba. The population of general microflora was found to be higher in the rhizosphere of Group B trees, more clearly reflected in terms of rhizosphere: soil (R:S) ratios. Contrary to this, per cent colonization and spore densities of AM fungi were higher in the rhizosphere of Group A trees as compared to the rhizosphere of Group B. AM fungal colonization was observed mostly in form of loose coils. All the spores detected, belonged to the genus Glomus with five different types. Presence of endophytes (both bacteria and fungi) was observed in the cortical cells of G. biloba roots, more profound in case of Group B trees. Data suggest that, while the species of Glomus dominated the rhizosphere of G. biloba, an inverse correlation exist between the colonization of general microflora and the colonization of AM fungi including endophytes.     

The Abundance and Structure of the Root-Associated Microbial Complexes of Two Greenhouse Rose Cultivars

The study of the root-associated microbial complexes of affected and healthy rose plants of two cultivars (Grand gala and Royal velvet) grown in a greenhouse showed that the biomass of eukaryotic microorganisms in the rhizoplane and rhizosphere of healthy rose plants and in the surrounding soil was considerably lower than in the same loci of affected plants. In contrast, the biomass of root-associated prokaryotic microorganisms was higher in the case of healthy than in the case of affected rose plants. The root-associated bacterial complexes of both affected and healthy rose plants were dominated by the genera Arthrobacter, Rhodococcus, and Myxobacterium and did not contain phytopathogenic bacteria. The root-associated fungal complex of healthy roses was dominated by fungi of the genus Trichoderma, whereas that of the affected rose plants was dominated by the species Aureobasidium microstictum. The affected cane cuttings and cankers occurring on affected canes were found to contain Coniothyrium fuckelii (the causal fungus of rose stem canker) and sclerotia of Botrytis cinerea (the causal fungus of gray rot). The micromycete complex of healthy rose plants was not so diverse as was the micromycete complex of affected rose plants.     

Molecular Profiling of Rhizosphere Microbial Communities Associated with Healthy and Diseased Black Spruce (Picea mariana) Seedlings Grown in a Nursery

Bacterial and fungal populations associated with the rhizosphere of healthy black spruce (Picea mariana) seedlings and seedlings with symptoms of root rot were characterized by cloned rRNA gene sequence analysis. Triplicate bacterial and fungal rRNA gene libraries were constructed, and 600 clones were analyzed by amplified ribosomal DNA restriction analysis and grouped into operational taxonomical units (OTUs). A total of 84 different bacterial and 31 different fungal OTUs were obtained and sequenced. Phylogenetic analyses indicated that the different OTUs belonged to a wide range of bacterial and fungal taxa. For both groups, pairwise comparisons revealed that there was greater similarity between replicate libraries from each treatment than between libraries from different treatments. Significant differences between pooled triplicate samples from libraries of genes from healthy seedlings and pooled triplicate samples from libraries of genes from diseased seedlings were also obtained for both bacteria and fungi, clearly indicating that the rhizosphere-associated bacterial and fungal communities of healthy and diseased P. mariana seedlings were different. The communities associated with healthy and diseased seedlings also showed distinct ecological parameters as indicated by the calculated diversity, dominance, and evenness indices. Among the main differences observed at the community level, there was a higher proportion of Acidobacteria, Gammaproteobacteria, and Homobasidiomycetes clones associated with healthy seedlings, while the diseased-seedling rhizosphere harbored a higher proportion of Actinobacteria, Sordariomycetes, and environmental clones. The methodological approach described in this study appears promising for targeting potential rhizosphere-competent biological control agents against root rot diseases occurring in conifer nurseries.     

黑老虎内生真菌及根际土壤真菌的群落结构与生态功能分析

为探讨黑老虎(Kadsura coccinea)根际土壤和组织内生真菌菌群的组成及其生态功能，该研究采用ITS高通量测序技术对成熟黑老虎(根、茎、叶)内生真菌及根际土壤真菌群落结构、多样性和生态功能进行了分析。结果表明：(1)从12个样品中共获得2 241个可操作分类单元(OTU),涉及10门、41纲、95目、212科、367属，内生真菌(根、茎、叶)和根际土壤真菌OTU数分别为386、536、258、1 435个，其中共有的OTU为18个。在门水平上，黑老虎内生真菌及根际土壤真菌优势群落均为子囊菌门和担子菌门，其中子囊菌门在叶和茎中占比分别高达96.99%和95.37%;在属水平上，黑老虎根际土壤真菌中腐生真菌被孢霉属占比较高(为13.5%),叶和茎等生长旺盛的组织中子囊菌门未分类属和痂囊腔菌属占比较高。(2)α多样性分析结果显示，黑老虎根际土壤真菌群落的丰度和多样性明显高于内生真菌，茎中内生真菌丰度显著高于根和叶，而根、茎和叶组织间内生真菌多样性差异不显著；PCoA分析结果显示，叶和茎的真菌群落结构相似性更高。(3)利用FUNGuild数据库进行的功能预测分析结果显示，黑老虎根际土...     

辽宁碱蓬根际土壤真菌多样性的季节变化及其耐盐性

采用稀释平板法和形态学鉴定法进行了辽宁碱蓬(Suaeda liaotungensis)根际土壤真菌的分离鉴定及季节、盐度对真菌多样性的影响分析。结果表明:春、夏、秋、冬四季辽宁碱蓬根际土壤中的真菌菌落数分别为6410、4180、5730和3340,种类分别为6属13种、9属16种、11属31种、6属12种,共分离鉴定出13属42种真菌;其根际土壤真菌的多样性指数、丰富度指数和均匀度指数,从大到小的次序均为秋夏春冬,3种指数均在秋季达到峰值;多样性指数和丰富度指数差异较大,均匀度指数波动较小,春季和冬季的均匀度指数相近。辽宁碱蓬根际土壤真菌种类明显多于无植被土壤,是无植被土壤的7倍,多样性指数和均匀度指数分别是无植被土壤的4—8倍和1—1.2倍;其根际土壤真菌优势种群包括青霉属(Penicillium)、葡萄穗霉属(Stachybotrys)、枝孢属(Cladosporium)、木霉属(Trichoderma)、曲霉属(Aspergillus)和镰孢属(Fusarium);4个季节的优势菌不尽相同,但均有一个共同的优势菌属青霉属。以Na Cl浓度梯度法制造盐胁迫生境,检测辽宁碱蓬根际土壤真菌对盐胁迫的响应,结果表明:60%左右的菌种能耐受5%以下盐度、15%左右的菌种能耐10%—20%盐度;筛选出了6株高度耐盐菌:细交链孢霉、草酸青霉、产黄青霉、烟曲霉、细极链格孢和赭曲霉,其中最高耐盐菌种赭曲霉可耐20%盐度。创新之处在于以盐度和季节为变量检测辽宁碱蓬根际土壤真菌的种群构成,为其深入研究奠定了理论基础。     

小蓬竹根际土壤微生物及内生真菌多样性分析

为了解喀斯特典型物种-小蓬竹根际土壤微生物及不同部位内生真菌多样性,采用沿等高线等距离取样法采集小蓬竹根际土壤及健康植株,通过可培养对根际土微生物及内生菌进行分离,利用分子技术对其进行鉴定,根据鉴定结果构建系统发育树,并计算小蓬竹根际土壤微生物和根茎叶内生真菌多样性。结果如下：（1）共从根际土壤、根、茎、叶分离得到139个真菌菌株,隶属于27属,其中根际土壤分离得到34个真菌菌株隶属于12属,根部分离得到的63个内生真菌菌株隶属于17个属,茎部分离得到的14个内生真菌菌株隶属于8个属,叶部分离得到28个内生真菌菌株隶属于9个属;（2）根际土壤共分离得到41株细菌菌株,隶属于7个属26个种,20株放线菌菌株,隶属于1属15种;从Shannon-Wiener多样性指数、均匀度指数、Simpson指数排序来看,真菌主要表现为根 > 根际土壤 > 茎 > 叶,细菌和放线菌多样性均较低。（3）按层次聚类分析可分别将真菌、细菌、放线菌聚为3支。小蓬竹根际土壤、根、茎和叶具有丰富的微生物多样性,不同部位菌群组成存在差异性（P<0.05）,且存在以假单胞菌属、芽孢杆菌属等为优势属的抗盐耐旱菌群,这有助于揭示小蓬竹对喀斯特生境的适应性,以及为微生物-植物群落之间相互关系提供一定基础数据,为后期寻找小蓬竹相关耐性功能菌奠定基础。     

Arbuscular mycorrhizal fungi associated with the Meliaceae on Hainan island, China

Species richness, spore density, frequency of occurrence, and relative abundance of AM fungi were determined in rhizosphere soil samples from nine tropical rainforest sites on Hainan island, south China, and the arbuscular mycorrhizal (AM) status of members of the Meliaceae was examined. All 28 plant taxa investigated (25 species including two varieties of 1 species and three varieties of another) were colonized by AM fungi. The mean proportion of root length colonized was 56% (range 10–95%). Vesicles were observed in 27 and hyphal coils in 26 of the 28 plant taxa. Mycorrhizas were of the Paris-type or intermediate-type, with no Arum-type mycorrhizas observed. Species richness of AM fungi varied from 3 to 15 and spore density from 46 to 1,499 per 100 g rhizosphere soil. Of 33 AM fungal taxa in five genera isolated and identified, 18 belonged to Glomus, 9 to Acaulospora, 1 to Entrophospora, 2 to Gigaspora, and 3 to Scutellospora. Acaulospora and Glomus were the dominant genera identified. Glomus claroideum was the taxon most commonly isolated, with a frequency of occurrence of 56.5% and relative abundance of 10.4%. A positive correlation was found between percentage of root length colonization and species richness. However, there was no correlation between spore density and percentage of root length colonized by AM fungi.     

Effect of foliar application of antibiotics and gibberellic acid on the rhizosphere microflora of pea,infected withVerticillium dahliae

A study was made of the effects of foliar spray of bacitracin, chloramphenicol and gibberellic acid on the rhizosphere microflora of pea seedlings (Pisum sativum L.) infected withVerticillium dahliae. The antibiotics increased fungus and actinomycete counts and reduced the bacterial populations in the rhizosphere. Gibberellic acid at 10 ppm concentration reduced all three groups of microorganisms while at 100 ppm fungi and actinomycetes increased slightly. Invariably the rhizosphere effect was as follows: bacteria > fungi > actinomycetes. Foliar sprays also affected the percentage occurrence of particular genera of fungi in the rhizosphere; for example,Trichoderma spp. were stimulated by all the treatments, the maximum being with 10 ppm gibberellic acid, even though the total fungus count was reduced. The disease severity was markedly reduced by foliar sprays.     

白马雪山云南黄芪与灰毛康定黄芪根际微生物物种多样性及抗生物膜活性菌株筛选

【背景】细菌生物膜是造成病原菌耐药性增强和持续感染的主要因素,但目前尚无针对抗菌膜的特效药物。特境植物根际微生物可产生大量具有提高宿主免疫功能的活性成分,极具抗生物膜药源开发潜力。【目的】了解滇西北高寒特境白马雪山分布的云南黄芪与灰毛康定黄芪植物根际微生物的物种多样性,并对可培养菌株进行抑菌与抗生物膜活性筛选。【方法】采用宏基因组技术结合传统微生物培养方法,对采自我国云南迪庆藏族自治州德钦县白马雪山的云南黄芪与灰毛康定黄芪的根际微生物进行物种多样性研究,并通过“孔板法”测定其可培养菌株发酵液乙酸乙酯粗浸膏的抗菌、抗生物膜活性。【结果】宏基因组测序结果显示,云南黄芪根际土壤样本中的微生物来自6门7纲8目8科9属10种,其中栖热菌属为优势菌群;灰毛康定黄芪根际土壤样本中的微生物来自6门8纲10目11科14属15种,其中慢生根瘤菌属为优势菌群。通过纯培养共获得145株可培养菌株,包括112株细菌和33株真菌。其中,云南黄芪根际细菌59株,共计16属35种,优势属为假单胞菌属和链霉菌属;根际真菌19株,共计4属5种,优势属为曲霉属;灰毛康定黄芪根际细菌53株,归属于16属29种,优势属为芽孢杆菌属与寡养单胞菌属;根际真菌14株,归属于3属4种,优势属为曲霉属。从不同种水平上选择51株细菌和7株真菌为代表菌株进行抗生素药源评估,发现5株细菌及1株真菌发酵液的乙酸乙酯粗浸膏具有中等至较强的抗革兰阳性菌活性,而且其中4株具有抗MRSA生物膜活性,最终确定了链霉属放线菌Streptomyces fulvissimus KTA1和曲霉属真菌Aspergillus fumigatus YNF5为潜力活性菌株。【结论】首次报道了滇西北地区高寒特境黄芪属植物根际微生物具有较好的物种多样性,而且具有一定的抗生素药用资源开发潜力。本研究对滇西北高寒特境特色植物来源的微生物资源开发利用与保护具有重要的借鉴意义。     

不同生境黑果枸杞根际与非根际土壤微生物群落多样性

研究典型生境黑果枸杞根际与非根际土壤微生物群落多样性及其与土壤理化性质间的关系,为进一步研究黑果枸杞抗逆性提供理论数据。采集新疆精河县艾比湖地区(EB)盐碱地、乌苏市(WS)路旁荒地、五家渠市(WQ)人工林带的黑果枸杞根际与非根际土壤,利用Illumina-MiSeq高通量测序技术分析细菌和真菌群落组成和多样性。结果表明:根际土壤细菌多样性高于非根际土壤(WQ除外),而根际真菌多样性低于非根际土壤。WQ非根际土壤细菌和真菌多样性均高于EB和WS;根际细菌多样性排序为EBWSWQ,根际真菌多样性排序为WSEBWQ。根际土壤优势细菌门依次是变形菌门、拟杆菌门、放线菌门、酸杆菌门,真菌优势门为子囊菌门、担子菌门。根际土壤细菌变形菌门、拟杆菌门、酸杆菌门的相对丰度高于非根际土壤,而厚壁菌在根际土壤中的丰度显著降低,真菌优势门丰度在根际土和非根际土中的变化趋势因地区而异; Haliea、Gp10、Pelagibius、Microbulbifer、假单胞菌属、Thioprofundum、Deferrisoma是根际土壤细菌优势属;多孢子菌属、支顶孢属、Corollospora、Cochlonema是根际真菌优势属。细菌、真菌优势类群(门、属)的组成以及丰富度存在地区间差异,厚壁菌门在EB地区的丰富度显著高于含盐量较低的WS、WQ;盐碱生境EB中根际土壤嗜盐细菌的丰度高于非盐碱生境(WQ、WS),如盐单胞菌属、动性球菌属、Geminicoccu、Pelagibius、Gracilimonas、Salinimicrobium等。小囊菌属是EB根际真菌的最优势属,Melanoleuca是WQ和WS的最优势属,地孔菌属、Xenobotrytis、Brachyconidiellopsis、多孢子菌属等在EB根际土壤中的丰度显著高于WQ和WS。非盐碱生境(WS和WQ)的微生物群落之间的相似性较高,并且高于与盐碱环境(EB)之间的相似性,表明土壤含盐量对微生物群落组成丰度具有重要的影响。     

Rhizosphere micro-organisms in relation to the apple replant problem

Summary One of the factors giving rise to soil sickness in apple orchards is the rhizosphere microflora. The composition of the microbial coenosis in the rhizosphere changes with increasing age of the apple trees. An increase in the counts of micromycetes and actinomycetes and a decrease in bacterial counts was found in agreement with the decreasing pH of the rhizosphere soil. The presence of fluorescent pseudomonads in the rhizosphere of old apple trees was rare, but the planting of apple seedling into sick soil induced their proliferation. The relative proportion of individual genera of micromycetes changed according to the tree age; fungi of the genus Mucor were more often found in the rhizosphere of younger trees than in that of older ones while fungi of the genus Penicillium had an opposite trend. Biological tests showed that Penicillium fungi form the majority of the phytotoxic microflora. The amount of phytotoxic micromycetes was higher in ‘sick’ soil as compared with control soil in which apple trees had not been grown for at least 15 years. Higher numbers of phytotoxic micromycetes were isolated also from the rhizosphere of apple seedlings grown in ‘sick’ soil as compared with those growing in control soil. An increase in the amount of phytotoxic micromycetes in apple tree rhizosphere could be induced by mere addition of 5% (w/w) ‘sick’ soil to the soil in which apple trees were grown for the first time. By adding sterilized ‘sick’ soil, the amount of phytotoxic micromycetes in the apple seedling rhizosphere was not affected. Increased numbers of phytotoxic micromycetes affected negatively the growth of apple trees and the morphology of apple tree roots. This demonstrated the possibility of transfer of a factor participating in the etiology of soil sickness in apple orchards.     

Comparative identification by fatty acid analysis of soil,rhizosphere, and geocarposphere bacteria of peanut (Arachis hypogaea L.)

Bacterial isolates were collected from the geocarposphere, rhizosphere, and root-free soil of field grown peanut (Arachis hypogaea L.) at three sample dates, and the isolates were identified by analysis of fatty acid methyl-esters to determine if qualitative differences exist among the bacterial microflora of these zones. Five bacterial genera were associated with isolates from soil, while pod and root isolates constituted 16 and 13 genera, respectively, indicating that bacterial diversity was higher in the rhizosphere and geocarposphere than in soil. The dominant (most frequently identified) genus across all three samples dates was Flavobacterium, for pods, Pseudomonas for roots, and Bacillus, for root-free soil. Sixteen bacterial taxa were only isolated from the geocarposphere, 7 only from the rhizosphere, and 5 only from soil. These results show that specific bacterial taxa are preferentially adapted to colonization of the geocarposphere and suggest that the soil, rhizosphere, and geocarposphere constitute three distinct ecological niches. Bacteria which colonize the geocarposphere should be examined as potential biological control agents for pod-invading fungi such as the toxigenic strains of Aspergillus flavus and A. parasiticus.     

Development of rhizosphere and rhizoplane microflora of Aristida coerulescens in the Libyan desert

Summary Microflora of rhizosphere soil, rhizoplane and macerated root-portions of Aristida coerulescens, naturally occurring in the Libyan desert, were different in count and isolates, in the different root zones. A rhizosphere effect characteristic of each zone is shown. The root base contained the lowest numbers of microflora (bacteria and fungi) whilst the root tip included the highest counts. Distribution of most of the individual fungal species in the different root zones and root-surfaces is given in text.     

Outbreak of Phytophthora cinnamomi causing severe decline of avocado trees in southern Turkey

Since the summer 2017, severe decline symptoms have been observed on 10- to 25-year-old avocado trees in almost all commercial orchards planted in the Mediterranean coastal region of Turkey. Young, newly planted trees in infected orchards were also affected by the disease. Affected trees showed wilting, leaf discoloration, defoliation and severe dieback. Some trees were completely desiccated. Although fine roots of symptomatic trees usually were decayed, reddish brown cankers also occurred on taproots and lateral roots of heavily infected trees. The pathogens were isolated from necrotic root and soil samples of symptomatic trees, using selective medium and soil baiting, and were identified based on morphological features and DNA sequences of the internal transcribed spacer (ITS) region. One isolate each of Phytophthora cryptogea and P. palmivora was identified, while all other isolates were P. cinnamomi. In addition, a subcortical fan-shaped mycelium, characteristic of Armillaria spp., was observed in the stem base of a symptomatic tree and identified as Armillaria gallica by DNA sequences of the internal transcribed spacer (ITS) and the translational elongation factor 1-α (EF 1-α) gene regions. Pathogenicity of Phytophthora isolates was tested by stem inoculation on one-year-old avocado seedlings. Two months after inoculation, canker lesions developed on stems of seedlings inoculated by any of the three Phytophthora spp. In contrast, collenchyma callus formed over the wound points on control plants over the same time period. This is the first report of P. cinnamomi, P. cryptogea, P. palmivora and A. gallica causing root rot of avocado trees in Turkey. In addition, P. cryptogea and A. gallica are reported for the first time associated with disease on this host. Due to the severe symptoms and widespread occurrence, P. cinnamomi should be considered a potential threat to avocado cultivation and natural ecosystems of this region of Turkey.     

南疆四种盐生植物根际土壤真菌群落结构特征

盐生植物特殊的生境孕育了独特的根际微生物群落。为了解南疆干旱区不同盐生植物根际土壤真菌群落结构特征,探讨影响真菌群落结构的土壤环境因子,选取南疆伽师县同一盐碱地盐爪爪（Kalidium foliatum）、黑果枸杞（Lycium ruthenicum）、花花柴（Karelinia caspia）和旱生芦苇（Phragmites australis）四种优势盐生植物,采用Illumina NovaSeq高通量测序技术分析根际土壤真菌群落结构和多样性,并探究其与土壤理化因子的相关性。结果表明,四种盐生植物根际土壤理化特征不尽相同,土壤pH均超过8.0,电导率（EC）由高到低为旱生芦苇 > 盐爪爪 > 花花柴 > 黑果枸杞,黑果枸杞根际土壤的有机质（SOM）、全氮（TN）、全磷（TP）、全钾（TK）、速效氮（AN）和速效磷（AP）含量均最高,旱生芦苇根际土壤的SOM、TN、TP、TK和AN值均最低,但土壤水分含量（SWC）和EC值最高。四种盐生植物共有的操作分类单元（OTUs）数量为153个,各自特有的OTUs数量不尽相同。根际土壤真菌群落丰富度（ACE、Chao 1指数）依次为盐爪爪 > 旱生芦苇 > 黑果枸杞 > 花花柴,Shannons指灵第和Simpson指数大小依次为盐爪爪 > 黑果枸杞 > 旱生芦苇 > 花花柴。从四种盐生植物根际共检测到真菌8门、21纲、44目、89科、124属,子囊菌门（Ascomycota）在四种盐生植物根际土壤中占绝对优势地位。镰孢菌属（Fusarium）、支顶孢属（Acremonium）、曲霉属（Aspergillus）和青霉菌属（Penicillium）是四种盐生植物根际土壤共有优势属,非优势属数量多但相对丰度因植物种类而异。典范对应分析（CCA）显示,土壤速效钾、AP、pH和EC是影响根际土壤真菌群落结构变化的主要驱动因子。研究表明南疆四种盐生植物根际土壤真菌群落具有相似性,但优势菌属丰度差异明显,具有植物种类特异性。     

PCR Primers That Amplify Fungal rRNA Genes from Environmental Samples

Two PCR primer pairs were designed to amplify rRNA genes (rDNA) from all four major phyla of fungi: Ascomycota, Basidiomycota, Chytridomycota, and Zygomycota. PCRs performed with these primers showed that both pairs amplify DNA from organisms representing the major taxonomic groups of fungi but not from nonfungal sources. To test the ability of the primers to amplify fungal rDNA from environment samples, clone libraries from two avocado grove soils were constructed and analyzed. These soils possess different abilities to inhibit avocado root rot caused by Phythophthora cinnamomi. Analysis of the two rDNA clone libraries revealed differences in the two fungal communities. It also revealed a markedly different depiction of the soil fungal community than that generated by a culture-based analysis, confirming the value of rDNA-based approaches for identifying organisms that may not readily grow on agar media. Additional evidence of the usefulness of the primers was obtained by identifying fungi associated with avocado leaves. In both the soil and leaf analyses, no nonfungal rDNA sequences were identified, illustrating the selectivity of these PCR primers. This work demonstrates the ability of two newly developed PCR primer sets to amplify fungal rDNA from soil and plant tissue, thereby providing unique tools to examine this vast and mostly undescribed community of organisms.