Presence and distribution of insect-associated and entomopathogenic fungi in a temperate pine forest soil: An integrated approach

For decades entomopathogenic fungi have garnered interest as possible alternatives to chemical pesticides. However, their ecology outside of agroecosystems demands further study. We assessed the diversity and abundance of entomopathogenic and insect-associated fungi at a loblolly pine forest in North Carolina, USA using culture-dependent and next-generation sequencing libraries. Fungi were isolated using Galleria mellonella larvae, as well as from soil dilutions plated on a selective medium. Isolates were identified using Sanger sequencing of the ITS and LSU rRNA gene regions, and represented 36 OTUs including Metarhizium, Lecanicillium, and Paecilomyces. Additionally, we assessed the chitinolytic potential of isolates and found widespread, variable ability to degrade chitin within and between genera. Phylogenetic analyses resolved several isolates to genus, with some forming clades with other insect-associated taxa, as well as with fungi associated with plant tissues. Saprophytes were widely distributed in soil, while entomopathogens were less abundant and present primarily in the top two cm of the soil. The similarity between culture-dependent and next-generation sequencing results demonstrates that both methods can be used concurrently in this system to study the ecology of entomopathogenic fungi.     

Specificity and localised distribution of mycorrhizal fungi in the soil may contribute to co-existence of orchid species

Co-occurring orchid species tend to occupy different areas and associate with different mycorrhizal fungi, suggesting that orchid mycorrhizal (OrM) fungi may be unevenly distributed within the soil and, therefore, impact the aboveground spatial distribution of orchids. To test this hypothesis, we investigated spatial variations in the community of potential OrM associates within the roots of three co-habitating orchid species (Anacamptis morio, Gymnadenia conopsea, and Orchis mascula) and the surrounding soil in an orchid-rich calcareous grassland in Southern Belgium using 454 amplicon pyrosequencing. Putative OrM fungi were broadly distributed in the soil, although variations in community composition were strongly related to the proximal host plant. The diversity and frequency of sequences corresponding to OrM fungi in the soil declined with increasing distance from orchid plants, suggesting that the clustered distribution of orchid species may to some extent be explained by the localised distribution of species-specific mycorrhizal associates.     

高通量测序分析环保肥料增效剂对马铃薯根际土壤真菌多样性变化影响

【目的】高通量测序技术对研究环境样品中微生物群落组成具有很大的应用价值。土壤微生物群落结构和多样性及其变化在一定程度上反映了土壤的质量。旨在从微生物群落结构的角度阐述环保肥料增效剂对马铃薯根际土壤主要真菌类群结构的影响。【方法】通过高通量测序结果对比分析应用增效剂前后马铃薯根际真菌宏基因组ITS1区,并依据RDP中设置的分类阈值对序列进行物种分类。【结果】测序结果经过质量控制,共获得有效条带437 375条,依据97%的序列相似性做聚类分析,获得全部样品的可分类操作单元(OTUs)共633个。子囊菌的总体数量在所有样品中最多(相对丰度在56.95%-97.23%之间),且处理后呈增加趋势(HY除外),而担子菌门数量在处理后呈下降的趋势。基于真菌ITS1高通量测序结果获得的α指数显示,样品内部处理和对照之间真菌物种多样性有差别。基于真菌ITS1高通量测序获得的β指数显示,处理组与对照组的真菌多样性之间没有差别,这表明真菌多样性之间的差异更多地取决于样品采集地点。【结论】土壤特性是影响真菌种群的重要因素之一,环保肥料增效剂显著改善了土壤真菌的种群结构。     

放线菌制剂对人参生长及根域土壤微生物区系的影响

以小兴安岭地区人参为研究对象,探索放线菌制剂对人参的促生效应及对人参根区、根表土壤微生物区系的影响.结果表明： 经放线菌制剂Streptomyces pactum（Act12）处理后,人参药用部分产量增加,品质改善;叶片诱导酶活性提高,根系活力增强;土壤中细菌、放线菌的数量和比例显著增加,真菌的数量和比例减少.与对照相比,土壤微生物区系结构改变:优势菌荧光假单胞菌、韩国假单胞菌和氧化微杆菌在根区、根表土壤中的数量大幅提高;病原真菌烟色织孢霉在根区土壤中减少,在根表土壤中消失.表明施用放线菌制剂Act12能够改善土壤微生物区系,提高人参植株的抗性和根系活力,增加产量并改善品质.     

Diversity and community structure of ectomycorrhizal mycorrhizal fungi in roots and rhizosphere soil of Abies koreana and Taxus cuspidata in Mt. Halla

In this study, the roots and rhizosphere soil of Abies koreana and Taxus cuspidata were collected from sites at two different altitudes on Mt. Halla. Ectomycorrhizal fungi (EMF) were identified by Illumina MiSeq sequencing. The proportion of EMF from the roots was 89% in A. koreana and 69% in T. cuspidata. Among EMF in rhizosphere soils, the genus Russula was the most abundant in roots of A. koreana (p < 0.05). The altitude did not affect the biodiversity of EMF communities but influenced fungal community composition. However, the host plants had the most significant effect on EMF communities. The result of the EMF community analysis showed that even if the EMF were isolated from the same altitudes, the EMF communities differed according to the host plant. The community similarity index of EMF in the roots of A. koreana was higher than that of T. cuspidata (p < 0.05). The results show that both altitude and host plants influenced the structure of EMF communities. Conifers inhabiting harsh sub-alpine environments rely strongly on symbiotic relationships with EMF. A. koreana is an endangered species with a higher host specificity of EMF and climate change vulnerability than T. cuspidata. This study provides insights into the EMF communities, which are symbionts of A. koreana, and our critical findings may be used to restore A. koreana.     

不同连作年限草莓根际细菌和真菌多样性变化

【目的】以不同连作年限草莓根际土壤为材料,探讨不同连作年限土壤细菌、真菌多样性的变化,以期为草莓连作障碍调控提供理论依据。【方法】将没有连作(CK)、连作1年(1Y)、连作8年(8Y)的土壤分别与珍珠岩以3:1的体积充分混匀装盆,"宁玉"草莓苗于2016年9月10日定植于盆栽土壤中,定植后50 d时(开花前)进行土壤取样,提取基因组DNA,通过PCR扩增建立文库,利用Mi Seq平台Illumina第二代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16S rRNA基因V4+V5区域和真菌ITS1+ITS2区域的丰富度和多样性指数以及群落结构。【结果】从9个草莓根际土壤样本中获得3 192个细菌分类操作单元OTU和762个真菌OTU,其中,变形菌门、厚壁菌门、放线菌门、拟杆菌门和蓝藻门为草莓根际土壤的优势菌种,在CK、1Y、8Y草莓根际土壤中分别占细菌总数的87.86%、64.83%和61.79%;子囊菌门和担子菌门为优势真菌,在CK、1Y、8Y草莓根际土壤中分别占真菌群落的69.17%、69.06%和76.18%。从门的分类水平看,酸杆菌门、放线菌门、拟杆菌门、厚壁菌门、绿弯菌门、蓝菌门、浮霉菌门、芽单胞菌门、变形菌门以及真菌担子菌门、接合菌门、壶菌门的比例在不同连作年限的草莓根际土壤中显著改变(P0.05)。属水平的分析也表明,共有29个细菌属和19个真菌属的比例发生改变(P0.05)。【结论】随连作年限的延长,草莓根际土壤生态系统中细菌和真菌群落各门类组成的比例会发生显著变化。     

滴灌对苜蓿根际土壤细菌多样性和群落结构的影响

【背景】细菌作为土壤微生物中的重要类群,能够有效促进土壤物质循环和能量流动,细菌多样性以及群落结构能够反映土壤的质量状况。【目的】了解滴灌条件下苜蓿根际土壤细菌群落结构及多样性变化,探讨土壤环境因子对细菌群落结构的影响。【方法】基于细菌16Sr RNAV3-V4区高通量测序技术,分析比较滴灌与自然降雨两种模式下生长的苜蓿根际与非根际土壤中细菌多样性和群落分布规律,然后采用冗余分析(Redundancy analysis,RDA)探讨土壤环境因子与细菌多样性的关系。【结果】苜蓿根际土壤中细菌多样性丰富,滴灌根际土壤中细菌多样性显著高于自然降雨根际土壤;土壤样品中共检测到细菌46门53纲116目220科469属,主要的优势菌门为变形菌门(Proteobacteria,25.27%-34.42%),其中α-变形菌纲(Alphaproteobacteria,11.41%-18.97%)为优势亚群,鞘氨醇单胞菌属(Sphingomonas,1.00%-4.54%)为优势属。相较于自然降雨,滴灌条件下苜蓿根际土壤细菌的6个门和16个属的群落结构发生显著变化;此外,RDA分析表明,不同环境因子对微生物群落的影响不同,滴灌根际土壤中9个细菌属的丰度与全磷、全钾、有效磷、碱解氮、有机质、土壤中性磷酸酶以及土壤脲酶的含量显著正相关。【结论】滴灌作为新型节水技术,在促进植物生长、提高产量、节约成本的基础上增加了植物根际土壤中细菌多样性和丰度,该结果为新型灌溉体制的改革以及土壤微生物资源的开发利用提供科学数据。     

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...     

Growth promotion ability of zoysiagrass rhizosphere fungi in consecutive plantings of wheat and soybean

Several isolates ofPhoma sp., certain nonsporulating fungi, as well asPenicillium andTrichoderma, all isolated from zoysiagrass rhizosphere, promoted growth of wheat and soybean under greenhouse conditions. However, the ability of these rhizosphere fungi to enhance plant growth varied with the crop tested. For example, most of the fungi effectively promoted the growth of wheat, whereas only a few fungi were effective on soybean. In consecutive plantings of wheat and soybean grown in soil previously infested with these zoysiagrass rhizosphere fungi, the growth promotion ability of the fungi was lowered. However, addition of fresh potting medium appeared to restore their growth-promotive effects. It appears that the activation of plant growth-promoting fungi in soil might depend on the availability of organic substrates to colonize, as evidenced by the promotion of plant growth.     

不同生境重楼内生真菌及土壤真菌多样性比较

【背景】植物内生菌的群落组成随物种基因型及不同部位、生长地和环境条件不同而发生变化,而植物相关微生物会影响植物的生长、代谢、化学成分的积累与合成,并且可能对药用植物有效成分的合成造成影响。【目的】通过对不同生长环境重楼内生及土壤真菌群落多样性的研究,寻找园区环境与野生环境的差异真菌,为野生药用植物的人工栽培提供理论基础。【方法】测定两种环境中土壤的理化指标,并采用Illumina HiSeq高通量测序技术分别对野生环境及园区环境中重楼内生及土壤真菌群落转录间隔区ITS进行扩增子序列测定,对不同土壤环境真菌群落组成与多样性差异进行生物信息学分析。【结果】对重楼相关土壤及内生真菌的多样性分析结果显示,野生环境中的真菌丰度及多样性均高于园区环境。其中野生环境与园区环境中根际土壤真菌的丰度有显著性差异(P0.05)。真菌群落注释结果表明,两种环境中分别注释到8个门;在属水平上,野生环境注释到82个属,园区环境注释到78个属;不同土壤环境对微生物群落丰度有着较为显著的差异。野生环境中有4.04%特有的OTU,园区环境中有4.84%特有的OTU。通过LEfSe分析找到了园区环境与野生环境具有显著性差异的生物标记物,Ttracladium marohalianum、Thelephorales和Thelephoraceae为区别两种环境的生物标记物。【结论】野生环境较园区环境在土壤条件上更为疏松,并具有较高的含水量,矿质元素含量也更高,其真菌群落多样性及丰度较园区环境更为丰富。该研究结果将对重楼引种栽培后的质效变迁研究提供理论基础。     

Effects of transgenic potatoes with an altered starch composition on the diversity of soil and rhizosphere bacteria and fungi

The aim of this study was to investigate potential effects on the composition of the bacterial and fungal diversity in rhizosphere and soil of a transgenic potato line (SIBU S1) which was modified in its starch composition by RNA anisensing, compared to the non-transgenic parental cultivar (SIBU) at the flowering stage in 2000. Furthermore a second non-transgenic cultivar (SOLANA) was included in the study. To avoid artefacts derived from cultivation depending approaches, molecular techniques based on 16S-(bacteria) and 18S-(fungi) rDNA respectively were used to describe the microbial community structure. Comparing 16S- and 18S-rDNA DGGE fingerprints from the different bulk soil samples, it could be shown that no significant differences between the two cultivars and the transgenic line were found. Similar results were obtained for the rhizosphere samples using the eubacterial, α-and β-proteobacterial and fungal specific primers with the exception of, the eubacterial DGGE patterns obtained for the rhizosphere of SOLANA. These patterns revealed that the relative abundance of one band was enhanced compared with the patterns of SIBU and SIBU S1 and the sequence of the differentiating band showed the highest similarity with Enterobacter amnigenus. When Pseudomonas specific primers were used, relevant differences were found between the rhizosphere patterns of the transgenic potato line (SIBU S1) and the parental cultivar (SIBU). However, clear effects of the cultivar SOLANA on the structure of the Pseudomonas community compared to SIBU were also detected.     

黄瓜连作土壤酚酸类物质积累对土壤微生物和酶活性的影响

伴随连作年限的增加,日光温室黄瓜连作土壤中酚酸类物质(对羟基苯甲酸、阿魏酸、苯甲酸)明显积累,连作5～9年的土壤酚酸类物质含量显著高于连作1～3年的土壤.伴随外源酚酸类物质处理浓度的增加,黄瓜根区土壤中细菌、放线菌和微生物总量以及N生理群均呈先升后降趋势,在80 μg·g^-1处理浓度下细菌、放线菌数量最多,处理浓度在120μg·g^-1以下土壤真菌数量(包括尖孢镰刀菌、疫霉)急剧增长;多酚氧化酶、过氧化氢酶、蔗糖酶、脲酶和蛋白酶活性也同样呈先升后降趋势,但其峰值对应的浓度不同.     

荒漠油蒿（Artemisia ordosica）根围AM真菌分布与土壤酶活性

于2007年10月在油蒿(Artemisia ordosica)集中分布区选取4个典型样地,分别从0～10cm、10～20cm、20～30cm、30～40cm和40～50cm土层采集根围土样,分离其丛枝菌根(arbuscular mycorrhiza,AM)真菌并测定了3种土壤酶活性.结果表明,在各样地0～50cm土层中油蒿根系AM真菌菌丝定殖率均很高,有典型的泡囊与丛枝结构.菌丝定殖率与泡囊定殖率呈显著正相关,但菌丝、泡囊和丛枝定殖率与AM真菌孢子密度和土壤酶活性之间都无相关性;孢子密度在不同样地及采样深度间差异明显,与土壤有机质、速效磷和速效氮含量都呈一定正相关关系.孢子密度与脲酶和酸性磷酸酶活性有显著或极显著相关关系,与碱性磷酸酶活性之间的相关性受到土壤pH的显著影响.孢子密度的峰值出现在0～10cm表层土,并随土壤剖面深度增加而降低.土壤酶活性在土壤垂直剖面显示与孢子密度同样的规律.不同样地间AM真菌分布及油蒿根系定殖率的差异表明,油蒿与AM真菌之间有良好共生性,对维护荒漠生态环境系统结构的完整性具有重要意义.     

传统绍兴黄酒酿造真菌群落与多样性发生机制

为了揭示绍兴黄酒酿造真菌群落与多样性的发生机制,依托Illumina MiSeq PE300高通量测序平台对淋饭酒母、麦曲与发酵醪液样本中的真菌资源进行研究。从淋饭酒母、麦曲、初醪、熟醪样本中分别获得136000、215283、166150、143624条优化序列,聚类后分别得到32、133、107、84个操作分类单元(OUT),酿造真菌多样性与丰富度表现出麦曲＞初醪＞熟醪＞淋饭酒母的趋势。从门水平来看,所有样本中的子囊菌均占有绝对优势;在属水平上,酵母在淋饭酒母中表现出明显的优势,曲霉为麦曲中的优势真菌,初醪与熟醪中的优势真菌主要是酵母和曲霉。随着黄酒酿造时间的延长,醪液中酵母的比例逐渐升高,而包括曲霉在内的其他真菌类群表现出逐渐减少的趋势。主坐标分析(PCoA)和相似性聚类分析结果表明,淋饭酒母、初醪和熟醪中的菌群结构更为相近,而麦曲中的真菌资源与其他样本差异很大。对落缸发酵的初始醪液中真菌群落特征进行分析发现,淋饭酒母与麦曲对绍兴黄酒酿造真菌群落的形成具有重要影响。在酿造真菌群落构建过程中,源于淋饭酒母、麦曲等糖化发酵剂的优势种类发挥主导作用。     

贵州草海流域三种不同植被类型根际土壤真菌结构组成和多样性

为了解草海流域3种不同植被类型（湿地、草坡和灌木林地）根际土壤真菌结构组成和多样性的差异,采集3种植被根际土壤,利用第二代高通量测序技术,对土壤真菌结构组成及其多样性进行分析,测定土壤理化性质和酶活性,并与土壤真菌多样性进行相关性分析。结果表明：3种植被类型根际土壤真菌物种隶属于4个门、34纲、109目、250科、537属,其中灌木林地和湿地的真菌结构组成相似性较高,优势种群均为座囊菌纲Dothideomycetes,分别占21.18%和11.13%,而草坡优势类群为属级未确定的真菌,占29.26%;土壤真菌的多样性表现为灌木林地>草坡>湿地;植被根际土壤酶活性随土层加深逐渐降低,其中灌木林地根际土壤酶活性最高,湿地根际土壤酶活性最低,植被根际土壤真菌多样性和土壤全N、全K和速效K显著相关。3种不同植被类型根际土壤理化性质和酶活性与土壤真菌组成、丰度和多样性显著相关（P<0.05）,不同植被类型凋落物归还土壤并改变根际土壤理化性质和酶活性,进而影响根际土壤真菌组成结构和多样性。     

Illumina高通量测序揭示艾比湖湿地6种盐生植物根际土壤真菌群落组成及多样性

【目的】通过研究新疆艾比湖湿地不同盐生植物根际土壤真菌的多样性和群落结构,为艾比湖湿地退化恢复工作和真菌深入研究提供理论支持。【方法】利用高通量测序技术对真菌扩增子ITS1区进行测定,从而分析艾比湖湿地6种盐生植物根际土壤真菌群落多样性,并结合相关土壤理化因子分析环境与真菌群落多样性和丰富度的关联。【结果】艾比湖湿地6种盐生植物根际土壤真菌群落多样性及丰富度存在差异,碱蓬根际土壤真菌多样性最高,芦苇根际土壤真菌群落丰富度最高。真菌群落组成分析表明,土壤样品中真菌菌落主要隶属于子囊菌门(Ascomycota)和担子菌门(Basidiomycota),其中子囊菌门为主要优势菌门;链格孢霉属(Alternaria)是6种植物共有的优势菌属,但是其在不同植物之间的丰度存在差异,在戟叶鹅绒藤中的丰度最高,在准噶尔大戟中的丰度最低。pH与真菌多样性呈显著负相关,全磷(TP)与真菌群落丰富度呈显著正相关,pH、电导率(EC)和有机质(OM)对优势菌属的影响最大。【结论】艾比湖湿地6种盐生植物根际土壤真菌群落组成及多样性具有显著差异,碱蓬和芦苇根际土壤真菌的多样性和丰度高于其他植物,子囊菌门和链格孢霉...     

盐渍化土壤根际微生物群落及土壤因子对AM真菌的影响

为探明盐渍化土壤影响下AM真菌与根际土壤间的关系,试验选取宁夏碱化龟裂土、草甸盐土、盐化灌淤土3种类型4个样地上典型植被群落,测定了植物根际土壤养分含量、微生物群落结构、AM真菌侵染率以及孢子密度。结果显示:盐渍化土壤根际微生物碳源利用类型显著不同,对芳香类化合物的代谢能力整体较弱;红寺堡草甸盐土上微生物优势群落为氨基酸代谢类群,惠农盐化灌淤土为多聚化合物代谢群,西大滩碱化龟裂土为碳水化合物代谢群。AM真菌孢子密度与微生物碳源代谢群间的关系比较复杂。其中,惠农样点根际土壤孢子密度与多聚化合物微生物代谢群呈显著正相关,西大滩地区孢子密度与碳水化合物微生物代谢群呈显著正相关。土壤有机质、全盐、全氮、碱解氮等土壤肥力因子及土壤中的HCO-3、Na+、Cl-等盐基离子含量能解释AM真菌孢子密度与土壤环境因子之间相互关系的大部分信息。较高的HCO-3浓度促进了AM真菌侵染率的提高,但高盐浓度下Na+和Cl-降低了菌根侵染率。土壤对AM真菌孢子密度、侵染率的影响因土壤盐分组成类型的不同而异。研究结果为深入了解AM真菌多样性,促进宁夏盐碱地的合理开发与利用提供了理论依据。     

不同改良措施对太子参根际土壤酚酸含量及特异菌群的影响

太子参为石竹科多年生草本植物,以根部入药,药用价值高,市场需求大,但其栽培过程中存在严重的连作障碍问题,连作导致产量品质严重下降,病虫害猖獗,探索太子参连作障碍形成的机制及其消减措施意义重大.本研究以广泛栽培的“柘参2号”为试验材料,采用土壤农化分析、高效液相色谱(HPLC)分析、荧光定量PCR(qPCR)等技术对不同连作年限或不同消减措施下太子参根际土壤的主要养分、酚酸含量动态及特异菌群变化进行分析.结果显示: 太子参连作导致产量显著下降,与正茬相比产量下降达43.5%,而水旱轮作和微生物菌肥处理可不同程度缓解其障碍效应.太子参连作土壤主要养分(如速效氮、速效磷、速效钾、全钾)未降反升,但导致土壤酸化.HPLC分析表明,太子参连作下或者随生长时期的推进,土壤酚酸类物质并未表现出明显的积累效应,膨大中期和收获期重茬土壤中大部分酚酸含量甚至低于正茬土壤.进一步荧光定量PCR分析发现,太子参连作导致前期分离筛选到的几类病原菌(如尖孢镰刀菌、踝节霉菌、Kosakonia sacchari)的绝对含量显著上升,而水旱轮作和微生物菌肥处理可有效降低3类病原菌的含量,改善微生态结构.综上认为,太子参连作障碍的发生并非由土壤养分匮乏或者酚酸持续积累直接造成,可能主要与连作下土传病原微生物的大量繁殖爆发有关,本研究结果为深入揭示太子参连作障碍的形成机制及消减修复机制提供了理论依据.     

Effects of nicosulfuron on the abundance and diversity of arbuscular mycorrhizal fungi used as indicators of pesticide soil microbial toxicity

The key role of arbuscular mycorrhizal (AM) fungi in maintaining soil fertility and ecosystem functioning and their general sensitivity to pesticides make them good candidate bioindicators in pesticide soil microbial toxicity assessment. We investigated the impact of the herbicide nicosulfuron on mycorrhizal colonization and community structure of AM fungi via a pot-to-field experimental approach. This allowed the assessment of nicosulfuron toxicity (i) at extreme exposure schemes (pot experiment, Tier I) invoked by the repeated application of a range of dose rates (x0, x10, x100, x1000 the recommended dose) and (ii) under realistic exposure scenarios (x0, x1, x2, x5 the recommended dose) in the field (Tier II). In the pot experiment, the x100 and x1000 dose rates significantly reduced plant biomass, mycorrhizal colonization and AM fungal richness as determined by DGGE. This coincided with the progressive accumulation of herbicide concentrations in soil. In contrast, no effects on AM fungi were observed at the nicosulfuron dose rates tested in the field. Clone libraries showed that the majority of AM fungi belonged to the Glomus group and were sensitive to the high levels of nicosulfuron accumulated in soil at the latter culture cycles. In contrast, a Paraglomeraceae and a Glomus etunicatum ribotype were present in maize roots in all cycles and dose rates implying a tolerance to nicosulfuron-induced stress. Overall, the deleterious effects of nicosulfuron on AM fungi induced by the highest dose rates in the pot experiment could be attributed either to fungal-driven toxicity or to plant-driven effects which have subsequent implications for mycorrhizal symbiosis. We suggest that the tiered pot-to-field experimental approach followed in our study combined with classic and standardized molecular tools could provide a realistic assessment of the toxicity of pesticides onto AM fungi as potential bioindicators.     

水旱轮作对滩涂绿花椰菜根际土壤真菌群落结构的影响

本研究比较轮作和连作下滨海滩涂地绿花椰菜根际土壤真菌群落结构和多样性的变化特征,以探索不同轮作方式减缓绿花椰菜连作田连作障碍的效果。采用Illumina Miseq高通量测序技术,分析单季稻-绿花椰菜水旱轮作、南瓜-绿花椰菜旱旱轮作和绿花椰菜连作3种耕作方式对绿花椰菜根际土壤真菌群落结构和多样性的影响。结果表明,在浙江省台州湾滨海滩涂地绿花椰菜种植区,水旱轮作、旱旱轮作和连作绿花椰菜根际土壤共获得11门、27纲、52目、94科、155属真菌。在门水平上相对丰度较高的为子囊菌门Ascomycota、油壶菌门Olpidiomycota、担子菌门Basidiomycota、接合菌门Zygomycota、壶菌门Chytridiomycota。与连作相比,轮作显著提高了绿花椰菜根际土壤中子囊菌门、壶菌门菌群的相对丰度,而以水旱轮作对其相对丰度的提高幅度更大。轮作显著降低了绿花椰菜根际土壤中油壶菌门、接合菌门菌群的相对丰度,而以水旱轮作对这2门菌群相对丰度的降低幅度更大。在属水平上轮作显著降低了绿花椰菜根际土壤中油壶菌属Olpidium、镰刀菌属Fusarium等的相对丰度,显著提高了绿花椰菜根际土壤中格孢腔菌属Pleospora的相对丰度,而水旱轮作更提高了格孢腔菌属的相对丰度,降低了油壶菌属、镰刀菌属的相对丰度。因此,轮作能改善台州湾滨海滩涂地上绿花椰菜根际土壤真菌群落结构,但水旱轮作更能增加绿花椰菜根际土壤真菌有益菌丰度,减少绿花椰菜根际土壤真菌有害菌群丰度,提高根际土壤真菌多样性指数,改良绿花椰菜根际土壤生态环境。