Illumina Amplicon Sequencing of 16S rRNA Tag Reveals Bacterial Community Development in the Rhizosphere of Apple Nurseries at a Replant Disease Site and a New Planting Site

We used a next-generation, Illumina-based sequencing approach to characterize the bacterial community development of apple rhizosphere soil in a replant site (RePlant) and a new planting site (NewPlant) in Beijing. Dwarfing apple nurseries of ‘Fuji’/SH6/Pingyitiancha trees were planted in the spring of 2013. Before planting, soil from the apple rhizosphere of the replant site (ReSoil) and from the new planting site (NewSoil) was sampled for analysis on the Illumina MiSeq platform. In late September, the rhizosphere soil from both sites was resampled (RePlant and NewPlant). More than 16,000 valid reads were obtained for each replicate, and the community was composed of five dominant groups (Proteobacteria, Acidobacteria, Bacteroidetes, Gemmatimonadetes and Actinobacteria). The bacterial diversity decreased after apple planting. Principal component analyses revealed that the rhizosphere samples were significantly different among treatments. Apple nursery planting showed a large impact on the soil bacterial community, and the community development was significantly different between the replanted and newly planted soils. Verrucomicrobia were less abundant in RePlant soil, while Pseudomonas and Lysobacter were increased in RePlant compared with ReSoil and NewPlant. Both RePlant and ReSoil showed relatively higher invertase and cellulase activities than NewPlant and NewSoil, but only NewPlant soil showed higher urease activity, and this soil also had the higher plant growth. Our experimental results suggest that planting apple nurseries has a significant impact on soil bacterial community development at both replant and new planting sites, and planting on new site resulted in significantly higher soil urease activity and a different bacterial community composition.     

Rootstock genotype and orchard replant position rather than soil fumigation or compost amendment determine tree growth and rhizosphere bacterial community composition in an apple replant soil

Apple replant disease (ARD) is a complex soilborne disease syndrome that often causes problems when renovating old orchard sites. Soil fumigants sometimes control ARD, but biological and cultural alternatives are needed. In this study the growth of two widely used clonal apple (Malus domestica) rootstocks (M7 and M26) were compared to three new rootstocks from the Cornell-Geneva series (CG16, CG30 and CG210 (a.k.a. CG6210)) in an orchard site with a history of ARD, in Ithaca, NY. Trees were planted in two distinguishable positions – the previous tree rows versus the old inter-row grass lanes. Additionally, we compared the effects of compost amendment and fumigation with dichloropropene plus chloropicrin on tree growth on each replant rootstock. Rhizosphere bacteria and actinobacteria were assessed using PCR-DGGE for the rootstocks M7, M26, CG30 and CG210. Tree growth on the rootstocks M7, M26 and CG16 was suppressed in the old tree rows relative to grass lanes, while trees on CG30 and CG210 rootstocks grew equally well in both positions. The species composition of rhizosphere bacteria and actinobacteria differed significantly between the planting positions and between the rootstocks M7 and M26 compared to CG30 and CG210. In contrast, the preplant compost or fumigation soil treatments had no effect on tree growth and little impact on rhizosphere bacterial community composition. Abbreviations: ARD – apple replant disease; DGGE – denaturing gradient gel electrophoresis; PCR – polymerase chain reaction.     

甲壳素对连作条件下平邑甜茶幼苗生长及土壤环境的影响

研究在苹果连作土壤中添加甲壳素对苹果幼苗生长、土壤酶及土壤真菌群落结构的影响,探讨甲壳素缓解苹果连作障碍的可能性,为防控苹果连作障碍提供依据。盆栽条件下,以平邑甜茶幼苗为试材,在苹果连作土壤中分别添加0,0.5,1.0和2.5g/kg的甲壳素,测定了连作土壤中添加不同量的甲壳素后,幼苗生物量、根系保护酶活性、土壤主要酶(蔗糖酶、脲酶、磷酸酶等)活性以及土壤中真菌群落结构的变化。9月份结果表明,与对照相比,1.0 g/kg的甲壳素处理连作土,可显著提高平邑甜茶幼苗株高和干鲜重,分别比对照增加了36.8%、82.1%和100.8%;甲壳素处理能增加幼苗根系保护酶活性,其中1.0 g/kg甲壳素处理SOD、POD和CAT活性最高,其次为0.5 g/kg,而2.5 g/kg甲壳素处理显著抑制了幼苗根系保护酶活性。1.0 g/kg甲壳素处理可提高土壤中细菌/真菌值,并且提高了土壤中蔗糖酶、脲酶、蛋白酶、磷酸酶、过氧化氢酶和多酚氧化酶活性,分别比对照提高了8.6%、40.5%、81.1%、15.3%、18.7%和49.8%,2.5 g/kg甲壳素处理则降低土壤酶活性或者使土壤酶活性与对照相当。根据T-RFLP的图谱中OUT的数量、种类及丰度,分别计算了不同处理土壤的真菌多样性,发现1.0 g/kg甲壳素处理的连作土具有最高的多样性、均匀度和丰富度指数,分别比对照增加了52.2%、8.0%和87.1%。主成分分析(PCA)结果显示,不同剂量甲壳素处理的连作土壤中真菌被PC2分成了两部分,其中0.5 g/kg和1.0 g/kg的甲壳素添加量分布在PC2的负方向上,而CK和2.5g/kg的甲壳素处理分布在PC2的正方向上,这说明添加不同量的甲壳素对连作土壤真菌群落多样性有显著影响,添加量太多或者太少均会造成土壤真菌多样性下降,只有适量的甲壳素可提高真菌群落结构多样性。实验结果表明1.0 g/kg的甲壳素可提高连作平邑甜茶幼苗生物量,改善连作土壤环境,有效缓解平邑甜茶的连作障碍。     

Linking Soil Biotic and Abiotic Factors to Apple Replant Disease: a Greenhouse Approach

Apple replant disease (ARD) is a frequently occurring plant disease, which causes retarded growth and mortality of young apple trees in replanted orchards. The aetiology is not well understood, but soil‐borne micro‐organisms are often discussed as primary causal agents of the replant problem. A greenhouse study was conducted in Laimburg, Italy, with orchard soils from the region, with the aim of obtaining information about the influence of soil biotic and abiotic factors on the aetiology of the disease. Apple rootstocks (M9) were planted into soils cultivated with apple trees that were either fumigated with chloropicrin or not fumigated, as well as mixtures of fumigated and non‐fumigated soils. In addition, uncultivated soils (from the inter‐row, from a fallow plot and from a meadow) were taken as controls. Various parameters were measured after 62 days in a controlled pot assay. Soils fumigated with chloropicrin resulted in higher apple shoot growth and lower microbial biomass carbon than non‐fumigated soils. Uncultivated soils had generally the highest microbial biomass carbon and the highest ergosterol contents. No considerable differences between basal respiration, ergosterol content, pH, electrical conductivity, and most nutrient and metal contents were observed between fumigated and non‐fumigated soils. Denaturing gradient gel electrophoresis gels of DNA extracted from the soils revealed differences in the fungal, bacterial and actinobacterial communities of the different soils, indicating significant shifts in microbial community composition after chloropicrin treatment. This study indicates biotic factors in soil to be a causal agent of apple replant disease.     

Long-term orchard groundcover management systems affect soil microbial communities and apple replant disease severity

Apple replant disease (ARD) is a soil-disease syndrome of complex etiology that affects apple tree roots in replanted orchards, resulting in stunted tree growth and reduced yields. To investigate whether different groundcover management systems (GMSs) influence subsequent ARD severity, we grew apple seedlings in an outdoor nursery in pots containing orchard soil from field plots where four GMSs had been maintained for 14 years in an orchard near Ithaca, NY, USA. The GMS treatments were: (1) pre-emergence herbicide (Pre-H), bare soil strips maintained by applying tank-mixed glyphosate, norflurazon and diuron herbicides annually; (2) post-emergence herbicide (Post-H), sparse weed cover maintained by applying glyphosate in May and July each year; (3) mowed sod grass (Mowed Sod); and (4) bark mulch (Mulch). Soils were also sampled from the grass drive lane maintained between the trees in the orchard (Grass Lane). Sampled soils (Orchard soil) were either pasteurized or left untreated, placed into 4-L pots, and planted with one apple seedling per pot. After 3 months of growth, soil (Bioassay soil) and apple tree roots (Bioassay roots) were sampled from each pot and microbial populations colonizing samples were characterized. Seedling growth was reduced in soils sampled from all four GMS treatments compared to the Grass Lane soils. Among the GMS treatments, seedling biomass was greater in Pre-H than in the Post-H soil. Soil microbial communities and nutrient availability differed among all four GMS treatments and the Grass Lane. Root-lesion (Pratylenchus sp.) nematode populations were higher in the Mowed Sod than in the other GMS treatments. Soil bacterial and fungal community composition was assessed in Orchard and Bioassay soils and Bioassay roots with a DNA fingerprinting method (T-RFLP). Redundancy analysis indicated that soils sampled from the different GMS treatments differentially influenced seedling biomass. A clone library of 267 soil bacteria was developed from sampled Orchard soils and Bioassay roots. These communities were dominated by Acidobacteria (25% of sequences), Actinobacteria (19%), δ-Proteobacteria (12%), β-Proteobacteria (10%), and these ratios differed among the GMS soils. Members of the family Comamonadaceae were detected only in tree-row soil, not in the Grass Lanes. The dominant sequences among 145 cloned fungi associated with apple seedling roots were Fusarium oxysporum (16% of sequences), an uncultured soil fungus submitted under DQ420986 (12%), and Rhodotorula mucilaginosa (9%). In a redundancy analysis, factors including fungal and oomycete community compositions, soil respiration rates, population sizes of culturable bacteria and fungi, soil organic matter content, and nutrient availability, were not significant predictors of apple seedling biomass in these soils. Different GMS treatments used by apple growers may influence subsequent ARD severity in replanted trees, but edaphic factors commonly associated with soil fertility may not reliably predict tree-root health and successful establishment of replanted orchards.     

甘肃省中部沿黄灌区轮作和连作马铃薯根际土壤真菌群落的结构性差异评估

甘肃省中部沿黄灌区是西北地区乃至全国重要的加工型马铃薯生产基地,然而因集约化种植带来的连作障碍问题已经严重影响到当地马铃薯种植业的可持续发展。采用大田试验与PCR-DGGE(Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis)技术相结合的方法,并通过真菌的18S r DNA序列分析,评估轮作(未连作)和连作条件下马铃薯根际土壤真菌群落在组成结构上的差异,以期为甘肃省中部沿黄灌区马铃薯连作的土壤障碍机理研究提供新证据。结果表明,同轮作相比,连作显著降低了马铃薯块茎产量和植株生物量,并且随着连作年限的延长,连作障碍也愈加严重。长期连作(6a)也导致马铃薯根冠比显著增加和植株收获指数的显著下降。在根际土壤真菌的种群数量和多样性上,连作和轮作间无显著差异,但在群落组成结构上差异明显。真菌18S r DNA测序分析进一步表明,马铃薯连作较轮作相比增加了Fusarium sp.和Fusarium solani以及Verticillium dahliae的种群或个体数量,而这些真菌是导致马铃薯土传病害的主要致病菌类型。根际土壤真菌群落组成结构的改变特别是与土传病害有关的致病菌滋生可能是导致当地马铃薯连作障碍的重要原因。     

Effects of Nitrogen and Phosphorus on the Growth of Microorganisms Associated with Apple Replant Disease and on Apple Seedlings Grown in Soil Infested with these Microorganisms*

The growth of fungi causing apple replant disease (ARD) was inhibited by the addition of N and P to the growing medium. The population of bacteria antagonistic to ARD-causing fungi was significantly increased in the growing medium supplemented with N 400 P 400 mg/l or greater. The application of nitrogen alone or in combination with phosphorus to soil infested with fungi or bacteria that cause apple replant disease significantly increased seedling height. The addition of P alone to these infested soils did not have any effect on seedling height. Significant increases in seedling height occurred with N applications when seedlings were grown in soil to which bacteria that are antagonistic to fungi causing ARD had been added. These results suggest that the application of N, with or without P, to apple replant soils may suppress the growth of ARD-causing fungi or bacteria and promote the growth of antagonistic bacteria.     

环渤海湾地区连作苹果园土壤中酚酸类物质变化

分析了山东昌邑、栖霞、蓬莱,辽宁大连、抚宁、绥远,河北昌黎、青县等地苹果园连作土壤中酚酸物质的组成和含量,结果表明:连作障碍发生的苹果园土壤中酚酸类物质的组成和含量在不同地区、不同土层厚度间存在显著差异。苹果园土壤中酚酸类物质的含量从春季到秋季随时间的延长逐渐减少,但过程缓慢。且非连作园土壤中酚酸类物质含量显著少于连作土。连作土壤中酚酸物质的组成和含量在不同季节间差异显著,尤其春季土壤中酚酸物质的种类与夏季和秋季显著不同。这可能是因为植物在不同季节分泌的酚酸类物质种类和含量有差别.不同土层中酚酸物质的分布因季节不同而有显著差异。夏季土壤中酚酸物质主要分布在浅层土壤中,而秋季则主要分布在深层土壤中。这可能是由于浇水等果园管理措施和自然降水对土壤中的酚酸物质产生的淋溶作用,使得大量的酚酸物质向深层土壤运动,最终造成了秋季果园连作土中随土层加深而酚酸物质含量增加的现象。不同地区苹果园连作土中,对羟基苯甲酸、(+)-儿茶素、咖啡酸、阿魏酸含量与非连作土无显著差异,而焦性没食子酸、绿原酸和根皮苷显著高于非连作土。焦性没食子酸、绿原酸和根皮苷可能是引起山东、辽宁、河北地区苹果园连作障碍的关键酚酸物质。     

Rootstock genotype succession influences apple replant disease and root-zone microbial community composition in an orchard soil

Apple replant disease (ARD) is a soil-borne disease complex that affects young apple trees in replanted orchards, resulting in stunted growth and reduced yields. Newly developed rootstock genotypes with tolerance to ARD may help to control this disease. We determined the effects of rootstock genotype rotations during orchard renovation, by investigating root-zone soil microbial consortia and the relative severity of ARD on seven rootstock genotypes (M.9, M.26, G.30, G.41, G.65, G.935, and CG.6210) planted in soil where trees on four of those same rootstocks (M.9, M.26, G.30 and CG.6210) had grown for the previous 15 years. Rootstock genotyping indicated that genetic distances among rootstocks were loosely correlated with their differential responses to ARD. Root-zone fungal and bacterial community composition, assessed by DNA fingerprinting (T-RFLP), differed between M.26 and CG.6210. Soil bacterial communities were influenced most by which rootstock had grown in the soil previously, while fungal communities were influenced more by the current replanted rootstock. In a clone library of bacteria from M.26 and CG.6210 root-zone soil, β-Proteobacteria was the most abundant phylum (25% of sequences). Sequences representing the Burkholderia cepacia complex were obtained only from CG.6210 soil. Rootstock genotypes that were grown in the orchard soil previously affected subsequent ARD severity, but replanting with the same or closely related rootstocks did not necessarily exacerbate this disease problem. Our results suggest that genotype-specific interactions with soil microbial consortia are linked with apple rootstock tolerance or susceptibility to ARD.     

连作苹果园土壤真菌的T-RFLP分析

为探讨连作苹果园不同土壤空间真菌群落结构,应用T-RFLP(Terminal Restriction Fragment Length Polymorphism)技术,比较了3个连作园不同取样位置(行间、原穴、株间)和不同土层(0—30 cm、30—60 cm)的土壤真菌多样性,并结合不同样品TRFLP图谱的差异,采用多样性指数分析、聚类分析和主成分分析(PCA),分析了3个连作园土壤真菌群落结构特征。结果表明,磁窑、道朗和金城连作园的土壤真菌多样性存在差异,各采样地区的Shannon多样性指数在0.43—2.47之间,Pielou均匀度指数在0.17—0.85之间,Simpson优势度指数在0.12—0.81之间,Margalef丰富度指数最高的是金城树穴0—30 cm土层(R=4.55),最低的是磁窑行间30—60 cm土层(R=0.77)。在调查的不同取样位置、不同土层中,原树穴具有最高的多样性指数、均匀度指数、丰富度指数和最低的优势度指数;0—30 cm土层的土壤真菌多样性指数、均匀度指数、丰富度指数均高于30—60cm土层,而优势度指数的趋势正好相反;PCA和聚类分析结果显示磁窑、道朗和金城连作园的土壤真菌群落结构均有明显差异,3个连作园的土壤真菌各自构成一个独立的群落结构,这些群落能够适应各自的土壤环境并成为环境的优势群落。     

Does apple replant disease affect the soil patch selection behaviour and population growth of Collembolans?

Apple replant disease (ARD) is common to all major apple-growing regions in the world. It occurs when new apple trees are replanted on sites where previously the same or closely related crop species were grown. Biotic (fungi, bacteria and nematodes) and abiotic soil factors (poor soil structure, nutrition) contribute to the development and severity of ARD. However, the aetiology of ARD and effects on higher trophic levels are still unknown. In that sense, Collembola might play an important role, since they are one of the dominant mesofauna groups in many soils. They act as decomposer, fungivores and predators, representing different trophic levels in soil food webs. Therefore, any effect of ARD on the occurrence of Collembola could have ecological impacts on the soil quality and health. Here, we examined the colonization behaviour of two Collembolan species, Folsomia candida and Sinella curviseta, in choice tests and population growth tests using Apple Replant Diseased soil (ARD) and non-ARD soil samples from different field sites and standardized laboratory bioassays. Additionally, Collembola behaviour was quantified by continuous video observations to investigate short-term behavioural changes. Results showed that both Collembolan species significantly preferred colonization of the non-ARD soils compared with ARD soils, independent of the origin of the soil samples or specific disinfection treatments. Moreover, the detailed video analysis of the foraging behaviour indicates rapid colonization of soil samples and low dispersal rates. Most likely, volatile compounds and to a lesser extent feeding stimulants play a vital role for the colonization process for both Collembolan species. Finally, results showed negative effects of ARD on population growth of both Collembolan species already after an 8-week period, implying strong nutritional deficiencies in ARD affected soils. The hypothesis that ARD causing microorganisms directly affected orientation, colonization and population development of Collembola is discussed.     

黄河三角洲不同类型盐生植物土壤真菌群落结构特征

盐生植物种类及其所具有的不同耐盐调节方式影响着根际微生物群落的结构与组成。为明确不同类型盐生植物根际与非根际土壤中真菌群落结构与组成的差异及其与土壤环境间的相互关系,该研究采集了黄河三角洲地区芦苇、盐地碱蓬、獐毛3种不同类型盐生植物0～20 cm土层的根际和非根际土壤,通过高通量测序对其真菌群落多样性和结构进行了分析,以探究真菌群落特征与土壤理化因子间的关系。结果表明：(1)3种不同类型盐生植物根际土壤真菌群落丰富度显著大于各自非根际土,且獐毛根际土壤真菌群落丰富度显著大于芦苇和盐地碱蓬的根际土。(2)距离热图分析表明,芦苇和盐地碱蓬非根际土壤真菌群落间的相似性最大。(3)土壤真菌多样性和丰富度与土壤总碳、总氮、有效磷、pH呈正相关关系,与土壤盐分含量呈负相关关系。(4)3种不同类型盐生植物的根际与非根际土壤中,球囊菌门(Glomeromycota)均为绝对优势门,盾巨孢囊霉属(Scutellospora)为优势属。(5)RDA分析表明,土壤盐分含量是影响土壤真菌群落结构的重要因子,球囊菌门丰度与土壤总氮、总碳、有效磷、有机碳、pH呈正相关关系,与盐分呈负相关关系。(6)植物土壤真菌群...     

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。     

Early rhizosphere microbiome composition is related to the growth and Zn uptake of willows introduced to a former landfill

Although plants introduced for site restoration are pre‐selected for specific traits (e.g. trace element bioaccumulation, rapid growth in poor soils), the in situ success of these plants likely depends on the recruitment of appropriate rhizosphere microorganisms from their new environment. We introduced three willow (Salix spp.) cultivars to a contaminated landfill, and performed soil chemical analyses, plant measurements, and Ion Torrent sequencing of rhizospheric fungal and bacterial communities at 4 and 16 months post‐planting. The abundance of certain dominant fungi was linked to willow accumulation of Zn, the most abundant trace element at the site. Interestingly, total Zn accumulation was better explained by fungal community structure 4 months post‐planting than 16 months post‐planting, suggesting that initial microbial recruitment may be critical. In addition, when the putative ectomycorrhizal fungi Sphaerosporella brunnea and Inocybe sp. dominated the rhizosphere 4 months post‐planting, Zn accumulation efficiency was negatively correlated with fungal diversity. Although field studies such as this rely on correlation, these results suggest that the soil microbiome may have the greatest impact on plant function during the early stages of growth, and that plant–fungus specificity may be essential.     

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...     

Evaluation of Bacillus subtilis for Potential Control of Apple Replant Disease

Apple seedlings (ev. Melntosh were stunted when grown in pasteurized field soil to which four fungi, previously isolated from apple replant disease (ARD) soil, were added. Twenty-one isolates of Bacillus sulnths produced inhibition zones in vitro against 20 ARD fungi. Isolate BACT-1 ol B. subtilis showed significant antagonism against 17 fungi isolated from ARD soil on potato dexirose agar and apple plant height was increased in its presence in pesteurized and fertilized soil in glasshouse pot tests. Plant height also increased in the presence of isolate F.BW-4 of B.subtilis.in pasteurized and fertilized soil in glasshouse pot tests but EBW-4 liid not show antagonism against most of the ARD tungi in vitro. The growth of apple seedlings was stunted in the presence of 12 isolates of B. subtilis. However, fertilization of non pasteurized soil with NPK 1l-55-0 reversed the stunting effect. Under field conditions, BACT-1 and EBW-4 isolates. led to increased shoot growth in unfertilized and unppasteurized ARD soil.     

Growth rate of fine root systems influences rootstock tolerance to replant disease

Aims

Plant tolerance to herbivory has often been linked to plant growth rate, with faster growing plants that present high tissue turnover rates expected to be more tolerant than slower-growing plants. We tested whether this relationship also holds for rootstock growth rate and tolerance to apple replant disease (ARD).

Methods

An ARD susceptible rootstock, M.26 and ARD tolerant rootstock, CG.6210 were grown in soil from an apple replant site (FS) and in pasteurized soil (PS) from the same site. Total below ground biomass production was determined by harvesting a subset of plants per soil treatment and rootstock at 11, 17, and 23 weeks after planting. Root samples were collected prior to each harvesting date to determine root respiration and total carbon (C) and nitrogen (N) content. Root dynamics were tracked during the growing season by digitally photographing root observation windows.

Results

Total root biomass, first and second order roots, and second-to-first order root ratio were higher in CG.6210 than in M.26 in both soil treatments. Roots of CG.6210 were thinner and had lower N concentration than those of M.26. Roots of M.26 had longer lifespans than those of CG.6210, and the mortality risk of M.26 roots was 56 % that of CG.6210 roots.

Conclusion

Our study indicates that rootstocks with faster growing root systems can tolerate ARD infection by investing fewer resources in individual root construction that can be shed more readily.     

Artificial induction of the apple replant problem byPenicillium claviforme inoculation

Inoculation of apple seedling roots withPenicillium claviforme reduced plant growth and resulted in morphological changes of roots resembling those found in seedlings growing in ‘apple replant problem’ soil (apple-sick soil). The introduced fungus persisted in the rhizosphere throughout the 30-month test period. The numbers of colony-forming units (CFU) ofPenicillium claviforme, as well as the ‘total’ number of CFU of micromycetes, were higher in the rhizoplane of the inoculated seedling in comparison with uninoculated plants. The numbers of CFU of phytotoxic micromycetes in the rhizosphere soil of inoculated seedlings were also proportionally higher. Apple tree growth was also reduced when seedlings were inoculated with the flucrescent bacteriumPseudomonas putida; however, no morphological changes were observed in the roots. Both micro-organisms introduced into the apple seedling rhizosphere caused changes in the microbial community. Inoculation withPenicillium claviforme andPseudomonas putida caused a decrease in the number of mycolytic bacteria in the rhizoplane of apple seedlings.     

土壤熏蒸剂棉隆和生物菌肥对草莓连作土壤真菌多样性的影响

【背景】草莓是我国和世界上重要的园艺作物,主要采用一年一栽的栽培模式,导致连作现象普遍存在,其中真菌性病害引起的土传病害是草莓栽培中面临的一个主要问题。【目的】以草莓连作土壤为材料,探讨土壤熏蒸剂棉隆加生物菌肥对草莓连作土壤微生物真菌多样性的影响,以期为草莓连作障碍治理提供理论依据。【方法】分别采集棉隆消毒前(A)、棉隆消毒后(B)、棉隆消毒后未添加生物菌肥(C1)和添加生物菌肥(C2)初花期的草莓连作土壤(或根际土壤)样本,提取DNA,通过PCR扩增建立文库,利用Hi Seq 2500平台Illumina第二代高通量测序技术并结合相关生物信息学,分析土壤真菌ITS1区域的丰富度、多样性以及群落结构。【结果】从4个不同处理草莓连作土壤样本中获得了723个真菌OTU(Operational taxonomic unit),其中子囊菌门和担子菌门均为优势真菌。多样性和丰富度研究发现,棉隆处理降低草莓连作土壤微生物丰富度和多样性;添加生物菌肥增加根际土壤生物丰富度,并降低其多样性。从门水平看,棉隆处理担子菌门真菌比例减少,子囊菌门真菌比例增加;添加生物菌肥处理两者比例均有增加。优势真菌群落分析表明棉隆消毒减少了枝顶孢属、曲霉属、管柄囊霉属、镰刀菌属、踝节菌属、链格孢属等真菌比例,增加了马拉色氏霉菌属、线孢虫草菌属、侧耳属等真菌比例;添加生物菌肥减少了管柄囊霉属、镰刀菌属、被孢菌属、轮枝菌属、隔指孢属等真菌比例,增加了曲霉属、青霉菌属、踝节菌属、简单壳菌属等真菌比例。【结论】采用棉隆消毒和生物菌肥处理草莓连作土壤可降低微生物真菌群落的多样性,并减少或灭杀土壤中的大部分致病菌属,增加有益菌属,起到有效防治草莓土传病害的作用。     

云南文山健康三七种植年限对根际微生物群落的影响

【背景】三七的连作障碍与微生物区系失衡有一定关联,了解种植年限对三七微生物区系的影响,判断根际微生物区系是否失衡,对三七连作障碍的防治具有重要意义。【目的】探究文山不同种植年限健康三七根际土壤细菌和真菌群落结构、多样性差异,以及与种植年限之间的关联,为后续缓解三七连作障碍提供理论依据。【方法】采集一年生、二年生和三年生健康三七根际土壤,采用16S rRNA基因和ITS序列进行高通量测序。对测序结果进行分析,比较一年生、二年生和三年生健康三七根际土壤细菌和真菌的多样性和群落分布规律及与种植年限的关联。【结果】一年生健康三七根际细菌和真菌群落的α指数均显著高于二年生和三年生。通过β多样性分析发现,一年生健康三七根际土壤中真菌群落结构与其他年限的真菌群落结构有较大差异。相关性分析结果表明,真菌的均匀度和香农指数与种植年限呈现显著的负相关性,其余指数不显著相关。三七根际丰富度前三的门和属,除Acidobacteria和Aquicella丰富度随着三七种植年限的增加而显著增加外,其余群落丰度都与种植年限呈负相关。细菌中Bacteroidetes、Proteobacteria和真菌中Ascomycota、Basidiomycota、Chytridiomycota的丰富度随着种植年限增加而降低。在属水平上,细菌中Opitutus和真菌中Mortierella、Clitopilus、Pholiota的丰富度随着三七种植年限的增加而降低。在二年生三七根际土壤中病原真菌Alternaria、Cylindrocarpon、Fusarium和Pestalotiopsis的丰富度低于一年生和三年生,而有益真菌Mucor和Bacillus的丰富度却呈相反趋势。同时,有益细菌Flavobacterium和有益真菌Myxocephala、Aspergillus的丰富度随着种植年限的增加呈下降趋势。【结论】三七种植年限的增加,会引起三七根际微生物区系失衡,增加根际病原真菌属的丰度,同时降低一些有益真菌属的丰度。