应用克隆文库构建法分析马铃薯块茎形成期根系AMF菌群组成

为弄清马铃薯生产中丛枝菌根真菌的作用,随机采取马铃薯块茎形成期的根系样品,以DNA提取产物为基础,Nested-PCR扩增目的片段,利用该产物构建AMF部分18S rRNA基因克隆文库,运用ARDRA筛选、DNA测序和系统发育树等方法分析丛枝菌根真菌的结构组成。结果表明:文库Coverage C值为89.7%,但Rarefaction曲线不够饱和;获得的8个AMF类型均与免培养的球囊霉属克隆序列相似度较高,其中Seq2、Seq3、Seq5和Seq8代表的摩西球囊霉(Glomus mosseae)、幼套球囊霉(Glomus etunicatum)、近明球囊霉(Glomus claroideum)和地表球囊霉(Glomus versifome)分类地位较为清晰,其余序列可能代表的球囊霉属较新种类。     

土壤中丛枝菌根真菌对宿主植物磷吸收作用机制综述

由于贫瘠土壤不能供给植物足够磷素,而丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)在促进植物生长和吸收利用磷方面发挥着重要的作用。评述了国内外丛枝菌根真菌吸收土壤磷、根际环境与AMF共同作用对土壤磷的影响、AMF储磷机理、磷从菌丝到根系转移和植物吸收利用磷机制的研究进展,为今后应用AMF改善土壤肥力和深刻了解接菌土壤中磷的迁移转化规律奠定基础。     

丛枝菌根真菌多样性的研究方法进展

介绍几种AMF多样性研究的常用方法及近年来应用较广的分子生物学技术,综述这些方法在AMF研究中的应用现状,指出在AMF生物多样性的研究中应将形态学方法和分子生物学技术有效结合,更好地揭示自然生态系统中AMF的多样性及生态功能。     

Trapping of phosphate solubilizing bacteria on hyphae of the arbuscular mycorrhizal fungus Rhizophagus irregularis DAOM 197198

A simple method is described for trapping phosphate solubilizing bacteria (PSB) strongly attached to the hyphae of the arbuscular mycorrhizal fungus (AMF) Rhizophagus irregularis (Ri). Bacteria were isolated from the hyphosphere of mycorrhizal leek plants growing on Turface previously inoculated with soil suspensions, obtained from the mycorrhizosphere of mycorrhizal plants growing in agricultural settings or maple forests in Quebec, Canada. Among the best PSB strongly attached to the hyphae of Ri, 26 isolates belonged to Burkholderia spp. and one was identified as Rhizobium miluonense. Four hyphobacteria exhibiting high potential of inorganic and organic P mobilization were further compared with four equivalent mycorrhizobacteria directly isolated from mycorrhizospheric soils sampled. In general, hyphobacteria were superior in mobilizing P from hydroxyapatite and from a low reactivity igneous phosphate rock from Quebec. Release of gluconic acid or the product of its oxidation 2-ketogluconic acid, are the main mechanisms involved in P solubilization. In a two compartments Petri plate system, Ri extraradical hyphal exudates, supported PSB growth and activity. In the absence of PSB Ri showed a negligible P solubilization activity. In the presence of PSB a substantial increase in P mobilization was observed, and the superiority of hyphobacterial activity was also observed under this system. Our results suggest that in developing a bioinoculant based on selected PSB, their interaction with AMF hyphae should not be overlooked.     

Enhancement of arbuscular mycorrhizal fungus (Glomus versiforme) on the growth and Cd uptake by Cd-hyperaccumulator Solanum nigrum

Arbuscular mycorrhizal fungus (AMF) can enhance plant growth and resistance to toxicity produced by heavy metals (HMs), affect the bioavailability of HMs in soil and the uptake of HMs by plants, and thus has been emerged as the most prominent symbiotic fungus for contribution to phytoremediation. A greenhouse pot experiment was conducted to assess the effect of Glomus versiforme BGC GD01C (Gv) on the growth and Cd accumulation of Cd-hyperaccumulator Solanum nigrum in different Cd-added soils (0, 25, 50, 100 mg Cd kg⁻¹ soil). Mycorrhizal colonization rates were generally high (from 71% to 82%) in Gv-inoculated treatments at all Cd levels. Gv colonization enhanced soil acid phosphatase activity, and hence elevated P acquisition and growth of S. nigrum at all Cd levels. Moreover, the presence of Gv significantly increased DTPA-extractable (phytoavailable) Cd concentrations in 25 and 50 mg Cd kg⁻¹ soils, but did not affect phytoavailable Cd in 100 mg Cd kg⁻¹ soil. Similarly, inoculation with Gv significantly increased Cd concentrations of S. nigrum in 25 and 50 mg Cd kg⁻¹ soils, but decreased Cd concentrations of the plants in 100 mg Cd kg⁻¹ soil. Overall, inoculation with Gv greatly improved the total Cd uptakes in all plant tissues at all Cd levels. The present results indicated that S. nigrum associated with Gv effectively improved the Cd uptake by plant and would be a new strategy in microbe-assisted phytoremediation for Cd-contaminated soils.     

接种丛枝菌根真菌和施用磷石膏对烤烟生长及砷累积的影响

为了探讨丛枝菌根真菌（Arbuscular mycorrhizalfungi,AMF）对烤烟生长及磷石膏农用安全的影响,通过盆栽模拟试验研究了磷石膏（Phosphogypsum,PG）不同添加量[0、40mg·g^-1（PG0、PG40）1和接种两种AMF[Glomus massecle（GM）、G．aggregatum（GA）】对烤烟（KRK26）苗期生长及其磷（P）、硫（S）、砷（As）吸收的影响。试验结果表明：无论是否接种AMF,磷石膏的添加均显著增加了KRK26地上部生物量及其植株s含量、吸收量及吸收效率;除不接种处理（NM）的烤烟根系外,PG40处理显著增加了KRK26植株P含量、吸收量及吸收效率,并显著降低了NM处理的地上部As含量及吸收量,进而增加了磷砷吸收比。相同PG添加水平下,与不接种相比,接种GM和GA均显著增加了KRK26植株的生物量。除PGO处理的烤烟根系外,接种GM显著增加了KRK26植株P、S含量与吸收量及吸收效率,以及植株As含量及吸收量,并显著增加了PG40处理的植株磷砷吸收比;接种GA也显著增加了KRK26植株P、S含量及吸收量,并显著降低了PG0处理地上部As含量及吸收量。所有复合处理,以添加磷石膏40mg·g^-1和接种GA处理对KRK26的生长促进效果较好,对磷石膏施用造成的As污染有一定抵御作用。     

丛枝菌根真菌对茶树耐盐性的影响

【目的】研究盐胁迫下接种丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)对茶树根系侵染率、生长、耐盐性及茶树根际土壤酸性磷酸酶活性的影响,探索AMF提高茶树植株耐盐性的生理机理。【方法】以1年生茶树品种‘乌牛早’扦插幼苗为试验材料,采用盆栽试验,向盆土中接种丛枝菌根真菌,以接种灭菌接种物为对照,在幼苗移栽后2个月进行5,10,15,20,25,30mmol/kg 6个盐胁迫处理,分别于处理后50和100d测定茶树株高、根系菌根侵染率及叶片游离脯氨酸、丙二醛含量,100d时测定地径、主根长度、生物量、茶树叶片膜透性及根际土壤酸性磷酸酶活性,并运用模糊综合评判法对各处理的茶树生长质量进行鉴评。【结果】随着盐胁迫浓度的增加,AMF对茶树根系的侵染率逐渐降低,茶树株高、地径、生物量显著降低,生长质量下降,叶片游离脯氨酸、丙二醛含量和膜透性显著增加;与对照相比,在6个盐浓度下接种AMF后,茶树的生长量均有所增加,生长质量明显提高,叶片游离脯氨酸、丙二醛含量和膜透性均有所降低,茶树根际土壤酸性磷酸酶活性均明显提高,但其随着盐浓度的升高而下降。【结论】盐胁迫能抑制茶树的生长,而AMF可通过显著促进茶树体内渗透调节物质的积累和根际土壤酸性磷酸酶活性的提高,有效降低植株体内膜脂的过氧化水平,从而缓解盐胁迫对茶树的伤害,促进茶树在盐胁迫环境中的生长。     

Effects of non-chemical soil fumigant treatments on root colonisation with arbuscular mycorrhizal fungi and strawberry fruit production

The effects of biofumigation and soil heating on arbuscular mycorrhizal fungi (AMF) colonisation, strawberry growth and strawberry yield in pot experiments compared with untreated soil and chemical fumigation with dazomet were tested. Three different Brassicaceae species (Brassica juncea, Eruca sativa, Sinapis alba) were used as biofumigant plant green manure and soil heating was applied to simulate soil solarisation. Half of the plants were inoculated with indigenous arbuscular mycorrhizal fungi inoculum. With one exception (E. sativa) among the uninoculated plants, the treatments significantly decreased the mycorrhizal colonisation parameters compared with the untreated control. Dazomet displayed the greatest inhibitory effects on AMF establishment. In addition, the intensity and number of bands corresponding to Glomus spp. obtained with temporal temperature-gradient gel electrophoresis were lower for strawberry plants from biofumigant treatments than from the control. For the inoculated plants, there were almost no significant differences among the mycorrhizal colonisation parameters. The mass of leaves for the uninoculated and inoculated plants was higher for almost all non-chemical soil fumigant treatments compared with the control, except for heating of the uninoculated treatments. The number of strawberry fruits for the uninoculated biofumigant treatments was the highest, being higher than the values observed for the heating treatments, the chemical disinfection treatments and the control. There were no significant differences among the inoculated treatments. Biofumigation with Brassicaceae species resulted in higher soil organic matter and mineral nutrients and had a relatively small effect on AMF colonisation (F% = 59.0, 80.3, 47.3 for Bj, Es and Sa, respectively) compared with uninoculated controls (F% = 84.3). Despite the reduced AMF colonisation, biofumigation resulted in a higher fruit number and mass of leaves. Therefore, it represents a non-chemical soil fumigation method that should be applied in sustainable strawberry production.     

Plant growth stage, fertiliser management and bio-inoculation of arbuscular mycorrhizal fungi and plant growth promoting rhizobacteria affect the rhizobacterial community structure in rain-fed wheat fields

The goal of this study was first to assess the dynamics of the bacterial community during a growing season in three Indian rain-fed wheat fields which differ mainly through their fertilizer management and yield and then to study the effects of PGPR/AMF bio-inoculations on the bacterial community structure and wheat growth. The bacterial community structure of the rhizosphere soil (RS) and the rhizoplane/endorhizosphere (RE) was determined by PCR-denaturing gradient gel electrophoresis. Seed treatments consisted of consortia of two PGPR strains alone or combined with AMF or AMF alone. The PGPR strains were Pseudomonas spp. which included some or all of the following plant growth promoting properties: phosphate solubilisation and production of indole-3-acetic acid, siderophores, 1-aminocyclopropane-1-carboxylate deaminase and diacetyl-phloroglucinol. The mycorrhizal inoculum was an indigenous AMF consortium isolated from the field with the lowest level of fertilization and yield. Variation partitioning analysis of the DGGE data indicated a predominant effect of the wheat growth stage (30.4% of the variance, P=0.001) over the type of field (9.0%, P=0.027) on the bacterial community structure in the RE. The impact of plant age in the RS was less than in the RE and the bacterial community structure of the field with the highest input of fertilization was very different from the low input fields. The bio-inoculants induced a significant modification in the bacterial community structure. In the RS, the bacterial consortia explained 28.3% (P=0.001) and the presence of AMF 10.6% (P=0.02) of the variance and the same trend was observed in the RE. Plant yield or grain quality was either increased or remained unaffected. For example, protein content was significantly higher in the treated plants' grain compared to the control plants; maximum values were obtained when the PGPR were co-inoculated with the AMF. The percentage of root colonization by AMF was significantly higher in the treatments containing a mycorrhizal inoculum than in the untreated control and remained unaffected by the PGPR treatments. In conclusion, the wheat rhizobacterial community structure is highly dynamic and influenced by different factors such as the plant's age, the fertilizer input and the type of bio-inoculant. In addition, there is a distance-related effect of the root on the bacterial community. Finally, a combined bio-inoculation of diacetyl-phloroglucinol producing PGPR strains and AMF can synergistically improve the nutritional quality of the grain without negatively affecting mycorrhizal growth.     

干热河谷不同树龄攀枝花苏铁丛枝菌根真菌多样性研究

攀枝花苏铁是我国特有的古老残遗树种,为适应干热河谷气候,攀枝花苏铁与丛枝菌根真菌（arbuscular mycorrhizal fungi,AMF）建立特殊的共生体系。选用不同树龄的攀枝花苏铁为研究对象,采集攀枝花苏铁根系及根际土壤样品,采用醋酸墨水染色法观察根系的AMF定殖情况并计算定殖率;同时采用湿筛沉淀法对AMF孢子进行分离,通过形态学特征及分子生物学鉴定相结合的方法研究攀枝花苏铁AMF的种类、多样性及与树龄的关系。结果表明,14年生、8年生和3年生攀枝花苏铁根系AMF定殖率分别为36.13%、28.54%、21.10%;根际土壤AMF孢子密度分别为16.02、14.13、13.3个·g^-1,形态学鉴定到6属22种,分别为球囊霉属（Glomus）、无梗囊霉属（Acaulospora）、近明球囊霉属（Claroideoglomus）、斗管囊霉属（Funneliforms）、盾巨孢囊霉属（Scutellospora）和硬囊霉属（Sclerocystis）,球囊霉属和无梗囊霉属为优势属,相对多度分别为56.91%和28.23%。分子生物学鉴定到3属6种,分别为近明球囊霉（C.claroideum）、明球囊霉（G.clarum）、缩球囊霉（G.constrictum）、无梗囊霉属（A.srobiculata）、黑球囊霉（G.melanosporum）和地表球囊霉（G.versiforme）。统计分析表明,AMF定殖率与AMF孢子密度之间的相关性是0.873 9,呈显著的正相关关系。该研究为攀枝花苏铁的保育提供了理论依据。