生物炭、磷及AMF对Cd胁迫下柳枝稷生长及土壤性质的影响

为探讨添加磷、生物炭和接种丛枝菌根真菌对柳枝稷镉（Cd）耐性的作用，采用盆栽法研究土壤Cd浓度为20 mg·kg^-1时，不同处理［对照（CK）、4.5%生物炭（biochar，B）、60 mg·kg^-1磷（phosphorus，P₆₀）、4.5%生物炭+60 mg·kg^-1磷（B+P₆₀）］结合丛枝菌根真菌（AMF）对柳枝稷生长状况、矿质离子（P、Se）和Cd含量、土壤pH值、速效磷和酸性磷酸酶活性及土壤不同形态Cd含量的影响。结果表明：磷添加可显著提高柳枝稷根系侵染率，P₆₀处理下根系侵染率达到了56.9%。与CK相比，B、P₆₀、B+P₆₀处理对柳枝稷植株高度、叶片SPAD值、生物量无显著影响，各处理结合AMF的植株高度、叶片SPAD值、生物量均显著升高，但接菌后各处理间差异不显著。除了P₆₀处理下地上部Se和Cd含量及B+P₆₀处理下地上部Cd含量高于CK外， B、P₆₀、B+P₆₀处理下P、Se、Cd与CK无显著差异；接种AMF后CK和B处理下地上部Se和Cd含量均高于未接菌处理，且B+AM处理地上部Cd含量显著高于其他处理；但接菌后P₆₀处理地上部Se和Cd含量均低于未接菌处理。此外，无论是否接种AMF，B和B+P₆₀处理根系Cd含量均显著高于CK和P₆₀，土壤速效磷含量高于CK，土壤酸性提取态Cd含量低于CK和P₆₀。接种AMF后CK、B和B+P₆₀处理的土壤残渣态Cd含量高于CK和对应未接菌处理，但接菌P₆₀处理的土壤残渣态Cd含量低于未接菌处理。由此可见，B或B+P₆₀处理提高了根系Cd和土壤速效磷含量，降低了土壤酸性提取态Cd含量；二者结合AMF提高了柳枝稷生物量和地上部Se和Cd含量及土壤残渣态Cd含量。因此，AMF结合生物炭或生物炭/磷添加提高了柳枝稷生物量、Cd吸收量，降低了土壤中重金属的生物活性，可以在重度Cd污染土壤中应用。     

梵净山不同海拔丛枝菌根真菌多样性

采集梵净山国家级自然保护区5个海拔高度(500~600、900~1000、1300~1400、1700~1800、2100~2200 m)植被根际土壤,利用湿筛倾析法分离丛枝菌根真菌孢子,通过孢子形态进行物种鉴定和物种多样性分析。结果表明:梵净山丛枝菌根(AM)真菌物种资源丰富,现确定6属23种;海拔显著影响AM真菌的多样性和分布,随海拔降低,AM真菌物种多样性指数显著下降;AM真菌优势群落,依次为无梗囊霉属、球囊霉属、硬囊霉属。     

混合接种摩西球囊霉和根瘤菌对紫花苜蓿耐碱能力的影响

紫花苜蓿是多年生优质豆科牧草,对改善生态环境和农牧作物结构调整有重要作用。然而,土地盐碱化严重影响苜蓿正常的生长发育。丛枝菌根真菌(AMF)和根瘤菌是土壤中较为常见的共生菌,可与紫花苜蓿等豆科植物共生,并能与苜蓿形成AMF-苜蓿-根瘤菌共生系统。本研究利用摩西球囊霉菌和中华根瘤菌单接种、混合接种紫花苜蓿,比较单接种和混接种对其耐碱能力的影响。结果表明:在正常情况下,单接种和混合接种与未接种(NI)相比生长量均有所提高,主要表现在叶绿素含量更高,植株的根长、株高相对较大,分枝数较多,生长更加茂密。碱胁迫处理后,各组植株生长均受到不同程度的抑制,而混合接菌组则表现出更高的耐碱性,与NI组相比,主要表现在有较低的MDA含量和相对电导率,有较高的细胞相对含水量, 并且表现出较高的抗氧化酶的活性(SOD、POD、CAT)。综合以上指标分析,单接种和混合接种均能提高植株的渗透调节能力和抗氧化能力,降低细胞膜的损伤,但混合接种的效果更佳。     

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。     

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.     

丛枝菌根真菌在不同基质下对枳砧容器苗生长

为促进AM真菌在柑桔砧木容器苗生产中的高效应用，本研究以枳砧为供试植物，选取腐叶土、麦糠和菌渣为主要栽培基质，以摩西球囊霉（Glomus mosseae）、地表球囊霉(Glomus versiforme)、地球囊霉(Glomus geosporum)和透光球囊霉（Glomus diaphanum）为供试菌种，采用容器育苗方法，研究了7种丛枝菌根真菌及其组合处理在三种基质下对枳砧木容器苗生长及其氮、磷吸收的影响，以筛选出适合枳砧木苗生长的菌根种类及基质组合。试验结果表明自然接种的侵染率不超过40%，而人工接种的侵染率可高达80%；混合接种的侵染率显著高于单一接种;枳砧木苗以地表球囊霉（GV）接种和腐叶土或麦糠组合侵染率为最高，侵染率分别为80.7%和78.7%；以GV接种和麦糠组合促生效果最为显著，其菌根依赖性达到342.4%，生物量比对照高出278%；该组合促进植株氮素吸收效果也最为显著，其吸氮总量比对照高482%，而以GV接种和腐叶土组合促进枳砧苗磷素吸收效果最为显著，其吸磷总量比对照高466%。适合枳砧容器苗生长的最佳AM真菌和基质组合是GV和麦糠。     

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.     

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.     

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。