The bacterial community of tomato rhizosphere is modified by inoculation with arbuscular mycorrhizal fungi but unaffected by soil enrichment with mycorrhizal root exudates or inoculation with Phytophthora nicotianae

Arbuscular mycorrhizal (AM) fungi have been shown to induce the biocontrol of soilborne diseases, to change the composition of root exudates and to modify the bacterial community structure of the rhizosphere, leading to the formation of the mycorrhizosphere. Tomato plants were grown in a compartmentalized soil system and were either submitted to direct mycorrhizal colonization or to enrichment of the soil with exudates collected from mycorrhizal tomato plants, with the corresponding negative controls. Three weeks after planting, the plants were inoculated or not with the soilborne pathogen Phytophthora nicotianae growing through a membrane from an adjacent infected compartment. At harvest, a PCR-Denaturing gradient gel electrophoresis analysis of 16S rRNA gene fragments amplified from the total DNA extracted from each plant rhizosphere was performed. Root colonization with the AM fungi Glomus intraradices or Glomus mosseae induced significant changes in the bacterial community structure of tomato rhizosphere, compared to non-mycorrhizal plants, while enrichment with root exudates collected from mycorrhizal or non-mycorrhizal plants had no effect. Our results support that the effect of AM fungi on rhizosphere bacteria would not be mediated by compounds present in root exudates of mycorrhizal plants but rather by physical or chemical factors associated with the mycelium, volatiles and/or root surface bound substrates. Moreover, infection of mycorrhizal or non-mycorrhizal plants with P. nicotianae did not significantly affect the bacterial community structure suggesting that rhizosphere bacteria would be less sensitive to the pathogen invasion than to mycorrhizal colonization. Of 96 unique sequences detected in the tomato rhizosphere, eight were specific to mycorrhizal fungi, including two Pseudomonas, a Bacillus simplex, an Herbaspirilium and an Acidobacterium. One Verrucomicrobium was common to rhizospheres of mycorrhizal plants and of plants watered with mycorrhizal root exudates.     

Mycorrhiza-induced resistance in banana acts on nematode host location and penetration

Mycorrhiza-induced resistance has been observed against a broad range of mainly soil-borne pathogens, including plant-parasitic nematodes, but the modes of action involved remain unclear. In this study the role of mycorrhiza-induced resistance was investigated during the pre-infectional phase of nematode host finding and penetration. Banana plants were colonized by Glomus mosseae or Glomus intraradices, two arbuscular mycorrhizal fungi. The plant-parasitic nematode Radopholus similis was inoculated after establishment of the mycorrhizal colonization. Nematode attraction and penetration were assessed within a 12-day period. In root exudate experiments, root exudates collected from both control and mycorrhizal plants were added both to control and mycorrhizal plants to assess their direct impact on the nematode penetration. In an in vitro chemotaxis bio-assay, the chemotactic behavior of R. similis was determined towards isolated root exudates of control and mycorrhizal plants. The penetration experiments clearly showed lower nematode penetration in mycorrhizal plants and the important contribution of differential root exudation by mycorrhizal plants was demonstrated in the exudate experiments as well as in the in vitro chemotaxis bio-assay, with the largest impact on juveniles. The root exudate experiments and in vitro chemotaxis bio-assay point towards a reduced attraction of the nematodes to the mycorrhizal plant roots. The results demonstrate that a water-soluble compound in mycorrhizal root exudates is at least partially responsible for the mycorrhiza-induced resistance at the pre-infectional level of R. similis infection.     

Can the arbuscular mycorrhizal fungus Glomus intraradices actively mobilize P from rock phosphates?

Plants colonized by arbuscular mycorrhizal (AM) fungi have been shown to respond positively to the application of insoluble forms of inorganic phosphorus (P) such as rock phosphates (RPs). The mechanism(s) underlying such responses remain(s) unknown and although it has been hypothesized, there is no experimental support for the production of chelating agents by AM fungal hyphae. Here we investigate whether AM fungi can solubilize P from RPs and transfer it to plant roots. Using root-organ cultures of Daucus carrota L. inoculated or not with Glomus intraradices Schenk & Smith and containing P from different RP sources, we predicted that: (1) roots inoculated with G. intraradices would take up more P than those uninoculated; that (2) the amount of P taken up by roots through G. intraradices would be positively correlated with the RP reactivity; and that (3) G. intraradices would have access to RP through localized alterations of pH and/or by the production of organic acid anions that may act as chelating agents. The RP reactivity was positively correlated with P uptake. However, mycorrhizal roots grew initially slower and did not respond differently to any P treatment than those uninoculated. There was no evidence of localized changes in pH in proximity of G. intraradices hyphae, indicating that responses to RP by mycorrhizal plants observed in previous studies do not appear to result from the release of H⁺ ions alone or in combination with organic acid anions.     

Improved growth and Cu tolerance of Cu excess-stressed white clover after inoculation with arbuscular mycorrhizal fungi

The effects of arbuscular mycorrhizal (AM) fungus, Glomus intraradices, on growth and copper (Cu) tolerance of white clover (Trifolium repens) were investigated in soils with different Cu amounts. The AM inoculation increased plant biomass and the total or bound Cu concentrations in shoots and roots but decreased the total Cu in soils and the exchangeable Cu in shoots, roots and soils at all Cu levels. Mycorrhizal plants had higher levels of root phosphorus and shoot zinc (Zn) at lower Cu levels and more nitrogen and Zn in roots and potassium, calcium and magnesium in shoots and roots at all Cu addition levels. Additionally, AM inoculation enhanced urease, acid phosphatase and catalase activities in rhizosphere soils and mycorrhizal roots showed higher levels of peroxidase, catalase, proline and soluble sugar at all Cu addition levels. These results indicate that mycorrhizal white clover is potentially suitable for Cu phytoremediation based on greenhouse studies.     

铅胁迫下小花南芥与玉米间作对根系分泌物 有机酸的影响

为了揭示Pb胁迫对间作和单作的超累积植物和作物根系分泌低分子有机酸的影响,研究设置400 mg·L?1Pb胁迫,采用水培曝气法试验,以玉米和小花南芥单作为对照处理,研究Pb胁迫下玉米和小花南芥间作对植物根系形态、根系分泌有机酸及Pb吸收的影响。结果表明:与单作相比,间作小花南芥情况下,玉米根系分泌物检测到乳酸;玉米分根条数、根表面积和根密度与单作相比分别增加60%、15%和42%,地下部和地上部干重生物量分别增加108%和75%,玉米地下部Pb含量下降44%;与单作相比,间作玉米条件下,小花南芥根系分泌物检测到乙酸和乳酸,小花南芥根系分泌物量与单作相比增加103%~1 700%,小花南芥地下部和地上部Pb累积量分别比单作增加49%和75%,转运系数增加22%。相关分析结果表明,单作小花南芥只有地上部Pb累积量与草酸显著相关,而间作小花南芥地下部和地上部Pb累积量与草酸、柠檬酸和苹果酸显著相关。研究表明超富集植物小花南芥与玉米间作体系,根系分泌的有机酸改变了Pb在小花南芥和玉米体内的累积特征,促进超累积植物小花南芥累积Pb,减少农作物玉米植株体内Pb含量。Pb胁迫下超累积植物小花南芥与玉米间作是一种可行的修复模式。     

Growth and Nutrient Content of Trifoliate Orange Seedlings Influenced by Arbuscular Mycorrhizal Fungi Inoculation in Low Magnesium Soil

A pot experiment was conducted to examine the effects of arbuscular mycorrhizal fungi, Glomus versiforme, G. mosseae, and G. intraradices on growth and nutrition of trifoliate orange (Poncirus trifoliata) seedlings under magnesium (Mg)-nontreated and Mg-treated conditions. Whether treated with Mg or not, G. versiforme inoculation significantly enhanced the growth and concentrations of Mg, phosphorus, calcium, potassium, zinc, and copper in shoots or roots, and activities of acid phosphatase, catalase, invertase, and urease in rhizosphere soils. Additionally, there were higher levels of chlorophyll, proline, soluble sugar and protein in leaves, root viability, superoxide dismutase, peroxidase and catalase in leaves and roots, but lower malondialdehyde content in leaves and roots of mycorrhizal seedlings than non-mycorrhizal ones. Data demonstrated that G. versiforme-inoculated citrus seedlings exhibited higher levels of soil enzymes, osmoregulation, and antioxidant matters, leading to improvement of growth and nutrition of seedlings in low Mg soil.     

Effect of the arbuscular mycorrhizal fungus <Emphasis Type="BoldItalic">Glomus intraradices</Emphasis> on the false root-knot nematode <Emphasis Type="BoldItalic">Nacobbus aberrans</Emphasis> in tomato plants

Information about the interaction between arbuscular mycorrhizal fungi (AMF) and the false root-knot nematode Nacobbus aberrans (Thorne, 1935) Thorne & Allen, 1944 is scarce. The effect of Glomus intraradices Schenk & Smith on tomato (Lycopersicon esculentum L.) cv. Platense inoculated with nematode juveniles from Lisandro Olmos (Argentina) was studied under greenhouse conditions. Six treatments with five replications were performed. After 80 days, nematode reproduction and percentage of AMF colonization in roots were estimated. Some plant growth parameters were also measured. In general, plants with AMF and AMF plus nematodes grew as well as the control without AMF and without nematodes. Furthermore, G. intraradices was beneficial in reducing nematode-induced damage in roots (lower number of galls) as well as in having a suppressive effect on parasite reproduction. This is the first study on the use of G. intraradices as a possible strategy in the control of N. aberrans in tomato.     

Effects of arbuscular mycorrhizal fungi on the growth and zinc uptake of trifoliate orange (Poncirus trifoliata) seedlings grown in low-zinc soil

The effects of the arbuscular mycorrhizal (AM) fungi, Glomus intraradices and G. versiforme, on growth and zinc (Zn) uptake were investigated in trifoliate orange (Poncirus trifoliata) seedlings exposed to low-Zn soil. Low-Zn decreased growth, levels of leaf chlorophyll, soluble protein and sugar, and soil enzymatic activities, and pH in 0–2 cm rhizosphere soil. Low-Zn soil also decreased mineral nutrients (including Zn) concentrations in the shoots and roots. Glomus intraradices especially, significantly enhanced plant biomass, leaf soluble protein and sugar concentrations, root viability, acid phosphatase, catalase, invertase and urease activities, and easily extractable glomalin content in 0–2 cm and 2–4 cm rhizosphere soil. It also increased concentrations of Zn, phosphorus, potassium and magnesium in the shoots and roots, while decreased the soil pH. Arbuscular mycorrhizal fungi, especially G. intraradices, has the potential to improve growth and Zn uptake of triofoliate orange seedlings grown in low-Zn soil.     

Pre-inoculation with arbuscular mycorrhizal fungi suppresses root knot nematode (<Emphasis Type="Italic">Meloidogyne incognita</Emphasis>) on cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis>)

A pot experiment was conducted to evaluate the influence of pre-inoculation of cucumber plants with each of the three arbuscular mycorrhizal (AM) fungi Glomus intraradices, Glomus mosseae, and Glomus versiforme on reproduction of the root knot nematode Meloidogyne incognita. All three AM fungi tested significantly reduced the root galling index, which is the percentage of total roots forming galls. Numbers of galls per root system were significantly reduced only in the G. intraradices + M. incognita treatment. The number of eggs per root system was significantly decreased by AM fungus inoculation, no significant difference among the three AM fungal isolates. AM inoculation substantially decreased the number of females, the number of eggs g⁻¹ root and of the number of eggs per egg mass. The number of egg masses g⁻¹ root was greatly reduced by inoculation with G. mosseae or G. versiforme. By considering plant growth, nutrient uptake, and the suppression of M. incognita together, G. mosseae and G. versiforme were more effective than G. intraradices.     

Rhodotorulic acid enhances root colonization of tomato plants by arbuscular mycorrhizal (AM) fungi due to its stimulatory effect on the pre-symbiotic stages of the AM fungi

Exudates of Rhodotorula mucilaginosa, a yeast commonly found in the rhizosphere, increased hyphal length of the arbuscular mycorrhizal (AM) fungi Gigaspora rosea and Gigaspora margarita. Rhodotorulic acid (RA), a siderophore compound obtained from R. mucilaginosa exudates, increased hyphal length and branching. Thus, the increase in the number of entry points and the higher AM root colonization of tomato plants in the presence of RA can at least partially be explained by the positive effect of RA on the pre-symbiotic stages of the AM fungi.     

Root exudate-stimulated RNA accumulation in the arbuscular mycorrhizal fungus Gigaspora rosea

Plant root exudates induce the transition from asymbiosis to presymbiosis in arbuscular mycorrhizal fungi. In order to get an insight into this developmental switch, two libraries of Gigaspora rosea and one library of Gigaspora gigantea were screened for fragments of genes that show enhanced RNA accumulation 1 h after addition of a semi-purified exudate fraction of carrot roots. Among 150 clones, 40 seemed to contain inserts of root exudate-induced genes. One of the genes, GrosRbp1, putatively encoding an RNA binding protein involved in developmental control showed RNA accumulation which correlates to the extent of stimulation of presymbiotic hyphal branching.     

Mycorrhizal association of Agrostis capillaris and Glomus intraradices under heavy metal stress: Combination of plant clones and fungal isolates from contaminated and uncontaminated substrates

The present study aimed to compare the impact of arbuscular mycorrhizal (AM) fungi on plant growth and heavy-metal (HM)-uptake when both plant and fungal symbionts originated either from contaminated or uncontaminated sites. HM-tolerance of six clones of the grass Agrostis capillaris and three isolates of the AM fungus Glomus intraradices of different origin was first tested separately. Plant clones from the vicinity of a lead smelter showed a consistently high HM-tolerance, whereas the control clones varied in their ability to cope with HMs. The AM isolate from the contaminated substrate also performed better under HM-stress than the isolates from uncontaminated soils. All A. capillaris clones were then grown in a contaminated substrate, uninoculated or inoculated either with a tolerant or non-tolerant G. intraradices isolate. Clones from the uncontaminated site accumulated considerably more HMs in their shoots and roots, regardless of inoculation. The effect of AM inoculation on plant growth and HM-uptake depended on the particular combination of plant clone and fungal isolate, without clear differences between tolerant and non-tolerant clones.     

Root herbivory by Tipula paludosa larvae increases colonization of Agrostis capillaris by arbuscular mycorrhizal fungi

The objective of this study was to determine if root-feeding by insect larvae affects subsequent colonization of roots by arbuscular mycorrhizal fungi. We investigated grazing by larvae of Tipula paludosa on colonization of Agrostis capillaris by two species of fungi, Glomus mosseae and G. intraradices. Host plants were subjected to 7 days of grazing only, continuous grazing for 42 days, or no herbivory. Those plants with no herbivory had significantly lower levels of colonization by arbuscules and hyphae compared to plants which were grazed for 7 or 42 days. The effect only occurred in the upper parts of the root system, where larvae were active. We suggest that this effect was most likely mediated by a change in quantity and composition of root exudates.     

Phosphorus uptake and growth of barley as affected by soil temperature and mycorrhizal infection

A glasshouse study was conducted to investigate the effects of soil temperatures of 20, 15 and 10°C on growth and phosphorus (P) uptake of barley (Hordeum vulgare L. cv. Galleon) inoculated with Glomus intraradices Schenck & Smith. Vesicular‐arbuscular (VA) mycorrhiza formation was significantly reduced as the soil temperature decreased. Plant growth depression due to temperature stress was more pronounced in mycorrhizal plants than in non‐mycorrhizal plants. The lower the soil temperature, the higher was the root‐shoot ratio. The ratio was also higher in non‐mycorrhizal plants than in mycorrhizal plants. Concentration of P in roots was influenced by mycorrhiza. Significant interaction between mycorrhiza and soil temperature was observed for root dry matter and specific P uptake (P uptake per unit weight of root). Compared to non‐mycorrhizal plants, specific P uptake in mycorrhizal plants was higher.     

Response of maize to mycorrhizal colonization at varying levels of zinc and phosphorus

A greenhouse experiment was conducted in a red sandy loam soil (Alfisol) to study the responses of arbuscular mycorrhizal (AM) fungus Glomus intraradices Schenck & Smith inoculated (M+) and uninoculated (M−) maize (Zea mays L) plants exposed to various levels of P (15 and 30 mg kg⁻¹) and Zn (0, 1.25, and 2.5 mg kg⁻¹). Roots and shoots were sampled at 55 and 75 days after sowing and assessed for their nutritional status, root morphology, and root cation exchange capacity (CEC) besides grain quality. Mycorrhizal plants had longer and more extensive root systems than nonmycorrhizal plants, indicating that M+ plants are nutritionally rich, especially with P, which directly assisted in the proliferation of roots. Further, root CEC of M+ plants were consistently higher than those of M− plants, suggesting that mycorrhizal colonization assists in the acquisition of nutrients from soil solution. Mycorrhizal inoculated plants had significantly (P ≤ 0.01) higher P and Zn concentrations in roots, shoots, and grains, regardless of P or Zn levels. The available Zn and P status of AM fungus-inoculated soils were higher than unioculated soils. The data suggest that mycorrhizal symbiosis improves root morphology and CEC and nutritional status of maize plants by orchestrating the synergistic interaction between Zn and P besides enhancing soil available nutrient status that enables the host plant to sustain zinc-deficient conditions.     

Soil inoculation with the biocontrol agent Clonostachys rosea and the mycorrhizal fungus Glomus intraradices results in mutual inhibition, plant growth promotion and alteration of soil microbial communities

Interactions between the biocontrol fungus Clonostachys rosea IK 726 and a tomato/Glomus intraradices BEG87 symbiosis were examined with and without wheat bran, which served as a food base for C. rosea. In soil without wheat bran amendment, inoculation with C. rosea increased plant growth and altered shoot nutrient content resulting in an increase and decrease in P and N content, respectively. Inoculation with G. intraradices had no effect on plant growth, but increased the shoot P content. Dual inoculation with G. intraradices and C. rosea followed the pattern of C. rosea in terms of plant growth and nutrient content. Wheat bran amendment resulted in marked plant growth depressions, which were counteracted by both inoculants and dual inoculation increased plant growth synergistically. Amendment with wheat bran increased the population density of C. rosea and reduced mycorrhizal fungus colonisation of roots. The inoculants were mutually inhibitory, which was shown by a reduction in root colonisation with G. intraradices in treatments with C. rosea and a reduction in colony-forming units (cfu) of C. rosea in treatments with G. intraradices, irrespective of wheat bran amendment. Moreover, both inoculants markedly influenced soil microbial communities examined with biomarker fatty acids. Inoculation with G. intraradices increased most groups of microorganisms irrespective of wheat bran amendment, whereas the influence of C. rosea on other soil microorganisms was affected by wheat bran amendment. Overall, inoculation with C. rosea increased and decreased most groups of microorganisms without and with wheat bran amendment, respectively. In conclusion, despite mutual inhibition between the two inoculants this interaction did not impair their observed plant growth promotion. Both inoculants also markedly influenced other soil microorganisms, which should be further studied in relation to their plant growth-promoting features.     

Growth and nitrate reductase activity in Juniperus oxycedrus subjected to organic amendments and inoculation with arbuscular mycorrhizae

The effectiveness of reforestation programs on degraded soils in the Mediterranean region is frequently limited by a low soil availability and a poor plant uptake and assimilation of nutrients. While organic amendments can improve the nutrient supply, inoculation with mycorrhizal fungi can enhance plant nutrient uptake. A pot experiment was conducted in 2004 to study the influence of inoculation with an arbuscular mycorrhizal (AM) fungus (Glomus intraradices Schenck & Smith) or with a mixture of three AM fungi (G. intraradices, G. deserticola Trappe, Bloss. & Menge, and G. mosseae (Nicol & Gerd.) Gerd. & Trappe) and of an addition of composted sewage sludge or Aspergillus niger–treated dry‐olive‐cake residue on plant growth, nutrient uptake, mycorrhizal colonization, and nitrate reductase (NR) activity in shoot and roots of Juniperus oxycedrus L. Six months after planting, the inoculation of the seedlings with G. intraradices or a mixture of three AM fungi was the most effective treatment for stimulating growth of J. oxycedrus. There were no differences between the two mycorrhizal treatments. All treatments increased plant growth and foliar N and P contents compared to the control plants. Mycorrhizal inoculation and organic amendments, particularly fermented dry olive cake, increased significantly the NR activity in roots.     

Interactions between a vesicular-arbuscular mycorrhizal fungus (Glomus intraradices) and Streptomyces coelicolor and their effects on sorghum plants grown in soil amended with chitin of brawn scales

 The effect of the interaction between a vesicular-arbuscular (VA)-mycorrhiza (Glomus intraradices no. LAP8) and Streptomyces coelicolor strain no. 2389 on the growth response, nutrition and metabolic activities of sorghum (Sorghum bicolor) plants grown in non-sterilized soil amended with chitin waste was studied in a greenhouse over 8 weeks. Chitin amendment resulted in an increase in the microbial population and chitinase activity in soils. Growth of mycorrhizal G. intraradices no. LAP8 and non-mycorrhizal sorghum plants increased as compared with other treatments either in the presence or absence of S. coelicolor strain 2389. VA-mycorrhizal inoculation significantly increased the growth, photosynthetic pigments, total soluble protein and nutrient contents of sorghum compared to non-mycorrhizal sorghum. Such increases were related to increased mycorrhizal colonization. Inoculation with S. coelicolor 2389 significantly increased the intensity of mycorrhizal root colonization and arbuscular formation, but the levels of mycorrhizal infection and their beneficial effects were significantly reduced with the addition of chitin waste to the soil. Analysis of the content of total amino acids and ammonia in leaves on the basis of dry matter production showed that, in most instances, total amino acids of mycorrhizal plants were significantly higher than those of non-inoculated plants. The microflora of the rhizosphere was highly affected by mycorrhizal inoculation. Quantitative changes in acid and alkaline phosphatase activities of the roots in response to the mycorrhizal inoculation are discussed. Received: 11 August 1999     

黄绿木霉T1010对日光温室番茄耐冷生理指标的影响

采用黄绿木霉T1010(Trichoderma aureoviride1010)制剂处理山东寿光日光温室番茄连作土壤,通过人工控制日光温室内温度(白天16±2℃,夜间8±2℃),使供试番茄经过3d亚低温胁迫,分别测定其第3功能叶和根系的耐冷生理指标,探索黄绿木霉T1010对亚低温胁迫下番茄保护酶系统、非酶抗氧化物质类胡萝卜素、抗寒基因表达启动因素脱落酸(ABA),以及细胞内渗透调节保护物质脯氨酸和可溶性糖等耐冷生理指标的影响。结果表明:与常规生产区(对照)相比,黄绿木霉T1010处理番茄功能叶超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)及多酚氧化酶(PPO)活力分别提高266.33%、16.85%、20.89%、632.89%,类胡萝卜素、脱落酸(ABA)、脯氨酸、葡萄糖、可溶性淀粉含量分别提高50.53%、119.40%、55.38%、128.57%和23.40%;番茄根系SOD、CAT、POD及PPO活力分别提高274.10%、69.06%、8.26%、164.14%,脯氨酸、葡萄糖、可溶性淀粉含量分别提高49.81%、66.00%和86.76%。1/2黄绿木霉T1010处理番茄功能叶以上耐冷生理指标除可溶性糖含量比对照有所降低外,其他指标均有不同程度的提高;番茄根系的相关耐冷生理指标则有不同程度的降低。ANOVA分析显示,亚低温胁迫下不同处理间日光温室番茄以上耐冷生理指标差异均达极显著水平。由此可见,在土壤有机质比较充裕的条件下,黄绿木霉T1010对亚低温胁迫下日光温室番茄功能叶和根系保护酶系统、类胡萝卜素、ABA、脯氨酸和可溶性糖等耐冷生理指标均有不同程度的积极影响。