Viability of VA-mycorrhizal fungi following soil solarization and fumigation

Two field experiments were conducted to examine the effect of soil solarization on the survival of arbuscular mycorrhizal (AM) fungi and root colonization of three crops. The experiments were carried out in a loamy sand soil (Rehovot) and a silty soil (Bet She'an Valley). For both experiments, assessment of indigenous AM fungal populations by the most probable number (MPN) method indicated that populations were reduced to zero after 2 or 4 weeks of solarization treatment. However, Glomus intraradices inoculum applied to the soil prior to solarization remained viable even after 8 weeks of solarization. After soil fumigation with methyl bromide both indigenous and applied AM fungi were nondetectable. Percentage root colonization by the indigenous AM fungal populations, together with plant-growth parameters, were assessed for three crops: onion and wheat (Rehovot), and carrot (Bet She'an). When sown on solarized field plots, onion and carrot seedlings showed a plant growth retardation, whereas wheat showed an increased growth response. Root colonization by indigenous AM fungi was not evident until 6 weeks after seedling emergence. Fumigation with methyl bromide reduced root colonization by indigenous AM populations, and reduced onion and wheat plant development at early growth stages. In a laboratory experiment, a temperature of 45° C for up to 24 h did not affect AM spore viability, indicating that temperatures reached during the solarization treatment cannot solely account for the reduced AM fungi viability in the field. Apparently, soil solarization temporarily delays root colonization by indigenous AM fungi until 6-8 weeks after plant emergence.     

Fast track in vitro mycorrhization of potato plantlets allow studies on gene expression dynamics

Root colonization by arbuscular mycorrhizal (AM) fungi is a dynamic process involving major changes in plant gene expression. Here, the expression of a phosphate transporter gene (PT3) and several defense genes, already known to be involved in the various stages of AM establishment, were monitored in the mycelium donor plant (MDP) in vitro culture system associating potato plantlets with an AM fungus. This system allows fast and homogenous mycorrhization of seedlings at their early stage of development by growing the plantlets in active mycelial networks, but has never been validated for gene expression analysis. Here, QRT-PCR analyses were conducted in parallel to pre- (1 day), early (2 and 3 days), and late (6, 9, and 15 days) stages of root colonization. We observed the induction of a plant gene marker of AM root colonization (PT3) at the late stage and the induction of MAPK and PAL genes at the early and late stages of root colonization. We also demonstrated the induction of PR1 and PR2 genes at pre- and late stages and of GST1 and Lox genes at a late stage of root colonization. These results validated the MDP in vitro culture system as an optimal tool to study gene expression analysis during the AM fungi establishment. This system further opened the door to investigate gene networks associated with the plants–AM fungi symbiosis.     

Influence of Arbuscular Mycorrhizal (AM) Symbiosis on Phosphorus Leaching through Soil Cores

Two experiments with soil cores were carried out to investigate the effects of arbuscular mycorrhizal (AM) fungal colonization on mobility of phosphorus (P) during leaching of repacked columns of a soil with a loamy sand texture. Trifolium subterraneum plants inoculated with an AM fungus or not inoculated were grown in cores with low or high P concentrations for 8 or 10 weeks in the glasshouse. Cores were then irrigated with 2500 mL water and the leachate collected. Plant growth and the amounts of P removed by plants, remaining in soil as available P and removed dissolved in leachate were measured. Mycorrhizal fungal colonization and development of external hyphae were also determined. Inoculation and/or P application significantly increased plant growth and plant P removal and decreased P leaching. In low P soils AM fungal colonization significantly increased plant P uptake and decreased soil available P and total dissolved P in leachates. Lower P leaching from cores with AM plants under low P conditions was related to enhancement of plant growth and to scavenging and removal of P from the soil by roots and/or external hyphae. When P was applied AM effects were not observed and available P remaining in the soil after leaching was much higher, regardless of AM fungal colonization.     

磷有效性对大豆菌根侵染的调控及其与根构型、磷效率的关系

以30个不同根构型的大豆基因型为材料,通过盆栽试验,研究了生长介质磷有效性对大豆接种摩西球囊霉属丛枝菌根真菌的影响及其与根构型、磷效率的关系.结果表明：生长介质磷有效性显著地影响大豆菌根真菌的接种效果.低磷条件下接种菌根真菌效果明显,菌根侵染率较高,菌根对大豆磷吸收的贡献率较大;高磷条件下接种菌根真菌效果不显著,菌根侵染率较低,菌根对大豆磷吸收的贡献率较低.磷有效性和大豆根构型对菌根真菌接种的影响具有交互作用.低磷条件下,中间型和深根型大豆的菌根侵染率最高,浅根型最小.高磷条件下,根构型与菌根侵染率间的关系不明显.根构型和菌根侵染状况对大豆磷效率的贡献存在互利互补关系,磷效率高的大豆基因型一般具有较好的根构型或较高的菌根侵染率.     

Constitutive overexpression of the sucrose transporter SoSUT1 in potato plants increases arbuscular mycorrhiza fungal root colonization under high, but not under low, soil phosphorus availability

The sucrose transporter SUT1 functions in phloem loading of photoassimilates in solanaceous plant species. In the present study, wildtype and transgenic potato plants with either constitutive overexpression or antisense inhibition of SUT1 were grown under high or low phosphorus (P) fertilization levels in the presence or absence of the arbuscular mycorrhizal (AM) fungus Glomus intraradices. At a low soil P fertilization level, the extent of AM fungal root colonization was not different among the genotypes. In all plants, the AM symbiosis contributed significantly to P uptake under these conditions. In response to a high soil P fertilization level, all genotypes showed a decrease in AM fungal root colonization, indicating that the expression level of SUT1 does not constitute a major mechanism of control over AM development in response to the soil P availability. However, plants with overexpression of SUT1 showed a higher extent of AM fungal root colonization compared with the other genotypes when the soil P availability was high. Whether an increased symbiotic C supply, alterations in the phytohormonal balance, or a decreased synthesis of antimicrobial compounds was the major cause for this effect requires further investigation. In plants with impaired phloem loading, a low C status of plant sink tissues did apparently not negatively affect plant C supply to the AM symbiosis. It is possible that, at least during vegetative and early generative growth, source rather than sink tissues exert control over amounts of C supplied to AM fungi.     

Arbuscular mycorrhizal status and root phosphatase activities in vegetative <Emphasis Type="Italic">Carica papaya</Emphasis> L. varieties

The arbuscular mycorrhizal (AM) status and root phosphatase activities were studied in four vegetative Carica papaya L. varieties viz., CO-1, CO-2, Honey Dew and Washington. Standard techniques were used to ascertain information on spore density and species diversity of AM fungi. Although in case of estimation of root colonization and root phosphatase activities, the existing methods were slightly modified. Root colonization and spore density of AM fungi along with root phosphatase (acid and alkaline) activities varied significantly in four papaya varieties. The present study recorded higher acid root phosphatase activity when compared with alkaline root phosphatase activity under P-deficient, acidic soil conditions. The present study revealed that the root colonization of AM fungi influenced acid root phosphatase activity positively and significantly under P-deficient, acidic soil conditions. A total of 11 species of AM fungi belonging to five genera viz., Acaulospora, Dentiscutata, Gigaspora, Glomus and Racocetra were recovered from the rhizosphere of four papaya varieties.     

Shifts in symbiotic associations in plants capable of forming multiple root symbioses across a long‐term soil chronosequence

Changes in soil nutrient availability during long‐term ecosystem development influence the relative abundances of plant species with different nutrient‐acquisition strategies. These changes in strategies are observed at the community level, but whether they also occur within individual species remains unknown. Plant species forming multiple root symbioses with arbuscular mycorrhizal (AM) fungi, ectomycorrhizal (ECM) fungi, and nitrogen‐(N) fixing microorganisms provide valuable model systems to examine edaphic controls on symbioses related to nutrient acquisition, while simultaneously controlling for plant host identity. We grew two co‐occurring species, Acacia rostellifera (N₂‐fixing and dual AM and ECM symbioses) and Melaleuca systena (AM and ECM dual symbioses), in three soils of contrasting ages (c. 0.1, 1, and 120 ka) collected along a long‐term dune chronosequence in southwestern Australia. The soils differ in the type and strength of nutrient limitation, with primary productivity being limited by N (0.1 ka), co‐limited by N and phosphorus (P) (1 ka), and by P (120 ka). We hypothesized that (i) within‐species root colonization shifts from AM to ECM with increasing soil age, and that (ii) nodulation declines with increasing soil age, reflecting the shift from N to P limitation along the chronosequence. In both species, we observed a shift from AM to ECM root colonization with increasing soil age. In addition, nodulation in A. rostellifera declined with increasing soil age, consistent with a shift from N to P limitation. Shifts from AM to ECM root colonization reflect strengthening P limitation and an increasing proportion of total soil P in organic forms in older soils. This might occur because ECM fungi can access organic P via extracellular phosphatases, while AM fungi do not use organic P. Our results show that plants can shift their resource allocation to different root symbionts depending on nutrient availability during ecosystem development.     

The effect of soil phosphorus on the external mycelium growth of arbuscular mycorrhizal fungi during the early stages of mycorrhiza formation

The effects of soil P amendments and time of application on the formation of external mycelium by different arbuscular mycorrhizal (AM) fungi were studied. In the first experiment the external mycelium produced in the soil by the AM fungus Glomus etunicatum Beck. and Gerd., during the early stages of root colonization (7 and 14 days after inoculation), was quantified by the soil-agar film technique. A Brazilian Oxisol was used with three different phosphate levels, varying from deficient to supra-optimal for the plant. Significant differences were observed in the phosphate and inoculation treatments for plant dry weight, P content in the tissue, root length and root colonization, at fourteen days after planting. At 7 days, mycelium growth, root colonization and their relationship were reduced at supra-optimal P concentrations. Applications of P one week after planting reduced mycelium growth and root colonization more than when applied to the soil before planting. In a second experiment the arbuscular mycorrhizal (AM) fungi, Scutellospora heterogama (Nicol. and Gerd.) Walker and Sanders and E3 were tested and compared with Glomus etunicatum. For the species studied, the length of external hyphae per unit of colonized root length was affected by small P additions but no further significant differences were observed at high P levels. The three AM endophytes showed marked differences in their response to P in the soil: Scutellospora heterogama, although producing external mycelium more profusely than the Glomus spp., showed a higher sensitivity to soil P supply.     

Root colonization and spore abundance of arbuscular mycorrhizal fungi in distinct successional stages from an Atlantic rainforest biome in southern Brazil

The influence of plant functional groups and moderate seasonality on arbuscular mycorrhizal (AM) fungal status (root colonization and spore density) was investigated during 13 consecutive months in a chronosequence of succession in southern Brazil, consisting of grassland field, scrub vegetation, secondary forest and mature forest, in a region of transition from tropical to subtropical zones. AM root colonization and spore density decreased with advancing succession and were highest in early successional sites with grassland and scrub vegetation, intermediary in the secondary forest and lowest in the mature forest. They were little influenced by soil properties, but were sufficiently influenced by the fine root nutrient status and fine root traits among different functional plant groups. AM root colonization and spore density were higher during the favourable plant growth season (spring and summer) than during the less favourable plant growth season (autumn and winter). Spore density displayed significant seasonal variation at all sites, whilst root colonization displayed significant seasonal variation in grassland, scrub and secondary forest, but not in mature forest. The data suggest that (1) different plant functional groups display different relationships with AM fungi, influencing their abundance differentially; (2) plant species from early successional phases are more susceptible to AM root colonization and maintain higher AM sporulation than late successional species; (3) fine root traits and nutrient status influence these AM fungal attributes; and (4) higher AM spore production and root colonization is associated with the season of higher light incidence and temperature, abundant water in soil and higher plant metabolic activity.     

Arbuscular mycorrhiza improves plant adaptation to phosphorus deficiency through regulating the expression of genes relevant to carbon and phosphorus metabolism

Aims Arbuscular mycorrhizal (AM) symbiosis plays an important role in plant adaptation to phosphorus (P) deficiency. The mycorrhizal fungi can directly regulate P stress response of the host plants, and can also indirectly influence neighbor plants via AM exudates. This study aimed to reveal the regulation mechanisms of plant response to P deficiency by AM associations. Methods In a compartmentation cultivation experiment with Zea mays ‘B73’ and AM fungus Rhizophagus irregularis ‘DAOM197198’, we investigated mycorrhizal effects on plant P nutrition and the expression of plant and fungal genes related to P and carbon (C) metabolisms under both low P (10 mg?kg^-1) and high P (100 mg?kg^-1) conditions. The cultivation system consisted of three compartments, namely donor compartment, buffer compartment and receiver compartment divided by two pieces of microporous filters with pore size of 0.45 μm. Maize plant in donor compartment inoculated with AM fungus served as a source of AM exudates. The microporous filters could restrict the development of extraradical mycelium of AM fungi, but allow diffusion of AM exudates. Real-time PCR was performed to quantify the gene expression levels both in maize plants and AM fungi. Important findings The experimental results indicated that under low P conditions mycorrhizal colonization increased plant dry weight and P concentration in donor plants, and up-regulated plant genes encoding P transporters Pht1;2, Pht1;6, phosphoenolpiruvate carboxylase (PEPC), inorganic pyrophosphatase (TC289), glycerol-3-phosphate transporter (G3PT) and malate synthase (MAS1). The expression of AM fungal genes encoding P transporter (GiPT), GlcNAc transporter (NGT1), GlcNAc kinase (HXK1b), GlcNAc phosphomutase (AGM1), UDP GlcNAc pyrophosphorylase (UAP1), chitin synthase (CHS1), GlcNAc-6-phosphate deacetylase (DAC1) and glucosamine-6-phosphate isomerase (NAG1) was significantly higher under low P conditions compared with high P conditions. However, for the receiver plants, plant dry mass and P concentration were only significantly increased by higher P addition, while inoculation treatment significantly up-regulated the expression of P transporter genes Pht1;2 and Pht1;6, C metabolism related genes G3PT, PEPC, TC289 and MAS1. The study proved that AM exudates could potentially stimulate plant response to P deficiency by regulating functional genes relevant to P and C metabolisms in the mycorrhizal associations.     

Effects of co-inoculation with arbuscular mycorrhizal fungi and rhizobia on soybean growth as related to root architecture and availability of N and P

Soybean plants can form tripartite symbiotic associations with rhizobia and arbuscular mycorrhizal (AM) fungi, but little is known about effects of co-inoculation with rhizobia and AM fungi on plant growth, or their relationships to root architecture as well as nitrogen (N) and phosphorus (P) availability. In the present study, two soybean genotypes contrasting in root architecture were grown in a field experiment to evaluate relationships among soybean root architecture, AMF colonization, and nodulation under natural conditions. Additionally, a soil pot experiment in greenhouse was conducted to investigate the effects of co-inoculation with rhizobia and AM fungi on soybean growth, and uptake of N and P. Our results indicated that there was a complementary relationship between root architecture and AMF colonization in the field. The deep root soybean genotype had greater AMF colonization at low P, but better nodulation with high P supply than the shallow root genotype. A synergistic relationship dependent on N and P status exists between rhizobia and AM fungi on soybean growth. Co-inoculation with rhizobia and AM fungi significantly increased soybean growth under low P and/or low N conditions as indicated by increased shoot dry weight, along with plant N and P content. There were no significant effects of inoculation under adequate N and P conditions. Furthermore, the effects of co-inoculation were related to root architecture. The deep root genotype, HN112, benefited more from co-inoculation than the shallow root genotype, HN89. Our results elucidate new insights into the relationship between rhizobia, AM fungi, and plant growth under limitation of multiple nutrients, and thereby provides a theoretical basis for application of co-inoculation in field-grown soybean.     

Impact of soil sterilization and irrigation intervals on P and K acquisition by mycorrhizal onion (<Emphasis Type="Italic">Allium cepa</Emphasis>)

Drought is a world-spread problem seriously influencing crop production. Arbuscular mycorrhizal (AM) association and soil microorganisms can help plant growth under water stress condition by improvement of its nutrient and water uptake. In this experiment, onion plants (Allium cepa L. cv. Red Azar Shahr) were inoculated with three AM fungi species (Glomus versiforme, G. intraradices, G. etunicatum) or left un-inoculated as non-mycorrhizal plants, in a sterile or non-sterile sandy loam soil. Plants were irrigated at 7, 9 or 11-day intervals to keep the soil moisture content to field capacity at the irrigation time. Mycorrhizal root colonization decreased (p < 0.05) with an increase in irrigation interval, and the highest root colonization was achieved at 7-day irrigated onions in symbiosis with G. versiforme. Phosphorus content in plant tissue was significantly increased in mycorrhizal than non-mycorrhizal onions. Plants inoculated with G. versiforme at 9-day interval treatment had the highest leaf P content, while the lowest P was observed in non-mycorrhizal plants at all irrigation intervals. Onions inoculated by G. versiforme or G. etunicatum at 9-day irrigation interval had the highest K content. Results revealed that the inoculation of onion plant with G. versiforme or G. etunicatum and increasing irrigation interval up to 9 days, could improve P and K uptake.     

Mycorrhizal influence on protein and lipid of durum wheat grown at different soil phosphorus levels

 Root colonization by arbuscular mycorrhizal fungi (AMF) may affect protein and lipid composition of plants by altering P nutrition or by eliciting other metabolic responses in the host plant. This study was conducted to determine the effects of an AMF and soil P on seed protein and lipid contents and yield of two genotypes of durum wheat (Triticum durum L.). Plants were grown in a greenhouse using soil: sand mixes with different levels of P, and with or without the AMF Glomus mosseae [(Nicol. and Gerd.) Gerd. and Trappe]. Percentage AMF root colonization decreased as P added to soil increased. The wheat genotype CR057 had higher AMF root colonization but lower seed P and protein concentrations than CR006. Without added soil P, protein concentration was significantly lower and lipid concentration and seed dry weight higher in arbuscular mycorrhizal (AM) than in nonAM plants. Seed lipid and protein contents were highly correlated with P content of plants. In nonAM plants, seed lipid and protein contents were low with no added soil P and did not differ with added soil P. Seed protein/lipid (Pro/L) concentration ratios of AM plants were higher than those of nonAM plants only when no P was added to the soil. The data indicate different patterns of seed P accumulation and different relationships between seed P and protein and lipid in AM and nonAM plants. Thus, both the presence and degree of AMF root colonization affected seed lipid metabolism in these durum wheat genotypes. Accepted: 18 May 1999     

Arbuscular mycorrhizal colonization of semi-aquatic grasses along a wide hydrologic gradient

The effect of soil flooding on arbuscular-mycorrhizal (AM) fungal colonization of wetland plants was investigated using Panicum hemitomon and Leersia hexandra , two semi-aquatic grasses (Graminaceae) that grow along a wide hydrologic gradient in Carolina bay wetlands of the southeastern US coastal plain. Three related investigations were conducted along the dry-to-wet gradient in these wetlands; a field survey of AM fungal root colonization in eight wetlands, monthly monitoring of colonization patterns in P. hemitomon over a growing season, and an inoculum potential bioassay of soils collected along the gradient. The field survey showed that AM fungal colonization was strongly negatively correlated with water depth, but colonization was present in most root samples. The monthly assessment indicated that AM fungal colonization was lowest in plots that were consistently wet but rose as some plots underwent seasonal drying. The inoculum potential assay of dry, intermediate, and wet soils performed under both dry and saturated conditions showed that soils that were wet for >1 yr had the same ability to form mycorrhizas in bait plants as those that had remained dry. These findings suggested that the lower degree of colonization in wet areas observed in the field survey was because of the presence of surface water rather than low numbers of mycorrhizal propagules in the soil. Overall, the results of these investigations show that flooding is partially but not totally inhibitory to AM fungal colonization of wetland grasses.     

贵州栽培笃斯越橘DSE真菌、AM真菌与ERM真菌空间分布研究

为探明贵州省栽培笃斯越橘根系深色有隔内生真菌（dark septate endophytes,DSE）、丛枝菌根（arbuscular mycorrhiza,AM）真菌与欧石南菌根（ericoid mycorrhiza,ERM）真菌的定殖及地理分布情况,揭示共生真菌在栽培笃斯越橘生长中的地位,本研究在贵州省笃斯越橘主栽区麻江县、凤岗县和高坡乡分别选取主栽品种圆蓝、粉蓝、奥尼尔和莱格西的根样及根围土样,观测不同地区不同品种根样DSE真菌、AM真菌和ERM真菌的定殖结构和定殖率,并测定土样土壤理化性质,分析不同真菌与土壤因子相关性。结果表明,3个地区的4个笃斯越橘品种均有DSE真菌、AM真菌和ERM真菌定殖,栽培笃斯越橘能与3类真菌形成共生关系,平均定殖率分别为61.11%、25.55%和22.50%。DSE真菌定殖率：高坡（62.50%）>麻江（61.66%）>凤岗（59.16%）;AM真菌定殖率：凤岗（34.14%）>麻江（25.83%）>高坡（16.66%）;ERM真菌定殖率：高坡（35.00%）>凤岗（20.00%）>麻江（12.5%）。相关性分析表明,DSE真菌中的菌丝与微菌核的定殖率呈负相关,AM真菌的总定殖率及定殖强度与微菌核的定殖率呈极显著正相关,与DSE真菌菌丝的定殖率呈负相关。ERM真菌总定殖率与DSE真菌菌丝的定殖率及AM真菌定殖强度呈极显著正相关,与DSE真菌总定殖率呈显著正相关。土壤有效磷与DSE真菌和ERM真菌总定殖率呈显著正相关,与AM真菌定殖率呈显著负相关。土壤铵态氮与DSE真菌中微菌核结构定殖率及AM真菌定殖率呈极显著正相关,与ERM真菌定殖率呈极显著负相关。土壤pH值与DSE定殖强度呈显著负相关,与ERM定殖强度呈极显著正相关。本研究分析比较贵州省3个笃斯越橘种植基地不同品种栽培笃斯越橘DSE真菌、AM真菌和ERM真菌的定殖及其与土壤理化性质之间的相关性,为贵州省栽培笃斯越橘的管理和发展提供技术基础和理论依据。     

Symbiotic interactions between arbuscular mycorrhizal (AM) fungi and male papaya plants: Its status,role and implications

Experiments were conducted to study the arbuscular mycorrhizal (AM) status and its role in P-uptake through assay of root phosphatases activities in four varieties of male Carica papaya L. viz. CO-1, CO-2, Honey Dew and Washington during flowering stages. In the present study, mean total root colonization of AM fungi recorded peak increase in flowering stage-II while mean root phosphatase (acid and alkaline) activities recorded peak increase in flowering stage-I. Unlike root colonization and root phosphatase activities, spore density did not exhibit any definite patterns and recorded a narrow range of fluctuation during different flowering stages of male C. papaya. The study brought out the fact that root colonization and spore density of AM fungi along with root phosphatase activities varied significantly within the four varieties of male C. papaya plants during each flowering stage. The study also recorded consistently higher acid root phosphatase activity than alkaline root phosphatase activity under P-deficient, acidic soil conditions during all flowering stages of male C. papaya plants. Studies revealed that the root colonization of AM fungi influenced root phosphatase activities (acid and alkaline) positively and significantly during all flowering stages of male C. papaya plants. A total of twelve species of AM fungi belonging to five genera viz. Acaulospora, Dentiscutata, Gigaspora, Glomus, and Racocetra were recovered from the rhizosphere of male C. papaya plants.     

Rice root colonisation by mycorrhizal and endophytic fungi in aerobic soil

Arbuscular mycorrhizal (AM) fungi are ubiquitous root symbionts that form intimate associations with the majority of plants growing in aerobic soil; fungal endophytes live internally, either intercellularly or intracellularly, and asymptomatically within plant tissues. Their presence is correlated with an increased response to biotic and abiotic stress. The populations of AM and of endophytic fungi were studied in the roots of different rice varieties grown in aerobic condition, in experimental fields in Vercelli, North Italy. All the rice varieties resulted colonised by AM fungi with a percentage of arbuscularisation ranging between 4% and 28%. Preliminary molecular analyses on some rice varieties showed that the AM population was composed of fungi identified as Glomus intraradices , on the basis of 18S ribosomal gene. All the varieties analysed but one resulted in colonisation by endophytic fungi. About 300 fungal isolates were obtained, belonging mainly to the genera Neotyphodium , Stagonospora and Penicillium .